research article qualitative and quantitative analysis of
TRANSCRIPT
Research ArticleQualitative and Quantitative Analysis of the Major Constituentsin Chinese Medical Preparation Lianhua-Qingwen Capsule byUPLC-DAD-QTOF-MS
Weina Jia123 Chunhua Wang123 Yuefei Wang123 Guixiang Pan123 Miaomiao Jiang123
Zheng Li123 and Yan Zhu123
1 Tianjin Key Laboratory of Modern Chinese Medicine Tianjin University of Traditional Chinese Medicine Tianjin 300193 China2 Tianjin Key Laboratory of TCM Chemistry and Analysis Tianjin University of Traditional Chinese Medicine Tianjin 300193 China3 Research andDevelopmentCenter of Traditional ChineseMedicine Tianjin International JointAcademy of BiotechnologyampMedicineTianjin 300457 China
Correspondence should be addressed to Yan Zhu yanzhuharvardgmailcom
Received 7 July 2014 Revised 14 August 2014 Accepted 15 August 2014
Academic Editor Feng Sun
Copyright copy 2015 Weina Jia et alThis is an open access article distributed under the Creative CommonsAttribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited
Lianhua-Qingwen capsule (LQC) is a commonly used Chinese medical preparation to treat viral influenza and especially played avery important role in the fight against severe acute respiratory syndrome (SARS) in 2002-2003 in China In this paper a rapidultraperformance liquid chromatography coupled with diode-array detector and quadrupole time-of-flight mass spectrometry(UPLC-DAD-QTOF-MS)methodwas established for qualitative and quantitative analysis of themajor constituents of LQC A totalof 61 compounds including flavonoids phenylpropanoids anthraquinones triterpenoids iridoids and other types of compoundswere unambiguously or tentatively identified by comparing the retention times and accurate mass measurement with referencecompounds or literature data Among them twelve representative compounds were further quantified as chemical markersin quantitative analysis including salidroside chlorogenic acid forsythoside E cryptochlorogenic acid amygdalin swerosidehyperin rutin forsythoside A phillyrin rhein and glycyrrhizic acid The UPLC-DAD method was evaluated with linearity limitof detection (LOD) limit of quantification (LOQ) precision stability repeatability and recovery tests The results showed that thedeveloped quantitative method was linear sensitive and precise for the quality control of LQC
1 Introduction
Lianhua-Qingwen capsule (LQC) developed from the twoclassical traditional Chinese medicine (TCM) formulaeMaxing-Shigan-Tang and Yinqiao-San which have a longhistory of clinical application in the treatment of influenza[1] is a commonly used Chinese medical preparation to treatviral influenza and especially played an important role in thefight against severe acute respiratory syndrome (SARS) in2002-2003 in China [2] LQC is composed of 11 herbs includ-ing Fructus Forsythiae (Lianqiao) Flos Lonicerae Japoni-cae (Jinyinhua) Herba Ephedrae (Mahuang) Semen Arme-niacae Amarum (Kuxingren) Radix Isatidis (Banlangen)Rhizoma Dryopteridis Crassirhizomatis (Mianmaguanzh-ong)Herba Houttuyniae (Yuxingcao) Herba Pogostemonis
(Guanghuoxiang) Radix et Rhizoma Rhei (Dahuang) Radixet Rhizoma Rhodiolae Crenulatae (Hongjingtian) and Radixet Rhizoma Glycyrrhizae (Gancao) along with menthol anda traditional Chinese mineral medicine Gypsum Fibrosum(Shigao) According to previous reports LQC has a goodclinical effect on influenza with the symptoms of high feveraversion to cold headache pharyngalgia cough sneezingmuscle ache and so on [3] Modern pharmacological studieshave shown that LQC also has the antiviral antibacterial andanti-inflammatory activities [4 5] Recently the study on itsbioactive ingredients andmolecularmechanism of action hasbeen gradually reported as well [6]
Although some preliminary analytical methods havebeen developed for the quality control for LQC includ-ing thin layer chromatography (TLC) [7] high performance
Hindawi Publishing Corporatione Scientific World JournalVolume 2015 Article ID 731765 19 pageshttpdxdoiorg1011552015731765
2 The Scientific World Journal
liquid chromatography (HPLC) [8 9] micellar electrokineticcapillary chromatography (MEKC) [10] and liquid chro-matography tandem mass spectrometry (LC-MSMS) [11]no systematical and comprehensive study on the chemicalprofiling and quality control method for LQC has beenreported so far For a classical and complex Chinese medicalpreparation the comprehensive quality evaluation methodshould be based on its multiple chemical constituentsThere-fore it is necessary to develop a rapid and sensitive methodto identify and quantify the chemical constituents in LQCwhich will be beneficial to investigate the effectiveness andevaluate the quality of LQC
In this study a reliable sensitive and simple ultraper-formance liquid chromatography coupled with diode-arraydetector and quadrupole time-of-flight mass spectrometry(UPLC-DAD-QTOF-MS) method which was more system-atical and comprehensive than the earlier ones was estab-lished for characterization and quantification of the majorchemical constituents of LQC A total of 61 compoundswere unambiguously or tentatively identified by comparingthe retention times exact molecular masses and MSMSspectral data with reference compounds or literature dataFurthermore twenty-seven compounds were confirmed bycomparing with the standards Among them twelve repre-sentative compounds were quantified as chemical markers inquantitative analysis including salidroside chlorogenic acidforsythoside E cryptochlorogenic acid amygdalin swero-side hyperin rutin forsythoside A phillyrin rhein andglycyrrhizic acid This is the first systematical and compre-hensive study on the qualitative and quantitative analysis ofLQC
2 Experimental
21 Reagents Chemicals and Materials Methanol and ace-tonitrile (HPLC grade) were purchased from Sigma Aldrich(St Louis MO USA) Formic acid (HPLC grade) waspurchased from Tianjin Damao chemical reagent factory(Tianjin China)Water (HPLC grade) for UPLC analysis wasproduced by theMilli-Qwater purification system (MilliporeUSA) Salidroside chlorogenic acid forsythoside E cryp-tochlorogenic acid amygdalin sweroside hyperin rutinforsythoside A phillyrin rhein and glycyrrhizic acid werepurchased from Sigma Aldrich (St Louis MO USA) Thepurity of standard substances was above 98 Ten batches ofLQC were provided by Shijiazhuang Yiling PharmaceuticalCo Ltd (Shijiazhuang China)
22 UPLC Analysis The UPLC analysis was performed on aWaters ACQUITY UPLC instrument (Waters CorporationMA USA) coupled with a binary pump a sample manageran autosampler a column compartment and diode-arraydetector (DAD)The separation of samples was performed ona Waters ACQUITY UPLC BEH C
18(100 times 21mm 17 120583m)
column with the column temperature at 50∘C The analysiswas completed in 30min with a gradient elution of 01formic acid aqueous solution (A) andmethanol (B) at the flowrate of 03mLmin The gradient program was designed as
follows 0ndash11min 5ndash35 B 11ndash18min 35ndash55 B 18ndash22min55ndash75 B 22ndash24min 75ndash90 B 24-25min 90ndash100 Band 25ndash30min 100 B The injection volume was 5120583L Thedetection wavelengths of DAD were set at 210 nm 225 nmand 254 nm
23 UPLC-DAD-QTOF-MSAnalysis TheWaters ACQUITYUPLC instrument (Waters MA USA) coupled with WatersSynapt HDMS G1 (Waters Manchester UK) via an elec-trospray ionization (ESI) interface The UPLC analyticalconditions were the same as the UPLC analysis describedabove The full scan mass spectra data were acquired inpositive and negative ion modes Acquisition parametersare as follows capillary voltage was 3000V for ESI (+)and 2600V for ESI (minus) cone voltage was 45V the ESIsource temperature was 100∘C the desolvation tempera-ture was 350∘C the nitrogen (N
2) was used as desolva-
tion gas at flow rates of 600 Lh for both ESI (+) andESI (minus) and the range of full scan was set at mz 150ndash1000Da The version of analysis software was Mass LynxV41
24 Sample and Standard Solutions Preparation The powderof LQC (04 g) was accurately weighed and extracted with60 methanol-water (vv) solution (20mL) in an ultrasonicwater bath for 30min at room temperature The supernatantsolution was diluted with the same amount of water andthen centrifuged for 10min at 14000 rmin All the obtainedsolutions were filtered through 022 120583m syringe filter beforethe UPLC analysis
Twelve standards were accurately weighed and dissolvedin methanol to obtain stock solutions respectively A mixedstock solution of standards was prepared by adding a suitablevolume of each stock solution to a 5mL flask and dilutedwith 30 methanol-water solution at the concentration of678 120583gmL for salidroside 10965 120583gmL for chlorogenicacid 7764 120583gmL for forsythoside E 10647 120583gmL for cryp-tochlorogenic acid 6257 120583gmL for amygdalin 3196 120583gmLfor sweroside 321 120583gmL for hyperin 85 120583gmL for rutin6734 120583gmL for forsythoside A 4571120583gmL for phillyrin5549 120583gmL for rhein and 8435 120583gmL for glycyrrhizicacid respectively The mixed stock solution was then seri-ally diluted with 30 methanol-water solution to obtainfive appropriate concentrations used for plotting standardcurves The lowest concentration of the mixture stocksolution was further diluted to give a series of differ-ent concentrations for investigating the limits of detection(LODs) and limits of quantification (LOQs) of the 12 chem-ical constituents All solutions were stored at 4∘C untilanalysis
25 Validation of the Quantitative Analysis TheUPLC-DADmethod was evaluated with linearity LOD LOQ precisionstability repeatability and recovery tests The calibrationcurves were constructed with five different concentrations ofchemical markers in triplicate The LODs and LOQs weremeasured under the UPLC analytical conditions at a signal-to-noise (SN) ratio of 3 and 10 respectively For intraday
The Scientific World Journal 3
Table 1 Quantitative results of 12 compounds in LQC extracted by different methods
Content (120583gg)
Methods30a 60 90 60 60 60 60 60 60
30minb 30min 30min 30min 30min 30min 15min 30min 45min1 100c 1 100 1 100 1 50 1 100 1 200 1 100 1 100 1 100
Salidroside 172628 170125 162202 152231 170125 165632 171154 170125 168854Chlorogenic acid 244497 249215 221689 228543 249215 228927 249262 249215 255217Forsythoside E 158393 162078 45191 146209 162078 140253 162701 162078 157963Cryptochlorogenic acid 186298 185164 66750 170305 185164 177179 183789 185164 185795Amygdalin 142411 145539 144256 126893 145539 129811 139597 145539 143138Sweroside 81619 81318 78932 74706 81318 77267 81251 81318 80824Hyperin 13510 15173 16726 14084 15173 14080 15272 15173 15767Rutin 12200 12117 11522 10611 12117 11667 11709 12117 11662Forsythoside A 248460 253634 266179 228576 253634 239635 254387 253634 252110Phillyrin 166026 152145 155159 141019 152145 139012 152301 152145 155152Rhein 80313 110206 137011 93763 110206 93205 95638 110206 105477Glycyrrhizic acid 153049 168043 159437 143740 168043 161999 167481 168043 166542aExtracting solvent 30 60 and 90 methanol-water solutionbUltrasonic time 15min 30min and 45mincExtraction solvent multiples 1 50 1 100 and 1 200 expressed 50 100 and 200 times per gram of sample
and interday precisions test the samples were analyzed bysix repetitive injections within one day and once a day forthree successive days respectively At room temperaturethe stability of sample solution was evaluated by replicateinjection at 0 1 2 4 6 8 10 14 24 and 48 h In order tocheck the repeatability six samples from the same sourcewereinvestigated Accurate amounts of the reference standardswere added to 020 g powder of sample in sextuplicate Theresultant sample solutions were then extracted and quantifiedwith the described method The relative standard deviation(RSD) was used to evaluate the results
3 Results and Discussion
31 Optimization of the Extraction and ChromatographicConditions A single-factor method was used to investigatethe extraction effect of the extraction solvent (30 60and 90 methanol-water solution) extraction solvent ratio(1 50 1 100 and 1 200 (wv)) and extraction time (15min30min and 45min) respectively By analyzing the extractionefficiency 60 methanol-water solution extraction solventratio at 1 100 and 30min of ultrasonic time were selected asthe eventual extraction conditions The results are describedin Table 1
Due to the existence of acidic constituents in samplesolutions formic acid was added into the mobile phasewhich could inhibit the ionization of these acidic ingredi-ents to improve the peak shape The mobile phase systems(methanol-formic acid aqueous solution and acetonitrile-formic acid aqueous solution) and column temperature (40∘Cand 50∘C) were investigated which showed that methanol-01 formic acid aqueous solution as mobile phase with
column temperature at 50∘C could obtain the best chro-matographic peak shape Because the maximum absorptionsof 12 reference compounds were different three detectionwavelengths were finally selected in order to achieve thegoal of high detection sensitivity and little interferenceForsythoside E (peak 7) cryptochlorogenic acid (peak 9)amygdalin (peak 33) and phillyrin (peak 39) had satisfactorysensitivity at 210 nm salidroside (peak 6) chlorogenic acid(peak 8) and rhein (peak 54) at 225 nm and sweroside(peak 13) hyperin (peak 26) rutin (peak 29) forsythosideA (peak 30) and glycyrrhizic acid (peak 57) at 254 nm Thechromatograms are presented in Figure 1
32 UPLC-DAD-QTOF-MS Analysis of Reference Compoundsand LQC Samples As shown in Table 2 a total of 61compounds were unambiguously or tentatively identifiedby comparing the retention times and accurate mass mea-surement with references or literature data These com-pounds were divided into six types according to their struc-tural characteristics including flavonoids phenylpropanoidsanthraquinones triterpenoids iridoids and other types Thestructures of identified compounds are listed in Figure 3Among them twenty-seven compounds were further con-firmed by comparing with standards The total ion chro-matograms are shown in Figure 2
321 Flavonoids Seventeen flavonoids (Figure 3(a)) in LQCincluding flavone aglycones and glycosides were identifiedThey were mainly obtained from Lianqiao Jinyinhua Gan-cao Hongjingtian and Mahuang Amongst them liquiritinapioside (22) ononin (25) hyperin (26) rutin (29) liquirit-igenin (35) isoliquiritin apioside (36) isoliquiritin (37) and
4 The Scientific World Journal
AU 0020406
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
minus06minus04minus02
687933
29
3039
(a)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
AU 00204
0
minus02
6879
29
30
39
54 57
13
(b)
AU
0004008012016
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
6
8
7
9
3329
30
39 54
57
13
26
(c)
AU
0002004006008
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
minus002
(d)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU
00204
minus06minus04minus02
minus1minus08
6
87933 29
30
39
57
26
(e)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU 00204
minus04
minus02
6
8
79
33 29
30
39
54 57
(f)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU
0004008012016
02
minus004
8
9
29
30
39
54
5713
26
(g)
Figure 1 UPLC-DAD chromatograms of standard solution of 210 nm (a) 225 nm (b) 254 nm (c) negative sample solution (d) and samplesolution of 210 nm (e) 225 nm (f) and 254 nm (g)
Time
()
0
100
()
0
100
2 4 6 8 10 12 14 16 18 20 22 24 26 28
Time2 4 6 8 10 12 14 16 18 20 22 24 26 28
1
2
34 5
67 8 9
101112
13
1415 16
1718
19
20
21
22
23
24
25
26
27
28
28
29
30
301317
31
32 33
34
35
3736
38
39
40
39
42
41
434445
46 47 484950
51
5253
54
5556
57
57
58
59
60
61
45
20130402-LHQW MSminus 1 TOF MS ESminusBPI
648e4
20130402-LHQW MS+1 TOF MS ES+
BPI813e4
Figure 2 UPLC-QTOF-MS chromatograms of sample solutionfrom negative ion mode and positive ion mode
formononetin (47) were unambiguously identified via thestandards
In negative and positive ion modes flavone aglyconesmainly gain fragment ions by the reverse Diels-Alder (RDA)reaction and the loss of CO (28Da) The characteris-tic fragmentation behavior of compound 35 is shown inFigure 4(a) with high abundant fragmentation [M+HndashVP (4-vinylphenol)]+ at mz 1370422 The abundance of fragmentions [M+HndashRL (resorcinol)]+ at mz 1470621 and [M+HndashRLndashCO]+ at mz 1190691 is relatively lower Compounds 2440 42 and 46were tentatively identified via comparing theirexact molecular masses MSMS spectra data and retentionbehaviors with literature data [13 26 39] Flavone glycosideshave the similar fragmentation pathways of simultaneous orsuccessive loss of glucose (162Da) rhamnose (146Da) orapiose (132Da) The fragmentation pathway of compound26 was exemplified in Figure 4(b) in negative ion modeCompounds 21 23 31 41 and 53 were tentatively identified
The Scientific World Journal 5
O
O
O
O
OH
OH
O
O
RO
HO
OR1
R1
R3
R2
R2
R4
R5R2OR1O
OR3
OR3
OCH3
21R1 = H R2 = H R3 = Glc 24 R1 = OH R2 = Rha (3rarr1) Glc R3 = OH R4 = H R5 = H 25 R = Glc22 R1 = H R2 = H R3 = Glc (2rarr4) Api 26 R1 = OGlc R2 = H R3 = H R4 = H R5 = OH 47 R = H35 R1 = H R2 = H R3 = H 29 R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = OH R5 = H41R1 = Glc (6rarr1) Rha R2 = OH R3 = CH3 31R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = H R5 = H
40 R1 = OH R2 = H R3 = H R4 = H R5 = OH42 R1 = OH R2 = H R3 = H R4 = H R5 = H53 R1 = H R2 = H R3 = H R4 = OCH3 R5 = OCH3
36 R1 = OH R2 = H R3 = Glc (2rarr4) Api37 R1 = H R2 = OH R3 = Glc46 R1 = H R2 = OH R3 = H
(a)
4 8 9 1618 R = OH
38
27 30 34
56
32 R1 = Glc R2 = H39 R1 = Glc R2 = CH3
O
O
O
O
O
O
O
O
O
OO O O
R
O
O
O
O
O
OO
O O O
OO
OO
O
O
O
O
OO
O
O
O
OO
OO
O
O
O
O
O
OH
OH
OHOH
OH
OHOH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
OHOH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
HO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
H3C
OR2
R1O
CH3
(b)
17R1 = Glc R2 = CH3 R3 = H R4 = H43 R1 = H R2 = CH3 R3 = H R4 = H45 R1 = H R2 = CH3 R3 = OH R4 = Glc49 R1 = H R2 = CH3 R3 = OCH3 R4 = Glc54 R1 = H R2 = COOH R3 = H R4 = H61R1 = H R2 = CH3 R3 = OH R4 = H
O
O
OR1
R3R2
OR4
(c)
Figure 3 Continued
6 The Scientific World Journal
6058 59
5550 52 R1 = GlcA (2rarr1) GlcA R2 = acetoxy57 R1 = GlcA (2rarr1) GlcA R2 = H
OO
OO
O
O
O
O
O
R2H
H
H
O
O
O
O
OO
O
O
O
O
OO
O
O
OO
O
O
O
OOO
OHOH
OH
OH
OH
OH
OH
OH OH
OHOH
OH
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
R1O
(d)
5 R1 = Glc R2 = H 13 2010R1 = Glc R2 = CH3
OH
HO
O O
O
HOO OO
OOH
H OO
OHOH
OHOH HO
HOHO
HOHOHO
OR1
OR2
(e)
1 2 3 6
15
7
11 33 51
O
O
O
O O
OO
OO
O O
OO
OOO
O
OO
N
OO
O
OHOH OH
OHOH
OH
OH
OH
OH
OHOHOH
OHOH
OH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
HOHO
HO HOHO
HOHO
HO
HOHO
HOHOHO
HOHO
HOHO HO
HOHO
HOHOHO
HO
HO
H3C
(f)
Figure 3 Chemical structures of the compounds identified from LQC (except for the 7 isomers)
by comparing the molecular mass and MSMS data withliterature data [25 45]
