European Reference Laboratory - FV
European Reference Laboratory - FV
Adding of new compounds to the routine methods from the
working document SANCO/12745/2013 of
21st – 22nd November 2017 rev. 9(1) 48 compounds by LC and GC, fully validated at 0.005 and 0.050 mg/kg in tomato, orange and avocado
Ametoctradin (LC) Novaluron (LC)
Anthraquinone (GC) Penflufen (LC)
BAC10 (LC) Penthiopyrad (LC)
BAC12 (LC) Phenthoate (GC)
BAC8 (LC) Picolinafen (GC)
Benalaxyl (GC) Propaquizafop (LC)
Chlorfluazuron (GC) Proquinazid (LC)
Clomazone (LC) Prothioconazole (LC)
Cyazofamid (LC) Prothioconazole-desthio (LC)
Cyflufenamid (GC) Prothiofos (GC)Etoxazole (LC) Pyridalil (LC)
Fenpyrazamine (LC) Pyridate (LC)
Flufenacet (LC) Pyriofenone (GC)
Fluopicolide (GC) Quinoclamine (LC)
Fluxapyrosad (LC) Quintozene (GC)
Heptachlor (GC) Rotenone (LC)
Heptachlor endo-epoxide (GC) Spinetoram (LC)
Heptachlor exo-epoxide (GC) Spirotetramat (LC)
Ioxynil (LC) Sulfoxaflor (LC)
Isopyrazam (GC) Tetramethrin (GC)
Isoxaflutole (LC) Triallate(GC)
Lufenuron (LC) Tricyclazole (LC)
Metconazole (LC) Triticonazole (LC)
Metrafenone (LC) Tritosulfuron (LC)
Molinate (GC)
European Reference Laboratory - FV
Molecular Components Map of representative matrices of commodity groups (48 matrices)
European Reference Laboratory - FV
Exact Mass database for LC-HRMS
(with 188 compounds)
Enlarge of the scope of the Accurate Mass Database by GC-HRMS (150
compounds)
European Reference Laboratory - FV
Validation of the MRM methods at low concentration levels
(0.002 mg/kg in GC-MS/MS and at
0.005 mg/kg in LC-MS/MS)
203 compounds by GC-MS/MS
195 compounds by LC
All of them, fully validated in tomato, orange, apple, avocado and tea
European Reference Laboratory - FV
SELECTIVITY SENSITIVITY
European Reference Laboratory - FV
Sensitivity
European Reference Laboratory - FV
Liquid-like density
Gas-like viscosity
Low critical point
High diffusivity
Widely available, safe to use
Capability to mix with a wide range of liquid solvents
Tc = 304.1 K
Pc = 73.8 bar
scCO2 AS MOBILE PHASE
European Reference Laboratory - FV
SFC-MS/MS Systems
Co-solventpump
Auxiliary
pump
ColumnBP
R
CO2
pump
MeOH
MeOH
Ammonium formate (5
mM)
Formic acid (0,1%)
Column flow1.5 mL/min
Post-column flow80 - 150 µL/min
MS
analyzer
ESI source
European Reference Laboratory - FV
BP
R
SUPERCRITICAL
CO2
(UNTIL CROSS THE
BPR DEVICE)
MS
ESI
SMALL AMOUNT OF ORGANIC SOLVENT REACHING THE SOURCE
European Reference Laboratory - FV
Absence of water
• Lower surface tension
• Lower polarity
CO2 evaporation
• Lower flow in the MS detector
• Reduced microdroplet size
Microdroplet in LC (containing
water)
Microdroplet in SFC (onlyMeOH)
IONIZATION EFFICIENCY
More analyte reaching the gas
phase and the mass analyzer.
