diapositiva 1 - agilent · european reference laboratory - fv amadeo r. fernández-alba – carmen...

European Reference Laboratory - FV

Amadeo R. Fernández-Alba – Carmen Ferrer

12-02-2014



S/N: 10.7 S/N: 16.9

Fenhexamid

Orange at 0.010 mg/kg LC-QQQ-MS/MS 6410

Orange at 0.010 mg/kg dilx10 LC-QQQ-MS/MS 6490

302 > 55

302 > 97

302 > 55

302 > 97

6410 / 6490 Comparison



S/N: 21.1

Aldicarb

213 > 89

213 > 116

213 > 89

213 > 116

S/N: 0.8

Orange at 0.010 mg/kg LC-QQQ-MS/MS 6410

Orange at 0.010 mg/kg dilx10 LC-QQQ-MS/MS 6490




S/N: - S/N: 16.9

Thiabendazole

Apple at 0.010 mg/kg LC-QQQ-MS/MS 6410

Apple at 0.010 mg/kg dilx10 LC-QQQ-MS/MS 6490

202 > 175

202 > 130

202 > 175

202 > 130




Fipronil 0.001 mg/kg Dilx5

LC-QQQ-MS/MS 6490

S/N: 8.7

Fipronil 0.005 mg/kg

LC-QQQ-MS/MS 6410

MRM2 only noise

Standards in baby food

Fipronil MRL: 0.004 mg/kg



Fipronil 0.001mg/kg Dilx5

S/N: 8.7

Standards in baby food

Fipronil MRL: 0.004 mg/kg

Fipronil-desulfinyl 0.001mg/kg Dilx5

S/N: 7.1



0.001mg/kg standard in baby food Dilx5

S/N: 24.2

Cadusafos MRL: 0.006 mg/kg



S/N: 11.3

0.001 mg/kg standard in baby food Dilx5

Ethoprophos MRL: 0.008 mg/kg



S/N: 5.7


Fensulfothion Shall not be used (Concentration level below 0.003 mg/kg)



S/N: 6.5


Omethoate Shall not be used (Concentration level below 0.003 mg/kg)



Dilution of black tea extract Results for fuberidazole

1:2

1:5

5 µg/kg

1:10

1:50

good S/N

even in 1:50

dilution

Fuberidazole

40 fg o.c. 200 fg o.c. 400 fg o.c. 1 pg o.c. 2 pg o.c.

increased

signal

October 23, 2012

WEBEX triggered MRM



Results for Fuberidazole Calibration in solvent and recoveries in different matrices

Matrix Concentration Response in %

Solvent 0.94

Black tea 0.25 24.6

Cucumber 0.95 94.7

Lemon 0.78 78.4

Pepper 0.97 97.0

Rocket 0.89 89.8

Tomato 1.04 104.1

Wheat flower 1.04 103.5 October 23, 2012

WEBEX triggered MRM



Results for Fuberidazole Recoveries for dilution in black tea

Matrix Concentration Response in %

Black tea 0.25 24.6

Black tea 1 in 2 0.22 44.9

Black tea 1 in 5 0.14 72.2

Black tea 1 in 10 0.078 77.6

October 23, 2012

WEBEX triggered MRM



Etoprofos at 0.010 mg/kg

LC-QQQ-MS/MS 6490 243 > 97

Dilution 5 times / without dilution comparison

Tomato without dilution

Tomato diluted 5 times 5.4

5



Dimethoate at 0.010 mg/kg

LC-QQQ-MS/MS 6490 230 > 199


Orange without dilution

Orange diluted 5 times

5

2.1



Pymetrozine at 0.010 mg/kg

LC-QQQ-MS/MS 6490 218 > 105



Tomato diluted 5 times 5

4.1



Pymetrozine at 0.010 mg/kg

LC-QQQ-MS/MS 6490 218 > 105




5

3.7



Dilution of black tea extract Response (%) after dilution

overall increase in recovery by dilution

0

20

40

60

80

100

120

140

< 20% 20% - 40% 40% - 60% 60% - 80% 80% -100%

100% -120%

120% -150%

150% -200%

> 200% not found

black tea spiked with 300 pesticides 1pg/µlrecovery of different diluted extracts compared to solvent