322 Phenylpropanoids Fourteen phenylpropanoids (Figure3(b)) including phenylpropionic acids lignans and coum-arins were identified in LQC They were mainly obtainedfrom Lianqiao Jinyinhua and Gancao Neochlorogenic acid
(4) chlorogenic acid (8) cryptochlorogenic acid (9) 35-dicaffeoylquinic acid (27) forsythoside A (30) 34-dicaffe-oylquinic acid (34) and phillyrin (39) were unambiguouslycharacterized by comparing with standards
In negative ion mode phenylpropionic acids have thesimilar fragmentation pathways of simultaneous or succes-sive loss of H
2O (18Da) CO (28Da) and CO
2(44Da)
The Scientific World Journal 7
OHO
OH
O
OHO
O
OHH
mz 1470621
mz 1190691
+H
mz 2570659mz 2570659
mz 1370422
2570659
172999816301591319873 1941022
23914082651331
3011509
1370422
1190691
1199948
13800271470621
1650917 16902931810699 2112177
(MSMS)
100 120 140 160 180 200 220 240 260 280 300
()
0
100
()
0
100
mzmz
120 130 140 150 160 170 180 190 200 210
(MS+)
(a)
OHO
OHO
OH
O
OH
OOH
HOOH
OH
mz 3010464
3010464
1370281
3020746
3027005 4652560 62322138921294
4630905
43109651450305 2210378
46409055310850 7371886 11080997 13072520 14077662
(MSMS)
()
0
100(
)
0
100
mz
200 400 600 800 1000 1200 1400mz
200 400 600 800 1000 1200 1400
(MSminus)
(b)
OHHO
OHHOOC
OHOHHO
O
OHHO OH
COOminus
COOminus COOminus
Cminus
mz 1910733mz 1270486
mz 1730583 mz 1090409
mz 930425 mz 850426
minusCO2
minusCO2
minus2H2O minusC2H2
minusCO
Ominus
minusH
minusH2O minusH2O
minusH2O
1910733
8504261270486930425 1090409 1350577
1610264 1730583 19207421938051
3530878
1910558
3510719192060770718303540916
70516854510630 5031459 7081906
(MSMS)
()
0
100
()
0
100
mzmz
80 90 100 110 120 130 140 150 160 170 180 190 200200 250 300 350 400 450 500 550 600 650 700 750 800
(MSminus)
(c)
O OO
O
OH
OHOH
HO
HO
HO
HO
O
O OHOH
mz 4611920 mz 1610328
CH3
6232001
3112033 6211844
6242032
6252064 124749349575357 13186510
1610328
62325071620342 4611920
(MSMS)
()
0
100
()
0
100
200 400 600 800 1000 1200 1400mzmz
100 200 300 400 500 600 700 800 900 1000
(MSminus)
(d)
Figure 4 Continued
8 The Scientific World Journal
3561547
13599643083514
3711826
37218625161718
5792105
1129871 4310958
58021345812158 111342197973569 11814053
(MSMS)
O
O
O
OO
OO
OH
HOHO
OH
[MndashHndashGlc]minus[MndashHndashGlcndashCH3]minus
()
0
100
()
0
100
mz
150 200 250 300 350 400 450 500 550 600 650 700 750mz
200 400 600 800 1000 1200 1400
(MSminus)
mz 3561547
mz 3711826
(e)
2250723
18204771810753
18006601540552
19706952240626
2410669 2690634
2420626 2700607
2690462
1129890 27004855610872 8234322 9694360 14023499
O O
O
O
O O
(MSMS)
()
0
100(
)
0
100
mz
160 180 200 220 240 260 280mz
200 400 600 800 1000 1200 1400
(MSminus)
OH
OH
OHOH OH OH OH
OHOHOH
OH
minusCO2
minusCO2minusCO
minusCO
minusHminusHminusH
minusHminusH
minusH
H3CH3C
H3C H3C
mz 1970695mz 2410669
mz 2250723mz 2690462
mz 1820477mz 1540552
minusCH3minusCO
(f)
4310988
11692794321016
4990790 8632153 11946926 14410397
2690673
2250753
18204861710837
1970771 241071927007752710783 3110845
(MSMS)
O
O
O OH
OHO
HOHO
OH
HO
[MndashHndashGlc]minus[MndashHndashGlcndashCO]minus
[MndashHndashGlcndashCO2]minus
[MndashHndashGlcndashCOndashCO2]minus
[MndashHndashGlcndashCO2ndashCH3ndashCO]minus()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400mz
140 160 180 200 220 240 260 280 300 320
(MSminus)
mz 3711826mz 2690673
CH3
(g)
8214583
35108091130354
19305102890758
645410258340994693705 7594542
8224637
8234628
(MSMS)
H
HO
HO
OOH
O
O
HOOCHO
HOOHO
HOOH
HOOC
[MndashHndash2GlcA]minus[MndashHndashGlcA]minus
[2GlcAndashH]minus
()
0
100
mz
100 200 300 400 500 600 700 800
8213977
1169246 3111153 8203445
8224006
8434348125530939433099 13258577
()
0
100
mz
200 400 600 800 1000 1200 1400
mz 4693705mz 3510809mz 6454102
(MSminus)
(h)
Figure 4 Continued
The Scientific World Journal 9
3751302
21307733761332
75126674731059 8093751 10383667 13123422
1250712
1210637
930423
650486 950155 1260535 151087824011931740320 1999976 2505933
(MSMS)
OH
H
HO
OH
O
O
O
HO
OHOH
OH
[MndashHndashGlc]minus
[14FndashH2O]minus [MndashHndashGlcndash2CO2]minus
14F
minusH2O
()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400
mz
60 80 100 120 140 160 180 200 220 240 260
(MSminus)
minusCO2
minusCO2
[MndashHndashGlcndashCO2ndashH2O]minus
(i)
Figure 4TheMS spectra and fragmentation pathway of compound 35 (a) compound 26 (b) compound 8 (c) compound 30 (d) compound39 (e) compound 61 (f) compound 45 (g) compound 57 (h) and compound 5 (i)
The fragmentation pathway of compound 8 [18] as the rep-resentative of phenylpropionic acids is shown in Figure 4(c)Compound 30 produced [MndashHndashCa (caffeoyl)]minus at mz4611920 and [CaffeicndashHndashH
2O]minus atmz 1610328 as displayed
in Figure 4(d) Lignans primarily generated [M+HCOO]minusin negative ion mode and further elimination of glucose(162Da) produced aglycone As shown in Figure 4(e) com-pound 39 produced characteristic fragments at mz 3711826and mz 3561547 corresponding to [MndashHndashGlc (glucose)]minusand [MndashHndashGlcndashCH
3]minus respectively Compounds 16 18 19
28 32 38 and 56 were tentatively identified on the basisof the exact molecular formulae matching fragmentationinformation and retention behaviors as well as literature data[23 24 33 36 47]
323 Anthraquinones Eight anthraquinones (Figure 3(c)) inLQC were definitely or tentatively identified All of themwere derived from Dahuang Chrysophanol glucoside (17)emodin-8-O-glucoside (45) rhein (54) and emodin (61)were confirmed via comparing with standard substances
The characteristic fragmentation behavior of anthra-quinones was the loss of CO
2(44Da) CH
3(15Da) and
CO (28Da) in negative ion mode Typical compound 61was used to explain the fragmentation pathway of anth-raquinones presented in Figure 4(f) Compound 45 asshown in Figure 4(g) produced characteristic fragments atmz 2690673 mz 2410719 mz 2250753 mz 1970771andmz 1820486 which corresponded to [MndashHndashGlc]minus [MndashHndashGlcndashCO]minus [MndashHndashGlcndashCO
2]minus [MndashHndashCOndashCO
2]minus and
[MndashHndashCO2ndashCH3ndashCO]minus respectively Compounds 12 43
44 and 49 were tentatively identified by comparing theiraccurate molecular masses and MSMS fragment data withliterature data [21 40 41]
324 Triterpenoids Eight triterpenoids (Figure 3(d)) inLQC were unambiguously or tentatively identified All ofthem were derived from Gancao Glycyrrhizic acid (57) wasconfirmed by comparing with standards In negative ion
mode representative compound 57 yielded [MndashHndashH2Ondash
CO2]minus at mz 7594524 [MndashHndashGlcA (glucuronic acid)]minus
at mz 6454102 [MndashHndash2GlcA]minus at mz 4693705 [2GlcAndashH]minus at mz 3510809 and [2GlcAndashHndashH
2OndashCO
2]minus at mz
2890758 as shown in Figure 4(h) Compounds 48 50 5255 58 59 and 60 were tentatively identified by comparingtheir exact molecular masses and MSMS spectral data withthe literature data [51]
325 Iridoids Four iridoids (Figure 3(e)) in LQC includingiridoid glycosides and secoiridoid glycosides were identifiedAll of them were derived from Jinyinhua Loganic acid (5)sweroside (13) and secoxyloganin (20) were unambiguouslyidentified via the standards
The fragment 14F (fragment generated from the fractureof 14 bonds of iridoids) in the negative ion mode was identi-fied as the characteristic fragment ion of iridoid glycosidesMeanwhile iridoid glycosides would lose the functionalgroups such as H
2O (18Da) CO
2(44Da) and glucose
(162Da) Typical compound 5 gave [MndashHndashGlcndashCO2ndashH2O]minus
at mz 1510878 [MndashHndashGlcndash2CO2]minus at mz 1250712 and
[14FndashH2O]minus at mz 930423 [52] as presented in Figure 4(i)
Compound 10 was tentatively identified by comparing itsexact molecular mass and MSMS spectral data with theliterature data [19]
326 Other Types of Compounds Compounds 1 2 11 14 15and 51 were tentatively identified by comparing their exactmolecularmasses andMSMS spectral datawith the literaturedata except gallic acid (3) salidroside (6) forsythoside E (7)and amygdalin (33) which were identified via the standards(Figure 3(f)) [12 53]
33 Methodological Validation of the Quantitative AnalysisAs shown in Table 3 twelve standards were of good lin-earity with high correlation coefficient values over 09993The LODs and LOQs were 0051ndash171 120583gmL and 016ndash569 120583gmL independently Twelve analytes in sample solu-tion were stable at room temperature within 48 h with
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
2 The Scientific World Journal
liquid chromatography (HPLC) [8 9] micellar electrokineticcapillary chromatography (MEKC) [10] and liquid chro-matography tandem mass spectrometry (LC-MSMS) [11]no systematical and comprehensive study on the chemicalprofiling and quality control method for LQC has beenreported so far For a classical and complex Chinese medicalpreparation the comprehensive quality evaluation methodshould be based on its multiple chemical constituentsThere-fore it is necessary to develop a rapid and sensitive methodto identify and quantify the chemical constituents in LQCwhich will be beneficial to investigate the effectiveness andevaluate the quality of LQC
In this study a reliable sensitive and simple ultraper-formance liquid chromatography coupled with diode-arraydetector and quadrupole time-of-flight mass spectrometry(UPLC-DAD-QTOF-MS) method which was more system-atical and comprehensive than the earlier ones was estab-lished for characterization and quantification of the majorchemical constituents of LQC A total of 61 compoundswere unambiguously or tentatively identified by comparingthe retention times exact molecular masses and MSMSspectral data with reference compounds or literature dataFurthermore twenty-seven compounds were confirmed bycomparing with the standards Among them twelve repre-sentative compounds were quantified as chemical markers inquantitative analysis including salidroside chlorogenic acidforsythoside E cryptochlorogenic acid amygdalin swero-side hyperin rutin forsythoside A phillyrin rhein andglycyrrhizic acid This is the first systematical and compre-hensive study on the qualitative and quantitative analysis ofLQC
2 Experimental
21 Reagents Chemicals and Materials Methanol and ace-tonitrile (HPLC grade) were purchased from Sigma Aldrich(St Louis MO USA) Formic acid (HPLC grade) waspurchased from Tianjin Damao chemical reagent factory(Tianjin China)Water (HPLC grade) for UPLC analysis wasproduced by theMilli-Qwater purification system (MilliporeUSA) Salidroside chlorogenic acid forsythoside E cryp-tochlorogenic acid amygdalin sweroside hyperin rutinforsythoside A phillyrin rhein and glycyrrhizic acid werepurchased from Sigma Aldrich (St Louis MO USA) Thepurity of standard substances was above 98 Ten batches ofLQC were provided by Shijiazhuang Yiling PharmaceuticalCo Ltd (Shijiazhuang China)
22 UPLC Analysis The UPLC analysis was performed on aWaters ACQUITY UPLC instrument (Waters CorporationMA USA) coupled with a binary pump a sample manageran autosampler a column compartment and diode-arraydetector (DAD)The separation of samples was performed ona Waters ACQUITY UPLC BEH C
18(100 times 21mm 17 120583m)
column with the column temperature at 50∘C The analysiswas completed in 30min with a gradient elution of 01formic acid aqueous solution (A) andmethanol (B) at the flowrate of 03mLmin The gradient program was designed as
follows 0ndash11min 5ndash35 B 11ndash18min 35ndash55 B 18ndash22min55ndash75 B 22ndash24min 75ndash90 B 24-25min 90ndash100 Band 25ndash30min 100 B The injection volume was 5120583L Thedetection wavelengths of DAD were set at 210 nm 225 nmand 254 nm
23 UPLC-DAD-QTOF-MSAnalysis TheWaters ACQUITYUPLC instrument (Waters MA USA) coupled with WatersSynapt HDMS G1 (Waters Manchester UK) via an elec-trospray ionization (ESI) interface The UPLC analyticalconditions were the same as the UPLC analysis describedabove The full scan mass spectra data were acquired inpositive and negative ion modes Acquisition parametersare as follows capillary voltage was 3000V for ESI (+)and 2600V for ESI (minus) cone voltage was 45V the ESIsource temperature was 100∘C the desolvation tempera-ture was 350∘C the nitrogen (N
2) was used as desolva-
tion gas at flow rates of 600 Lh for both ESI (+) andESI (minus) and the range of full scan was set at mz 150ndash1000Da The version of analysis software was Mass LynxV41
24 Sample and Standard Solutions Preparation The powderof LQC (04 g) was accurately weighed and extracted with60 methanol-water (vv) solution (20mL) in an ultrasonicwater bath for 30min at room temperature The supernatantsolution was diluted with the same amount of water andthen centrifuged for 10min at 14000 rmin All the obtainedsolutions were filtered through 022 120583m syringe filter beforethe UPLC analysis
Twelve standards were accurately weighed and dissolvedin methanol to obtain stock solutions respectively A mixedstock solution of standards was prepared by adding a suitablevolume of each stock solution to a 5mL flask and dilutedwith 30 methanol-water solution at the concentration of678 120583gmL for salidroside 10965 120583gmL for chlorogenicacid 7764 120583gmL for forsythoside E 10647 120583gmL for cryp-tochlorogenic acid 6257 120583gmL for amygdalin 3196 120583gmLfor sweroside 321 120583gmL for hyperin 85 120583gmL for rutin6734 120583gmL for forsythoside A 4571120583gmL for phillyrin5549 120583gmL for rhein and 8435 120583gmL for glycyrrhizicacid respectively The mixed stock solution was then seri-ally diluted with 30 methanol-water solution to obtainfive appropriate concentrations used for plotting standardcurves The lowest concentration of the mixture stocksolution was further diluted to give a series of differ-ent concentrations for investigating the limits of detection(LODs) and limits of quantification (LOQs) of the 12 chem-ical constituents All solutions were stored at 4∘C untilanalysis
25 Validation of the Quantitative Analysis TheUPLC-DADmethod was evaluated with linearity LOD LOQ precisionstability repeatability and recovery tests The calibrationcurves were constructed with five different concentrations ofchemical markers in triplicate The LODs and LOQs weremeasured under the UPLC analytical conditions at a signal-to-noise (SN) ratio of 3 and 10 respectively For intraday
The Scientific World Journal 3
Table 1 Quantitative results of 12 compounds in LQC extracted by different methods
Content (120583gg)
Methods30a 60 90 60 60 60 60 60 60
30minb 30min 30min 30min 30min 30min 15min 30min 45min1 100c 1 100 1 100 1 50 1 100 1 200 1 100 1 100 1 100
Salidroside 172628 170125 162202 152231 170125 165632 171154 170125 168854Chlorogenic acid 244497 249215 221689 228543 249215 228927 249262 249215 255217Forsythoside E 158393 162078 45191 146209 162078 140253 162701 162078 157963Cryptochlorogenic acid 186298 185164 66750 170305 185164 177179 183789 185164 185795Amygdalin 142411 145539 144256 126893 145539 129811 139597 145539 143138Sweroside 81619 81318 78932 74706 81318 77267 81251 81318 80824Hyperin 13510 15173 16726 14084 15173 14080 15272 15173 15767Rutin 12200 12117 11522 10611 12117 11667 11709 12117 11662Forsythoside A 248460 253634 266179 228576 253634 239635 254387 253634 252110Phillyrin 166026 152145 155159 141019 152145 139012 152301 152145 155152Rhein 80313 110206 137011 93763 110206 93205 95638 110206 105477Glycyrrhizic acid 153049 168043 159437 143740 168043 161999 167481 168043 166542aExtracting solvent 30 60 and 90 methanol-water solutionbUltrasonic time 15min 30min and 45mincExtraction solvent multiples 1 50 1 100 and 1 200 expressed 50 100 and 200 times per gram of sample
and interday precisions test the samples were analyzed bysix repetitive injections within one day and once a day forthree successive days respectively At room temperaturethe stability of sample solution was evaluated by replicateinjection at 0 1 2 4 6 8 10 14 24 and 48 h In order tocheck the repeatability six samples from the same sourcewereinvestigated Accurate amounts of the reference standardswere added to 020 g powder of sample in sextuplicate Theresultant sample solutions were then extracted and quantifiedwith the described method The relative standard deviation(RSD) was used to evaluate the results
3 Results and Discussion
31 Optimization of the Extraction and ChromatographicConditions A single-factor method was used to investigatethe extraction effect of the extraction solvent (30 60and 90 methanol-water solution) extraction solvent ratio(1 50 1 100 and 1 200 (wv)) and extraction time (15min30min and 45min) respectively By analyzing the extractionefficiency 60 methanol-water solution extraction solventratio at 1 100 and 30min of ultrasonic time were selected asthe eventual extraction conditions The results are describedin Table 1
Due to the existence of acidic constituents in samplesolutions formic acid was added into the mobile phasewhich could inhibit the ionization of these acidic ingredi-ents to improve the peak shape The mobile phase systems(methanol-formic acid aqueous solution and acetonitrile-formic acid aqueous solution) and column temperature (40∘Cand 50∘C) were investigated which showed that methanol-01 formic acid aqueous solution as mobile phase with
column temperature at 50∘C could obtain the best chro-matographic peak shape Because the maximum absorptionsof 12 reference compounds were different three detectionwavelengths were finally selected in order to achieve thegoal of high detection sensitivity and little interferenceForsythoside E (peak 7) cryptochlorogenic acid (peak 9)amygdalin (peak 33) and phillyrin (peak 39) had satisfactorysensitivity at 210 nm salidroside (peak 6) chlorogenic acid(peak 8) and rhein (peak 54) at 225 nm and sweroside(peak 13) hyperin (peak 26) rutin (peak 29) forsythosideA (peak 30) and glycyrrhizic acid (peak 57) at 254 nm Thechromatograms are presented in Figure 1
32 UPLC-DAD-QTOF-MS Analysis of Reference Compoundsand LQC Samples As shown in Table 2 a total of 61compounds were unambiguously or tentatively identifiedby comparing the retention times and accurate mass mea-surement with references or literature data These com-pounds were divided into six types according to their struc-tural characteristics including flavonoids phenylpropanoidsanthraquinones triterpenoids iridoids and other types Thestructures of identified compounds are listed in Figure 3Among them twenty-seven compounds were further con-firmed by comparing with standards The total ion chro-matograms are shown in Figure 2
321 Flavonoids Seventeen flavonoids (Figure 3(a)) in LQCincluding flavone aglycones and glycosides were identifiedThey were mainly obtained from Lianqiao Jinyinhua Gan-cao Hongjingtian and Mahuang Amongst them liquiritinapioside (22) ononin (25) hyperin (26) rutin (29) liquirit-igenin (35) isoliquiritin apioside (36) isoliquiritin (37) and
4 The Scientific World Journal
AU 0020406
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
minus06minus04minus02
687933
29
3039
(a)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
