joint research centre - policija guillou, fabiano reniero, hubert chassaigne, joana lobo vicente,...

Joint Research CentreThe European Commissions in-house science service

ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315

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Eurodat number 15040014Received on 13 April 2015PACKAGING in 1 glass vialRegistration date 23 April 2015Name of customer Service commun des laboratoires FranceCustomer's identification 2015-25717Customer's information 2015-25717 white powder, SCL Paris, GC-MSAgilent, FT-IRThermo Nicolet iS10, LC-MSBruker Impact LC-Q-TofCustomer's comments hypothetic molecular weight[MH]+ 393.21Hypothetic raw formulaC23H26FN4Ocannabinoid ?

There was/were identified following structure(s) in the sample:

NN

F

F

NH

O

NH2O

Data of identified compound(s)

Claude Guillou, Fabiano Reniero, Hubert Chassaigne, Joana Lobo Vicente, Veronica Holland, Kamil Kolar

European Commission, Joint Research CentreInstitute for Health and Consumer Protection (IHCP)Chemical Assessment and Testing Unitvia E. Fermi, 2749TP 281 I-21027 Ispra (VA) - ItalyPhone: +39 0332 785678Fax: +39 0332 [email protected]://ihcp.jrc.ec.europa.eu

ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315 BETWEENDG TAXATION AND CUSTOMS UNION (DG TAXUD) AND THE JOINT RESEARCH CENTRE (JRC) for fastrecognition of New Psychoactive Substances (NPS) and identification of unknown chemicalsThis report was generated automatically by Kamil Kolar on 27/05/2015 based on data from the European Customslaboratories and the Joint Research Centre. This report includes sample and molecular information, spectra data andassociated tables and figures. The chemical structure(s) was/were identified by BCIM group chemists on the basis ofanalytical data available. The assignments proposed below were performed by using ACD labs tools in agreementwith the chemical structure identified by analytical experts. Reported data are connected to the sample in thefollowing table:

FW 392.4428IUPAC Name (v.14.01) N-(1-amino-3-methyl-1-oxobutan-2-yl)-1-(5-fluoropentyl)-5-(4-fluorophenyl)-1H-pyrazole-3-carboxamide (named manually)InChI (v.1.04) InChI=1S/C20H26F2N4O2/c1-13(2)18(19(23)27)24-20(28)16-12-17(14-6-8-15(22)9-7-14)26(25-16)11-5-3-4-10-21/h6-9,

12-13,18H,3-5,10-11H2,1-2H3,(H2,23,27)(H,24,28)InChI Key (v.1.04) GSXRDTDYPSATDE-UHFFFAOYSA-NSMILES (v.14.01) NC(=O)C(NC(=O)c2cc(c1ccc(F)cc1)n(CCCCCF)n2)C(C)CFormula C20H26F2N4O2



of 14

NN

F

F

NH

O

NH2O

Data measured by SCL Paris:

Wavenumber (cm-1)3500 3000 2500 2000 1500

%R

efle

ctan

ce

0

8

16

24

32

40

48

56

64

72

80

88

534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19534.19

555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88555.88

597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34597.34

660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98660.98675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45675.45

701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48701.48

747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76747.76

804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17

818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15818.15

839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85839.85

866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36866.36

882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27882.27

958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45958.45

1001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.841001.84

1043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.781043.78

1135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.381135.38

1157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.081157.08

1216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.861216.86

1273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.271273.27

1293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.521293.52

1303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.161303.16

1390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.911390.91

1440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.081440.08

1456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.961456.96

1505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171505.171528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.311528.31

1544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.221544.22

1563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.51563.5

1640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.161640.16

1688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.37

1704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.761704.76

1714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.161748.16

2798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.692798.69

2869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.082869.08

2956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.822956.82

2970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.322970.32

3032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513032.513076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.393076.39

3192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583192.583276.46

3276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.463276.46

3362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.283362.28

Title 25717File Name \\s-jrciprna004p-fs-vihcp.jrc.it\ihcp∴I01\BIO_CHEMICAL_INTERACTION_METABONOMICS\CLEN2SAND\!PROPOSED-ST