(calculated by comparing with the appropriate level in solvent)

undiluted

1:2

1:10

October 23, 2012

WEBEX triggered MRM



Co-extracted matrix components of orange. LC-TOF-MS

8017 Matrix

compounds

2408 Matrix

compounds

Co-extracted matrix components of tomato. LC-TOF-MS

Isoproturon (100 µg ∙ kg-1)

LC-TOF-MS XIC of orange matrix compounds and

Isoproturon

LC-TOF-MS XIC of tomato matrix compounds and Isoproturon

Isoproturon (100 µg ∙ kg-1)



Co-extracted matrix components of orange. LC-TOF-MS

8017 Matrix compounds


No Dilution

Dilution 1/5



Azoxystrobyn at 0.010 mg/kg

LC-QQQ-MS/MS 6490 404 > 344

Tomato diluted 5 times

Pepper diluted 5 times



Pepper without dilution





Methomyl at 0.010 mg/kg




LC-QQQ-MS/MS 6490 163 > 106







Pirimicarb at 0.010 mg/kg

LC-QQQ-MS/MS 6490 239 > 72










Dimethoate at 0.010 mg/kg







LC-QQQ-MS/MS 6490 230 > 199




Avocado is an example of high oil commodity, it contains up to 30% of fat.

The major components of avocado fat are fatty acids (oleic, palmitic and linoleic)

and triglycerides.

Introduction

The main challenge is in obtaining an extract which contains the target analytes

but possibly low amount of fat. The most common methods for fat removal from

extract are:

• low temperature precipitation (freezing-out),

• gel permeation chromatography (GPC)

• adsorption (dispersive solid-phase extraction, solid-phase extraction).



QuEChERS Ethyl acetate

method

miniLuke

Average composition of avocado:

water (66.5%); lipids (23.5%); protein (1.9%); sugars (0.4%).

Introduction



Interaction of

phospholipid

molecule with

empty d-orbital

of zirconium atom.

Sorbent selection for QuEChERS



Selected compounds # Compound tR (min) SRM1 CE1 (V) SRM2 CE2 (V)