AU 00204
0
minus02
6879
29
30
39
54 57
13
(b)
AU
0004008012016
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
6
8
7
9
3329
30
39 54
57
13
26
(c)
AU
0002004006008
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
minus002
(d)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU
00204
minus06minus04minus02
minus1minus08
6
87933 29
30
39
57
26
(e)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU 00204
minus04
minus02
6
8
79
33 29
30
39
54 57
(f)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU
0004008012016
02
minus004
8
9
29
30
39
54
5713
26
(g)
Figure 1 UPLC-DAD chromatograms of standard solution of 210 nm (a) 225 nm (b) 254 nm (c) negative sample solution (d) and samplesolution of 210 nm (e) 225 nm (f) and 254 nm (g)
Time
()
0
100
()
0
100
2 4 6 8 10 12 14 16 18 20 22 24 26 28
Time2 4 6 8 10 12 14 16 18 20 22 24 26 28
1
2
34 5
67 8 9
101112
13
1415 16
1718
19
20
21
22
23
24
25
26
27
28
28
29
30
301317
31
32 33
34
35
3736
38
39
40
39
42
41
434445
46 47 484950
51
5253
54
5556
57
57
58
59
60
61
45
20130402-LHQW MSminus 1 TOF MS ESminusBPI
648e4
20130402-LHQW MS+1 TOF MS ES+
BPI813e4
Figure 2 UPLC-QTOF-MS chromatograms of sample solutionfrom negative ion mode and positive ion mode
formononetin (47) were unambiguously identified via thestandards
In negative and positive ion modes flavone aglyconesmainly gain fragment ions by the reverse Diels-Alder (RDA)reaction and the loss of CO (28Da) The characteris-tic fragmentation behavior of compound 35 is shown inFigure 4(a) with high abundant fragmentation [M+HndashVP (4-vinylphenol)]+ at mz 1370422 The abundance of fragmentions [M+HndashRL (resorcinol)]+ at mz 1470621 and [M+HndashRLndashCO]+ at mz 1190691 is relatively lower Compounds 2440 42 and 46were tentatively identified via comparing theirexact molecular masses MSMS spectra data and retentionbehaviors with literature data [13 26 39] Flavone glycosideshave the similar fragmentation pathways of simultaneous orsuccessive loss of glucose (162Da) rhamnose (146Da) orapiose (132Da) The fragmentation pathway of compound26 was exemplified in Figure 4(b) in negative ion modeCompounds 21 23 31 41 and 53 were tentatively identified
The Scientific World Journal 5
O
O
O
O
OH
OH
O
O
RO
HO
OR1
R1
R3
R2
R2
R4
R5R2OR1O
OR3
OR3
OCH3
21R1 = H R2 = H R3 = Glc 24 R1 = OH R2 = Rha (3rarr1) Glc R3 = OH R4 = H R5 = H 25 R = Glc22 R1 = H R2 = H R3 = Glc (2rarr4) Api 26 R1 = OGlc R2 = H R3 = H R4 = H R5 = OH 47 R = H35 R1 = H R2 = H R3 = H 29 R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = OH R5 = H41R1 = Glc (6rarr1) Rha R2 = OH R3 = CH3 31R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = H R5 = H
40 R1 = OH R2 = H R3 = H R4 = H R5 = OH42 R1 = OH R2 = H R3 = H R4 = H R5 = H53 R1 = H R2 = H R3 = H R4 = OCH3 R5 = OCH3
36 R1 = OH R2 = H R3 = Glc (2rarr4) Api37 R1 = H R2 = OH R3 = Glc46 R1 = H R2 = OH R3 = H
(a)
4 8 9 1618 R = OH
38
27 30 34
56
32 R1 = Glc R2 = H39 R1 = Glc R2 = CH3
O
O
O
O
O
O
O
O
O
OO O O
R
O
O
O
O
O
OO
O O O
OO
OO
O
O
O
O
OO
O
O
O
OO
OO
O
O
O
O
O
OH
OH
OHOH
OH
OHOH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
OHOH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
HO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
H3C
OR2
R1O
CH3
(b)
17R1 = Glc R2 = CH3 R3 = H R4 = H43 R1 = H R2 = CH3 R3 = H R4 = H45 R1 = H R2 = CH3 R3 = OH R4 = Glc49 R1 = H R2 = CH3 R3 = OCH3 R4 = Glc54 R1 = H R2 = COOH R3 = H R4 = H61R1 = H R2 = CH3 R3 = OH R4 = H
O
O
OR1
R3R2
OR4
(c)
Figure 3 Continued
6 The Scientific World Journal
6058 59
5550 52 R1 = GlcA (2rarr1) GlcA R2 = acetoxy57 R1 = GlcA (2rarr1) GlcA R2 = H
OO
OO
O
O
O
O
O
R2H
H
H
O
O
O
O
OO
O
O
O
O
OO
O
O
OO
O
O
O
OOO
OHOH
OH
OH
OH
OH
OH
OH OH
OHOH
OH
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
R1O
(d)
5 R1 = Glc R2 = H 13 2010R1 = Glc R2 = CH3
OH
HO
O O
O
HOO OO
OOH
H OO
OHOH
OHOH HO
HOHO
HOHOHO
OR1
OR2
(e)
1 2 3 6
15
7
11 33 51
O
O
O
O O
OO
OO
O O
OO
OOO
O
OO
N
OO
O
OHOH OH
OHOH
OH
OH
OH
OH
OHOHOH
OHOH
OH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
HOHO
HO HOHO
HOHO
HO
HOHO
HOHOHO
HOHO
HOHO HO
HOHO
HOHOHO
HO
HO
H3C
(f)
Figure 3 Chemical structures of the compounds identified from LQC (except for the 7 isomers)
by comparing the molecular mass and MSMS data withliterature data [25 45]
322 Phenylpropanoids Fourteen phenylpropanoids (Figure3(b)) including phenylpropionic acids lignans and coum-arins were identified in LQC They were mainly obtainedfrom Lianqiao Jinyinhua and Gancao Neochlorogenic acid
(4) chlorogenic acid (8) cryptochlorogenic acid (9) 35-dicaffeoylquinic acid (27) forsythoside A (30) 34-dicaffe-oylquinic acid (34) and phillyrin (39) were unambiguouslycharacterized by comparing with standards
In negative ion mode phenylpropionic acids have thesimilar fragmentation pathways of simultaneous or succes-sive loss of H
2O (18Da) CO (28Da) and CO
2(44Da)
The Scientific World Journal 7
OHO
OH
O
OHO
O
OHH
mz 1470621
mz 1190691
+H
mz 2570659mz 2570659
mz 1370422
2570659
172999816301591319873 1941022
23914082651331
3011509
1370422
1190691
1199948
13800271470621
1650917 16902931810699 2112177
(MSMS)
100 120 140 160 180 200 220 240 260 280 300
()
0
100
()
0
100
mzmz
120 130 140 150 160 170 180 190 200 210
(MS+)
(a)
OHO
OHO
OH
O
OH
OOH
HOOH
OH
mz 3010464
3010464
1370281
3020746
3027005 4652560 62322138921294
4630905
43109651450305 2210378
46409055310850 7371886 11080997 13072520 14077662
(MSMS)
()
0
100(
)
0
100
mz
200 400 600 800 1000 1200 1400mz
200 400 600 800 1000 1200 1400
(MSminus)
(b)
OHHO
OHHOOC
OHOHHO
O
OHHO OH
COOminus
COOminus COOminus
Cminus
mz 1910733mz 1270486
mz 1730583 mz 1090409
mz 930425 mz 850426
minusCO2
minusCO2
minus2H2O minusC2H2
minusCO
Ominus
minusH
minusH2O minusH2O
minusH2O
1910733
8504261270486930425 1090409 1350577
1610264 1730583 19207421938051
3530878
1910558
3510719192060770718303540916
70516854510630 5031459 7081906
(MSMS)
()
0
100
()
0
100
mzmz
80 90 100 110 120 130 140 150 160 170 180 190 200200 250 300 350 400 450 500 550 600 650 700 750 800
(MSminus)
(c)
O OO
O
OH
OHOH
HO
HO
HO
HO
O
O OHOH
mz 4611920 mz 1610328
CH3
6232001
3112033 6211844
6242032
6252064 124749349575357 13186510
1610328
62325071620342 4611920
(MSMS)
()
0
100
()
0
100
200 400 600 800 1000 1200 1400mzmz
100 200 300 400 500 600 700 800 900 1000
(MSminus)
(d)
Figure 4 Continued
8 The Scientific World Journal
3561547
13599643083514
3711826
37218625161718
5792105
1129871 4310958
58021345812158 111342197973569 11814053
(MSMS)
O
O
O
OO
OO
OH
HOHO
OH
[MndashHndashGlc]minus[MndashHndashGlcndashCH3]minus
()
0
100
()
0
100
mz
150 200 250 300 350 400 450 500 550 600 650 700 750mz
200 400 600 800 1000 1200 1400
(MSminus)
mz 3561547
mz 3711826
(e)
2250723
18204771810753
18006601540552
19706952240626
2410669 2690634
2420626 2700607
2690462
1129890 27004855610872 8234322 9694360 14023499
O O
O
O
O O
(MSMS)
()
0
100(
)
0
100
mz
160 180 200 220 240 260 280mz
200 400 600 800 1000 1200 1400
(MSminus)
OH
OH
OHOH OH OH OH
OHOHOH
OH
minusCO2
minusCO2minusCO
minusCO
minusHminusHminusH
minusHminusH
minusH
H3CH3C
H3C H3C
mz 1970695mz 2410669
mz 2250723mz 2690462
mz 1820477mz 1540552
minusCH3minusCO
(f)
4310988
11692794321016
4990790 8632153 11946926 14410397
2690673
2250753
18204861710837
1970771 241071927007752710783 3110845
(MSMS)
O
O
O OH
OHO
HOHO
OH
HO
[MndashHndashGlc]minus[MndashHndashGlcndashCO]minus
[MndashHndashGlcndashCO2]minus
[MndashHndashGlcndashCOndashCO2]minus
[MndashHndashGlcndashCO2ndashCH3ndashCO]minus()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400mz
140 160 180 200 220 240 260 280 300 320
(MSminus)
mz 3711826mz 2690673
CH3
(g)
8214583
35108091130354
19305102890758
645410258340994693705 7594542
8224637
8234628
(MSMS)
H
HO
HO
OOH
O
O
HOOCHO
HOOHO
HOOH
HOOC
[MndashHndash2GlcA]minus[MndashHndashGlcA]minus
[2GlcAndashH]minus
()
0
100
mz
100 200 300 400 500 600 700 800
8213977
1169246 3111153 8203445
8224006
8434348125530939433099 13258577
()
0
100
mz
200 400 600 800 1000 1200 1400
mz 4693705mz 3510809mz 6454102
(MSminus)
(h)
Figure 4 Continued
The Scientific World Journal 9
3751302
21307733761332
75126674731059 8093751 10383667 13123422
1250712
1210637
930423
650486 950155 1260535 151087824011931740320 1999976 2505933
(MSMS)
OH
H
HO
OH
O
O
O
HO
OHOH
OH
[MndashHndashGlc]minus
[14FndashH2O]minus [MndashHndashGlcndash2CO2]minus
14F
minusH2O
()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400
mz
60 80 100 120 140 160 180 200 220 240 260
(MSminus)
minusCO2
minusCO2
[MndashHndashGlcndashCO2ndashH2O]minus
(i)
Figure 4TheMS spectra and fragmentation pathway of compound 35 (a) compound 26 (b) compound 8 (c) compound 30 (d) compound39 (e) compound 61 (f) compound 45 (g) compound 57 (h) and compound 5 (i)
The fragmentation pathway of compound 8 [18] as the rep-resentative of phenylpropionic acids is shown in Figure 4(c)Compound 30 produced [MndashHndashCa (caffeoyl)]minus at mz4611920 and [CaffeicndashHndashH
2O]minus atmz 1610328 as displayed
in Figure 4(d) Lignans primarily generated [M+HCOO]minusin negative ion mode and further elimination of glucose(162Da) produced aglycone As shown in Figure 4(e) com-pound 39 produced characteristic fragments at mz 3711826and mz 3561547 corresponding to [MndashHndashGlc (glucose)]minusand [MndashHndashGlcndashCH
3]minus respectively Compounds 16 18 19
28 32 38 and 56 were tentatively identified on the basisof the exact molecular formulae matching fragmentationinformation and retention behaviors as well as literature data[23 24 33 36 47]
323 Anthraquinones Eight anthraquinones (Figure 3(c)) inLQC were definitely or tentatively identified All of themwere derived from Dahuang Chrysophanol glucoside (17)emodin-8-O-glucoside (45) rhein (54) and emodin (61)were confirmed via comparing with standard substances
The characteristic fragmentation behavior of anthra-quinones was the loss of CO
2(44Da) CH
3(15Da) and
CO (28Da) in negative ion mode Typical compound 61was used to explain the fragmentation pathway of anth-raquinones presented in Figure 4(f) Compound 45 asshown in Figure 4(g) produced characteristic fragments atmz 2690673 mz 2410719 mz 2250753 mz 1970771andmz 1820486 which corresponded to [MndashHndashGlc]minus [MndashHndashGlcndashCO]minus [MndashHndashGlcndashCO
2]minus [MndashHndashCOndashCO
2]minus and
[MndashHndashCO2ndashCH3ndashCO]minus respectively Compounds 12 43
44 and 49 were tentatively identified by comparing theiraccurate molecular masses and MSMS fragment data withliterature data [21 40 41]
324 Triterpenoids Eight triterpenoids (Figure 3(d)) inLQC were unambiguously or tentatively identified All ofthem were derived from Gancao Glycyrrhizic acid (57) wasconfirmed by comparing with standards In negative ion
mode representative compound 57 yielded [MndashHndashH2Ondash
CO2]minus at mz 7594524 [MndashHndashGlcA (glucuronic acid)]minus
at mz 6454102 [MndashHndash2GlcA]minus at mz 4693705 [2GlcAndashH]minus at mz 3510809 and [2GlcAndashHndashH
2OndashCO
2]minus at mz
2890758 as shown in Figure 4(h) Compounds 48 50 5255 58 59 and 60 were tentatively identified by comparingtheir exact molecular masses and MSMS spectral data withthe literature data [51]
325 Iridoids Four iridoids (Figure 3(e)) in LQC includingiridoid glycosides and secoiridoid glycosides were identifiedAll of them were derived from Jinyinhua Loganic acid (5)sweroside (13) and secoxyloganin (20) were unambiguouslyidentified via the standards
The fragment 14F (fragment generated from the fractureof 14 bonds of iridoids) in the negative ion mode was identi-fied as the characteristic fragment ion of iridoid glycosidesMeanwhile iridoid glycosides would lose the functionalgroups such as H
2O (18Da) CO
2(44Da) and glucose
(162Da) Typical compound 5 gave [MndashHndashGlcndashCO2ndashH2O]minus
at mz 1510878 [MndashHndashGlcndash2CO2]minus at mz 1250712 and
[14FndashH2O]minus at mz 930423 [52] as presented in Figure 4(i)
Compound 10 was tentatively identified by comparing itsexact molecular mass and MSMS spectral data with theliterature data [19]
326 Other Types of Compounds Compounds 1 2 11 14 15and 51 were tentatively identified by comparing their exactmolecularmasses andMSMS spectral datawith the literaturedata except gallic acid (3) salidroside (6) forsythoside E (7)and amygdalin (33) which were identified via the standards(Figure 3(f)) [12 53]
33 Methodological Validation of the Quantitative AnalysisAs shown in Table 3 twelve standards were of good lin-earity with high correlation coefficient values over 09993The LODs and LOQs were 0051ndash171 120583gmL and 016ndash569 120583gmL independently Twelve analytes in sample solu-tion were stable at room temperature within 48 h with
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
The Scientific World Journal 3
Table 1 Quantitative results of 12 compounds in LQC extracted by different methods
Content (120583gg)
Methods30a 60 90 60 60 60 60 60 60
30minb 30min 30min 30min 30min 30min 15min 30min 45min1 100c 1 100 1 100 1 50 1 100 1 200 1 100 1 100 1 100
Salidroside 172628 170125 162202 152231 170125 165632 171154 170125 168854Chlorogenic acid 244497 249215 221689 228543 249215 228927 249262 249215 255217Forsythoside E 158393 162078 45191 146209 162078 140253 162701 162078 157963Cryptochlorogenic acid 186298 185164 66750 170305 185164 177179 183789 185164 185795Amygdalin 142411 145539 144256 126893 145539 129811 139597 145539 143138Sweroside 81619 81318 78932 74706 81318 77267 81251 81318 80824Hyperin 13510 15173 16726 14084 15173 14080 15272 15173 15767Rutin 12200 12117 11522 10611 12117 11667 11709 12117 11662Forsythoside A 248460 253634 266179 228576 253634 239635 254387 253634 252110Phillyrin 166026 152145 155159 141019 152145 139012 152301 152145 155152Rhein 80313 110206 137011 93763 110206 93205 95638 110206 105477Glycyrrhizic acid 153049 168043 159437 143740 168043 161999 167481 168043 166542aExtracting solvent 30 60 and 90 methanol-water solutionbUltrasonic time 15min 30min and 45mincExtraction solvent multiples 1 50 1 100 and 1 200 expressed 50 100 and 200 times per gram of sample
and interday precisions test the samples were analyzed bysix repetitive injections within one day and once a day forthree successive days respectively At room temperaturethe stability of sample solution was evaluated by replicateinjection at 0 1 2 4 6 8 10 14 24 and 48 h In order tocheck the repeatability six samples from the same sourcewereinvestigated Accurate amounts of the reference standardswere added to 020 g powder of sample in sextuplicate Theresultant sample solutions were then extracted and quantifiedwith the described method The relative standard deviation(RSD) was used to evaluate the results
3 Results and Discussion
31 Optimization of the Extraction and ChromatographicConditions A single-factor method was used to investigatethe extraction effect of the extraction solvent (30 60and 90 methanol-water solution) extraction solvent ratio(1 50 1 100 and 1 200 (wv)) and extraction time (15min30min and 45min) respectively By analyzing the extractionefficiency 60 methanol-water solution extraction solventratio at 1 100 and 30min of ultrasonic time were selected asthe eventual extraction conditions The results are describedin Table 1
Due to the existence of acidic constituents in samplesolutions formic acid was added into the mobile phasewhich could inhibit the ionization of these acidic ingredi-ents to improve the peak shape The mobile phase systems(methanol-formic acid aqueous solution and acetonitrile-formic acid aqueous solution) and column temperature (40∘Cand 50∘C) were investigated which showed that methanol-01 formic acid aqueous solution as mobile phase with
column temperature at 50∘C could obtain the best chro-matographic peak shape Because the maximum absorptionsof 12 reference compounds were different three detectionwavelengths were finally selected in order to achieve thegoal of high detection sensitivity and little interferenceForsythoside E (peak 7) cryptochlorogenic acid (peak 9)amygdalin (peak 33) and phillyrin (peak 39) had satisfactorysensitivity at 210 nm salidroside (peak 6) chlorogenic acid(peak 8) and rhein (peak 54) at 225 nm and sweroside(peak 13) hyperin (peak 26) rutin (peak 29) forsythosideA (peak 30) and glycyrrhizic acid (peak 57) at 254 nm Thechromatograms are presented in Figure 1
32 UPLC-DAD-QTOF-MS Analysis of Reference Compoundsand LQC Samples As shown in Table 2 a total of 61compounds were unambiguously or tentatively identifiedby comparing the retention times and accurate mass mea-surement with references or literature data These com-pounds were divided into six types according to their struc-tural characteristics including flavonoids phenylpropanoidsanthraquinones triterpenoids iridoids and other types Thestructures of identified compounds are listed in Figure 3Among them twenty-seven compounds were further con-firmed by comparing with standards The total ion chro-matograms are shown in Figure 2
321 Flavonoids Seventeen flavonoids (Figure 3(a)) in LQCincluding flavone aglycones and glycosides were identifiedThey were mainly obtained from Lianqiao Jinyinhua Gan-cao Hongjingtian and Mahuang Amongst them liquiritinapioside (22) ononin (25) hyperin (26) rutin (29) liquirit-igenin (35) isoliquiritin apioside (36) isoliquiritin (37) and
4 The Scientific World Journal
AU 0020406
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
minus06minus04minus02
687933
29
3039
(a)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
AU 00204
0
minus02
6879
29
30
39
54 57
13
(b)
AU
0004008012016
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
6
8
7
9
3329
30
39 54
57
13
26
(c)
AU
0002004006008
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
minus002
(d)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU
00204
minus06minus04minus02
minus1minus08
6
87933 29
30
39
57
26
(e)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU 00204
minus04
minus02
6
8
79
33 29
30
39
54 57
(f)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU
0004008012016
02
minus004
8
9
29
30
39
54
5713
26
(g)
Figure 1 UPLC-DAD chromatograms of standard solution of 210 nm (a) 225 nm (b) 254 nm (c) negative sample solution (d) and samplesolution of 210 nm (e) 225 nm (f) and 254 nm (g)
Time
()
0
100
()
0
100
2 4 6 8 10 12 14 16 18 20 22 24 26 28
Time2 4 6 8 10 12 14 16 18 20 22 24 26 28
1
2
34 5
67 8 9
101112
13
1415 16
1718
19
20
21
22
23
24
25
26
27
28
28
29
30
301317
31
32 33
34
35
3736
38
39
40
39
42
41
434445
46 47 484950
51
5253
54
5556
57
57
58
59
60
61
45
20130402-LHQW MSminus 1 TOF MS ESminusBPI
648e4
20130402-LHQW MS+1 TOF MS ES+
BPI813e4
Figure 2 UPLC-QTOF-MS chromatograms of sample solutionfrom negative ion mode and positive ion mode
formononetin (47) were unambiguously identified via thestandards