RUCTURE\CLEN_NAS\15040014\2015-25717\IR.SPADate Stamp 23 Feb 2015 09:01:49 Date 23 Feb 2015 11:02:04Technique Infrared Spectral Region IRX Axis Wavenumber (cm-1) Y Axis %ReflectanceSpectrum Range 525.0251 - 3500.1670 Points Count 6172Data Spacing 0.4821



of 14

m/z50045040035030025020015010050

0

20000

40000

60000

80000

100000

120000

140000

160000

180000

200000

220000

240000

260000

280000

300000

320000

340000

360000

380000

69.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.100069.1000

132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000132.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000133.0000

134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000134.0000

189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000189.0000 249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000249.1000 277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000277.1000

348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000348.2000

EI+

Comment eb Count 394 Data Type CondensedDate 18 Feb 2015 22:59:16 Date Stamp 18 Feb 15 9:47 pmEstimated Molecular Ion 547.000 File Name 2015-25717_Centroid_EI+Inlet Model GC Ion Mode EI+ Mass Spec Model STUP1Plot Type Stick Retention Time 6.770 Sample Ds CH2Cl2/CH3OH :

2015-25717Scan 1726 Scan Mode Centroid Spectrum Assigned 41.0% [34-549/0-100]TIC 1074.62 Total Signal 4180201

NN

F

F

NH

O

NH2O



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Table of fragments

No. Fragment Structure Formula Label m/z Calc.TIC Calc.(%)

Difference(Da)

m/z Exp. RI Exp.(%)

TIC Exp.(%)

AQI RDBEOriginComment

1 1-5,12-18,21,23(+H)N

+

1

N2

4

5

3

21O23

1213

15

1716

14

F18

C10H6FN2O M - C10H20FN2O 189.046 10.499 -0.046 189.000 100.000 10.499 1.0009.0Manual

2 1-18N1

N2

C+

4

53

6

78

910

F11

1213

1517

16

14

F18

C14H15F2N2 M - C6H11N2O2 249.120 10.905 -0.020 249.100 99.506 10.905 1.0007.5Manual

3 1-5,12-18,20-23,26-28(+H) N1N

2

4

5 3

21O23

12 13

15

1716

14

NH22CH+

2026

27

F18

28

C14H15FN3O M - C6H11FNO 260.119 1.004 -0.019 260.100 9.105 1.004 1.0009.0Manual

4 1-18,21,23 N1N

2

4

5 3

6

7 8

9 10

F11

C+

21

O23

12 13

15

1716

14

F18

C15H15F2N2O M - C5H11N2O 277.115 11.006 -0.015 277.100 99.095 11.006 1.0008.5Manual

5 1-18,20-23,26-28 N1

N2

4

53

6

78

910

F1121O

23

1213

1517

16

14

NH22

CH+

2026 27

F18

28

C19H24F2N3O M - CH2NO 348.188 6.438 0.012 348.200 55.199 6.431 0.9998.5Manual

6 M N+

1N2

4

53

67

89

10

F1121O

23

1213

1517

16

14

NH22

2026

27

F18

19 O25

28

NH224

C20H26F2N4O2 M 392.202 1.365 -0.002 392.200 11.500 1.365 1.0009.0Manual

NN

F

F

NH

O

NH2O



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Data measured by BCIM group at IHCP JRC Ispra

NMR spectraAcquisition Time (sec) 2.7263 Comment 25718 (F) 10mg in 600 uL DMSO-d6Date 14 Apr 2015 17:49:42 Date Stamp 14 Apr 2015 17:49:42File Name \\s-jrciprna004p-fs-vihcp.jrc.it\ihcp∴I01\Bio_Chemical_Interaction_Metabonomics\CLEN2SAND\!KK-proposed-structure-

temp-dir\BCIM_NAS\BCIM_600\15040003\1\PDATA\1\1rFrequency (MHz) 600.13 Nucleus 1HNumber of Transients 16 Origin spectOriginal Points Count 32768 Owner nmrsuPoints Count 65536 Pulse Sequence zg30Receiver Gain 1.91 SW(cyclical) (Hz) 12019.23Solvent DMSO-d6 Spectrum Offset (Hz) 3706.0515Spectrum Type standard Sweep Width (Hz) 12019.05Temperature (degree C) 27.000