1 Acephate 3,11 184 / 143 5 184 / 125 15

2 Acetamiprid 10,99 223 / 126 20 223 / 56 15

3 Aldicarb 13,58 213 / 89 15 213 / 116 10

4 Azinphos-

methyl 20,17 318 / 132 8 318 / 261 0

5 Azoxystrobin 20,80 404 / 372 10 404 / 344 20

6 Bitertanol 21,81 338 / 269 5 338 / 99 10

7 Boscalid 21,06 343 / 307 16 343 / 272 32

8 Bromuconazole 20,26 ; 21, 28 378 / 159 20 378 / 70 20

9 Bupirimate 19,25 317 / 166 20 317 / 272 20

10 Buprofezin 24,59 306 / 201 10 306 / 116 15

11 Carbaryl 16,90 202 / 145 10 202 / 127 20

12 Carbendazim 3,31 192 / 160 15 192 / 132 20

13 Chlorantranilipr

ole 18,76 484 / 286 8 484 / 453 16

14 Chlorpyriphos 28,07 350 / 198 20 350 / 97 20

15 Chlorpyriphos-

methyl 27,47 322 / 125 16 322 / 290 14

16 Clofentezine 24,66 303 / 138 12 303 / 102 40

17 Cymoxanil 12,47 199 / 128 4 199 / 111 12

18 Cyproconazole 19,85 292 / 70 16 292 / 125 32

19 Cyprodinil 18,71 226 / 93 40 226 / 77 40

20 Cyromazine 2,28 167 / 85 20 167 / 125 15

21 Diazinon 25,03 305 / 169 15 305 / 153 20

22 Dicrotophos 5,53 238 / 112 8 238 / 72 28

23 Diethofencarb 20,31 268 / 226 5 268 / 180 15

24 Difenoconazole 23,45 406 / 251 20 406 / 337 15

25 Dimethoate 10,89 230 / 199 5 230 / 171 10

26 Dimethoate-d6 10,88 236 / 205 4 236 / 131 16

27 Dimethomorph 18,56 ; 18,99 388 / 301 20 388 / 165 20

28 Diniconazole 22,74 326 / 70 28 326 / 159 28

29 Diphenylamine 22,95 170 / 93 32 170 / 65 36

30 Dodine 17,91 228 / 57 20 228 / 60 20

31 Ethion 28,33 385 / 199 5 385 / 171 10

32 Ethirimol 6,62 210 / 140 20 210 / 43 52

33 Ethoprophos 21,57 243 / 131 15 243 / 97 30

34 Fenamidone 21,18 312 / 92 28 312 / 65 56

35 Fenamiphos 20,59 304 / 217 20 304 / 234 12

36 Fenarimol 20,25 331 / 268 20 331 / 259 20

37 Fenazaquin 27,47 307 / 161 15 307 / 147 15

38 Fenbuconazole 21,81 337 / 70 33 337 / 125 40

39 Fenoxycarb 22,37 302 / 88 20 302 / 116 5

# Compound tR (min) SRM1 CE1 (V) SRM2 CE2 (V)

40 Fenpropathrin 28,07 350 / 97 32 350 / 125 10

41 Fenpropimorph 15,84 304 / 147 30 304 / 130 25

42 Fenpyroximate 27,91 422 / 366 12 422 / 107 64

43 Fenthion 24,36 279 / 247 8 279 / 169 12

44 Fluazifop 20,09 328 / 282 15 328 / 254 20

45 Flufenoxuron 27,28 489 / 158 20 489 / 141 56

46 Fluquinconazole 21,22 376 / 307 24 376 / 108 56

47 Flusilazole 21,76 316 / 247 12 316 / 165 24

48 Flutriafol 16,72 302 / 70 16 302 / 95 56

49 Formetanate 2,97 222 / 65 52 222 / 165 8

50 Haloxyfop 22,18 362 / 316 12 362 / 288 24

51 Hexythiazox 28,04 353 / 228 10 353 / 168 20

52 Imazalil 13,69 297 / 159 20 297 / 255 15

53 Imidacloprid 9,98 256 / 209 15 256 / 175 15

54 Indoxacarb 25,94 528 / 203 45 528 / 150 20

55 Iprovalicarb 20,79 321 / 119 16 321 / 203 0

56 Isofenphos-

Methyl 25,14 231 / 121 15 231 / 199 15

57 Isoprocarb 18,45 194 / 95 15 194 / 152 5

58 Kresoxim-methyl 23,91 314 / 267 0 314 / 222 10

59 Linuron 20,35 249 / 160 20 249 / 133 36

60 Malathion 22,74 331 / 127 10 331 / 99 20

61 Malathion-d10 22,76 341 / 132 12 341 / 100 24

62 Mandipropamid 21,13 412 / 328 8 412 / 356 4

63 Metalaxyl 17,25 280 / 220 5 280 / 192 10

64 Metconazole 22,13 320 / 70 24 320 / 125 48

65 Methamidophos 3,07 142 / 94 10 142 / 125 10