In negative and positive ion modes flavone aglyconesmainly gain fragment ions by the reverse Diels-Alder (RDA)reaction and the loss of CO (28Da) The characteris-tic fragmentation behavior of compound 35 is shown inFigure 4(a) with high abundant fragmentation [M+HndashVP (4-vinylphenol)]+ at mz 1370422 The abundance of fragmentions [M+HndashRL (resorcinol)]+ at mz 1470621 and [M+HndashRLndashCO]+ at mz 1190691 is relatively lower Compounds 2440 42 and 46were tentatively identified via comparing theirexact molecular masses MSMS spectra data and retentionbehaviors with literature data [13 26 39] Flavone glycosideshave the similar fragmentation pathways of simultaneous orsuccessive loss of glucose (162Da) rhamnose (146Da) orapiose (132Da) The fragmentation pathway of compound26 was exemplified in Figure 4(b) in negative ion modeCompounds 21 23 31 41 and 53 were tentatively identified
The Scientific World Journal 5
O
O
O
O
OH
OH
O
O
RO
HO
OR1
R1
R3
R2
R2
R4
R5R2OR1O
OR3
OR3
OCH3
21R1 = H R2 = H R3 = Glc 24 R1 = OH R2 = Rha (3rarr1) Glc R3 = OH R4 = H R5 = H 25 R = Glc22 R1 = H R2 = H R3 = Glc (2rarr4) Api 26 R1 = OGlc R2 = H R3 = H R4 = H R5 = OH 47 R = H35 R1 = H R2 = H R3 = H 29 R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = OH R5 = H41R1 = Glc (6rarr1) Rha R2 = OH R3 = CH3 31R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = H R5 = H
40 R1 = OH R2 = H R3 = H R4 = H R5 = OH42 R1 = OH R2 = H R3 = H R4 = H R5 = H53 R1 = H R2 = H R3 = H R4 = OCH3 R5 = OCH3
36 R1 = OH R2 = H R3 = Glc (2rarr4) Api37 R1 = H R2 = OH R3 = Glc46 R1 = H R2 = OH R3 = H
(a)
4 8 9 1618 R = OH
38
27 30 34
56
32 R1 = Glc R2 = H39 R1 = Glc R2 = CH3
O
O
O
O
O
O
O
O
O
OO O O
R
O
O
O
O
O
OO
O O O
OO
OO
O
O
O
O
OO
O
O
O
OO
OO
O
O
O
O
O
OH
OH
OHOH
OH
OHOH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
OHOH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
HO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
H3C
OR2
R1O
CH3
(b)
17R1 = Glc R2 = CH3 R3 = H R4 = H43 R1 = H R2 = CH3 R3 = H R4 = H45 R1 = H R2 = CH3 R3 = OH R4 = Glc49 R1 = H R2 = CH3 R3 = OCH3 R4 = Glc54 R1 = H R2 = COOH R3 = H R4 = H61R1 = H R2 = CH3 R3 = OH R4 = H
O
O
OR1
R3R2
OR4
(c)
Figure 3 Continued
6 The Scientific World Journal
6058 59
5550 52 R1 = GlcA (2rarr1) GlcA R2 = acetoxy57 R1 = GlcA (2rarr1) GlcA R2 = H
OO
OO
O
O
O
O
O
R2H
H
H
O
O
O
O
OO
O
O
O
O
OO
O
O
OO
O
O
O
OOO
OHOH
OH
OH
OH
OH
OH
OH OH
OHOH
OH
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
R1O
(d)
5 R1 = Glc R2 = H 13 2010R1 = Glc R2 = CH3
OH
HO
O O
O
HOO OO
OOH
H OO
OHOH
OHOH HO
HOHO
HOHOHO
OR1
OR2
(e)
1 2 3 6
15
7
11 33 51
O
O
O
O O
OO
OO
O O
OO
OOO
O
OO
N
OO
O
OHOH OH
OHOH
OH
OH
OH
OH
OHOHOH
OHOH
OH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
HOHO
HO HOHO
HOHO
HO
HOHO
HOHOHO
HOHO
HOHO HO
HOHO
HOHOHO
HO
HO
H3C
(f)
Figure 3 Chemical structures of the compounds identified from LQC (except for the 7 isomers)
by comparing the molecular mass and MSMS data withliterature data [25 45]
322 Phenylpropanoids Fourteen phenylpropanoids (Figure3(b)) including phenylpropionic acids lignans and coum-arins were identified in LQC They were mainly obtainedfrom Lianqiao Jinyinhua and Gancao Neochlorogenic acid
(4) chlorogenic acid (8) cryptochlorogenic acid (9) 35-dicaffeoylquinic acid (27) forsythoside A (30) 34-dicaffe-oylquinic acid (34) and phillyrin (39) were unambiguouslycharacterized by comparing with standards
In negative ion mode phenylpropionic acids have thesimilar fragmentation pathways of simultaneous or succes-sive loss of H
2O (18Da) CO (28Da) and CO
2(44Da)
The Scientific World Journal 7
OHO
OH
O
OHO
O
OHH
mz 1470621
mz 1190691
+H
mz 2570659mz 2570659
mz 1370422
2570659
172999816301591319873 1941022
23914082651331
3011509
1370422
1190691
1199948
13800271470621
1650917 16902931810699 2112177
(MSMS)
100 120 140 160 180 200 220 240 260 280 300
()
0
100
()
0
100
mzmz
120 130 140 150 160 170 180 190 200 210
(MS+)
(a)
OHO
OHO
OH
O
OH
OOH
HOOH
OH
mz 3010464
3010464
1370281
3020746
3027005 4652560 62322138921294
4630905
43109651450305 2210378
46409055310850 7371886 11080997 13072520 14077662
(MSMS)
()
0
100(
)
0
100
mz
200 400 600 800 1000 1200 1400mz
200 400 600 800 1000 1200 1400
(MSminus)
(b)
OHHO
OHHOOC
OHOHHO
O
OHHO OH
COOminus
COOminus COOminus
Cminus
mz 1910733mz 1270486
mz 1730583 mz 1090409
mz 930425 mz 850426
minusCO2
minusCO2
minus2H2O minusC2H2
minusCO
Ominus
minusH
minusH2O minusH2O
minusH2O
1910733
8504261270486930425 1090409 1350577
1610264 1730583 19207421938051
3530878
1910558
3510719192060770718303540916
70516854510630 5031459 7081906
(MSMS)
()
0
100
()
0
100
mzmz
80 90 100 110 120 130 140 150 160 170 180 190 200200 250 300 350 400 450 500 550 600 650 700 750 800
(MSminus)
(c)
O OO
O
OH
OHOH
HO
HO
HO
HO
O
O OHOH
mz 4611920 mz 1610328
CH3
6232001
3112033 6211844
6242032
6252064 124749349575357 13186510
1610328
62325071620342 4611920
(MSMS)
()
0
100
()
0
100
200 400 600 800 1000 1200 1400mzmz
100 200 300 400 500 600 700 800 900 1000
(MSminus)
(d)
Figure 4 Continued
8 The Scientific World Journal
3561547
13599643083514
3711826
37218625161718
5792105
1129871 4310958
58021345812158 111342197973569 11814053
(MSMS)
O
O
O
OO
OO
OH
HOHO
OH
[MndashHndashGlc]minus[MndashHndashGlcndashCH3]minus
()
0
100
()
0
100
mz
150 200 250 300 350 400 450 500 550 600 650 700 750mz
200 400 600 800 1000 1200 1400
(MSminus)
mz 3561547
mz 3711826
(e)
2250723
18204771810753
18006601540552
19706952240626
2410669 2690634
2420626 2700607
2690462
1129890 27004855610872 8234322 9694360 14023499
O O
O
O
O O
(MSMS)
()
0
100(
)
0
100
mz
160 180 200 220 240 260 280mz
200 400 600 800 1000 1200 1400
(MSminus)
OH
OH
OHOH OH OH OH
OHOHOH
OH
minusCO2
minusCO2minusCO
minusCO
minusHminusHminusH
minusHminusH
minusH
H3CH3C
H3C H3C
mz 1970695mz 2410669
mz 2250723mz 2690462
mz 1820477mz 1540552
minusCH3minusCO
(f)
4310988
11692794321016
4990790 8632153 11946926 14410397
2690673
2250753
18204861710837
1970771 241071927007752710783 3110845
(MSMS)
O
O
O OH
OHO
HOHO
OH
HO
[MndashHndashGlc]minus[MndashHndashGlcndashCO]minus
[MndashHndashGlcndashCO2]minus
[MndashHndashGlcndashCOndashCO2]minus
[MndashHndashGlcndashCO2ndashCH3ndashCO]minus()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400mz
140 160 180 200 220 240 260 280 300 320
(MSminus)
mz 3711826mz 2690673
CH3
(g)
8214583
35108091130354
19305102890758
645410258340994693705 7594542
8224637
8234628
(MSMS)
H
HO
HO
OOH
O
O
HOOCHO
HOOHO
HOOH
HOOC
[MndashHndash2GlcA]minus[MndashHndashGlcA]minus
[2GlcAndashH]minus
()
0
100
mz
100 200 300 400 500 600 700 800
8213977
1169246 3111153 8203445
8224006
8434348125530939433099 13258577
()
0
100
mz
200 400 600 800 1000 1200 1400
mz 4693705mz 3510809mz 6454102
(MSminus)
(h)
Figure 4 Continued
The Scientific World Journal 9
3751302
21307733761332
75126674731059 8093751 10383667 13123422
1250712
1210637
930423
650486 950155 1260535 151087824011931740320 1999976 2505933
(MSMS)
OH
H
HO
OH
O
O
O
HO
OHOH
OH
[MndashHndashGlc]minus
[14FndashH2O]minus [MndashHndashGlcndash2CO2]minus
14F
minusH2O
()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400
mz
60 80 100 120 140 160 180 200 220 240 260
(MSminus)
minusCO2
minusCO2
[MndashHndashGlcndashCO2ndashH2O]minus
(i)
Figure 4TheMS spectra and fragmentation pathway of compound 35 (a) compound 26 (b) compound 8 (c) compound 30 (d) compound39 (e) compound 61 (f) compound 45 (g) compound 57 (h) and compound 5 (i)
The fragmentation pathway of compound 8 [18] as the rep-resentative of phenylpropionic acids is shown in Figure 4(c)Compound 30 produced [MndashHndashCa (caffeoyl)]minus at mz4611920 and [CaffeicndashHndashH
2O]minus atmz 1610328 as displayed
in Figure 4(d) Lignans primarily generated [M+HCOO]minusin negative ion mode and further elimination of glucose(162Da) produced aglycone As shown in Figure 4(e) com-pound 39 produced characteristic fragments at mz 3711826and mz 3561547 corresponding to [MndashHndashGlc (glucose)]minusand [MndashHndashGlcndashCH
3]minus respectively Compounds 16 18 19
28 32 38 and 56 were tentatively identified on the basisof the exact molecular formulae matching fragmentationinformation and retention behaviors as well as literature data[23 24 33 36 47]
323 Anthraquinones Eight anthraquinones (Figure 3(c)) inLQC were definitely or tentatively identified All of themwere derived from Dahuang Chrysophanol glucoside (17)emodin-8-O-glucoside (45) rhein (54) and emodin (61)were confirmed via comparing with standard substances
The characteristic fragmentation behavior of anthra-quinones was the loss of CO
2(44Da) CH
3(15Da) and
CO (28Da) in negative ion mode Typical compound 61was used to explain the fragmentation pathway of anth-raquinones presented in Figure 4(f) Compound 45 asshown in Figure 4(g) produced characteristic fragments atmz 2690673 mz 2410719 mz 2250753 mz 1970771andmz 1820486 which corresponded to [MndashHndashGlc]minus [MndashHndashGlcndashCO]minus [MndashHndashGlcndashCO
2]minus [MndashHndashCOndashCO
2]minus and
[MndashHndashCO2ndashCH3ndashCO]minus respectively Compounds 12 43
44 and 49 were tentatively identified by comparing theiraccurate molecular masses and MSMS fragment data withliterature data [21 40 41]
324 Triterpenoids Eight triterpenoids (Figure 3(d)) inLQC were unambiguously or tentatively identified All ofthem were derived from Gancao Glycyrrhizic acid (57) wasconfirmed by comparing with standards In negative ion
mode representative compound 57 yielded [MndashHndashH2Ondash
CO2]minus at mz 7594524 [MndashHndashGlcA (glucuronic acid)]minus
at mz 6454102 [MndashHndash2GlcA]minus at mz 4693705 [2GlcAndashH]minus at mz 3510809 and [2GlcAndashHndashH
2OndashCO
2]minus at mz
2890758 as shown in Figure 4(h) Compounds 48 50 5255 58 59 and 60 were tentatively identified by comparingtheir exact molecular masses and MSMS spectral data withthe literature data [51]
325 Iridoids Four iridoids (Figure 3(e)) in LQC includingiridoid glycosides and secoiridoid glycosides were identifiedAll of them were derived from Jinyinhua Loganic acid (5)sweroside (13) and secoxyloganin (20) were unambiguouslyidentified via the standards
The fragment 14F (fragment generated from the fractureof 14 bonds of iridoids) in the negative ion mode was identi-fied as the characteristic fragment ion of iridoid glycosidesMeanwhile iridoid glycosides would lose the functionalgroups such as H
2O (18Da) CO
2(44Da) and glucose
(162Da) Typical compound 5 gave [MndashHndashGlcndashCO2ndashH2O]minus
at mz 1510878 [MndashHndashGlcndash2CO2]minus at mz 1250712 and
[14FndashH2O]minus at mz 930423 [52] as presented in Figure 4(i)
Compound 10 was tentatively identified by comparing itsexact molecular mass and MSMS spectral data with theliterature data [19]
326 Other Types of Compounds Compounds 1 2 11 14 15and 51 were tentatively identified by comparing their exactmolecularmasses andMSMS spectral datawith the literaturedata except gallic acid (3) salidroside (6) forsythoside E (7)and amygdalin (33) which were identified via the standards(Figure 3(f)) [12 53]
33 Methodological Validation of the Quantitative AnalysisAs shown in Table 3 twelve standards were of good lin-earity with high correlation coefficient values over 09993The LODs and LOQs were 0051ndash171 120583gmL and 016ndash569 120583gmL independently Twelve analytes in sample solu-tion were stable at room temperature within 48 h with
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
4 The Scientific World Journal
AU 0020406
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
minus06minus04minus02
687933
29
3039
(a)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
AU 00204
0
minus02
6879
29
30
39
54 57
13
(b)
AU
0004008012016
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
6
8
7
9
3329
30
39 54
57
13
26
(c)
AU
0002004006008
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
minus002
(d)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU
00204
minus06minus04minus02
minus1minus08
6
87933 29
30
39
57
26
(e)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU 00204
minus04
minus02
6
8
79
33 29
30
39
54 57
(f)
(min)2 4 6 8 10 12 14 16 18 20 22 24 26 28 300
AU
0004008012016
02
minus004
8
9
29
30
39
54
5713
26
(g)
Figure 1 UPLC-DAD chromatograms of standard solution of 210 nm (a) 225 nm (b) 254 nm (c) negative sample solution (d) and samplesolution of 210 nm (e) 225 nm (f) and 254 nm (g)
Time
()
0
100
()
0
100
2 4 6 8 10 12 14 16 18 20 22 24 26 28
Time2 4 6 8 10 12 14 16 18 20 22 24 26 28
1
2
34 5
67 8 9
101112
13
1415 16
1718
19
20
21
22
23
24
25
26
27
28
28
29
30
301317
31
32 33
34
35
3736
38
39
40
39
42
41
434445
46 47 484950
51
5253
54
5556
57
57
58
59
60
61
45
20130402-LHQW MSminus 1 TOF MS ESminusBPI
648e4
20130402-LHQW MS+1 TOF MS ES+
BPI813e4
Figure 2 UPLC-QTOF-MS chromatograms of sample solutionfrom negative ion mode and positive ion mode
formononetin (47) were unambiguously identified via thestandards
In negative and positive ion modes flavone aglyconesmainly gain fragment ions by the reverse Diels-Alder (RDA)reaction and the loss of CO (28Da) The characteris-tic fragmentation behavior of compound 35 is shown inFigure 4(a) with high abundant fragmentation [M+HndashVP (4-vinylphenol)]+ at mz 1370422 The abundance of fragmentions [M+HndashRL (resorcinol)]+ at mz 1470621 and [M+HndashRLndashCO]+ at mz 1190691 is relatively lower Compounds 2440 42 and 46were tentatively identified via comparing theirexact molecular masses MSMS spectra data and retentionbehaviors with literature data [13 26 39] Flavone glycosideshave the similar fragmentation pathways of simultaneous orsuccessive loss of glucose (162Da) rhamnose (146Da) orapiose (132Da) The fragmentation pathway of compound26 was exemplified in Figure 4(b) in negative ion modeCompounds 21 23 31 41 and 53 were tentatively identified
The Scientific World Journal 5
O
O
O
O
OH
OH
O
O
RO
HO
OR1
R1
R3
R2
R2
R4
R5R2OR1O
OR3
OR3
OCH3
21R1 = H R2 = H R3 = Glc 24 R1 = OH R2 = Rha (3rarr1) Glc R3 = OH R4 = H R5 = H 25 R = Glc22 R1 = H R2 = H R3 = Glc (2rarr4) Api 26 R1 = OGlc R2 = H R3 = H R4 = H R5 = OH 47 R = H35 R1 = H R2 = H R3 = H 29 R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = OH R5 = H41R1 = Glc (6rarr1) Rha R2 = OH R3 = CH3 31R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = H R5 = H
40 R1 = OH R2 = H R3 = H R4 = H R5 = OH42 R1 = OH R2 = H R3 = H R4 = H R5 = H53 R1 = H R2 = H R3 = H R4 = OCH3 R5 = OCH3
36 R1 = OH R2 = H R3 = Glc (2rarr4) Api37 R1 = H R2 = OH R3 = Glc46 R1 = H R2 = OH R3 = H
(a)
4 8 9 1618 R = OH
38
27 30 34
56
32 R1 = Glc R2 = H39 R1 = Glc R2 = CH3
O
O
O
O
O
O
O
O
O
OO O O
R
O
O
O
O
O
OO
O O O
OO
OO
O
O
O
O
OO
O
O
O
OO
OO
O
O
O
O
O
OH
OH
OHOH
OH
OHOH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
OHOH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
HO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
H3C
OR2
R1O
CH3
(b)
17R1 = Glc R2 = CH3 R3 = H R4 = H43 R1 = H R2 = CH3 R3 = H R4 = H45 R1 = H R2 = CH3 R3 = OH R4 = Glc49 R1 = H R2 = CH3 R3 = OCH3 R4 = Glc54 R1 = H R2 = COOH R3 = H R4 = H61R1 = H R2 = CH3 R3 = OH R4 = H
O
O
OR1
R3R2
OR4
(c)
Figure 3 Continued
6 The Scientific World Journal
6058 59
5550 52 R1 = GlcA (2rarr1) GlcA R2 = acetoxy57 R1 = GlcA (2rarr1) GlcA R2 = H
OO
OO
O
O
O
O
O
R2H
H
H
O
O
O
O
OO
O
O
O
O
OO
O
O
OO
O
O
O
OOO
OHOH
OH
OH
OH
OH
OH
OH OH
OHOH
OH
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
R1O
(d)
5 R1 = Glc R2 = H 13 2010R1 = Glc R2 = CH3
OH
HO
O O
O
HOO OO
OOH
H OO
OHOH
OHOH HO
HOHO
HOHOHO
OR1
OR2
(e)
1 2 3 6
15
7
11 33 51
O
O
O
O O
OO
OO
O O
OO
OOO
O
OO
N
OO
O
OHOH OH
OHOH
OH
OH
OH
OH
OHOHOH
OHOH
OH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
HOHO
HO HOHO
HOHO
HO
HOHO
HOHOHO
HOHO
HOHO HO
HOHO
HOHOHO
HO
HO
H3C
(f)
Figure 3 Chemical structures of the compounds identified from LQC (except for the 7 isomers)
by comparing the molecular mass and MSMS data withliterature data [25 45]
322 Phenylpropanoids Fourteen phenylpropanoids (Figure3(b)) including phenylpropionic acids lignans and coum-arins were identified in LQC They were mainly obtainedfrom Lianqiao Jinyinhua and Gancao Neochlorogenic acid
(4) chlorogenic acid (8) cryptochlorogenic acid (9) 35-dicaffeoylquinic acid (27) forsythoside A (30) 34-dicaffe-oylquinic acid (34) and phillyrin (39) were unambiguouslycharacterized by comparing with standards
In negative ion mode phenylpropionic acids have thesimilar fragmentation pathways of simultaneous or succes-sive loss of H
2O (18Da) CO (28Da) and CO
2(44Da)
The Scientific World Journal 7
OHO
OH
O
OHO
O
OHH
mz 1470621
mz 1190691
+H
mz 2570659mz 2570659
mz 1370422
2570659
172999816301591319873 1941022
23914082651331
3011509
1370422
1190691
1199948
13800271470621
1650917 16902931810699 2112177
(MSMS)
100 120 140 160 180 200 220 240 260 280 300
()
0
100
()
0
100
mzmz
120 130 140 150 160 170 180 190 200 210
(MS+)
(a)
OHO
OHO
OH
O
OH
OOH
HOOH
OH
mz 3010464
3010464
1370281
3020746
3027005 4652560 62322138921294
4630905
43109651450305 2210378
46409055310850 7371886 11080997 13072520 14077662
(MSMS)
()
0
100(
)
0
100
mz
200 400 600 800 1000 1200 1400mz
200 400 600 800 1000 1200 1400
(MSminus)
(b)
OHHO
OHHOOC
OHOHHO
O
OHHO OH
COOminus
COOminus COOminus
Cminus
mz 1910733mz 1270486
mz 1730583 mz 1090409
mz 930425 mz 850426
minusCO2
minusCO2
minus2H2O minusC2H2
minusCO
Ominus
minusH
minusH2O minusH2O
minusH2O
1910733
8504261270486930425 1090409 1350577
1610264 1730583 19207421938051
3530878
1910558
3510719192060770718303540916
70516854510630 5031459 7081906
(MSMS)
()
0
100
()
0
100
mzmz
80 90 100 110 120 130 140 150 160 170 180 190 200200 250 300 350 400 450 500 550 600 650 700 750 800
(MSminus)
(c)
O OO
O
OH
OHOH
HO
HO
HO
HO
O
O OHOH
mz 4611920 mz 1610328
CH3
6232001
3112033 6211844
6242032
6252064 124749349575357 13186510
1610328
62325071620342 4611920
(MSMS)
()
0
100
()
0
100
200 400 600 800 1000 1200 1400mzmz
100 200 300 400 500 600 700 800 900 1000
(MSminus)
(d)
Figure 4 Continued
8 The Scientific World Journal
3561547
13599643083514
3711826
37218625161718
5792105
1129871 4310958
58021345812158 111342197973569 11814053
(MSMS)
O
O
O
OO
OO
OH
HOHO
OH
[MndashHndashGlc]minus[MndashHndashGlcndashCH3]minus
()
0
100
()
0
100
mz
150 200 250 300 350 400 450 500 550 600 650 700 750mz
200 400 600 800 1000 1200 1400
(MSminus)
mz 3561547
mz 3711826
(e)
2250723
18204771810753
18006601540552
19706952240626
2410669 2690634
2420626 2700607
2690462
1129890 27004855610872 8234322 9694360 14023499
O O
O
O
O O
(MSMS)
()
0
100(
)
0
100
mz
160 180 200 220 240 260 280mz
200 400 600 800 1000 1200 1400
(MSminus)
OH
OH
OHOH OH OH OH
OHOHOH
OH
minusCO2
minusCO2minusCO
minusCO
minusHminusHminusH
minusHminusH
minusH
H3CH3C
H3C H3C
mz 1970695mz 2410669
mz 2250723mz 2690462
mz 1820477mz 1540552
minusCH3minusCO
(f)
4310988
11692794321016
4990790 8632153 11946926 14410397
2690673
2250753
18204861710837
1970771 241071927007752710783 3110845
(MSMS)
O
O
O OH
OHO
HOHO
OH
HO
[MndashHndashGlc]minus[MndashHndashGlcndashCO]minus
[MndashHndashGlcndashCO2]minus
[MndashHndashGlcndashCOndashCO2]minus
[MndashHndashGlcndashCO2ndashCH3ndashCO]minus()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400mz
140 160 180 200 220 240 260 280 300 320
(MSminus)
mz 3711826mz 2690673
CH3
(g)
8214583
35108091130354