Chemical Shift (ppm)8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0

Nor

mal

ized

Inte

nsity

0

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

0.55

0.60

0.65

0.70

0.75

0.80

0.85

M04(s, 5)

M03(s, 24)

M15(m, 28)

M14(s, 27)

M01(d, 22) M09(dd, 20)

M08(td, 6) M11(m, 7)

M02(m, 14,13)

M05(m, 16,15)

M12(m, 9)

M06(s, 24)

M13(br s, 8)

M07(m, 10)

M10(m, 26)

N1

N2

3

4

5

6

7

8

9

10F11

12

13

14

15

16

17

F18

1920

21

NH22

O23

NH224O

2526

CH327

CH328



of 14

Table of assignments and zoomed parts:

No. Atom Exp. Shift(ppm)

Multiplet

1 27 0.93 M142 28 0.95 M153 8 1.32 M134 9 1.64 M125 7 1.80 M116 26 2.11 M107 20 4.27 M098 10 4.40 M079 6 4.50 M0810 24 7.12 M0611 16 7.27 M0512 15 7.27 M0513 5 7.43 M0414 24 7.49 M0315 14 7.83 M0216 13 7.83 M0217 22 8.18 M01

Chemical Shift (ppm)8.2 8.1 8.0 7.9 7.8 7.7 7.6 7.5 7.4 7.3

Nor

mal

ized

Inte

nsity

0

0.1

0.2

0.3

M04(s, 5)

M03(s, 24)M01(d, 22)

M02(m, 14,13)

M05(m, 16,15)

M06(s, 24)

N1

N2

3

4

5

6

7

8

9

10F11

12

13

14

15

16

17

F18

1920

21

NH22

O23

NH224O

2526

CH327

CH328

Chemical Shift (ppm)4.6 4.5 4.4

Nor

mal

ized

Inte

nsity

0

0.05

0.10

0.15M09(dd, 20)

M08(td, 6)

M07(m, 10)

Chemical Shift (ppm)2.0 1.5

Nor

mal

ized

Inte

nsity

0.25

0.50

M15(m, 28)

M14(s, 27)

M11(m, 7)M12(m, 9) M13(br s, 8)M10(m, 26)



of 14

Acquisition Time (sec) 0.9044 Comment 5 mm CPQCI 1H/19F-13C/15N/D Z-GRD Z114073/0012Date 14 Apr 2015 18:02:01 Date Stamp 14 Apr 2015 18:02:01File Name \\s-jrciprna004p-fs-vihcp.jrc.it\ihcp∴I01\Bio_Chemical_Interaction_Metabonomics\CLEN2SAND\!proposed-structure\BCI

M_NAS\BCIM_600\15040014\3\PDATA\1\1rFrequency (MHz) 150.90 Nucleus 13CNumber of Transients 512 Origin spectOriginal Points Count 32768 Owner nmrsuPoints Count 32768 Pulse Sequence jmodReceiver Gain 81.72 SW(cyclical) (Hz) 36231.88Solvent DMSO-d6 Spectrum Offset (Hz) 18108.3359Spectrum Type APT Sweep Width (Hz) 36230.78Temperature (degree C) 27.002

Chemical Shift (ppm)170 160 150 140 130 120 110 100 90 80 70 60 50

Nor

mal

ized

Inte

nsity

-1.00

-0.95

-0.90

-0.85

-0.80

-0.75

-0.70

-0.65

-0.60

-0.55

-0.50

-0.45

-0.40

-0.35

-0.30

-0.25

-0.20

-0.15

-0.10

-0.05

0

0.05

0.10

M12(s) M06(s)