Increase of sensitivity
European Reference Laboratory - FV
0
1
2
3
4
5
LC SFC
1 mg sample
0.4 mg sample
Lower injection volumes(matrix concentration 0.2 mg/uL)
Lower amount of sample
with no loss of sensitivity
Injection volume (µL)
INCREASE OF SENSITIVITY
Reduced matrixeffect
European Reference Laboratory - FV
164 pesticides (log KOW -0.8 to
6.9)Tomato High water content
FRUITS AND VEGETABLES
Orange Acidic matrix
Leek Interferingcompounds
Application of SFC-MS/MS
for the detection of
pesticides
in fruits and vegetables
European Reference Laboratory - FV
150µL/min
125µL/min
100µL/min
90µL/min
80µL/min
MAKE-UP FLOW
7,7
x1
08
8,5
x1
08
8,9
x1
08
9,4
x1
08
9,7
x1
08
I.D: 127µm I.D: 64 µm
PEEK DIAMETER
TOTAL AREA OF COMPOUNDS
European Reference Laboratory - FV
50
55
60
65
70
75
80
85
90
95
100
50
55
60
65
70
75
80
85
90
95
100
50
55
60
65
70
75
80
85
90
95
100100%
90%
80%
70%
60%
50%
5 ppb 10 ppb 20 ppb
98% 98%94%
SENSITIVITY - IDENTIFIED COMPOUNDS
European Reference Laboratory - FV
-1
0
1
2
3
4
5
6
7
0% 20% 40% 60% 80% 100%
log
Po
w
RT(%)Increasedpolarity LC SFC
ELUTION ORDER
End of chromatograp
hic run
European Reference Laboratory - FV
0
10
20
30
40
50
60
70
80
90
100
Irrelevant matrixeffect
(<20% suppression)
Low matrix effect (20-50%
suppression)
Significant matrix effect
(>50% suppression)
99% 87% 62%
12%
35%
1% 3%
MATRIX EFFECTEvaluation
Comparison between calibration curves built in matrix and solvent (considered as no suppression)
European Reference Laboratory - FV
Carbaryl
Tomato Orange Leek
0
500000
1000000
1500000
2000000
2500000
3000000
3500000
4000000
0 100 200 300 400 500
Profenofos
0
1000000
2000000
3000000
4000000
5000000
6000000
7000000
8000000
9000000
0 100 200 300 400 500
Matrix effect is more
intense in samples with a
high number of
interfering compounds.
Similar calibration curves
for these three different
matrices illustrate the low
matrix effect.
“
”
MATRIX EFFECT
European Reference Laboratory - FV
The use of high flow rates promotes the
coelution of matrix compounds and
analytes, which leads to modification of
ion ratios (>30%).
Tomato
Isoprocarb
Fenhexamid
Orange
Isoprocarb
Lufenuron
Epoxiconazole
Penthion-sulfone
Fenoxycarb
Flutriafol
Imazalil
Fenarimol
Phoxim
Trichlorfon
Leek
331>268 331>189331>110
Ion Ratio
53,2Ion Ratio
51,5
Ion Ratio
240,1Ion Ratio
54,1
ISOBARIC INTERFERENCES
European Reference Laboratory - FV
Reproducibility
5 replicates of each matrixextract
Linearity
Matrix-matched standards in the
range of 1-100 µg/L
(5-500 µg/kg in samples)
0,0E+0
1,0E+6
2,0E+6
3,0E+6
4,0E+6
5,0E+6
6,0E+6
7,0E+6
8,0E+6
0 100 200 300 400 500
Calibration curve (Profenofos –leek)
METHOD VALIDATION
7,00
6,00
5,00
4,00
3,00
2,00
1,00
x106
R2 = 0,99969
83
161
81
181
80