19305102890758
645410258340994693705 7594542
8224637
8234628
(MSMS)
H
HO
HO
OOH
O
O
HOOCHO
HOOHO
HOOH
HOOC
[MndashHndash2GlcA]minus[MndashHndashGlcA]minus
[2GlcAndashH]minus
()
0
100
mz
100 200 300 400 500 600 700 800
8213977
1169246 3111153 8203445
8224006
8434348125530939433099 13258577
()
0
100
mz
200 400 600 800 1000 1200 1400
mz 4693705mz 3510809mz 6454102
(MSminus)
(h)
Figure 4 Continued
The Scientific World Journal 9
3751302
21307733761332
75126674731059 8093751 10383667 13123422
1250712
1210637
930423
650486 950155 1260535 151087824011931740320 1999976 2505933
(MSMS)
OH
H
HO
OH
O
O
O
HO
OHOH
OH
[MndashHndashGlc]minus
[14FndashH2O]minus [MndashHndashGlcndash2CO2]minus
14F
minusH2O
()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400
mz
60 80 100 120 140 160 180 200 220 240 260
(MSminus)
minusCO2
minusCO2
[MndashHndashGlcndashCO2ndashH2O]minus
(i)
Figure 4TheMS spectra and fragmentation pathway of compound 35 (a) compound 26 (b) compound 8 (c) compound 30 (d) compound39 (e) compound 61 (f) compound 45 (g) compound 57 (h) and compound 5 (i)
The fragmentation pathway of compound 8 [18] as the rep-resentative of phenylpropionic acids is shown in Figure 4(c)Compound 30 produced [MndashHndashCa (caffeoyl)]minus at mz4611920 and [CaffeicndashHndashH
2O]minus atmz 1610328 as displayed
in Figure 4(d) Lignans primarily generated [M+HCOO]minusin negative ion mode and further elimination of glucose(162Da) produced aglycone As shown in Figure 4(e) com-pound 39 produced characteristic fragments at mz 3711826and mz 3561547 corresponding to [MndashHndashGlc (glucose)]minusand [MndashHndashGlcndashCH
3]minus respectively Compounds 16 18 19
28 32 38 and 56 were tentatively identified on the basisof the exact molecular formulae matching fragmentationinformation and retention behaviors as well as literature data[23 24 33 36 47]
323 Anthraquinones Eight anthraquinones (Figure 3(c)) inLQC were definitely or tentatively identified All of themwere derived from Dahuang Chrysophanol glucoside (17)emodin-8-O-glucoside (45) rhein (54) and emodin (61)were confirmed via comparing with standard substances
The characteristic fragmentation behavior of anthra-quinones was the loss of CO
2(44Da) CH
3(15Da) and
CO (28Da) in negative ion mode Typical compound 61was used to explain the fragmentation pathway of anth-raquinones presented in Figure 4(f) Compound 45 asshown in Figure 4(g) produced characteristic fragments atmz 2690673 mz 2410719 mz 2250753 mz 1970771andmz 1820486 which corresponded to [MndashHndashGlc]minus [MndashHndashGlcndashCO]minus [MndashHndashGlcndashCO
2]minus [MndashHndashCOndashCO
2]minus and
[MndashHndashCO2ndashCH3ndashCO]minus respectively Compounds 12 43
44 and 49 were tentatively identified by comparing theiraccurate molecular masses and MSMS fragment data withliterature data [21 40 41]
324 Triterpenoids Eight triterpenoids (Figure 3(d)) inLQC were unambiguously or tentatively identified All ofthem were derived from Gancao Glycyrrhizic acid (57) wasconfirmed by comparing with standards In negative ion
mode representative compound 57 yielded [MndashHndashH2Ondash
CO2]minus at mz 7594524 [MndashHndashGlcA (glucuronic acid)]minus
at mz 6454102 [MndashHndash2GlcA]minus at mz 4693705 [2GlcAndashH]minus at mz 3510809 and [2GlcAndashHndashH
2OndashCO
2]minus at mz
2890758 as shown in Figure 4(h) Compounds 48 50 5255 58 59 and 60 were tentatively identified by comparingtheir exact molecular masses and MSMS spectral data withthe literature data [51]
325 Iridoids Four iridoids (Figure 3(e)) in LQC includingiridoid glycosides and secoiridoid glycosides were identifiedAll of them were derived from Jinyinhua Loganic acid (5)sweroside (13) and secoxyloganin (20) were unambiguouslyidentified via the standards
The fragment 14F (fragment generated from the fractureof 14 bonds of iridoids) in the negative ion mode was identi-fied as the characteristic fragment ion of iridoid glycosidesMeanwhile iridoid glycosides would lose the functionalgroups such as H
2O (18Da) CO
2(44Da) and glucose
(162Da) Typical compound 5 gave [MndashHndashGlcndashCO2ndashH2O]minus
at mz 1510878 [MndashHndashGlcndash2CO2]minus at mz 1250712 and
[14FndashH2O]minus at mz 930423 [52] as presented in Figure 4(i)
Compound 10 was tentatively identified by comparing itsexact molecular mass and MSMS spectral data with theliterature data [19]
326 Other Types of Compounds Compounds 1 2 11 14 15and 51 were tentatively identified by comparing their exactmolecularmasses andMSMS spectral datawith the literaturedata except gallic acid (3) salidroside (6) forsythoside E (7)and amygdalin (33) which were identified via the standards(Figure 3(f)) [12 53]
33 Methodological Validation of the Quantitative AnalysisAs shown in Table 3 twelve standards were of good lin-earity with high correlation coefficient values over 09993The LODs and LOQs were 0051ndash171 120583gmL and 016ndash569 120583gmL independently Twelve analytes in sample solu-tion were stable at room temperature within 48 h with
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
The Scientific World Journal 5
O
O
O
O
OH
OH
O
O
RO
HO
OR1
R1
R3
R2
R2
R4
R5R2OR1O
OR3
OR3
OCH3
21R1 = H R2 = H R3 = Glc 24 R1 = OH R2 = Rha (3rarr1) Glc R3 = OH R4 = H R5 = H 25 R = Glc22 R1 = H R2 = H R3 = Glc (2rarr4) Api 26 R1 = OGlc R2 = H R3 = H R4 = H R5 = OH 47 R = H35 R1 = H R2 = H R3 = H 29 R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = OH R5 = H41R1 = Glc (6rarr1) Rha R2 = OH R3 = CH3 31R1 = OGlc (6rarr1) Rha R2 = H R3 = H R4 = H R5 = H
40 R1 = OH R2 = H R3 = H R4 = H R5 = OH42 R1 = OH R2 = H R3 = H R4 = H R5 = H53 R1 = H R2 = H R3 = H R4 = OCH3 R5 = OCH3
36 R1 = OH R2 = H R3 = Glc (2rarr4) Api37 R1 = H R2 = OH R3 = Glc46 R1 = H R2 = OH R3 = H
(a)
4 8 9 1618 R = OH
38
27 30 34
56
32 R1 = Glc R2 = H39 R1 = Glc R2 = CH3
O
O
O
O
O
O
O
O
O
OO O O
R
O
O
O
O
O
OO
O O O
OO
OO
O
O
O
O
OO
O
O
O
OO
OO
O
O
O
O
O
OH
OH
OHOH
OH
OHOH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
OHOH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
OH
HO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
HOHO
HO
HO
HO
HOHO
H3C
OR2
R1O
CH3
(b)
17R1 = Glc R2 = CH3 R3 = H R4 = H43 R1 = H R2 = CH3 R3 = H R4 = H45 R1 = H R2 = CH3 R3 = OH R4 = Glc49 R1 = H R2 = CH3 R3 = OCH3 R4 = Glc54 R1 = H R2 = COOH R3 = H R4 = H61R1 = H R2 = CH3 R3 = OH R4 = H
O
O
OR1
R3R2
OR4
(c)
Figure 3 Continued
6 The Scientific World Journal
6058 59
5550 52 R1 = GlcA (2rarr1) GlcA R2 = acetoxy57 R1 = GlcA (2rarr1) GlcA R2 = H
OO
OO
O
O
O
O
O
R2H
H
H
O
O
O
O
OO
O
O
O
O
OO
O
O
OO
O
O
O
OOO
OHOH
OH
OH
OH
OH
OH
OH OH
OHOH
OH
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
R1O
(d)
5 R1 = Glc R2 = H 13 2010R1 = Glc R2 = CH3
OH
HO
O O
O
HOO OO
OOH
H OO
OHOH
OHOH HO
HOHO
HOHOHO
OR1
OR2
(e)
1 2 3 6
15
7
11 33 51
O
O
O
O O
OO
OO
O O
OO
OOO
O
OO
N
OO
O
OHOH OH
OHOH
OH
OH
OH
OH
OHOHOH
OHOH
OH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
HOHO
HO HOHO
HOHO
HO
HOHO
HOHOHO
HOHO
HOHO HO
HOHO
HOHOHO
HO
HO
H3C
(f)
Figure 3 Chemical structures of the compounds identified from LQC (except for the 7 isomers)
by comparing the molecular mass and MSMS data withliterature data [25 45]
322 Phenylpropanoids Fourteen phenylpropanoids (Figure3(b)) including phenylpropionic acids lignans and coum-arins were identified in LQC They were mainly obtainedfrom Lianqiao Jinyinhua and Gancao Neochlorogenic acid
(4) chlorogenic acid (8) cryptochlorogenic acid (9) 35-dicaffeoylquinic acid (27) forsythoside A (30) 34-dicaffe-oylquinic acid (34) and phillyrin (39) were unambiguouslycharacterized by comparing with standards
In negative ion mode phenylpropionic acids have thesimilar fragmentation pathways of simultaneous or succes-sive loss of H
2O (18Da) CO (28Da) and CO
2(44Da)
The Scientific World Journal 7
OHO
OH
O
OHO
O
OHH
mz 1470621
mz 1190691
+H
mz 2570659mz 2570659
mz 1370422
2570659
172999816301591319873 1941022
23914082651331
3011509
1370422
1190691
1199948
13800271470621
1650917 16902931810699 2112177
(MSMS)
100 120 140 160 180 200 220 240 260 280 300
()
0
100
()
0
100
mzmz
120 130 140 150 160 170 180 190 200 210
(MS+)
(a)
OHO
OHO
OH
O
OH
OOH
HOOH
OH
mz 3010464
3010464
1370281
3020746
3027005 4652560 62322138921294
4630905
43109651450305 2210378
46409055310850 7371886 11080997 13072520 14077662
(MSMS)
()
0
100(
)
0
100
mz
200 400 600 800 1000 1200 1400mz
200 400 600 800 1000 1200 1400
(MSminus)
(b)
OHHO
OHHOOC
OHOHHO
O
OHHO OH
COOminus
COOminus COOminus
Cminus
mz 1910733mz 1270486
mz 1730583 mz 1090409
mz 930425 mz 850426
minusCO2
minusCO2
minus2H2O minusC2H2
minusCO
Ominus
minusH
minusH2O minusH2O
minusH2O
1910733
8504261270486930425 1090409 1350577
1610264 1730583 19207421938051
3530878
1910558
3510719192060770718303540916
70516854510630 5031459 7081906
(MSMS)
()
0
100
()
0
100
mzmz
80 90 100 110 120 130 140 150 160 170 180 190 200200 250 300 350 400 450 500 550 600 650 700 750 800
(MSminus)
(c)
O OO
O
OH
OHOH
HO
HO
HO
HO
O
O OHOH
mz 4611920 mz 1610328
CH3
6232001
3112033 6211844
6242032
6252064 124749349575357 13186510
1610328
62325071620342 4611920
(MSMS)
()
0
100
()
0
100
200 400 600 800 1000 1200 1400mzmz
100 200 300 400 500 600 700 800 900 1000
(MSminus)
(d)
Figure 4 Continued
8 The Scientific World Journal
3561547
13599643083514
3711826
37218625161718
5792105
1129871 4310958
58021345812158 111342197973569 11814053
(MSMS)
O
O
O
OO
OO
OH
HOHO
OH
[MndashHndashGlc]minus[MndashHndashGlcndashCH3]minus
()
0
100
()
0
100
mz
150 200 250 300 350 400 450 500 550 600 650 700 750mz
200 400 600 800 1000 1200 1400
(MSminus)
mz 3561547
mz 3711826
(e)
2250723
18204771810753
18006601540552
19706952240626
2410669 2690634
2420626 2700607
2690462
1129890 27004855610872 8234322 9694360 14023499
O O
O
O
O O
(MSMS)
()
0
100(
)
0
100
mz
160 180 200 220 240 260 280mz
200 400 600 800 1000 1200 1400
(MSminus)
OH
OH
OHOH OH OH OH
OHOHOH
OH
minusCO2
minusCO2minusCO
minusCO
minusHminusHminusH
minusHminusH
minusH
H3CH3C
H3C H3C
mz 1970695mz 2410669
mz 2250723mz 2690462
mz 1820477mz 1540552
minusCH3minusCO
(f)
4310988
11692794321016
4990790 8632153 11946926 14410397
2690673
2250753
18204861710837
1970771 241071927007752710783 3110845
(MSMS)
O
O
O OH
OHO
HOHO
OH
HO
[MndashHndashGlc]minus[MndashHndashGlcndashCO]minus
[MndashHndashGlcndashCO2]minus
[MndashHndashGlcndashCOndashCO2]minus
[MndashHndashGlcndashCO2ndashCH3ndashCO]minus()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400mz
140 160 180 200 220 240 260 280 300 320
(MSminus)
mz 3711826mz 2690673
CH3
(g)
8214583
35108091130354
19305102890758
645410258340994693705 7594542
8224637
8234628
(MSMS)
H
HO
HO
OOH
O
O
HOOCHO
HOOHO
HOOH
HOOC
[MndashHndash2GlcA]minus[MndashHndashGlcA]minus
[2GlcAndashH]minus
()
0
100
mz
100 200 300 400 500 600 700 800
8213977
1169246 3111153 8203445
8224006
8434348125530939433099 13258577
()
0
100
mz
200 400 600 800 1000 1200 1400
mz 4693705mz 3510809mz 6454102
(MSminus)
(h)
Figure 4 Continued
The Scientific World Journal 9
3751302
21307733761332
75126674731059 8093751 10383667 13123422
1250712
1210637
930423
650486 950155 1260535 151087824011931740320 1999976 2505933
(MSMS)
OH
H
HO
OH
O
O
O
HO
OHOH
OH
[MndashHndashGlc]minus
[14FndashH2O]minus [MndashHndashGlcndash2CO2]minus
14F
minusH2O
()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400
mz
60 80 100 120 140 160 180 200 220 240 260
(MSminus)
minusCO2
minusCO2
[MndashHndashGlcndashCO2ndashH2O]minus
(i)
Figure 4TheMS spectra and fragmentation pathway of compound 35 (a) compound 26 (b) compound 8 (c) compound 30 (d) compound39 (e) compound 61 (f) compound 45 (g) compound 57 (h) and compound 5 (i)
The fragmentation pathway of compound 8 [18] as the rep-resentative of phenylpropionic acids is shown in Figure 4(c)Compound 30 produced [MndashHndashCa (caffeoyl)]minus at mz4611920 and [CaffeicndashHndashH
2O]minus atmz 1610328 as displayed
in Figure 4(d) Lignans primarily generated [M+HCOO]minusin negative ion mode and further elimination of glucose(162Da) produced aglycone As shown in Figure 4(e) com-pound 39 produced characteristic fragments at mz 3711826and mz 3561547 corresponding to [MndashHndashGlc (glucose)]minusand [MndashHndashGlcndashCH
3]minus respectively Compounds 16 18 19
28 32 38 and 56 were tentatively identified on the basisof the exact molecular formulae matching fragmentationinformation and retention behaviors as well as literature data[23 24 33 36 47]
323 Anthraquinones Eight anthraquinones (Figure 3(c)) inLQC were definitely or tentatively identified All of themwere derived from Dahuang Chrysophanol glucoside (17)emodin-8-O-glucoside (45) rhein (54) and emodin (61)were confirmed via comparing with standard substances
The characteristic fragmentation behavior of anthra-quinones was the loss of CO
2(44Da) CH
3(15Da) and
CO (28Da) in negative ion mode Typical compound 61was used to explain the fragmentation pathway of anth-raquinones presented in Figure 4(f) Compound 45 asshown in Figure 4(g) produced characteristic fragments atmz 2690673 mz 2410719 mz 2250753 mz 1970771andmz 1820486 which corresponded to [MndashHndashGlc]minus [MndashHndashGlcndashCO]minus [MndashHndashGlcndashCO
2]minus [MndashHndashCOndashCO
2]minus and
[MndashHndashCO2ndashCH3ndashCO]minus respectively Compounds 12 43
44 and 49 were tentatively identified by comparing theiraccurate molecular masses and MSMS fragment data withliterature data [21 40 41]
324 Triterpenoids Eight triterpenoids (Figure 3(d)) inLQC were unambiguously or tentatively identified All ofthem were derived from Gancao Glycyrrhizic acid (57) wasconfirmed by comparing with standards In negative ion
mode representative compound 57 yielded [MndashHndashH2Ondash
CO2]minus at mz 7594524 [MndashHndashGlcA (glucuronic acid)]minus
at mz 6454102 [MndashHndash2GlcA]minus at mz 4693705 [2GlcAndashH]minus at mz 3510809 and [2GlcAndashHndashH
2OndashCO
2]minus at mz
2890758 as shown in Figure 4(h) Compounds 48 50 5255 58 59 and 60 were tentatively identified by comparingtheir exact molecular masses and MSMS spectral data withthe literature data [51]
325 Iridoids Four iridoids (Figure 3(e)) in LQC includingiridoid glycosides and secoiridoid glycosides were identifiedAll of them were derived from Jinyinhua Loganic acid (5)sweroside (13) and secoxyloganin (20) were unambiguouslyidentified via the standards
The fragment 14F (fragment generated from the fractureof 14 bonds of iridoids) in the negative ion mode was identi-fied as the characteristic fragment ion of iridoid glycosidesMeanwhile iridoid glycosides would lose the functionalgroups such as H
2O (18Da) CO
2(44Da) and glucose
(162Da) Typical compound 5 gave [MndashHndashGlcndashCO2ndashH2O]minus
at mz 1510878 [MndashHndashGlcndash2CO2]minus at mz 1250712 and
[14FndashH2O]minus at mz 930423 [52] as presented in Figure 4(i)
Compound 10 was tentatively identified by comparing itsexact molecular mass and MSMS spectral data with theliterature data [19]
326 Other Types of Compounds Compounds 1 2 11 14 15and 51 were tentatively identified by comparing their exactmolecularmasses andMSMS spectral datawith the literaturedata except gallic acid (3) salidroside (6) forsythoside E (7)and amygdalin (33) which were identified via the standards(Figure 3(f)) [12 53]
33 Methodological Validation of the Quantitative AnalysisAs shown in Table 3 twelve standards were of good lin-earity with high correlation coefficient values over 09993The LODs and LOQs were 0051ndash171 120583gmL and 016ndash569 120583gmL independently Twelve analytes in sample solu-tion were stable at room temperature within 48 h with
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
6 The Scientific World Journal
6058 59
5550 52 R1 = GlcA (2rarr1) GlcA R2 = acetoxy57 R1 = GlcA (2rarr1) GlcA R2 = H
OO
OO
O
O
O
O
O
R2H
H
H
O
O
O
O
OO
O
O
O
O
OO
O
O
OO
O
O
O
OOO
OHOH
OH
OH
OH
OH
OH
OH OH
OHOH
OH
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOHO
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
HOOC
R1O
(d)
5 R1 = Glc R2 = H 13 2010R1 = Glc R2 = CH3
OH
HO
O O
O
HOO OO
OOH
H OO
OHOH
OHOH HO
HOHO
HOHOHO
OR1
OR2
(e)
1 2 3 6
15
7
11 33 51
O
O
O
O O
OO
OO
O O
OO
OOO
O
OO
N
OO
O
OHOH OH
OHOH
OH
OH
OH
OH
OHOHOH
OHOH
OH
OH
OH
OH
OH
OH
OHOH
OH
OHOH
HOHO
HO HOHO
HOHO
HO
HOHO
HOHOHO
HOHO
HOHO HO
HOHO
HOHOHO
HO
HO
H3C
(f)
Figure 3 Chemical structures of the compounds identified from LQC (except for the 7 isomers)
by comparing the molecular mass and MSMS data withliterature data [25 45]
322 Phenylpropanoids Fourteen phenylpropanoids (Figure3(b)) including phenylpropionic acids lignans and coum-arins were identified in LQC They were mainly obtainedfrom Lianqiao Jinyinhua and Gancao Neochlorogenic acid
(4) chlorogenic acid (8) cryptochlorogenic acid (9) 35-dicaffeoylquinic acid (27) forsythoside A (30) 34-dicaffe-oylquinic acid (34) and phillyrin (39) were unambiguouslycharacterized by comparing with standards
In negative ion mode phenylpropionic acids have thesimilar fragmentation pathways of simultaneous or succes-sive loss of H
2O (18Da) CO (28Da) and CO
2(44Da)
The Scientific World Journal 7
OHO
OH
O
OHO
O
OHH
mz 1470621
mz 1190691
+H
mz 2570659mz 2570659
mz 1370422
2570659
172999816301591319873 1941022
23914082651331
3011509
1370422
1190691
1199948
13800271470621
1650917 16902931810699 2112177
(MSMS)
100 120 140 160 180 200 220 240 260 280 300
()
0
100
()
0
100
mzmz
120 130 140 150 160 170 180 190 200 210
(MS+)
(a)
OHO
OHO
OH
O
OH
OOH
HOOH
OH
mz 3010464
3010464
1370281
3020746
3027005 4652560 62322138921294
4630905
43109651450305 2210378
46409055310850 7371886 11080997 13072520 14077662
(MSMS)
()
0
100(
)
0
100
mz
200 400 600 800 1000 1200 1400mz
200 400 600 800 1000 1200 1400
(MSminus)
(b)
OHHO
OHHOOC
OHOHHO
O
OHHO OH
COOminus
COOminus COOminus
Cminus
mz 1910733mz 1270486
mz 1730583 mz 1090409
mz 930425 mz 850426
minusCO2
minusCO2
minus2H2O minusC2H2
minusCO
Ominus
minusH
minusH2O minusH2O
minusH2O
1910733
8504261270486930425 1090409 1350577
1610264 1730583 19207421938051
3530878
1910558
3510719192060770718303540916
70516854510630 5031459 7081906
(MSMS)
()
0
100
()
0
100
mzmz
80 90 100 110 120 130 140 150 160 170 180 190 200200 250 300 350 400 450 500 550 600 650 700 750 800
(MSminus)
(c)
O OO
O
OH
OHOH
HO
HO
HO
HO
O
O OHOH
mz 4611920 mz 1610328
CH3
6232001
3112033 6211844
6242032
6252064 124749349575357 13186510
1610328
62325071620342 4611920
(MSMS)
()
0
100
()
0
100
200 400 600 800 1000 1200 1400mzmz
100 200 300 400 500 600 700 800 900 1000
(MSminus)
(d)
Figure 4 Continued
8 The Scientific World Journal
3561547
13599643083514
3711826
37218625161718
5792105
1129871 4310958
58021345812158 111342197973569 11814053
(MSMS)
O
O
O
OO
OO
OH
HOHO
OH
[MndashHndashGlc]minus[MndashHndashGlcndashCH3]minus
()
0
100
()
0
100
mz
150 200 250 300 350 400 450 500 550 600 650 700 750mz
200 400 600 800 1000 1200 1400
(MSminus)
mz 3561547
mz 3711826
(e)
2250723
18204771810753
18006601540552
19706952240626
2410669 2690634
2420626 2700607
2690462
1129890 27004855610872 8234322 9694360 14023499