M04(s)

M03(s)

M02(s, 27)

M26(s, 21)M25(s, 3)

M24(s, 4)

M23(s)

M22(s, 12)

M20(s, 14,13)M19(s, 15)

M17(s, 5)

M10(s)

M29(s, 19)

M21(s)M14(s, 20)

M13(s, 6)

M11(s)

M08(s, 26)

M05(s)

M01(s, 28)

M30(m, 8)

M28(m, 9)

M09(dd)

M15(br d, 10)

M27(br d, 17)

N1

N2

3

4

5

6

7

8

9

10F11

12

13

14

15

16

17F18

1920

21

NH22

O23

NH224O

2526

CH327

CH328



of 14

Table of assignments and zoomed parts:

No. Atom Exp. Shift(ppm)

Multiplet

1 28 18.89 M012 27 19.84 M023 8 22.21 M304 9 29.78 M285 7 30.22 M076 26 30.54 M087 6 50.93 M138 20 58.61 M149 10 84.06 M1510 5 105.13 M1711 16 116.05 M1812 15 116.20 M1913 14 127.40 M2014 13 127.40 M2015 12 129.76 M2216 4 137.01 M2417 3 147.91 M2518 21 159.72 M2619 17 162.29 M2720 19 173.05 M29


Nor

mal

ized

Inte

nsity

-0.2

-0.1

0

M17(s, 5)

M23(s)

M22(s, 12)

M20(s, 14,13)

M29(s, 19)

M26(s, 21) M25(s, 3)M24(s, 4)

M19(s, 15)

M27(br d, 17)


Nor

mal

ized

Inte

nsity

-1.0

-0.5

M12(s)M07(s, 7)

M06(s) M04(s)

M03(s)

M02(s, 27)

M10(s)

M05(s)

M14(s, 20)

M13(s, 6)

M11(s)

M08(s, 26)

M30(m, 8)M28(m, 9)

M09(dd)

M15(br d, 10)

N1

N2

3

4

5

6

7

8

9

10F11

12

13

14

15

16

17F18

1920

21

NH22

O23

NH224O

2526

CH327

CH328



of 14

Acquisition Time (sec) (0.2840, 0.0206) Comment 5 mm CPQCI 1H/19F-13C/15N/D Z-GRD Z114073/0012Date 15 Apr 2015 09:34:54File Name \\s-jrciprna004p-fs-vihcp.jrc.it\ihcp∴I01\Bio_Chemical_Interaction_Metabonomics\CLEN2SAND\!proposed-structure\BCI

M_NAS\BCIM_600\15040014\6\PDATA\1\2rrFrequency (MHz) (600.13, 150.90) Nucleus (1H, 13C)Number of Transients 4 Origin spectOriginal Points Count (2048, 512) Owner nmrsuPoints Count (1024, 1024) Pulse Sequence hsqcedetgpsisp2.3Solvent DMSO-d6 Spectrum Type HSQCSweep Width (Hz) (7204.50, 24851.33) Temperature (degree C) 27.001

7.83, 127.43 (13, 13)(14, 14)

0.93, 18.87 (27, 27)

0.94, 19.83 (28, 28)

7.27, 116.15 (15, 15)(16, 16)

7.43, 105.10 (5, 5)

1.80, 30.21 (7, 7)

4.50, 50.91 (6, 6)

4.27, 58.59 (20, 20)

1.32, 22.20 (8, 8)

2.11, 30.54 (26, 26)

1.64, 29.77 (9, 9)

2.51, 40.21

4.38, 84.60 (10, 10)

8.18, 58.59

15040014.006.001.2rr.man-assign.esp

F2 Chemical Shift (ppm) 8 7 6 5 4 3 2 1 0 -1

F1 C

hem

ical

Shi

ft (p

pm)

24

32

40

48

56

64

72

80

88

96

104

112

120

128

136

144

13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14

5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16

6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8

28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27

13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14 13, 14

15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16 15, 16

5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5

10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10

20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20

6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6

26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 26 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9