191
0%
20%
40%
60%
80%
100%
RSD of compounds (5ppb)
<10 % 10-20 % >20 %
European Reference Laboratory - FV
SPICES
Application of SFC-MS/MS for the
detection of pesticides in spices
162 pesticides (log KOW -0.9 to 6.9)
Cayenne
Interferring matrixcomponents
Strong matrix effect
Black pepper
European Reference Laboratory - FV
MATRIX EFFECT
Low flow rate in the source
Reduced microdroplet size
Absence in water in the mobile
phase
Lower polarity and surface tension
Smaller amount of sample injected
Fewer interferences81 56
13
27
6
17
0
20
40
60
80
100
Cayenne Black pepper
Perc
enta
ge o
f com
pounds (
%)
Weak Medium Strong
European Reference Laboratory - FV
METHOD VALIDATION
Calibration curve (Isoprocarb in cayenne)
0
500000
1000000
1500000
2000000
2500000
0 100 200 300 400 500 600
Sig
nal
Concentration
R2 = 0.99995
0
20
40
60
80
100
RSD < 20% RSD < 10%
Perc
enta
ge o
f com
pou
nds
Cayenne Black Pepper
ReproducibilityLinearity
Matrix-matched standards in the range of 5-
500 µg/L
(50-5000 µg/kg in samples)
European Reference Laboratory - FV
REAL SAMPLES
0
2
4
6
8
10
12
14
16
18
20
Carb
en
dazi
m
Imid
acl
op
rid
Ace
tam
ipri
d
Azo
xyst
rob
in
Meta
laxy
l
Teb
uco
nazo
le
Ch
lorp
yrif
os
Pro
fen
ofo
s
Ch
lora
ntr
an
ilip
role
Flu
op
yram
Myc
lob
uta
nil
Tri
azo
ph
os
Eth
ion
2,4
-D
Deet
Dif
en
oco
nazo
le
Kre
soxi
m-m
eth
yl
Lin
uro
n
Pro
pic
on
azo
le
Meth
am
ido
ph
os
Flo
nic
am
id
Flu
azi
fop
Hexa
con
azo
le
Pyr
acl
ost
rob
in
Th
iacl
op
rid
Th
iam
eth
oxa
m
Bo
scalid
Man
dip
rop
am
id
Fen
pyro
xim
ate
Flu
sila
zole
Ind
oxa
carb
Tri
flo
xyst
rob
in
Eto
fen
pro
x
Meth
om
yl
Pir
imip
ho
s-m
eth
yl
Pro
parg
ite
Pro
piz
am
ide
Bro
mu
con
azo
le
Bu
pro
fezi
n
Dic
roto
ph
os
Eth
op
rop
ho
s
Fen
pro
path
rin
Imaza
lil
Pyr
idab
en
Pyr
ipro
xyfe
n
Teb
ufe
no
zid
e
Nu
mb
er
of
dete
ctio
ns
Above the MRLBelow the MRL
Total number of samples: 47
European Reference Laboratory - FV
FAPAS (2017)
Pesticide Residues in pear purée
JRC-GEEL (2017)
Fipronil in eggs
1,2
0,9
1,2
1,6
0,6
-2
-1,5
-1
-0,5
0
0,5
1
1,5
2
Z-S
co
re
0,2 0,2 0,3
-2
-1,5
-1
-0,5
0
0,5
1
1,5
2
Z-s
co
re
PROFICIENCY TESTS
European Reference Laboratory - FV
CONCLUSIONS
SFC provides a high sensitivity of the analysis
even with small amounts of sample injected:
- Absence of water.
- Evaporation of CO2 in the mobile phase.
The use of SFC-MS/MS allowed the
identification of the majority of 164 pesticides
in tomato (98%), orange (98%) and leek (94%)
at 5 µg/kg. Good recoveries (70-120%) were
obtained in cayenne (89%) and black pepper
(93%) at 50 µg/kg.Some polar and acidic compounds underwent
an increase of their signals. Their peak shapes
could also be improved.
Very low matrix effect was observed for most
compounds in all matrices studied.