O O
O
O
O O
(MSMS)
()
0
100(
)
0
100
mz
160 180 200 220 240 260 280mz
200 400 600 800 1000 1200 1400
(MSminus)
OH
OH
OHOH OH OH OH
OHOHOH
OH
minusCO2
minusCO2minusCO
minusCO
minusHminusHminusH
minusHminusH
minusH
H3CH3C
H3C H3C
mz 1970695mz 2410669
mz 2250723mz 2690462
mz 1820477mz 1540552
minusCH3minusCO
(f)
4310988
11692794321016
4990790 8632153 11946926 14410397
2690673
2250753
18204861710837
1970771 241071927007752710783 3110845
(MSMS)
O
O
O OH
OHO
HOHO
OH
HO
[MndashHndashGlc]minus[MndashHndashGlcndashCO]minus
[MndashHndashGlcndashCO2]minus
[MndashHndashGlcndashCOndashCO2]minus
[MndashHndashGlcndashCO2ndashCH3ndashCO]minus()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400mz
140 160 180 200 220 240 260 280 300 320
(MSminus)
mz 3711826mz 2690673
CH3
(g)
8214583
35108091130354
19305102890758
645410258340994693705 7594542
8224637
8234628
(MSMS)
H
HO
HO
OOH
O
O
HOOCHO
HOOHO
HOOH
HOOC
[MndashHndash2GlcA]minus[MndashHndashGlcA]minus
[2GlcAndashH]minus
()
0
100
mz
100 200 300 400 500 600 700 800
8213977
1169246 3111153 8203445
8224006
8434348125530939433099 13258577
()
0
100
mz
200 400 600 800 1000 1200 1400
mz 4693705mz 3510809mz 6454102
(MSminus)
(h)
Figure 4 Continued
The Scientific World Journal 9
3751302
21307733761332
75126674731059 8093751 10383667 13123422
1250712
1210637
930423
650486 950155 1260535 151087824011931740320 1999976 2505933
(MSMS)
OH
H
HO
OH
O
O
O
HO
OHOH
OH
[MndashHndashGlc]minus
[14FndashH2O]minus [MndashHndashGlcndash2CO2]minus
14F
minusH2O
()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400
mz
60 80 100 120 140 160 180 200 220 240 260
(MSminus)
minusCO2
minusCO2
[MndashHndashGlcndashCO2ndashH2O]minus
(i)
Figure 4TheMS spectra and fragmentation pathway of compound 35 (a) compound 26 (b) compound 8 (c) compound 30 (d) compound39 (e) compound 61 (f) compound 45 (g) compound 57 (h) and compound 5 (i)
The fragmentation pathway of compound 8 [18] as the rep-resentative of phenylpropionic acids is shown in Figure 4(c)Compound 30 produced [MndashHndashCa (caffeoyl)]minus at mz4611920 and [CaffeicndashHndashH
2O]minus atmz 1610328 as displayed
in Figure 4(d) Lignans primarily generated [M+HCOO]minusin negative ion mode and further elimination of glucose(162Da) produced aglycone As shown in Figure 4(e) com-pound 39 produced characteristic fragments at mz 3711826and mz 3561547 corresponding to [MndashHndashGlc (glucose)]minusand [MndashHndashGlcndashCH
3]minus respectively Compounds 16 18 19
28 32 38 and 56 were tentatively identified on the basisof the exact molecular formulae matching fragmentationinformation and retention behaviors as well as literature data[23 24 33 36 47]
323 Anthraquinones Eight anthraquinones (Figure 3(c)) inLQC were definitely or tentatively identified All of themwere derived from Dahuang Chrysophanol glucoside (17)emodin-8-O-glucoside (45) rhein (54) and emodin (61)were confirmed via comparing with standard substances
The characteristic fragmentation behavior of anthra-quinones was the loss of CO
2(44Da) CH
3(15Da) and
CO (28Da) in negative ion mode Typical compound 61was used to explain the fragmentation pathway of anth-raquinones presented in Figure 4(f) Compound 45 asshown in Figure 4(g) produced characteristic fragments atmz 2690673 mz 2410719 mz 2250753 mz 1970771andmz 1820486 which corresponded to [MndashHndashGlc]minus [MndashHndashGlcndashCO]minus [MndashHndashGlcndashCO
2]minus [MndashHndashCOndashCO
2]minus and
[MndashHndashCO2ndashCH3ndashCO]minus respectively Compounds 12 43
44 and 49 were tentatively identified by comparing theiraccurate molecular masses and MSMS fragment data withliterature data [21 40 41]
324 Triterpenoids Eight triterpenoids (Figure 3(d)) inLQC were unambiguously or tentatively identified All ofthem were derived from Gancao Glycyrrhizic acid (57) wasconfirmed by comparing with standards In negative ion
mode representative compound 57 yielded [MndashHndashH2Ondash
CO2]minus at mz 7594524 [MndashHndashGlcA (glucuronic acid)]minus
at mz 6454102 [MndashHndash2GlcA]minus at mz 4693705 [2GlcAndashH]minus at mz 3510809 and [2GlcAndashHndashH
2OndashCO
2]minus at mz
2890758 as shown in Figure 4(h) Compounds 48 50 5255 58 59 and 60 were tentatively identified by comparingtheir exact molecular masses and MSMS spectral data withthe literature data [51]
325 Iridoids Four iridoids (Figure 3(e)) in LQC includingiridoid glycosides and secoiridoid glycosides were identifiedAll of them were derived from Jinyinhua Loganic acid (5)sweroside (13) and secoxyloganin (20) were unambiguouslyidentified via the standards
The fragment 14F (fragment generated from the fractureof 14 bonds of iridoids) in the negative ion mode was identi-fied as the characteristic fragment ion of iridoid glycosidesMeanwhile iridoid glycosides would lose the functionalgroups such as H
2O (18Da) CO
2(44Da) and glucose
(162Da) Typical compound 5 gave [MndashHndashGlcndashCO2ndashH2O]minus
at mz 1510878 [MndashHndashGlcndash2CO2]minus at mz 1250712 and
[14FndashH2O]minus at mz 930423 [52] as presented in Figure 4(i)
Compound 10 was tentatively identified by comparing itsexact molecular mass and MSMS spectral data with theliterature data [19]
326 Other Types of Compounds Compounds 1 2 11 14 15and 51 were tentatively identified by comparing their exactmolecularmasses andMSMS spectral datawith the literaturedata except gallic acid (3) salidroside (6) forsythoside E (7)and amygdalin (33) which were identified via the standards(Figure 3(f)) [12 53]
33 Methodological Validation of the Quantitative AnalysisAs shown in Table 3 twelve standards were of good lin-earity with high correlation coefficient values over 09993The LODs and LOQs were 0051ndash171 120583gmL and 016ndash569 120583gmL independently Twelve analytes in sample solu-tion were stable at room temperature within 48 h with
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
The Scientific World Journal 7
OHO
OH
O
OHO
O
OHH
mz 1470621
mz 1190691
+H
mz 2570659mz 2570659
mz 1370422
2570659
172999816301591319873 1941022
23914082651331
3011509
1370422
1190691
1199948
13800271470621
1650917 16902931810699 2112177
(MSMS)
100 120 140 160 180 200 220 240 260 280 300
()
0
100
()
0
100
mzmz
120 130 140 150 160 170 180 190 200 210
(MS+)
(a)
OHO
OHO
OH
O
OH
OOH
HOOH
OH
mz 3010464
3010464
1370281
3020746
3027005 4652560 62322138921294
4630905
43109651450305 2210378
46409055310850 7371886 11080997 13072520 14077662
(MSMS)
()
0
100(
)
0
100
mz
200 400 600 800 1000 1200 1400mz
200 400 600 800 1000 1200 1400
(MSminus)
(b)
OHHO
OHHOOC
OHOHHO
O
OHHO OH
COOminus
COOminus COOminus
Cminus
mz 1910733mz 1270486
mz 1730583 mz 1090409
mz 930425 mz 850426
minusCO2
minusCO2
minus2H2O minusC2H2
minusCO
Ominus
minusH
minusH2O minusH2O
minusH2O
1910733
8504261270486930425 1090409 1350577
1610264 1730583 19207421938051
3530878
1910558
3510719192060770718303540916
70516854510630 5031459 7081906
(MSMS)
()
0
100
()
0
100
mzmz
80 90 100 110 120 130 140 150 160 170 180 190 200200 250 300 350 400 450 500 550 600 650 700 750 800
(MSminus)
(c)
O OO
O
OH
OHOH
HO
HO
HO
HO
O
O OHOH
mz 4611920 mz 1610328
CH3
6232001
3112033 6211844
6242032
6252064 124749349575357 13186510
1610328
62325071620342 4611920
(MSMS)
()
0
100
()
0
100
200 400 600 800 1000 1200 1400mzmz
100 200 300 400 500 600 700 800 900 1000
(MSminus)
(d)
Figure 4 Continued
8 The Scientific World Journal
3561547
13599643083514
3711826
37218625161718
5792105
1129871 4310958
58021345812158 111342197973569 11814053
(MSMS)
O
O
O
OO
OO
OH
HOHO
OH
[MndashHndashGlc]minus[MndashHndashGlcndashCH3]minus
()
0
100
()
0
100
mz
150 200 250 300 350 400 450 500 550 600 650 700 750mz
200 400 600 800 1000 1200 1400
(MSminus)
mz 3561547
mz 3711826
(e)
2250723
18204771810753
18006601540552
19706952240626
2410669 2690634
2420626 2700607
2690462
1129890 27004855610872 8234322 9694360 14023499
O O
O
O
O O
(MSMS)
()
0
100(
)
0
100
mz
160 180 200 220 240 260 280mz
200 400 600 800 1000 1200 1400
(MSminus)
OH
OH
OHOH OH OH OH
OHOHOH
OH
minusCO2
minusCO2minusCO
minusCO
minusHminusHminusH
minusHminusH
minusH
H3CH3C
H3C H3C
mz 1970695mz 2410669
mz 2250723mz 2690462
mz 1820477mz 1540552
minusCH3minusCO
(f)
4310988
11692794321016
4990790 8632153 11946926 14410397
2690673
2250753
18204861710837
1970771 241071927007752710783 3110845
(MSMS)
O
O
O OH
OHO
HOHO
OH
HO
[MndashHndashGlc]minus[MndashHndashGlcndashCO]minus
[MndashHndashGlcndashCO2]minus
[MndashHndashGlcndashCOndashCO2]minus
[MndashHndashGlcndashCO2ndashCH3ndashCO]minus()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400mz
140 160 180 200 220 240 260 280 300 320
(MSminus)
mz 3711826mz 2690673
CH3
(g)
8214583
35108091130354
19305102890758
645410258340994693705 7594542
8224637
8234628
(MSMS)
H
HO
HO
OOH
O
O
HOOCHO
HOOHO
HOOH
HOOC
[MndashHndash2GlcA]minus[MndashHndashGlcA]minus
[2GlcAndashH]minus
()
0
100
mz
100 200 300 400 500 600 700 800
8213977
1169246 3111153 8203445
8224006
8434348125530939433099 13258577
()
0
100
mz
200 400 600 800 1000 1200 1400
mz 4693705mz 3510809mz 6454102
(MSminus)
(h)
Figure 4 Continued
The Scientific World Journal 9
3751302
21307733761332
75126674731059 8093751 10383667 13123422
1250712
1210637
930423
650486 950155 1260535 151087824011931740320 1999976 2505933
(MSMS)
OH
H
HO
OH
O
O
O
HO
OHOH
OH
[MndashHndashGlc]minus
[14FndashH2O]minus [MndashHndashGlcndash2CO2]minus
14F
minusH2O
()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400
mz
60 80 100 120 140 160 180 200 220 240 260
(MSminus)
minusCO2
minusCO2
[MndashHndashGlcndashCO2ndashH2O]minus
(i)
Figure 4TheMS spectra and fragmentation pathway of compound 35 (a) compound 26 (b) compound 8 (c) compound 30 (d) compound39 (e) compound 61 (f) compound 45 (g) compound 57 (h) and compound 5 (i)
The fragmentation pathway of compound 8 [18] as the rep-resentative of phenylpropionic acids is shown in Figure 4(c)Compound 30 produced [MndashHndashCa (caffeoyl)]minus at mz4611920 and [CaffeicndashHndashH
2O]minus atmz 1610328 as displayed
in Figure 4(d) Lignans primarily generated [M+HCOO]minusin negative ion mode and further elimination of glucose(162Da) produced aglycone As shown in Figure 4(e) com-pound 39 produced characteristic fragments at mz 3711826and mz 3561547 corresponding to [MndashHndashGlc (glucose)]minusand [MndashHndashGlcndashCH
3]minus respectively Compounds 16 18 19
28 32 38 and 56 were tentatively identified on the basisof the exact molecular formulae matching fragmentationinformation and retention behaviors as well as literature data[23 24 33 36 47]
323 Anthraquinones Eight anthraquinones (Figure 3(c)) inLQC were definitely or tentatively identified All of themwere derived from Dahuang Chrysophanol glucoside (17)emodin-8-O-glucoside (45) rhein (54) and emodin (61)were confirmed via comparing with standard substances
The characteristic fragmentation behavior of anthra-quinones was the loss of CO
2(44Da) CH
3(15Da) and
CO (28Da) in negative ion mode Typical compound 61was used to explain the fragmentation pathway of anth-raquinones presented in Figure 4(f) Compound 45 asshown in Figure 4(g) produced characteristic fragments atmz 2690673 mz 2410719 mz 2250753 mz 1970771andmz 1820486 which corresponded to [MndashHndashGlc]minus [MndashHndashGlcndashCO]minus [MndashHndashGlcndashCO
2]minus [MndashHndashCOndashCO
2]minus and
[MndashHndashCO2ndashCH3ndashCO]minus respectively Compounds 12 43
44 and 49 were tentatively identified by comparing theiraccurate molecular masses and MSMS fragment data withliterature data [21 40 41]
324 Triterpenoids Eight triterpenoids (Figure 3(d)) inLQC were unambiguously or tentatively identified All ofthem were derived from Gancao Glycyrrhizic acid (57) wasconfirmed by comparing with standards In negative ion
mode representative compound 57 yielded [MndashHndashH2Ondash
CO2]minus at mz 7594524 [MndashHndashGlcA (glucuronic acid)]minus
at mz 6454102 [MndashHndash2GlcA]minus at mz 4693705 [2GlcAndashH]minus at mz 3510809 and [2GlcAndashHndashH
2OndashCO
2]minus at mz
2890758 as shown in Figure 4(h) Compounds 48 50 5255 58 59 and 60 were tentatively identified by comparingtheir exact molecular masses and MSMS spectral data withthe literature data [51]
325 Iridoids Four iridoids (Figure 3(e)) in LQC includingiridoid glycosides and secoiridoid glycosides were identifiedAll of them were derived from Jinyinhua Loganic acid (5)sweroside (13) and secoxyloganin (20) were unambiguouslyidentified via the standards
The fragment 14F (fragment generated from the fractureof 14 bonds of iridoids) in the negative ion mode was identi-fied as the characteristic fragment ion of iridoid glycosidesMeanwhile iridoid glycosides would lose the functionalgroups such as H
2O (18Da) CO
2(44Da) and glucose
(162Da) Typical compound 5 gave [MndashHndashGlcndashCO2ndashH2O]minus
at mz 1510878 [MndashHndashGlcndash2CO2]minus at mz 1250712 and
[14FndashH2O]minus at mz 930423 [52] as presented in Figure 4(i)
Compound 10 was tentatively identified by comparing itsexact molecular mass and MSMS spectral data with theliterature data [19]
326 Other Types of Compounds Compounds 1 2 11 14 15and 51 were tentatively identified by comparing their exactmolecularmasses andMSMS spectral datawith the literaturedata except gallic acid (3) salidroside (6) forsythoside E (7)and amygdalin (33) which were identified via the standards(Figure 3(f)) [12 53]
33 Methodological Validation of the Quantitative AnalysisAs shown in Table 3 twelve standards were of good lin-earity with high correlation coefficient values over 09993The LODs and LOQs were 0051ndash171 120583gmL and 016ndash569 120583gmL independently Twelve analytes in sample solu-tion were stable at room temperature within 48 h with
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
8 The Scientific World Journal
3561547
13599643083514
3711826
37218625161718
5792105
1129871 4310958
58021345812158 111342197973569 11814053
(MSMS)
O
O
O
OO
OO
OH
HOHO
OH
[MndashHndashGlc]minus[MndashHndashGlcndashCH3]minus
()
0
100
()
0
100
mz
150 200 250 300 350 400 450 500 550 600 650 700 750mz
200 400 600 800 1000 1200 1400
(MSminus)
mz 3561547
mz 3711826
(e)
2250723
18204771810753
18006601540552
19706952240626
2410669 2690634
2420626 2700607
2690462
1129890 27004855610872 8234322 9694360 14023499
O O
O
O
O O
(MSMS)
()
0
100(
)
0
100
mz
160 180 200 220 240 260 280mz
200 400 600 800 1000 1200 1400
(MSminus)
OH
OH
OHOH OH OH OH
OHOHOH
OH
minusCO2
minusCO2minusCO
minusCO
minusHminusHminusH
minusHminusH
minusH
H3CH3C
H3C H3C
mz 1970695mz 2410669
mz 2250723mz 2690462
mz 1820477mz 1540552
minusCH3minusCO
(f)
4310988
11692794321016
4990790 8632153 11946926 14410397
2690673
2250753
18204861710837
1970771 241071927007752710783 3110845
(MSMS)
O
O
O OH
OHO
HOHO
OH
HO
[MndashHndashGlc]minus[MndashHndashGlcndashCO]minus
[MndashHndashGlcndashCO2]minus
[MndashHndashGlcndashCOndashCO2]minus
[MndashHndashGlcndashCO2ndashCH3ndashCO]minus()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400mz
140 160 180 200 220 240 260 280 300 320
(MSminus)
mz 3711826mz 2690673
CH3
(g)
8214583
35108091130354
19305102890758
645410258340994693705 7594542
8224637
8234628
(MSMS)
H
HO
HO
OOH
O
O
HOOCHO
HOOHO
HOOH
HOOC
[MndashHndash2GlcA]minus[MndashHndashGlcA]minus
[2GlcAndashH]minus
()
0
100
mz
100 200 300 400 500 600 700 800
8213977
1169246 3111153 8203445
8224006
8434348125530939433099 13258577
()
0
100
mz
200 400 600 800 1000 1200 1400
mz 4693705mz 3510809mz 6454102
(MSminus)
(h)
Figure 4 Continued
The Scientific World Journal 9
3751302
21307733761332
75126674731059 8093751 10383667 13123422
1250712
1210637
930423
650486 950155 1260535 151087824011931740320 1999976 2505933
(MSMS)
OH
H
HO
OH
O
O
O
HO
OHOH
OH
[MndashHndashGlc]minus
[14FndashH2O]minus [MndashHndashGlcndash2CO2]minus
14F
minusH2O
()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400
mz
60 80 100 120 140 160 180 200 220 240 260
(MSminus)
minusCO2
minusCO2
[MndashHndashGlcndashCO2ndashH2O]minus
(i)
Figure 4TheMS spectra and fragmentation pathway of compound 35 (a) compound 26 (b) compound 8 (c) compound 30 (d) compound39 (e) compound 61 (f) compound 45 (g) compound 57 (h) and compound 5 (i)
The fragmentation pathway of compound 8 [18] as the rep-resentative of phenylpropionic acids is shown in Figure 4(c)Compound 30 produced [MndashHndashCa (caffeoyl)]minus at mz4611920 and [CaffeicndashHndashH
2O]minus atmz 1610328 as displayed
in Figure 4(d) Lignans primarily generated [M+HCOO]minusin negative ion mode and further elimination of glucose(162Da) produced aglycone As shown in Figure 4(e) com-pound 39 produced characteristic fragments at mz 3711826and mz 3561547 corresponding to [MndashHndashGlc (glucose)]minusand [MndashHndashGlcndashCH
3]minus respectively Compounds 16 18 19
28 32 38 and 56 were tentatively identified on the basisof the exact molecular formulae matching fragmentationinformation and retention behaviors as well as literature data[23 24 33 36 47]
323 Anthraquinones Eight anthraquinones (Figure 3(c)) inLQC were definitely or tentatively identified All of themwere derived from Dahuang Chrysophanol glucoside (17)emodin-8-O-glucoside (45) rhein (54) and emodin (61)were confirmed via comparing with standard substances
The characteristic fragmentation behavior of anthra-quinones was the loss of CO
2(44Da) CH
3(15Da) and
CO (28Da) in negative ion mode Typical compound 61was used to explain the fragmentation pathway of anth-raquinones presented in Figure 4(f) Compound 45 asshown in Figure 4(g) produced characteristic fragments atmz 2690673 mz 2410719 mz 2250753 mz 1970771andmz 1820486 which corresponded to [MndashHndashGlc]minus [MndashHndashGlcndashCO]minus [MndashHndashGlcndashCO
2]minus [MndashHndashCOndashCO
2]minus and
[MndashHndashCO2ndashCH3ndashCO]minus respectively Compounds 12 43
44 and 49 were tentatively identified by comparing theiraccurate molecular masses and MSMS fragment data withliterature data [21 40 41]
324 Triterpenoids Eight triterpenoids (Figure 3(d)) inLQC were unambiguously or tentatively identified All ofthem were derived from Gancao Glycyrrhizic acid (57) wasconfirmed by comparing with standards In negative ion
mode representative compound 57 yielded [MndashHndashH2Ondash
CO2]minus at mz 7594524 [MndashHndashGlcA (glucuronic acid)]minus
at mz 6454102 [MndashHndash2GlcA]minus at mz 4693705 [2GlcAndashH]minus at mz 3510809 and [2GlcAndashHndashH
2OndashCO
2]minus at mz
2890758 as shown in Figure 4(h) Compounds 48 50 5255 58 59 and 60 were tentatively identified by comparingtheir exact molecular masses and MSMS spectral data withthe literature data [51]
325 Iridoids Four iridoids (Figure 3(e)) in LQC includingiridoid glycosides and secoiridoid glycosides were identifiedAll of them were derived from Jinyinhua Loganic acid (5)sweroside (13) and secoxyloganin (20) were unambiguouslyidentified via the standards
The fragment 14F (fragment generated from the fractureof 14 bonds of iridoids) in the negative ion mode was identi-fied as the characteristic fragment ion of iridoid glycosidesMeanwhile iridoid glycosides would lose the functionalgroups such as H