8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 28 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27 27

N1N

2

3

4

5

67

89

10F11

12

13

14

15

16

17F18

1920

21

NH22

O23

NH224O2526

CH3 27CH328



of 14

Table of assignments:No. F2 Atom F1 Atom F2

(ppm)F1

(ppm)1 5 5 7.43 105.102 6 6 4.50 50.913 7 7 1.80 30.214 8 8 1.32 22.205 9 9 1.64 29.776 9 9 1.32 29.877 10 10 4.38 84.608 13 13 7.83 127.439 14 14 7.83 127.4310 15 15 7.27 116.1511 16 16 7.27 116.1512 20 20 4.27 58.5913 26 26 2.11 30.5414 27 27 0.93 18.8715 28 28 0.94 19.83



of 14

NN

F

F

NH

O

NH2O

MS spectra

Retention Time: 0.823 Ion Mode: ESI+

m/z50045040035030025020015010050

0

1090000

2180000

3270000

4360000

5450000

6540000

7630000

8720000

9810000

10900000

11990000

13080000

14170000

15260000

16350000

17440000

18530000

19620000

20710000

21800000

22890000

M+K

M+Na

M+H84.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.748484.7484 168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660168.3660180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505180.0505

193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711193.0711

198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657198.0657

212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849212.0849

250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449250.1449

339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554339.0554

393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115393.2115

431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676431.1676ESI+

Combine <84-87> Count 409Data Source Name 15040014_25717_MS mode_50_1000_RES30000_1Data Type p Date 24 Apr 2015Estimated Molecular Ion 996.921File Name \\s-jrciprna004p-fs-vihcp.jrc.it\ihcp∴I01\Bio_Chemical_Interaction_Metabonomics\CLEN2SAND\!proposed-structure\B

CIM_NAS\BCIM_Orbi\15040014_25717\15040014_25717_MS mode_50_1000_RES30000.rawIon Mode ESI+ Mass Spec Model LTQ Orbitrap/LTQ OrbitrapPlot Type Profile Retention Time 0.823Scan 85 Scan Filter FTMS + p ESI Full ms [50.00-1000.00]Scan Mode ms Scan Type FullSpectrum Assigned 27.8% [50-997/0-100] TIC 421.47Total Signal 95607024

No. Fragment Structure Formula Label m/z Calc. TIC Calc.(%)

Difference(Da)

m/z Exp. RI Exp.(%)

TIC Exp.(%)

AQI RDBE Origin Comment

1 M(+H) NH+

1N2

4

53

67

89

10

F1121O

23

1213

1517

16

14

NH22

2026

27

F18

19 O25

28

NH224

C20H27F2N4O2 M + H 393.210 29.502 0.002 393.212 97.507 28.402 0.963 8.5 Manual



of 14

NN

F

F

NH

O

NH2O

Retention Time: 0.978 Ion Mode: ESI+

m/z380360340320300280260240220200180160

0

60000

120000

180000

240000

300000

360000

420000

480000

540000

600000

660000

251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667251.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667260.1667

277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500277.2500 288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667288.1667

305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500305.2500328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333328.3333

348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500348.2500

373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333373.3333

376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667376.1667393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500393.2500

ESI+

Count 1915 Data Type pDate 20 Apr 2015 Estimated Molecular Ion 393.500File Name \\s-jrciprna004p-fs-vihcp.jrc.it\ihcp∴I01\Bio_Chemical_Interaction_Metabonomics\CLEN2SAND\!proposed-structure\BC

IM_NAS\BCIM_Orbi\15040014_25717\15040014_25717_MSMS mode_393_CE 20_105_400.rawIon Mode ESI+ Mass Spec Model LTQ Orbitrap/LTQ OrbitrapPlot Type Profile Retention Time 0.978Scan 120 Scan Filter ITMS + p ESI Full ms2 [email protected] [105.00-400.00]Scan Mode ms2 Scan Type FullSpectrum Assigned 17.5% [149-399/0-100] TIC 654.02Total Signal 4466182.5