European Reference Laboratory - FV
LC/MSGC/MS
European Reference Laboratory - FV
0
50
100
150
200
250
300
350
0 5 10 15 20
T °C
Time (min)
Oven temperature program with
different runtime methods
9,412,4 19,3Ramp 1
30ºC/min
Ramp 2
20ºC/min
Ramp 3
10ºC/min
European Reference Laboratory - FV
2,4'-DDE
2-Phenylphenol
4,4'-DDD
4,4'-DDE
4,4'-DDT
Acrinathrin
Alachlor
Aldrin
Ametryn
Atrazine
Azoxystrobin
Benalaxyl
Bifenox
Bifenthrin
Biphenyl
Bixafen
Boscalid
Bromopropylate
Bupirimate
Buprofezin
Butralin
Butylate
Cadusafos
Carbofuran
Carbophenothio
n
Chinomethionat
e
Chlorbromuron
Chlordane
Chlorfenapyr
Chlorfenvinphos
Chlorobenzilate
Chlorothalonil
Chlorpropham
Chlorpyrifos
Chlorpyrifos-methyl
Chlorthal-dimethyl
Chlozolinate
Coumaphos
Cyfluthrin
Cypermethrin
Cyproconazole
Cyprodinil
Deltamethrin
Desmethyl-pirimicarb
Diazinon
Dichlofluanid
Dichloran
Dichlorobenzopheno
ne
Dichlorvos
Diclobutrazol
Dicofol
Dieldrin
Diethofencarb
Dimethenamid
Dimethipin
Diphenylamine
Disulfoton
Disulfoton-sulfoxide
Dodemorph
Endosulfan sulfate
Endosulfan-alpha
Endosulfan-beta
Endrin
EPN
Epoxiconazole
Ethion
Ethofenprox
Ethofumesate
Ethoprophos
Ethoxyquin
Etrimfos
Fenamidone
Fenarimol
Fenazaquin
Fenbuconazole
Fenchlorphos
Fenhexamid
Fenitrothion
Fenpropathrin
Fenpropidin
Fenpropimorph
Fenthion
Fenvalerate
Fipronil
Fipronil sulfone
Fipronil-desulfinil
Flamprop-
isopropyl
Flamprop-methyl
Fluacrypyrim
Fluazifop-p-butyl
Flucythrinate
Fludioxonil
Fluopicolide
Fluopyram
Fluquinconazole
Flusilazole
Flutolanil
Flutriafol
Fluvalinate-tau
Fonofos
Formothion
Fosthiazate
HCB
HCH-alpha
HCH-beta
Heptachlor
Heptachlor endo-
epoxide
Heptachlor exo-epoxide
Heptenophos
Hexaconazole
Indoxacarb
Iprodione
Iprovalicarb
Isazofos
Isocarbophos
Isofenphos
Isofenphos-methyl
Isoprothiolane
Isopyrazam
Kresoxim-methyl
-Cyhalothrin
Lindane
Malaoxon
Malathion
Mecarbam
Mepanipyrim
Merphos
Metalaxyl
Metazachlor
Metconazole
Methidathion
Methiocarb
Methoxychlor
Metolachlor
Mevinphos
Molinate
Myclobutanil
Napropamide
Nuarimol
Ofurace
Oxadixyl
Paclobutrazol
Paraoxon-methyl
Parathion
Parathion-methyl
Penconazole
Pendimethalin
Pentachloroanili
ne
Permethrin
Phenothrin
Phenthoate
Phorate
Phorate sulfone
Phosmet
Phthalimide
Picolinafen
Picoxystrobin
Pirimicarb
Pirimiphos-
methyl
Procymidone
Profenofos
Prometon
Prometryn
Propaphos
Propazine
Propiconazole
Propyzamide
Prosulfocarb
Prothiofos
Pyraclostrobin
Pyrazophos
Pyridaben
Pyrifenox
Pyrimethanil
Pyriproxyfen
Quinalphos
Quinoxyfen
Quintozene
Secbumeton
Spirodiclofen
Spiromesifen
Sulfotep
Sulprofos
Tebuconazole
Tebufenpyrad
Tecnazene
Tefluthrin
Terbufos
Terbumeton
Terbutryn
Tetrachlorvinphos
Tetraconazole
Tetradifon
THPI
Tetramethrin
Thiobencarb
Tolclofos-methyl
Tolylfluanid
Triadimefon
Triazophos
Trifloxystrobin
Trifluralin
Vinclozolin
203 GC compounds
European Reference Laboratory - FV
Azoxystrobin (data points per peak with the 5 runtime methods)
16 data points per peak
38
38
42
4219.3 min
17.6 min
15.3 min
12.4 min
9.4 min
European Reference Laboratory - FV
European Reference Laboratory - FV
12.4 min9.4 min
Cyproconaz
ole
Chlorobenzilate