66 Methidathion 20,39 303 / 145 0 303 / 85 15

67 Methiocarb 19,99 226 / 169 5 226 / 121 12

68 Methoxyfenozide 22,11 369 / 149 15 369 / 133 20

69 Metobromuron 18,26 259 / 170 15 259 / 148 10

70 Monocrotophos 4,66 224 / 193 5 224 / 127 10

71 Myclobutanil 20,93 289 / 70 15 289 / 125 20

72 Nitenpyram 4,38 271 / 225 10 271 / 99 10

73 Oxamyl 4,87 237 / 72 10 237 / 90 5

74 Oxydemeton-

methyl 3,97 247 / 109 24 247 / 169 8

75 Paclobutrazole 19,42 294 / 70 16 294 / 125 36

76 Pencycuron 25,15 329 / 125 24 329 / 89 60

77 Pendimethalin 27,98 282 / 212 4 282 / 194 16

78 Phenthoate 24,73 321 / 247 4 321 / 79 44

# Compound tR (min) SRM1 CE1 (V) SRM2 CE2 (V)

79 Phosalone 25,47 368 / 182 8 368 / 111 44

80 Phosmet 20,74 318 / 160 8 318 / 133 26

81 Phoxim 25,59 299 / 129 4 299 / 77 24

82 Pirimicarb 6,61 239 / 72 20 239 / 182 15

83 Pirimiphos-

methyl 25,04 306 / 164 20 306 / 108 20

84 Prochloraz 19,29 376 / 308 10 376 / 266 15

85 Profenofos 25,95 375 / 305 15 375 / 347 5

86 Propamocarb 3,07 189 / 102 15 189 / 144 10

87 Propiconazole 22,81 342 / 159 32 342 / 69 16

88 Propoxur 16,06 210 / 168 0 210 / 153 0

89 Propyzamide 21,65 256 / 190 10 256 / 173 20

90 Pymetrozine 2,36 218 / 105 20 218 / 51 60

91 Pyraclostrobin 24,47 388 / 194 8 388 / 163 20

92 Pyrethrins 29,14 329 / 161 5 329 / 143 20

93 Pyridaben 29,34 365 / 309 10 365 / 147 20

94 Pyrimethanil 15,33 200 / 107 20 200 / 183 20

95 Pyriproxyfen 27,13 322 / 96 10 322 / 185 20

96 Quinoxyfen 25,64 308 / 197 35 308 / 272 25

97 Rotenone 22,30 395 / 213 20 395 / 192 20

98 Spinosyn A 17,36 732 / 142 20 732 / 98 20

99 Spinosyn D 18,16 746 / 142 20 746 / 98 20

100 Spirodiclofen 30,19 411 / 71 15 411 / 313 5

101 Spiromesifen 30,08 371 / 273 5 371 / 255 20

102 Tebuconazole 21,41 308 / 70 20 308 / 125 20

103 Tebufenozide 23,49 353 / 133 15 353 / 297 5

104 Tebufenpyrad 26,03 344 / 145 20 344 / 171 20

105 Tetraconazole 21,58 372 / 159 36 372 / 70 20

106 Trichlorfon 8,30 257/221 4 257/109 12

107 Thiabendazole 3,50 202 / 175 30 202 / 131 40

108 Thiacloprid 12,97 253 / 126 20 253 / 186 10

109 Thiamethoxam 7,12 292 / 211 10 292 / 181 20

110 Thiophanate-

methyl 15,30 343 / 151 20 343 / 93 56

111 TPP 24,35 327 / 77 35 327 / 152 30

112 Triadimefon 21,35 294 / 197 10 294 / 225 10

113 Triazophos 22,57 314 / 162 20 314 / 286 10

114 Triflumuron 23,82 359 / 156 8 359 / 139 32

115 Triticonazole 19,55 318 / 70 33 318 / 125 41

116 Zoxamide 24,65 336 / 187 16 336 / 159 44



Agilent Zorbax Eclipse XDB C8 150×4.6mm; d.i.:5 µm

Injection volumen: 10µl

Flow: 0.6 ml/min

Mobile phase: Acetonitrile and water with 0.1 % formic acid

Gradient: 20% AcN isocratic mode for 3 min. Lineal increasing until el 100% in 27 min. and 3 min. more on isocratic mode at 100% AcN

Agilent 6490 Triple Quadrupole

Gas Temp 120ºC

Gas Flow 15 l/min Nebulizer 35 psi

Sheat Gas Temp 375ºC Sheat Gas Flow 12 l/min Capillary 3500 V

Nozzle voltaje 300 V

iFunnel Parameters High Pressure RF: 150 V Low Pressure RF: 60 V

Source Ailent Jet Stream: ESI (+) and ESI (-)

0

20

40

60

80

100

120

0 10 20 30 40

%B

Time (min)