No. Fragment Structure Formula Label m/z Calc.TIC Calc.(%)

Difference(Da)

m/z Exp. RI Exp.(%)

TIC Exp.(%)

AQI RDBEOriginComment

1 1-5,12-18,20-23,26-28(+H) N1N

2

4

5 3

21O23

12 13

15

1716

14

NH22CH+

2026

27

F18

28

C14H15FN3O M - C6H11FNO 260.119 0.427 -0.036 260.083 2.356 0.367 0.861 9.0 Manual

2 1-18,21,23 N1N

2

4

5 3

6

7 8

9 10

F11

C+

21

O23

12 13

15

1716

14

F18

C15H15F2N2O M - C5H11N2O 277.115 0.538 -0.031 277.083 2.946 0.472 0.878 8.5 Manual

3 1-18,20-23,26-28 N1

N2

4

53

6

78

910

F1121O

23

1213

1517

16

14

NH22

CH+

2026 27

F18

28

C19H24F2N3O M - CH2NO 348.188 17.513 -0.022 348.167 91.391 14.355 0.820 8.5 Manual

4 1-23,25-28 N1

N2

4

53

6

78

910

F1121O

23

1213

1517

16

14

NH22

20

26 27

F18

C+

19O25

28

C20H24F2N3O2 M - H2N 376.183 1.503 -0.016 376.167 7.737 1.222 0.813 9.5 Manual

5 M(+H) NH+

1N2

4

53 6

7 89

10

F1121O

23

1213

1517

16

14

NH22

2026

27

F18

19 O25

28

NH224

C20H27F2N4O2 M + H 393.210 1.693 0.040 393.250 8.683 1.382 0.816 8.5 Manual



of 14

Fragmentation of the observed molecule and the virtual MS and MSMS spectra predictedby Thermo Scientific Mass Frontier

FNN

F

NH

O

N 2HO

FNN

F

NH

O

N 2HO

H

m/z 393.21

FNN

F

NH

O

N 2HO

FNN

F

NH

O

N 3HO

m/z 393.21

+H

O

NH

ON

N

F

F

m/z 376.18

-NH3

FNN

F

NH

O

N 2HO

FNN

F

NH

OH

N 2HO

m/z 393.21

+H

NH

ON

N

F

F

m/z 348.19

rHC

FNN

F

NH

O

N 2HO

FNN

F

N2H

O

N 2HO

m/z 393.21

+H

ON

N

F

F

m/z 277.11

i

FNN

F

NH

O

N 2HO

FNN

F

NH

OH

N 2HO

m/z 393.21

+H

NH

ON

N

F

F

m/z 348.19

rHCNH

NNH

O

F

m/z 260.12

rHR



of 14

3060

9012

015

018

021

024

027

030

033

036

039

042

045

048

051

054

057

060

063

066

069

072

075

078

081

084

087

090

093

096

099

0

013253850637588100

393.2

118

250.1

4474 250.1

4497

393.2

115

394.2

151

393.2

121

212.0

8436

193.0

7016

251.1

4821

338.3

434

251.1

4799

415.1

937

149.0

2361

100.1

1212

373.2

055

462.0

785

943.0

146

570.4

663

721.9

965

821.4

725

511.9

969

639.5

547

758.1

659

675.6

817

886.0

710

312.0

101

979.7

163

605.3

137

FN

N

F

NHO

N2

HO

H

170

180

190

200

210

220

230

240

250

260

270

280

290

300

310

320

330

340

350

360

370

380

390

013253850637588100

348.1

4

376.0

9739

3.20

277.0

8426

0.093

328.1

628

8.01

305.1

925

1.11

382.8

0920

9.00

231.0

9018

8.998

221.0

1435

6.115

317.1

524

3.110

337.2

7929

4.29

372.6

436

5.28

181.0

7226

9.00

NHO

NN

F

F

O

NHO

NN

F

F

FN

N

F

NHO

N2

HO

H

ON

N

F

FN

HN

NHO

F

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