2O (18Da) CO
2(44Da) and glucose
(162Da) Typical compound 5 gave [MndashHndashGlcndashCO2ndashH2O]minus
at mz 1510878 [MndashHndashGlcndash2CO2]minus at mz 1250712 and
[14FndashH2O]minus at mz 930423 [52] as presented in Figure 4(i)
Compound 10 was tentatively identified by comparing itsexact molecular mass and MSMS spectral data with theliterature data [19]
326 Other Types of Compounds Compounds 1 2 11 14 15and 51 were tentatively identified by comparing their exactmolecularmasses andMSMS spectral datawith the literaturedata except gallic acid (3) salidroside (6) forsythoside E (7)and amygdalin (33) which were identified via the standards(Figure 3(f)) [12 53]
33 Methodological Validation of the Quantitative AnalysisAs shown in Table 3 twelve standards were of good lin-earity with high correlation coefficient values over 09993The LODs and LOQs were 0051ndash171 120583gmL and 016ndash569 120583gmL independently Twelve analytes in sample solu-tion were stable at room temperature within 48 h with
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
The Scientific World Journal 9
3751302
21307733761332
75126674731059 8093751 10383667 13123422
1250712
1210637
930423
650486 950155 1260535 151087824011931740320 1999976 2505933
(MSMS)
OH
H
HO
OH
O
O
O
HO
OHOH
OH
[MndashHndashGlc]minus
[14FndashH2O]minus [MndashHndashGlcndash2CO2]minus
14F
minusH2O
()
0
100
()
0
100
mz
200 400 600 800 1000 1200 1400
mz
60 80 100 120 140 160 180 200 220 240 260
(MSminus)
minusCO2
minusCO2
[MndashHndashGlcndashCO2ndashH2O]minus
(i)
Figure 4TheMS spectra and fragmentation pathway of compound 35 (a) compound 26 (b) compound 8 (c) compound 30 (d) compound39 (e) compound 61 (f) compound 45 (g) compound 57 (h) and compound 5 (i)
The fragmentation pathway of compound 8 [18] as the rep-resentative of phenylpropionic acids is shown in Figure 4(c)Compound 30 produced [MndashHndashCa (caffeoyl)]minus at mz4611920 and [CaffeicndashHndashH
2O]minus atmz 1610328 as displayed
in Figure 4(d) Lignans primarily generated [M+HCOO]minusin negative ion mode and further elimination of glucose(162Da) produced aglycone As shown in Figure 4(e) com-pound 39 produced characteristic fragments at mz 3711826and mz 3561547 corresponding to [MndashHndashGlc (glucose)]minusand [MndashHndashGlcndashCH
3]minus respectively Compounds 16 18 19
28 32 38 and 56 were tentatively identified on the basisof the exact molecular formulae matching fragmentationinformation and retention behaviors as well as literature data[23 24 33 36 47]
323 Anthraquinones Eight anthraquinones (Figure 3(c)) inLQC were definitely or tentatively identified All of themwere derived from Dahuang Chrysophanol glucoside (17)emodin-8-O-glucoside (45) rhein (54) and emodin (61)were confirmed via comparing with standard substances
The characteristic fragmentation behavior of anthra-quinones was the loss of CO
2(44Da) CH
3(15Da) and
CO (28Da) in negative ion mode Typical compound 61was used to explain the fragmentation pathway of anth-raquinones presented in Figure 4(f) Compound 45 asshown in Figure 4(g) produced characteristic fragments atmz 2690673 mz 2410719 mz 2250753 mz 1970771andmz 1820486 which corresponded to [MndashHndashGlc]minus [MndashHndashGlcndashCO]minus [MndashHndashGlcndashCO
2]minus [MndashHndashCOndashCO
2]minus and
[MndashHndashCO2ndashCH3ndashCO]minus respectively Compounds 12 43
44 and 49 were tentatively identified by comparing theiraccurate molecular masses and MSMS fragment data withliterature data [21 40 41]
324 Triterpenoids Eight triterpenoids (Figure 3(d)) inLQC were unambiguously or tentatively identified All ofthem were derived from Gancao Glycyrrhizic acid (57) wasconfirmed by comparing with standards In negative ion
mode representative compound 57 yielded [MndashHndashH2Ondash
CO2]minus at mz 7594524 [MndashHndashGlcA (glucuronic acid)]minus
at mz 6454102 [MndashHndash2GlcA]minus at mz 4693705 [2GlcAndashH]minus at mz 3510809 and [2GlcAndashHndashH
2OndashCO
2]minus at mz
2890758 as shown in Figure 4(h) Compounds 48 50 5255 58 59 and 60 were tentatively identified by comparingtheir exact molecular masses and MSMS spectral data withthe literature data [51]
325 Iridoids Four iridoids (Figure 3(e)) in LQC includingiridoid glycosides and secoiridoid glycosides were identifiedAll of them were derived from Jinyinhua Loganic acid (5)sweroside (13) and secoxyloganin (20) were unambiguouslyidentified via the standards
The fragment 14F (fragment generated from the fractureof 14 bonds of iridoids) in the negative ion mode was identi-fied as the characteristic fragment ion of iridoid glycosidesMeanwhile iridoid glycosides would lose the functionalgroups such as H
2O (18Da) CO
2(44Da) and glucose
(162Da) Typical compound 5 gave [MndashHndashGlcndashCO2ndashH2O]minus
at mz 1510878 [MndashHndashGlcndash2CO2]minus at mz 1250712 and
[14FndashH2O]minus at mz 930423 [52] as presented in Figure 4(i)
Compound 10 was tentatively identified by comparing itsexact molecular mass and MSMS spectral data with theliterature data [19]
326 Other Types of Compounds Compounds 1 2 11 14 15and 51 were tentatively identified by comparing their exactmolecularmasses andMSMS spectral datawith the literaturedata except gallic acid (3) salidroside (6) forsythoside E (7)and amygdalin (33) which were identified via the standards(Figure 3(f)) [12 53]
33 Methodological Validation of the Quantitative AnalysisAs shown in Table 3 twelve standards were of good lin-earity with high correlation coefficient values over 09993The LODs and LOQs were 0051ndash171 120583gmL and 016ndash569 120583gmL independently Twelve analytes in sample solu-tion were stable at room temperature within 48 h with
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
10 The Scientific World Journal
Table2Ch
aracteriz
ationof
compo
unds
inLQ
Cby
UPL
C-DAD-Q
TOF-MS
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
10817
C 7H
12O
61910
555(minus05)
Quinica
cid
[12]
20982
223287
C 6H
8O7
1910
198(31)
17300
99[M
ndashHndashH
2O]minus
1110
088[M
ndashHndash2H
2OndashC
O2]minus
Citricacid
[13]
3a15
45216271
C 7H
6O5
1690
142(30)
1250241
[MndashH
ndashCO
2]minus
Gallic
acid
[14]
4a3010
325
C 16H
17O
9
3530878
(14)
3750695
[Mndash2H+
Na]minus70718
30[2MndashH
]minus
1910
497[M
ndashHndashC
a]minus
1790
312[C
affeica
cidndash
H]minus
1350359
[Caffeica
cidndash
HndashC
O2]minus
Neochlorogenic
acid
[13]
5a3639
254
C 16H
24O
103751302
(29)
7512
667[2MndashH
]minus
2130773
[MndashH
ndashGlc]minus
1510
878
[MndashH
ndashGlcndashC
O2ndashH
2O]minus
1250721
[MndashH
ndashGlcndash2CO
2]minus
Loganica
cid
[15]
6a404
8224
278
C 14H
20O
72991
133(07)
1790
447[M
ndashHndashC
8H8O
]minus
1190246[M
ndashHndashG
lcndashH
2O]minus
Salid
rosid
e[16]
7a4557
C 20H
30O
124611674(33)
3151348
[MndashH
ndashRha]minus
2971589[M
ndashHndashR
handashH
2O]minus
15306
44[M
ndashHndashR
handashG
lc]minus
1350573
[MndashH
ndashRhandashG
lcndashH
2O]minus
4631220
(minus43)
4851568
1370
599[M
+HndashR
handashG
lcndashH
2O]+
Forsytho
sideE
[17]
8a4749
327
C 16H
17O
93530880
(20)
1910
556[M
ndashHndashC
a]minus
1790
224[C
affeica
cidndash
H]minus
3551028
(minus03)
3770
857
Chlorogenica
cid
[18]
9a5281
325
C 16H
17O
93530880
(20)
1790
348[C
affeica
cidndash
H]minus
17304
61[Q
uinica
cidndash
HndashH
2O]minus
1910
417[M
ndashHndashC
a]minus
Cryptochlorogenic
acid
[18]
106435
270
C 17H
26O
103891
096(31)
3912
176
(minus08)
4131067
3952354
[M+
NandashH
2O]+
2290
835[M
+HndashG
lc]+
Loganin
[19]
116749
C 14H
23O
93350776
(27)
1730459
[MndashH
ndashGlc]minus
1610
373[G
lcndashHndashH
2O]minus
13304
08[G
lcndashH
ndashH2O
ndashCO]minus
3370
883
(53)
3590
686
1630
419[G
lc+
HndashH
2O]+
Reng
ynic
acid-11015840-O
-120573-D
-glucoside
[20]
1272
22C 2
1H20O
94151261
(51)
4391
207
2550879
[M+HndashG
lc]+
Isom
erof
chrysoph
anol
glucoside
[21]
13a
7428
245
C 16H22O9
3571208(62)4031263
[M
+HCO
O]minus
3930938
[M
+Cl]minus
3591
337
(minus14
)3811154
Sweroside
[15]
1478
59C 1
4H23O
93350871
(42)
1930506161037713304
233370
774
(09)
3590
225
1630
416[G
lc+
HndashH
2O]+
Isom
erof
reng
ynic
acid-11015840-O
-120573-D
-glucoside
158106
306
C 20H
22O
83891458(26)4351519
[M+HCO
O]minus
2270
943[M
ndashHndashG
lc]minus
Polydatin
[22]
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
The Scientific World Journal 11
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
168222
C 29H
36O
166392
219(minus17
)4771894[M
ndashHndashG
lc]minus
1790
546[M
ndashRhandashC
17H
14O
6]minus
1610
328[G
lcndashHndashH
2O]minus
R-suspensasid
e[23]
17a
8320
C 21H
20O
94151253
(31)
2530542
[MndashH
ndashGlc]minus
4171394
(minus07)
4391
200
2550864
[M+HndashG
lc]+
Chrysoph
anol
glucoside
[21]
188874
C 29H
36O
166392
002(minus27)
1610
328[G
lcndashHndashH
2O]minus
1790
349[M
ndashRhandashC
17H
14O
6]minus
S-suspensasid
e[24]
1990
54C 2
9H36O
156231999
(minus12
)3112
176
6252137
(08)
6471942
4791
5624711479
32509251630394
Isom
erof
forsytho
sideA
20a
9470
235
C 17H
24O
114031261
(52)
4051331
(minus17
)4271203
2430871
[M+HndashG
lc]+
Secoxyloganin
[19]
2195
52276313
C 21H
22O
94171205(46)8352331
[2MndashH
]minus2550652
[MndashH
ndashGlc]minus
4411132
2570
811[M
+HndashG
lc]+
Liqu
iritin
[25]
22a
9886
277312
C 26H
30O
135491
627(35)
5512
823
(47)
5731737
2570
822[M
+HndashA
pindashG
lc]+
Liqu
iritin
apiosid
e[26]
2399
95C 2
6H30O
135491
629(38)
5513
403
(minus51)
5731735
2570
815[M
+HndashA
pindashG
lc]+
Isom
erof
liquiritin
apiosid
e[26]
2410088
C 27H
30O
1660
91844(minus18
)6331712
Rhod
iosin
[27]
25a
10384
250300
C 22H
22O
94291410(30)
453117
62690
974[M
+HndashG
lc]+
Ono
nin
[2829]
26a
10722
360
C 21H
20O
124630905
(28)
3010
464[M
ndashHndashG
lc]minus
4651033
(52)
4870
333
3030515
[M+HndashG
lc]+
Hyperin
[24]
27a
10819
326
C 25H
24O
125151205
(29)
3530751
[MndashH
ndashCa]minus
5171296
(17)
5391
156
4991
187[M
+HndashH
2O]+
1630
395[M
+HndashC
andashQuinica
cid]
+
35-Dicaffeoylqu
inic
acid
[30]
2810931
C 29H
36O
156231999
(minus19
)46113781610
185
6471941
47114973250925
1630
415
Isom
erof
forsytho
sideA
29a
1110
256354
C 27H
30O
1660
91465(15)
6111573
(33)
6331404
4650982
[M+HndashR
ha]+
3030495
[M+HndashR
ut]+
Rutin
[31]
30a
11220
280
C 29H
36O
156232001
(minus16
)4611920[M
ndashHndashC
a]minus
1610
328[C
affeica
cidndash
HndashH
2O]minus
6471927
1630386
[Caffeica
cid+
HndashH
2O]+
Forsytho
sideA
[32]
3111589
283
C 27H
30O
155931534
(minus13
)2850677
[MndashH
ndashRut]minus
5951694
(minus07)
2870
936[M
+HndashR
ut]+
Kaem
pferol-3-O
-rutin
oside
[13]
3211692
C 19H
35O
165191
884(35)5651953
[M+HCO
O]minus
3571366[M
ndashHndashG
lc]minus
5431855
3591454[M
+HndashG
lc]+
3411411[M
+HndashG
lcndashH
2O]+
(+)-Pino
resin
ol-120573-
D-glucosid
e[33]
33a
12213
210262
C 20H
27NO
114561523
(37)
4581641
(minus46)
2961124
[M+HndashG
lc]+
1620862
[M+
HndashG
lcndashH
CT]+
Amygdalin
[13]
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
12 The Scientific World JournalTa
ble2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
34a
12659
326
C 25H
24O
12515116
2(minus54)
3530786
[MndashH
ndashCa]minus
5171348
(04)
5391
193
4991
248[M
+HndashH
2O]+1630387[M
+HndashC
andashQuinica
cid]
+
34-
Dicaffeoylqu
inic
acid
[30]
35a
13082
276
C 15H
12O
42550654
(minus12
)2570
659
(minus08)
1470
621[M
+HndashR
L]+
1370
422[M
+HndashV
P]+
1190691[M
+HndashR
LndashCO
]+Liqu
iritig
enin
[34]
36a
13251
243372
C 26H
30O
135491
621(24)
5511725
(34)
5731563
4191
360[M
+HndashA
pi]+
2570
814[M
+HndashA
pindashG
lc]+
Isoliquiritin
apiosid
e[35]
37a
13300
242362
C 21H
22O
94171198(29)
2550518
[MndashH
ndashGlc]minus
4191
314
(minus67)
4411150
2570
823[M
+HndashG
lc]+
2291
357[M
+HndashG
lcndashC
O]+
Isoliquiritin
[26]
3813479
280
C 27H
34O
115792
103[M
+HCO
O]minus
5571987
5522463
[M+NH
4]+
3551516
[M+
HndashG
lcndashH
2O]+
Arctiin
[36]
39a
1416
1277
C 27H
34O
115792
106[M
+HCO
O]minus
3711826[M
ndashHndashG
lc]minus
3561547
[MndashH
ndashGlcndashH
2O]minus
5522444
[M
+NH
4]+5571991
3731563
[M+HndashG
lc]+
3551544
[M+
HndashG
lcndashH
2O]+
Phillyrin
[3738]
4014678
372255
C 15H
10O
73010
355(23)
1510
137[M
ndashHndashC
8H6O
3]minus
3030520
(49)
Quercetin
[39]
4115039
284
C 28H
34O
1560
91522(13)
3010
582[M
ndashHndashR
ut]minus
Hesperid
in[13]
4215402
265366
C 15H
10O
628504
06(25)
2870
721
(45)
15304
69[M
+HndashH
EP]+
Kaem
pferol
[13]
4316627
255
C 15H
10O
42530507
(24)
2250741
[MndashH
ndashCO]minus
2100494
[MndashH
ndashCOndashC
H3]minus
1820542
[MndashH
ndash2CO
ndashCH
3]minus
1540553
[MndashH
ndash3CO
ndashCH
3]minus
Chrysoph
anol
[40]
4417035
C 15H
10O
42530512
(43)
2250752210047418204
85
1540531
Isom
erof
chrysoph
anol
[40]
45a
17347
254
287426
C 21H
20O
104310
989(26)
2690
673[M
ndashHndashG
lc]minus
2410
719[M
ndashHndashG
lcndashC
O]minus
2250753
[MndashH
ndashGlcndashC
O2]minus
2100578
[MndashH
ndashGlcndashC
O2ndashCH
3]minus
1970
771[MndashH
ndashGlcndashC
OndashC
O2]minus
18204
86[M
ndashHndashG
lcndashC
O2ndashCH
3ndashCO
]minus
4551010
2710
715[M
+HndashG
lc]+
Emod
in-8-O
-glucoside
[41]
4617396
240
330395
C 15H
12O
42550639
(minus71)
2570
812
(minus08)
Isoliquiritigenin
[26]
47a
18385
250304
C 16H
12O
42670
653(minus15
)2690
827
(48)
2910
751
Form
onon
etin
[42]
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
The Scientific World Journal 13
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
4818804
89540
82(28)
7193
849[M
ndashHndashG
lcA]minus
3510
630[2GlcAndash
H]minus
8974
107
(12)
9193
925
22120573-Acetoxy
licorices
apon
inB2
uralsa
poninF
[43]
491919
4271421
C 22H22O10
445116
8(minus56)
2830632
[MndashH
ndashGlc]minus
4691
128
2850775
[M+HndashG
lc]+
Physcion
-8-O
-120573-D
-glucop
yranoside
[41]
5019603
248
C 42H
60O
168193
843(06)
3510
699[2GlcAndash
H]minus
8213
994
(minus01)
8433996
6453612
[M+
HndashG
lcA]+
4693
302[M
+Hndash2GlcA]+
4513
211[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inE2
[35]
5119704
C 16H
24O
73272
180(28)
1649664
[MndashH
ndashGlc]minus
3512
133
Rhod
odendrol-41015840-
O-120573-D
-glucop
yranoside
[44]
5220000
254
C 44H
64O
188794
062(13)
3510
806[M
ndashHndash22A
G]minus
8814
170
(minus01)
7053897
[M+
HndashG
lcA]+
5293
588[M
+Hndash2GlcA]+
5113
395[M
+Hndash2GlcA
ndashH2O
]+
22-
Acetoxyglycyrrhizin
[42]
5320573
352
C 17H
14O
73292
341(39)
3532280
301144
3[M
+HndashC
H2O
]+
2872
002[M
+HndashC
O2]
+ 2591
248[M
+HndashC
O2ndashCO
]+
Tricin
[45]
54a
2119
9260
C 15H
8O6
2830262
(67)
2390
359[M
ndashHndashC
O2]minus
2850515
(minus130)
Rhein
[46]
5521540
253
8373977(12)
8194
404[M
ndashHndashH
2O]minus
6614
100[M
ndashHndashG
lcA]minus
3510
815[2GlcAndash
H]minus
2890
840
[2GlcAndash
HndashH
2OndashC
O2]minus
8394
061
(minus05)
8613
890
6633868
[M+
HndashG
lcA]+
4873395[M
+Hndash2GlcA]+
4693
286[M
+Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inG2
[42]
5621626
236
327370
384
C 21H
20O
63671208(71)
3091
566[M
ndashHndashC
OndashC
H2O
]minus
2650635
[MndashH
ndashCOndashC
H2O
ndashCO
2]minus
2210
221
[MndashH
ndashCOndashC
H2O
ndash2CO
2]minus
3691
369
(minus76
)3911176
Glycycoum
arin
[47]
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
14 The Scientific World Journal
Table2Con
tinued
Peak
number119879119877 (min)120582max
(nm)
Form
ula
ESminus(119898119911)
ES+(119898119911)
Identifi
catio
nRe
ference
[MndashH
]minus(ppm
)MS2
b[M
+H]+
(ppm
)[M
+Na]
+MS2
b
5722087
249
C 42H
62O
168213
977(17)
8034501
[MndashH
ndashH2O
]minus
7594
542[M
ndashHndashH
2OndashC
O2]minus
6454102
[MndashH
ndashGlcA]minus
6274
000[M
ndashHndashG
lcAndash
H2O
]minus
4693
705[M
ndashHndash2GlcA]minus
3510
809[2GlcAndash
H]minus
2890
758[2GlcAndash
HndashH
2OndashC
O2]minus
8234128
(15)
8454557
6473797[M
+HndashG
lcA]+
4713
480[M
+Hndash2GlcA]+
4533
365[M
+Hndash2GlcA
ndashH2O
]+
Glycyrrhizica
cid
[48]
582218
8252
C 42H
62O
168213
986(minus11)
3510
768[M
ndashHndashG
A]minus
8234134
(minus21)
8454127
6473
823[M
+HndashG
lcA]+
4713
365[M
+Hndash2GlcA]+
4533324
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inH2
[26]
5922389
248
C 42H
64O
158074
197(minus06)
74546
01[M
ndashHndashG
lc]minus
6314
210[M
ndashHndashG
lcA]minus
3510
699[M
ndashHndashG
A]minus
8094
410
(minus09)
8314
138
Licoric
esapon
inB2
[49]
6022623
252
C 42H
62O
168213
997(02)
3510
837[M
ndashHndashG
A]minus
8234105
(minus13
)8453900
6473
838[M
+HndashG
lcA]+
4713
288[M
+Hndash2GlcA]+
4533279
[M+
Hndash2GlcA
ndashH2O
]+
Licoric
esapon
inK2
[50]
61a
23230
288439
C 15H
10O
52690
462(45)
2410
669[M
ndashHndashC
O]minus
2250723
[MndashH
ndashCO
2]minus
21004
27[M
ndashHndashC
O2ndashCH
3]minus
1970
695[M
ndashHndashC
OndashC
O2]minus
1820477
[MndashH
ndashCO
2ndashCH
3ndashCO
]minus
1540552
[MndashH
ndashCO
2ndashCH
3ndash2C
O]minus
Emod
in[40]
a Com
paredwith
referencec
ompo
unds
b 22A
G2
2-acetoxyglycyrrhizin
ApiapioseC
acaffeoylGAg
lycyrrhetin
icacidG
lcg
lucoseG
lcAg
lucuronicacidH
CT4
-(hydroxym
ethyl)c
yclobu
tane-123-tr
iolHEP
4-(hydroxyethynyl)ph
enolR
ha
rham
noseR
LresorcinolR
utrutinoseVP
4-vinylpheno
l
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
The Scientific World Journal 15
Table3Linear
regressio
nLO
Dsa
ndLO
Qsintraday
andinterday
precision
srepeatabilitystabilityandrecovery
for12compo
unds
Com
poun
dRe
gressio
nequatio
na(119899=3)
1198772
Linear
range
(120583gmL)
LOD
b
(120583gmL)
LOQ
c
(120583gmL)
Intraday
(RSD
)
(119899=6)
Interday
(RSD
)
(119899=3)
Repeatability(119899=6)Stability
(RSD
)
Recovery
(119899=6)
Mean
(120583gg)
RSD
()
Orig
inal
(120583g)
Spiked
(120583g)
Detected
(120583g)
Recovery
()
RSD
()
Salid
rosid
e119910=
13980119909minus11265
09999
527ndash8
435
106
350
012
026177955plusmn
1092
061
138
35591
35420
7075
09928
124
Chlorogenica
cid
119910=
26510119909+36733
09998
685ndash10956
171
569
077
212247371plusmn
1274
051
254
49474
49385
9599
09419
242
Forsytho
sideE
119910=
11793119909+50168
09995
485ndash776
412
1404
067
225175403plusmn
1872
107
267
3518
13510
56772
69299
354
Cryptochlorogenic
acid
119910=
21498119909minus50189
09997
665ndash10647
166
553
110
163202949plusmn
2065
102
246
40590
40660
78400
9310
143
Amygdalin
119910=
10530119909minus34647
09998
391ndash6
257
098
326
079
093148538plusmn
2585
174
248
2970
829862
57639
9358
332
Sweroside
119910=
20593119909minus60034
09999
200ndash319
6025
080
011
033
79806plusmn
293
037
048
1596
11595
132543
10395
041
Hyperin
119910=
27773119909minus87941
09999
020ndash321
0051
016
078
062
9203plusmn
080
087
083
1841
1824
3707
10229
182
Rutin
119910=
30768119909minus37837
09993
053ndash850
0072
021
041
224
18415plusmn
306
166
157
3683
3681
7250
9690
234
Forsytho
sideA
119910=
85466119909minus60245
09999
421ndash6
734
030
105
020
292248499plusmn
1843
074
195
4970
049595
99733
10089
149
Phillyrin
119910=
30390119909+52828
09999
286ndash4
571
071
236
079
163157780plusmn
1011
064
276
31556
31672
62399
10025
160
Rhein
119910=
74736119909minus51727
09996
347ndash5549
087
288
040
028
68192plusmn
1632
239
097
13638
13627
27348
10061
161
Glycyrrhizica
cid
119910=
10296119909minus65313
09999
527ndash8
435
0092
033
034
209162275plusmn