139>11
1
Overlapping peaks
100>72
9.3 min 12.4 min
Thiobencarb
Interference
Interference
+changement in
peaks order
European Reference Laboratory - FV
Extraction MethodCitrate buffered QuEChERS
Centrifuge 5 minutes at 3700 rpm
10 g of Sample + 10 ml of AcN
Acidify with 10 uL formic acid 5 %
per mL of extract
4 g MgSO4 + 1 g NaCl +
1 g NaCit·2H2O +0.5 g NaCit·1.5H2O
Centrifuge 5 minutes at 3700 rpm
Shake 4 min
Take 5 mL + 750 mg MgSO4
+ 125 mg PSA
Shake 4 min
Analysis
1 g Sample/mL
Solvent exchange (EtAc)
European Reference Laboratory - FV
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
2 µg.kg-1 10 µg.kg-1 50 µg.kg-1
Tomato
Apple
Orange
LOQ
European Reference Laboratory - FV
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
Weak Moderate Strong
75%
25%
0%
86%
13%
0%
Apple/Tomato
Orange/Tomato
Matrix effect %
-20 to 20 % -20 to -50
%
and
20 to 50 %
<-50 and >50
%
European Reference Laboratory - FV
Samples with no pesticide residues
Samples with detections > 2 µg.kg-1
Samples with detections > 10 µg.kg-1
12%
88 %
European Reference Laboratory - FV
European Reference Laboratory - FV
Extraction MethodModified Citrate buffered QuEChERS
Mix and centrifuge for 5 minutes at 3700 rpm
2 g of Sample + 7 ml H20 (30 min soaking time)
Transfer 5 mL of the supernatant into Polish tubes (0.4 g NaCl+ 1.6 g MgSO4)
Add 4 g MgSO4 + 1 g NaCl +
1 g NaCit·2H2O +0.5 g NaCit·1.5H2O
Shake 7 min
Centrifuge 5 minutes at 3700 rpm
Shake 7 min
Activate EMR-Lipid by adding 5 mL of water
Transfer 5 mL of the supernatant into EMR-Lipid tubes
Analysis of
0,04 g Sample/mL
Add 10 mL ACN 5 times dilution
Mix and centrifuge for 5 minutes at 3700 rpm
Solvent Exchange (EtAc) 5 times dilution
European Reference Laboratory - FV
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
50 µg.kg-1 200 µg.kg-1 >200 µg.kg-1
2 µg.kg-1 8 µg.kg-1 >8 µg.kg-1
Black Pepper
Cayenne Pepper
LOQM/LOQI
European Reference Laboratory - FV
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Weak Moderate Strong
97%
12%5%
55%
89%
4%
Black pepper/tomato
Cayenne/tomato
Matrix effect
European Reference Laboratory - FV
0%
5%
10%
15%
20%
25%
30%
No detectedpesticides
1 detectedpesticide
2 detectedpesticides
3 detectedpesticides
4 detectedpesticides
> 5 detectedpesticides
5% 5…
23%
27%
18%
14%
Total of 50 samples
Percentage of samples
European Reference Laboratory - FV
European Reference Laboratory - FV
Injection
volume
5 mg
1 mg
LC: 5 µL
GC: 1 µL
2.5 mg
0.5 mg
0.5 mg
0.1 mg
1 mg
0.2 mg
0.25 mg
0.05 mg
Matrix injected in column
Dilx01 g/mL
Dilx20.5 g/mL
Dilx50.2 g/mL
Dilx100.1 g/mL
Dilx200.05 g/mL
Green
Tea
European Reference Laboratory - FV
0
10
20
30
40
50
60
No Dilution Dil x 5 Dil x 10 Dil x 20
% P
es
tic
ide
s S
up
pre
ss
ion
>50
%
Leek
Orange
Broccoli
Tomato
Pepper
Grape
Cucumber
5 mg 1 mg 0.5 mg 0.25 mg
LC-TOF-MS
EFFECT OF DILUTION IN “EASY-MEDIUM” COMPLEX MATRICES
European Reference Laboratory - FV
RESOLUTION
European Reference Laboratory - FV
50% of hight
Resolution at FWHM - full-width half maximum(Full Width of the peak at Half its Maximum height)
200
0.04= 5000
200.