Gradient



34%

42% 47% 46%

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

QuEChERSZ-Sep+

QuEChERSPSA/C18

QuEChERSSiO2

QuEChERSZ-Sep

Pre

cen

tag

e o

f to

tal n

um

ber

of

pesti

cid

es

QuEChERS – recoveries with different clean-up sorbents


evaporation of 0.50 mL, reconstitution in 0,50 mL of H2O (10% AcN)

Percentage of pesticides with recoveries in the 70-120% range



34%

42% 47% 46%

54% 59%

68%

95%

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

QuEChERSZ-Sep+

QuEChERSPSA/C18

QuEChERSSiO2

QuEChERSZ-Sep

Pre

cen

tag

e o

f to

tal n

um

be

r o

f p

esti

cid

es

QuEChERS – recoveries with different clean-up sorbents

evaporation of 0.50 mL, reconstitution in 0,50 mL of H2O (10% AcN)

evaporation of 0.50 mL, reconstitution in 1 mL of H2O (30% AcN)





Z-Sep

PSA/C18

Z-Sep provides cleaner QuEChERS extracts than combination of PSA and C18




PSA+C18 4067 matrix compounds

Z-Sep 2807 matrix compounds 1757 497 2310

Absolute height ≥ 10000 counts

Co-extracted matrix components of avocado with Z-Sep and PSA+C18. LC-TOF/MS

PSA+C18

Z-Sep

Mass Profiler Professional 12.1.

Agilent Tech.



Add 20mL of Acetone

Mix

30s by Polytron

Mix

30s by Polytron

10 g of sample

Add 30 g Na2SO4 and 40 mL

Petroleum ether:

Dichloromethane (1:1)

Centrifugue 5 min.

at 3500 r.p.m.

Analysis

Evaporate to dryness

and reconstitute

miniLuke method

Addition of Dilution Internal

Standard-Linuron-d6

Addition of Injection Internal

Standard-Dimethoate-d6



Add 10 mL ethyl acetate and

Shake automatically

in Agitax 15 min

Shake 3 s by hand

10 g of sample

1.5 g of NaCl and 8 g of

MgSO4

Centrifugue 5 min.

3700 r.p.m.

Analysis

Ethyl acetate multiresidue method

Addition of Dilution

Internal Standard

Linuron-d6 Addition of Injection Internal




Modified QuEChERS method

Add 10 mL acetonitrile

Shake automatically

in Agitax

Take 5 mL aliquot

Shake 30 second

in Vortex

Evaporate to dryness

and reconstitute

Shake automatically

in Agitax

Add 4 g MgSO4 + 1 g NaCl + 1 g

NaCitrate·2 H2O + 0.5 g

NaCitrate·3/2 H2O

10 g of sample

Centrifugue 5 min.

3700 r.p.m.

Clean-up with 175 mg

Z-Sep + 750 mg MgSO4

Centrifugue 5 min.

3700 r.p.m.

Analysis

Addition of Dilution Internal

Standard-Linuron-d6

Addition of Injection Internal




Fat content in the final avocado extracts

QuEChERS

with Z-Sep miniLuke

Ethyl acetate

method

Comparison of different MRMs



Comparison of different MRMs

42%

54% 62%

70% 76%

96%

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

MiniLuke Z+ Fat 150 mg silica EtAc Z-Sep

Pre

ce

nta

ge

of

tota

l n

um

be

r o

f p

esticid

es

QuEChERS

Z-Sep+

QuEChERS

PSA/C18

QuEChERS

150 mg silica

QuEChERS

Z-Sep miniLuke Ethyl

acetate




18%

26% 57% 0%-20%

21%-50%

51%<

44%

26%

30% 0%-20%

21%-50%

51%<

44% 39%

17% 0%-20%

21%-50%

51%<

Suppression in particular extracts of avocado

Ethyl acetate

method miniLuke

QuEChERS

with Z-Sep

Matrix effects



0.E+00

5.E+07

1.E+08

2.E+08

2.E+08

3.E+08

3.E+08

4.E+08

0 50 100 150 200 250 300

Concentration [ppb]

Calibration plots of diazinon (304.6 > 169.0)