1759
108
215
32455
323939
65216
1013
1327
a 119910isthep
eakarea119909
isthec
oncentratio
nof
stand
ardsolutio
ns
b LODrefersto
thelim
itsof
detection119878119873=3
c LOQrefersto
thelim
itsof
quantity119878119873=10
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
16 The Scientific World Journal
Table4Con
tentso
fthe
12compo
unds
in10
batches
Sample
Com
poun
d(m
eanplusmnSD
)(120583gg)
Total
RSD(
)Salid
rosid
eCh
lorogenic
acid
Forsytho
sideE
Cryptochlorogenic
acid
Amygdalin
Sweroside
Hyperin
Rutin
Forsytho
sideA
Phillyrin
Rhein
Glycyrrhizic
acid
Lot1174057plusmn3070215911plusmn1247188899plusmn795146488plusmn1022210985plusmn312783023plusmn8446706plusmn06815622plusmn111316384plusmn3314159920plusmn122957070plusmn223102845plusmn4211677910
1003
Lot2169704plusmn1254181069plusmn1173142467plusmn3804116741plusmn835162314plusmn251465095plusmn7216807plusmn13710685plusmn282249734plusmn2721122059plusmn355755426plusmn82783716plusmn11651365817
Lot3145636plusmn2208194191plusmn4494147793plusmn2966123230plusmn2988164125plusmn343168153plusmn6357266plusmn20511031plusmn216208922plusmn3609131499plusmn312161791plusmn107090475plusmn15731354112
Lot4152075plusmn891199338plusmn5086150109plusmn4260140915plusmn632214701plusmn603774398plusmn18006256plusmn12810086plusmn239265253plusmn6703113947plusmn240869642plusmn187698560plusmn22851492580
Lot5159312plusmn3500208189plusmn5077162662plusmn4829152311plusmn4322228276plusmn461977798plusmn4187589plusmn16912106plusmn241307210plusmn4489147718plusmn254868635plusmn164498479plusmn29131630285
Lot6182134plusmn4855243669plusmn3100202528plusmn4331149476plusmn1265237704plusmn636486029plusmn133710080plusmn18714577plusmn294368072plusmn9285199109plusmn182964886plusmn391105729plusmn30411863993
Lot7147432plusmn3685222261plusmn3827137297plusmn3685172736plusmn1640259475plusmn506785430plusmn19317725plusmn21415690plusmn465298754plusmn3676116812plusmn127055126plusmn518122774plusmn20241641512
Lot8159866plusmn1951218354plusmn1511158979plusmn1971172126plusmn1136249481plusmn196182471plusmn4478126plusmn06917462plusmn086316455plusmn3681127714plusmn110471694plusmn574122087plusmn9891704815
Lot9153578plusmn3036196172plusmn2233178955plusmn2812124734plusmn3659182487plusmn356669640plusmn7087333plusmn04613623plusmn251282309plusmn1519149585plusmn177662510plusmn114489145plusmn10601510071
Lot10183641plusmn2914221636plusmn818150075plusmn806167072plusmn2319228174plusmn143280941plusmn3137533plusmn08513419plusmn124301034plusmn1264120492plusmn282689101plusmn999117672plusmn6681680790
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
The Scientific World Journal 17
the RSD less than 276 The RSD values of intraday andinterday precisions were less than 110 and 292 res-pectively The RSD of repeatability was less than 239The average recovery rates of 12 compounds ranged from9299 to 10395 with the RSD less than 354 All theresults showed that the assay was satisfactory with highaccuracy good reproducibility and high sensitivity whichwere beneficial to the analytical investigation and qualitycontrol for LQC
34 Sample Analysis Twelve representative compounds in10 batches of LQC were quantified through the developedUPLC-DAD analytical method described above The resultsare summarized in Table 4 which showed that the totalconcentrations of 12 quantitative compounds in differentbatches of the LQC varied narrowly moreover the 12components differed greatly in their contents which may beaffected by the source of medicinal materials the quality ofthe plant material or the preparation technology Amongthem forsythoside A showed the highest amount (316455ndash208922120583gg) followed by amygdalin (259475ndash162314120583gg)and hyperin had the lowest amount at 10080ndash6256120583gg
4 Conclusion
LQC is a commonly used Chinese medical preparation totreat viral influenza To date there has not been a systematicaland comprehensive study on the chemical profiling andquality control method for LQC Therefore an accuratesensitive and reliable quality control procedure for LQC isin urgent need to be established In our study the chemicalprofile of LQC was thoroughly and systematically investi-gated by UPLC-DAD-QTOF-MS for the first time Sixty-onecompounds were unambiguously or tentatively identifiedBased on the qualitative analysis a UPLC-DAD method wasestablished for quantitative analysis of 12 representative com-pounds in LQC which has been demonstrated to be effectivefor the analysis of 10 batches of LQCThis developed methodcould be applied as an effective quality control procedure forLQC In addition this study would be a powerful referencefor the identification of similar compounds presented heresuch as flavonoids phenylpropanoids anthraquinones triter-penoids and iridoids by MS spectra
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Weina Jia and Chunhua Wang contributed equally to thiswork
Acknowledgments
Financial support from Tianjin Applied Basic and Cutting-Edge Technology Research Programs (nos 13JCZDJC28600
and 13JCYBJC42000) Project of New Drug Discovery Plat-form (no 2012ZX09304007) and State Key DevelopmentProgram for Basic Research of China (ldquo973rdquo Program no2012CB723504) is gratefully acknowledged
References
[1] C Y Liu X Q Li and S Q Cai ldquoResearch progress onthe pharmacological and clinical study of Lianhua Qingwencapsulerdquo Pharmacology and Clinics of Chinese Materia Medicavol 26 no 6 pp 84ndash85 2010
[2] X Yi ldquoThe antiviral report of Lianhua Qingwen capsulerdquo TheChinese Health Care no 9 pp 13ndash14 2012
[3] Z-P Duan Z-H Jia J Zhang et al ldquoNatural herbal medicinelianhuaqingwen capsule anti-influenza a (H1N1) trial a ran-domized double blind positive controlled clinical trialrdquo Chi-nese Medical Journal vol 124 no 18 pp 2925ndash2933 2011
[4] H Kiyohara C Ichino Y Kawamura T Nagai N Sato and HYamada ldquoPatchouli alcohol in vitro direct anti-influenza virussesquiterpene in Pogostemon cablin Benthrdquo Journal of NaturalMedicines vol 66 no 1 pp 55ndash61 2012
[5] L Dong J-W Xia Y Gong et al ldquoEffect of Lianhuaqingwencapsules on airway inflammation in patients with acute exac-erbation of chronic obstructive pulmonary diseaserdquo Evidence-Based Complementary and Alternative Medicine vol 2014Article ID 637969 11 pages 2014
[6] G Chen H M Guang L Li et al ldquoBiological network analysisonmechanism of LianhuaQingwen granulescapsules forH1N1influenzaArdquo Journal of Traditional ChineseMedicine vol 55 no8 pp 703ndash707 2014
[7] S H Zhao H H Xu X Y Li et al ldquoTLC and quantitativedetermination by HPLC for Lianhua Qingwen capsulerdquo DrugStandards of China vol 8 no 1 pp 55ndash59 2007
[8] Q F Qin ldquoDetermination of chlorogenic acid forsythin rheinemodin and chrysophanol in Lianhua Qingwen capsules byHPLCrdquo Chinese Journal of Modern Applied Pharmacy vol 31no 2 pp 210ndash213 2014
[9] Y Ling Y Y Wang G T Chen and X Y Wang ldquoSimultaneousdetermination of five components from radixet rhizoma rhei inLianhua Qingwen capsule by HPLCrdquo China Journal of ChineseMedicine vol 27 no 172 pp 1157ndash1158 2012
[10] P Huo S Qin L H Huang and Y J Xu ldquoSimultaneousdetermination of six effective components in Lianhuaqingwencapsule by micellar electrokinetic capillary chromatographyrdquoJournal of Instrumental Analysis vol 30 no 4 pp 396ndash4002011
[11] R Shi W J Jiang S Qiao M Y Liu and Q Wang ldquoDeter-mination of nine constituents in Lianhua Qingwen capsules byLC-MSMS combined with ion switching technologyrdquo ChineseJournal of Pharmaceuticals vol 42 no 10 pp 777ndash780 2011
[12] Y Ogawa M Kawai A Kinoshita and T Konishi ldquoNitrogen-containing compounds from Hemerocallis fulva var semper-virensrdquoChemistry of Natural Compounds vol 49 no 6 pp 991ndash995 2014
[13] L Zhang L Zhu YWang et al ldquoCharacterization and quantifi-cation of major constituents of Xue Fu Zhu Yu by UPLC-DAD-MSMSrdquo Journal of Pharmaceutical and Biomedical Analysisvol 62 pp 203ndash209 2012
[14] W Lan L Bian X Zhao et al ldquoLiquid chromatogra-phyquadrupole time-of-flight mass spectrometry for identi-fication of in vitro and in vivo metabolites of bornyl gallate
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
18 The Scientific World Journal
in ratsrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 473649 10 pages 2013
[15] S Deng B J West and C J Jensen ldquoUPLC-TOF-MS char-acterization and identification of bioactive iridoids in Cornusmas fruitrdquo Journal of Analytical Methods in Chemistry vol 2013Article ID 710972 7 pages 2013
[16] N Guo ZW Hu X X Fan et al ldquoSimultaneous determinationof Salidroside and its aglycone metabolite p-tyrosol in ratplasma by liquid chromatography-tandem mass spectrometryrdquoMolecules vol 17 no 4 pp 4733ndash4754 2012
[17] L H Jian Q Hu J Q Zhong Q Meng K Wang and S JildquoDetermination of forsythoside E in Tanreqing injection by LC-MSMSrdquoChinese Journal of Pharmaceutical Analysis vol 33 no3 pp 435ndash438 2013
[18] Y-J Li J Chen and P Li ldquoIdentification and quantification offree radical scavengers in the flower buds of Lonicera species byonline HPLC-DPPH assay coupled with electrospray ionizationquadrupole time-of-flight tandemmass spectrometryrdquoBiomed-ical Chromatography vol 26 no 4 pp 449ndash457 2012
[19] Z-M Qian H-J Li P Li M-T Ren and D Tang ldquoSimul-taneous qualitation and quantification of thirteen bioactivecompounds in Flos Lonicerae by high-performance liquid chro-matography with diode array detector and mass spectrometryrdquoChemical and Pharmaceutical Bulletin vol 55 no 7 pp 1073ndash1076 2007
[20] Y Liu Studies on the Chemical Constituents of the Extractwith Water from Forsythia Suspensa (Thunb) Vahl ShenyangPharmaceutical University Shenyang China 2003
[21] H Q Jiang R Rong and Q H Lv ldquoIdentification of chemicalcomposition in Rhubarb by high performance liquid chro-matography with mass spectrometryrdquo Lishizhen Medicine andMateria Medica Research vol 22 no 7 pp 1075ndash1076 2011
[22] G Qian S-Y Leung L U Guanghua and K S-Y LeungldquoDifferentiation of Rhizoma et Radix Polygoni Cuspidati fromclosely related herbs by HPLC fingerprintingrdquo Chemical andPharmaceutical Bulletin vol 54 no 8 pp 1179ndash1186 2006
[23] H Guo A-H Liu L Li and D-A Guo ldquoSimultaneousdetermination of 12 major constituents in Forsythia suspensaby high performance liquid chromatography-DAD methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 43 no3 pp 1000ndash1006 2007
[24] Y Cui Q Wang X Shi X Zhang X Sheng and L ZhangldquoSimultaneous quantification of 14 bioactive constituents inForsythia suspensa by liquid chromatography-electrosprayionisation-mass spectrometryrdquo Phytochemical Analysis vol 21no 3 pp 253ndash260 2010
[25] H Liu J Su X Liang et al ldquoIdentification and determinationof the major constituents in traditional Chinese medicineLongdan Xiegan Pill by HPLC-DAD-ESI-MSrdquo Journal of Phar-maceutical Analysis vol 1 no 1 pp 1ndash7 2011
[26] Q W Yin P Wang A H Zhang H Sun X H Wu andXWang ldquoUltra-performance LC-ESIquadrupole-TOFMS forrapid analysis of chemical constituents of Shaoyao-Gancaodecoctionrdquo Journal of Separation Science vol 36 no 7 pp 1238ndash1246 2013
[27] J L Huo J H Wang Z J Wu and G L Zhang ldquoAnalysisof chemical constituents of Rhodiola crenulata using high per-formance liquid chromatography-mass spectrometryrdquo NaturalProduct Research and Development vol 24 pp 1405ndash1407 2012
[28] J Dong Y Zhu X Gao Y Chang M Wang and P ZhangldquoQualitative and quantitative analysis of the major constituents
in Chinese medicinal preparation Dan-Lou tablet by ultrahigh performance liquid chromatographydiode-array detec-torquadrupole time-of-flight tandem mass spectrometryrdquoJournal of Pharmaceutical and Biomedical Analysis vol 80 pp50ndash62 2013
[29] M I Kusaikin A M Zakharenko S P Ermakova et al ldquoDeg-lycosylation of isoflavonoid glycosides fromMaackia amurensiscell culture by 120573-D-glucosidase from Littorina sitkana hep-atopancreaserdquo Chemistry of Natural Compounds vol 47 no 2pp 197ndash200 2011
[30] C P Wan Y Y Yu S R Zhou S G Tian and S WCao ldquoIsolation and identification of phenolic compounds fromGynura divaricata leavesrdquo Pharmacognosy Magazine vol 7 no26 pp 101ndash108 2011
[31] X Yao G S Zhou Y P Tang et al ldquoA UPLC-MSMS methodfor qualification of quercetin-3-O-120573-D- glucopyranoside -(4 rarr 1)-120572-L- rhamnoside in rat plasma and application topharmacokinetic studiesrdquo Molecules vol 18 no 3 pp 3050ndash3059 2013
[32] H Guo A-H Liu M Ye M Yang and D-A Guo ldquoChar-acterization of phenolic compounds in the fruits of Forsythiasuspensa by high-performance liquid chromatography coupledwith electrospray ionization tandemmass spectrometryrdquo RapidCommunications in Mass Spectrometry vol 21 no 5 pp 715ndash729 2007
[33] G L Yan A H Zhang H Sun et al ldquoAn effective methodfor determining the ingredients of Shuanghuanglian for-mula in blood samples using high-resolution LC-MS coupledwith background subtraction and a multiple data processingapproachrdquo Journal of Separation Science vol 36 no 19 pp 3191ndash3199 2013
[34] M Ye W Z Yang K D Liu et al ldquoCharacterizationof flavonoids in Millettia nitida var hirsutissima byHPLCDADESI-MS119899rdquo Journal of Pharmaceutical Analysis vol2 no 1 pp 35ndash42 2012
[35] W-J Miao Q Wang T Bo et al ldquoRapid characteri-zation of chemical constituents and rats metabolites ofthe traditional Chinese patent medicine Gegen-Qinlian-Wanby UHPLCDADqTOF-MSrdquo Journal of Pharmaceutical andBiomedical Analysis vol 72 pp 99ndash108 2013
[36] F He D-Q Dou Y Sun L Zhu H-B Xiao and T-G KangldquoPlasma pharmacokinetics and tissue distribution of arctiin andits main metabolite in rats by HPLC-UV and LC-MSrdquo PlantaMedica vol 78 no 8 pp 800ndash806 2012
[37] Q Su Q-F Liu Y Wang K-S Bi and G-A Luo ldquoRapid andsensitive assay of the main components in Shuanghuanglianpreparations by UPLC-TOF-MSrdquo Chemical Research in ChineseUniversities vol 25 no 1 pp 122ndash124 2009
[38] QWuM-H Bang J-G Cho et al ldquoPhenolic compounds fromthe roots of Brassica rapa ssp campestrisrdquo Chemistry of NaturalCompounds vol 49 no 5 pp 852ndash856 2013
[39] N Li C H Liu S Q Mi et al ldquoSimultaneous determinationof oleanolic acid p-coumaric acid ferulic acid kaemperol andquercetin in rat plasma by LC-MS-MS and application to apharmacokinetic study of oldenlandia diffusa extract in ratsrdquoJournal of Chromatographic Science vol 50 no 10 pp 885ndash8922012
[40] J Z Zhang W Y Gao Z Liu and Z D Zhang ldquoIdentificationand simultaneous determination of twelve active components inthe methanol extract of traditional medicine Weichangrsquoan pillby HPLC-DAD-ESI-MSMSrdquo Iranian Journal of Pharmaceuti-cal Research vol 12 no 1 pp 15ndash24 2013
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
The Scientific World Journal 19
[41] Z T Liang H B Chen Z L Yu and Z Z Zhao ldquoComparisonof raw and processed Radix Polygoni Multiflori (Heshouwu) byhigh performance liquid chromatography and mass spectrom-etryrdquo Chinese Medicine vol 5 article 29 2010
[42] W-WHuangM-YWang H-M Shi et al ldquoComparative studyof bioactive constituents in crude and processed Glycyrrhizaeradix and their respective metabolic profiles in gastrointestinaltract in vitro by HPLC-DAD and HPLC-ESIMS analysesrdquoArchives of Pharmacal Research vol 35 no 11 pp 1945ndash19522012
[43] W W Tao J A Duan J M Guo et al ldquoSimultaneous deter-mination of triterpenoid saponins in dog plasma by a validatedUPLC-MSMS and its application to a pharmacokinetic studyafter administration of total saponin of licoricerdquo Journal ofPharmaceutical and Biomedical Analysis vol 75 pp 248ndash2552013
[44] W Zhao Studies on the Chemical Constituents of Ephedra sinicaPeking Union Medical College Beijing China 2009
[45] J Sun Y D Yue F Tang X F Guo and J Wang ldquoFlavonoidsfrom the leaves ofNeosinocalamus affinisrdquo Chemistry of NaturalCompounds vol 49 pp 822ndash825 2013
[46] M L Hou L W Chang C H Lin L C Lin and T H TsaildquoDetermination of bioactive components in Chinese herbalformulae and pharmacokinetics of rhein in rats by UPLC-MSMSrdquoMolecules vol 19 pp 4058ndash4075 2014
[47] L Zhou Y-P Tang L Gao X-S Fan C-M Liu and D-KWu ldquoSeparation characterization and dose-effect relationshipof the PPAR120574-activating bio-active constituents in the chineseherb formulation lsquoSan-ao decoctionrsquordquo Molecules vol 14 no 10pp 3942ndash3951 2009
[48] S Tsuruda T Sakamoto and K Akaki ldquoSimultaneous deter-mination of twelve sweeteners and nine preservatives in foodsby solid-phase extraction and LC-MSMSrdquo Shokuhin EiseigakuZasshi vol 54 no 3 pp 204ndash212 2013
[49] I KitagawaKHori T Taniyama J-L Zhou andMYoshikawaldquoSaponin and sapogenol XLVIIOn the constituents of the rootsof Glycyrrhiza uralensis Fisher from Northeastern China (1)Licorice-saponins A3 B2 and C2rdquo Chemical and Pharmaceu-tical Bulletin vol 41 no 1 pp 43ndash49 1993
[50] I Kitagawa K Hori M Sakagami J-L Zhou and MYoshikawa ldquoSaponin and sapogenol XLVIII On the con-stituents of the roots of Glycyrrhiza uralensis FISCHER fromNortheastern China (2) Licorice-saponins D3 E2 F3 G2 H2J2 and K2rdquo Chemical and Pharmaceutical Bulletin vol 41 no8 pp 1337ndash1345 1993
[51] X-D Yang X-Y Tang and L Sang ldquoRapid identificationof micro-constituents in monoammonium glycyrrhizinate rawmaterials by high-pressure solid phase extraction-high per-formance liquid chromatography-mass spectrometryrdquo ChinaJournal of ChineseMateriaMedica vol 37 no 22 pp 3416ndash34212012
[52] C-M Li X-M Liang and X-Y Xue ldquoElectrospray ionizationquadrupole time-of-flight tandemmass spectrometry of iridoidglucosides in positive ion moderdquo Chemical Journal of ChineseUniversities vol 34 no 3 pp 567ndash572 2013
[53] F Wang J Y Zhang Q Wang Y Liu Z J Wang and J QLu ldquoIdentification of phenolic acids from Kudiezi injection byHPLC-HR-MSsimnrdquo Central South Pharmacy vol 11 no 8 pp561ndash565 2013
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Inorganic ChemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
International Journal ofPhotoenergy
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Carbohydrate Chemistry
International Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Physical Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2014
Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
SpectroscopyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Chromatography Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Applied ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Theoretical ChemistryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Spectroscopy
Analytical ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Quantum Chemistry
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Organic Chemistry International
ElectrochemistryInternational Journal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
CatalystsJournal of