0.004= 50000
For example:
m
Δm
R =m
∆m
200
0.4= 500
European Reference Laboratory - FV
Effects of increased resolution
5 00010 00020 00050 000
100 000
199.90 199.92 199.94 199.96 199.98 200.00 200.02 200.04 200.06 200.08 200.10
m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rela
tive
Ab
un
da
nce
European Reference Laboratory - FV
Resolution: 20 000
200.0000 200.0100 200.0000 200.0040
Resolution: 50 000
Effects of increased resolution
200.0000 200.0200 200.0000 200.0080
European Reference Laboratory - FV
RESOLUTION
(30,-50,000)
+MASS ACCURACY
(5ppm)
Nominal Mass----Exact Mass
European Reference Laboratory - FV
INSTRUMENTATION:• Time of flight• Orbitrap• Fourier transform ion cyclotron• (electric/magnetic) sector instrument
European Reference Laboratory - FV
Full Scan or Full
Scan?
That is the NO
question
GC-EI-HRMS
European Reference Laboratory - FV
Thiobencarb0.01 mg/kgin Tomato
GC-Intuvo MS/MS (100>72)
9.3 minInterference +shifts in peaks order
Thiobencarb
Interference
GC-Orbitrap MS
m/z 100.0757m/z 125.0153
European Reference Laboratory - FV
Diethofencarb
0.010 mg/kg in Tomato
GC-Intuvo-MS/MS
SRM1: 151>122SRM2: 207>151
GC-Orbitrap MS
m/z 124.0393m/z 158.0291
European Reference Laboratory - FV
Fluquinconazole
0.005 mg/kg in Tomato
7250 GC-QToF-MS GC-Orbitrap-MS
m/z 340.0396m/z 313.0287
European Reference Laboratory - FV
Full scan MS60k
GC-EI-QExative
Real Samples: Fluopyram
Orbitrap GC-EI-MS System
50 µg/kg Std in Tomato Real Sample: Pear
C = 26 µg/kg
Ratio Ion 2/1: 50%
Ratio Ion 3/1: 30%
Ratio Ion 4/1: 35%
Ratio Ion 2/1: 52%
Ratio Ion 3/1: 29%
Ratio Ion 4/1: 34%
The ion ratios are within ± 4% with
respect to the standard and mass
accuracy below 0.2 mDaIon 2 (145.0260): 0.14 mDa
Ion 3 (195.0057): -0.08 mDa
Ion 4 (223.0244): 0.02 mDa
Ion 1 (173.0209): 0.01 mDa
European Reference Laboratory - FV
SENSITIVITY
(10 ug/kg)
LINEARITY (2
orders) +
REPRODUCIILITY
(< 20%)
European Reference Laboratory - FV
MS MS2
targeted SIMNarrow quadrupole window
with or without triggering
non-targeted Full Scan Wide quadrupole window(s)
All Ion
Fragmentation or
NOT AIF?