Solvent

QuEChERS with Z-Sep

miniLuke

Ethyl acetate

Matrix effects



0.E+00

2.E+07

4.E+07

6.E+07

8.E+07

1.E+08

1.E+08

1.E+08

0 50 100 150 200 250 300

Concentration [ppb]

Calibration plots of cyprodinil (225.8 > 93.0)

Solvent

QuEChERS with Z-Sep

miniLuke

Ethyl acetate

Matrix effects



Calibration plots of pymetrozine (118.1 > 105.0)

Matrix effects

0.E+00

5.E+06

1.E+07

2.E+07

2.E+07

3.E+07

0 50 100 150 200 250 300

Concentration [ppb]

Solvent

QuEChERS with Z-Sep

miniLuke

Ethyl acetate



Validation of QuEChERS with Z-Sep

in

avocado and almond



Sample amount: 10 g

Water addition: no

Evaporation: 0.5 mL

Reconstitution: 0.3 mL ACN + 0.7 H2O

Sample amount: 5 g

Water addition: 5 mL

Evaporation: 0.5 mL

Reconstitution: 0.4 mL ACN + 0.6 H2O

Dilution factor: 2 Dilution factor: 4

Validation



0%

10%

20%

30%

40%

50%

Perc

en

tag

e o

f to

tal

nu

mb

er

of

pesti

cid

es

0%

10%

20%

30%

40%

50%

Avocado

Almond

Recoveries

0.010 mg/kg 0.050 mg/kg

Percentage of pesticides with

recoveries in the 70-120% range:

-avocado 95%

-almond 81%

Percentage of pesticides with

recoveries in the 70-120% range:

-avocado 95%

-almond 83%

Validation



0%

70%

0 20 40 60 80 100

Reco

very

120%

Almond, fat content around 50%

0%

70%

0 20 40 60 80 100

120%

Water solubility [mg/L] Water solubility [mg/L]

Avocado, fat content up to 30%

Validation



LOQs

0

10

20

30

40

50

60

70

80

90

100

0.010 mg/kg 0.050 mg/kg > 0.050 mg/kg

Perc

en

tag

e o

f to

tal n

um

ber

of

pe

sti

cid

es

Avocado

Almond



0%

20%

40%

60%

80%

100%

1-5% 6-10% 11-15% 16-20%

Avocado

Almond

0%

20%

40%

60%

80%

100%

1-5% 6-10% 11-15% 16-20%

Perc

en

tag

e o

f to

tal n

um

ber

of

pesti

cid

es

Validation

Relative standard deviation (n = 5)

0.010 mg/kg 0.050 mg/kg



0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

< 20% 21% - 50% 50% <

Suppression

Avocado

Almond

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

1-500 2 - 500 1 - 200 2 - 200 1 - 50 2 - 50 10 - 500

Perc

en

tag

e o

f to

tal n

um

ber

of

pe

sti

cid

es

Linearity range (ppb)

Linearity Matrix effects

Validation



60%

70%

80%

90%

100%

110%

120%

0 5 10 15 20

Re

cove

ry

Days

Malathion-d10

TPP



0%

2%

4%

6%

8%

10%

12%

14%

0 5 10 15 20

STD

Days

Dimethoate-d6

Malathion-d10

TPP



Collaborative study with tea



Co-extracted matrix components of green tea. LC-TOF-MS

AOAC method

Modified QuEChERs with CaCl2

2641 compounds

Mass Profiler Professional 12.1.

Agilent Tech.

4398 compounds




5g tea sample

Add 15 mL acetonitrile

Add IS Heptachlor epoxide for GC/MS-SIM, GC/MS/MS or Chlorpyrifos-methyl for LC-MS/MS. Evaporate to dryness with nitrogen at 35º

Cleanert TPT Cartridge (2g, 12mL) or Envi-Carb/PSA(0.5g/0.5g,6mL) for clean up. Elute with 25 mL acetonitrile + toluene (3+1), concentrated to about 0.5 mL

Make up to 1.5 mL with hexane Make up to 1.5 mL with

acetonitrile + water (3+2)

GC/MS Determination

LC-MS/MS Determination

Homogenize at 13500r/min for 1min, and centrifuge at 4200r/min for 5 min before the supernatants are taken, while the dregs are re-extracted and the extracts are combined and then concentrated to about 1 mL