That is the question
LC-HR-ESI-MS/MS
European Reference Laboratory - FV
Carbendazim in Carrot
0.01 mg/kg 0.1 mg/kg
Mass accuracy precursor: -1.0 ppm
Mass accuracy fragment: -4.9 ppm
Ion ratio: 0.14
Mass accuracy precursor: -0.1 ppm
Mass accuracy fragment: -4.4 ppm
Ion ratio: 0.17
Precursor Ion: 192.0768
Fragment Ion: 160.0495
MRM-LC-HR-MS/MS, Isolation Mass Window: 0.7 Da
Ratio Difference: -18%
European Reference Laboratory - FV
SENSITIVITY
(10 ug/kg)
LINEARITY (2
orders) +
REPRODUCIILITY
(< 20%)
European Reference Laboratory - FV
INSTRUMENTAL ISSUES ASSOCIATED
WITH LC-HRMS
WORKING IN ALL ION FRAGMENTATION
European Reference Laboratory - FV
Aubergine
Flufenoxuron – AIF 1 window
Broccoli Lemon
Ratio difference within ± 65 %
0.010 mg/kg
0.050 mg/kg
[M+H]+
F1
[M+H]+
F1
[M+H]+
F1
[M+H]+
F1
[M+H]+
F1
[M+H]+
F1
European Reference Laboratory - FV
Mass Peticides - tomato matrix compounds
2408 Matrix
compounds
1Da
1 min
European Reference Laboratory - FV
Mass
Pesticides - orange matrix compounds
8017 Matrix
compounds
1Da
1 min
European Reference Laboratory - FV
Mass
Pesticides - orange matrix compounds
8017 Matrix
compounds
1 min
European Reference Laboratory - FV
Mass
Pesticides - orange matrix compounds
8017 Matrix
compounds
80Da
1 min
European Reference Laboratory - FV
Aubergine
Flufenoxuron – AIF 10 windows
Broccoli Lemon
Ratio difference within ± 18 %
0.010 mg/kg
0.050 mg/kg
[M+H]+
F1
[M+H]+
F1
[M+H]+
F1
[M+H]+
F1
[M+H]+
F1
[M+H]+
F1
European Reference Laboratory - FV
BUT……….
European Reference Laboratory - FV
Cycle Time
LC-QTOF-MS X500R (AIF -20 Windows)
LC-TQ-MS
LC-QOrbitrap-MS (AIF)
70k 70k
0.27 s 0.6 s
0.2 s 1.2 s
32k32
k
32
k
32
k
32
k
32
k
32
k
32
k
32
k
32
k
32
k
32
k
0.05 s x 20 Windows = 1 s
MS1
MS2
0.38 s
LC-QTOF-MS X500R (AIF -10 Windows)
0.2 s 0.7 s
32k32
k
32
k
32
k
32
k
32
k
32
k0.05 s x 10 Windows = 0.5 s
32
k
32
k
European Reference Laboratory - FV
“Old” or very rare detected
compounds
OR
“Very expensive” analytical
standards
Compounds “produced” during
the analysis
Retrospective Analysis combined
with the use of databases and
libraries.?
European Reference Laboratory - FV
SELECTIVITY SENSITIVITY
European Reference Laboratory - FV
Green Tea
Acquisition Time (min)10 10.1 10.2 10.3 10.4 10.5
Co
un
ts 2x10
0
1
2
3
4
5
6
7
8
318.0 -> 132.1 , 318.0 -> 261.0
Ratio = 50.9
Azinphos- methyl at 0.010 mg kg-1
Injected amount:0.2 mg sample
LC-QqQ-MS/MS LC-Q-Exactive-Orbitrap-MS/MS
RT: 5.77 - 11.73
6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5
Time (min)
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lative
Ab
un
da
nce
NL: 1.24E4
Base Peak m/z=
318.0115-318.0147 F:
FTMS + p ESI Full ms
[138.0000-910.0000] MS
Te_verde_10pbb_dil25_5mi
cro
EIC 318.0131 ± 5ppm
European Reference Laboratory - FV
CONCLUSIONS
GC and LC-HRMS provide very accurate
results improving the identification
confidence ( 10 ug/kg). In cases of
limitation in achieving analytical
standards prediction of presence/ no
presence can be provided.
In special GC-EI-HRMS can be easily
implemented in the laboratories very
effectively.
However, an increase of the sensitivity
around 5-10 times should be necessary
to be applied with the same workflows
than triple quadrupoles
European Reference Laboratory - FV
www.eurl-pesticides.eu