3.33 times

concentration

Tea Extraction method



Chromatographic and acquisition parameters

Brand name(type) Agilent (1200)

Column temperature 40ºC

Mobile phase A: H2O 0.1% formic

acid - B: AcN

Flow rate(mL/min) 0.4

Gradient program (min-mobile phase

A%)

Min - A(%)

0 - 99

3 - 70

6 - 60

9 - 60

15 - 40

19 - 1

23 - 1

Injection volume 10 µL

Brand name(type) Agilent Zorbax C8

Specification 5um, 4.6x150 mm

Compounds Quantifying transition Qualifying

transition

Chlorpyrifos-methyl 321.9>125 321.9>289.9

Acetochlor 270.2>224.2 270.2>148.1

Benalaxyl 326>148.1 326>294

Bensulide 398.1>158.1 398.1>314.5

Butralin 296.2>240.2 296.2>222.2

Chlorpyriphos 349.9>198 349.9>97

Clomazone 240.1>125 240.1>89

Diazinon 305>169 305>153

Ethoprophos 243.1>173.1 243.1>214.9

Flutolanil 324.1>262.1 324.1>282.2

Imidacloprid 256>209 256>175

Indoxacarb 528.1>218.2 528.1>150

Kresoxim methyl 314.1>267 314.1>206

Monolinuron 215>126 215>148.1

Picoxystrobin 368.1>205.2 368.1>145

Pirimiphos-methyl 306.2>164.2 306.2>108.2

Propoxur 210.1>111.1 210.1>168.1

Quinoxyfen 308.1>196.9 308.1>271.9

Tebufenpyrad 334.2>117 334.2>147.1

Triadimefon 294.2>197.1 294.2>69.1

Trifloxystrobin 409.2>186.2 409.2>206.2

Brand name(type) Agilent QQQ 6490

Source mode ESI

Ion spray voltage 3500

Dry gas temperature 120ºC

Dry gas flow rate 15 l/min

Sheat Gas gas temperature 375ºC

Sheat gas pressure 12 l/min

MS/MS Parameters

HPLC Parameters



TIC Green Tea Spiked with 50uL working solution + Chlorpyrifos methyl

TIC Oolong Tea Spiked with 50uL working solution + Chlorpyrifos methyl



Green tea validation data

No. Pesticides Retention time/ min Calibration curve Spike levels (µg/kg) RSD/%

R2 Slope Intercept Recovery/%

IS Chlorpyrifos-methyl 17.8 1.9

1 Acetochlor 14.8 0.9996 1253.8 -27358.0 133.3 88.2 2.4

2 Benalaxyl 16.2 0.9970 11728.2 -77541.1 66.7 87.3 2.8

3 Bensulide 17.2 0.9969 382.0 8104.6 200.0 75.6 2.9

4 Butralin 19.9 0.9953 204.5 -1931.4 66.7 88.9 3.8

5 Chlorpyriphos 19.6 0.9940 80.6 7077.6 666.7 76.4 1.5

6 Clomazone 10.3 0.9983 9091.7 -70141.8 66.7 83.1 1.8

7 Diazinon 17.0 0.9992 12293.5 -120027.4 66.7 85.6 2.1

8 Ethoprophos 13.2 0.9988 3031.8 -18414.9 66.7 79.8 2.1

9 Flutolanil 15.0 0.9987 10241.3 -73393.8 66.7 85.4 2.3

10 Imidacloprid 6.0 0.9955 197.7 2278.2 150.0 64.0 1.7

11 Indoxacarb 18.6 0.9960 481.1 2642.7 66.7 72.7 1.4

12 Kresoxim methyl 16.4 0.9884 865.7 260188.1 666.7 70.0 1.7

13 Monolinuron 8.9 0.9962 1378.6 7321.0 66.7 72.9 1.2

14 Picoxystrobin 16.8 0.9977 11715.6 -7683.8 66.7 80.7 2.6

15 Pirimiphos-methyl 16.8 0.9988 10652.5 -80579.9 66.7 83.2 1.8

16 Propoxur 8.0 0.9931 2339.7 53776.6 166.7 74.2 1.4

17 Quinoxyfen 18.4 0.9890 1719.6 120653.2 333.3 60.6 2.4

18 Tebufenpyrad 18.7 0.9971 1388.6 1425.2 66.7 81.9 1.7

19 Triadimefon 13.4 0.9994 1946.2 -22800.5 66.7 85.8 2.3

20 Trifloxystrobin 18.7 0.9968 8149.4 28719.2 66.7 77.4 1.7



Oolong tea validation data

No. Pesticides Retention time/ min Calibration curve Spike levels (µg/kg) RSD/%

R2 Slope Intercept Recovery/%

IS Chlorpyrifos-methyl 17.8 1.7

1 Acetochlor 14.8 0.9972 1092.4 -17506.8 133.3 77.6 1.6

2 Benalaxyl 16.2 0.9987 12861.6 -33911.2 66.7 75.0 2.3

3 Bensulide 17.2 0.9985 345.4 -1802.1 200.0 74.7 1.1

4 Butralin 19.9 0.9979 116.0 -1347.7 66.7 65.0 2.5

5 Chlorpyriphos 19.6 0.9970 48.7 -3927.6 666.7 68.6 2.1

6 Clomazone 10.3 0.9958 5994.8 -64502.7 66.7 75.2 0.9

7 Diazinon 17.0 0.9973 11378.9 -91525.9 66.7 78.5 2.3

8 Ethoprophos 13.2 0.9981 2563.4 -11951.3 66.7 74.3 2.3

9 Flutolanil 15.0 0.9974 8846.7 -51232.7 66.7 73.2 1.3

10 Imidacloprid 6.0 0.9979 194.9 -2534.2 150.0 73.7 2.1

11 Indoxacarb 18.6 0.9956 342.4 -3154.1 66.7 73.9 0.8

12 Kresoxim methyl 16.4 0.9988 943.3 55578.1 666.7 72.3 1.0

13 Monolinuron 8.9 0.9959 906.3 -5784.4 66.7 65.7 0.4

14 Picoxystrobin 16.8 0.9977 12459.7 -23007.8 66.7 74.3 1.7

15 Pirimiphos-methyl 16.8 0.9967 9237.3 -80485.6 66.7 78.2 2.6

16 Propoxur 8.0 0.9960 1648.7 -34433.4 166.7 74.5 1.3

17 Quinoxyfen 18.4 0.9954 1285.9 -19053.4 333.3 62.3 2.0

18 Tebufenpyrad 18.7 0.9956 931.1 -2984.3 66.7 63.7 0.8

19 Triadimefon 13.4 0.9975 1622.5 -8205.6 66.7 77.2 3.0

20 Trifloxystrobin 18.7 0.9961 6346.3 -19126.7 66.7 64.4 1.0



Samples

Butralin in green tea at 0.050 mg/kg Chlorpyrifos in oolong tea at 0.050 mg/kg



Pooling Samples



Azinphos-methyl 0.010 mg/kg in Tomato

LC-QQQ-MS/MS 6490 318 >132

Tomato without dilution Pooling samples (Ratio 1:1:1) Pooling samples (Ratio 1:1:1 diluted 5 times) 10 ppb

3.3 ppb

0.8 ppb

Pooling samples (Tomato, Cucumber and Lettuce)



Methomyl 0.010 mg/kg in Tomato

LC-QQQ-MS/MS 6490 163 >106

Tomato without dilution Pooling samples (Ratio 1:1:1) Pooling samples (Ratio 1:1:1 diluted 5 times)

Pooling samples (Tomato, Cucumber and Lettuce)

10 ppb

3.4 ppb

0.7 ppb



Co-extracted matrix components. LC-TOF-MS


Cucumber

4559 compounds

Cucumber+Lettuce+Tomato


Tomato

3775 compounds

Lettuce

3718 compounds

diapositiva 1 - agilent · european reference laboratory - fv amadeo r. fernández-alba – carmen...

Documents