Learn Pharma Sci Issue 8

Table of contents
Advances in GC-MS
Analysis of Pesticides
Video Introduction
Laura Bush
NCI, SRM, and LVI

By Renata Raina-Fulton
Accurate-Mass Database
Noelia Belmonte, Samanta Ucls,
Miguel Gamn, Carmen Ferrer,
Milagros Mezcua, and
Amadeo R. FernndezAlba
Rapid Sample Prep
Yelena Sapozhnikova
Welcome
Improving Sensitivity and Selectivity in

Pesticide Analysis with GCMS and
GCMS-MS
An Accurate-Mass Database for Screening

Pesticide Residues in Fruits and Vegetables
by Gas ChromatographyTime-of-Flight Mass
Spectrometry
Rapid Sample Preparation and Fast GCMS-MS

for the Analysis of Pesticides and Environmental
Contaminants in Fish
CONTENTS
TOC
INTRODUCTION
Welcome
NCI, SRM, AND LVI
Improving Sensitivity
and Selectivity in
Pesticide Analysis with
GCMS and GCMS-MS
By R
enata RainaFulton
The selection of analytical methods for gas chromatography (GC)-amenable

pesticides is often based on requirements for sensitivity and selectivity for
regulatory needs or other monitoring requirements. Methods with both electron
ionization (EI) and negative chemical ionization (NCI) are often required to cover
the full range of GCamenable pesticides at trace levels. Pesticides fragment
easily in EI and CI sources such that the molecular ion is often low in abundance.
NCI can provide added selectivity and sensitivity over EI methods. NCI is most
commonly used in selected-ion monitoring mode. The lack of availability of parent
ions for collision-induced dissociation for tandem mass spectrometry (MS) can
limit the feasibility of GCMS-MS for pesticides that significantly fragment in the
ion source. Options for improving sensitivity by using of large-volume cold oncolumn or programmable temperature vaporizer injections are presented.
Gas chromatography (GC) continues to be a viable technique for the analysis of a range of chemical
classes of pesticides including herbicides (chloroacetanilides, triazines, and selected dinitroanilines and
thiocarbamates), insecticides (organochlorines, organophosphorus pesticides, and pyrethroids), and
fungicides (selected triazoles, oxazoles, imidazole, dicarboximides, and phthalimides). Several approaches
have been used to improve detection limits or reduce matrix interferences for GC analysis and extend the
range of pesticides that can be analyzed with GC. These approaches include the use of negative chemical
NCI, SRM, AND LVI

SPONSORED
Pesticide Analysis in Herbal
Products with ASE and GC
Triple-Quadrupole MS-MS
Multi-Residue Pesticide Analysis in Herbal
Products Using Accelerated Solvent
Extraction with a Triple Quadrupole
GC-MS/MS System
Hans-Joachim Huebschmann, Joachim Gummersbach, Thermo Fisher Scientific, Dreieich, Germany
Nicole Rueckert, Johann Kirchner, Elmar Hfner, Phytolab GmbH & Co KG, Vestenbergsgreuth, Germany
A ppli cati o n N o te 52291
Click to
view PDF
Key Words
Pesticides, Tea, Herbal products, ASE, SRM, MRM, Multi-residue analysis,

TSQ 8000 GC-MS/MS
Introduction
The residue analysis of pesticides has developed in recent
years into a comprehensive methodology for the detection
of many hundreds of potential contaminating compounds.
A multi-residue method for herbal products and teas is
faced with additional challenges from the worldwide
origin of the products and the complex matrix of the
dried materials. In the due quality control of raw
materials, the unknown or undeclared local plant
protection treatments must be taken into account with
a wide variety of potential pesticide contaminations.
Dried leaves, fruits or seeds and other herbal products of
medical use deliver highly complex extracts from the
sample preparation due to the rich content of active
ingredients, essential oils and the typical high boiling
natural polymer compounds from broken cells, leaves or
fruit skins. A thorough clean up of the extracted sample
can lead to losses of critical analytes of interest. A
complete characterization of pesticide, and other residue,
contamination is done by both LC and GC-MS/MS to cover
the complete range of functional groups.
This application report describes the
methodology used for the multi-residue
pesticide analysis of herbal products
using accelerated solvent extraction
(ASE) and gel permeation
chromatography (GPC)
sample preparation with
detection and
quantitation by the
Thermo Scientific
TSQ 8000 GC-MS/MS
system.
A routine screening method for more than 200 pesticide

compounds was applied to a wide variety of different
sample types, ranging from regular black tea or sage
leaves, to seeds like fennel and herbs of medical and
fragrance use like thyme and chamomile. The data
processing and reporting was achieved by using the
Thermo Scientific TraceFinder quantitation software suite.
The sensitivity requirement for this analysis was
determined by the regulatory background. The analysis of
pesticide residues in tea and herbal products follows the
regulations of the European Directorate General for
Health and Consumer Affairs (SANCO) for Method
Validation and Quality Control Procedures for Pesticide
Residue Analysis in Food and Feed [1]. The sensitivity
requirements for these products as referenced in the
Codex Alimentarius [2] result in maximum residue levels
of 0.01 mg/kg for most of the pesticide compounds.
ionization (NCI) rather than electron ionization (EI), tandem mass spectrometry
(MS), and large-volume injections (LVI) using cold on-column (COC) injection or
a programmable-temperature vaporizer (PTV). When detection limits are not the
most critical factor, multiresidue pesticide analysis is frequently accomplished by
GC methods with EI. This approach is cost effective, easy to set up based on the
availability of an MS library, and for a few chemical classes (chloroacetanilides,
triazines, and some fungicides) provides the best GCMS sensitivity. But even
though EI provides detection for the largest number of GCamenable pesticides,
it is more prone to matrix interferences and thus requires additional sample
cleanup, and sensitivity may not be sufficient to meet regulatory or monitoring
needs.
Negative Chemical Ionization

Chemical ionization can be used in positive- or negative-ion mode. In positiveion chemical ionization mode, however, background and matrix interferences are
observed and the analytes of interest are often coeluted with matrix compounds,
as occurs with EI. As a result, positive-ion chemical ionization is not commonly
used in GCMS pesticides analysis (16). Pesticides that contain halogen, NO2,
or P ester groups are electron-capturing and thus have potential to exhibit
enhanced response up to several orders of magnitude with NCI (116). The
response of hydrocarbons, which are a common source of matrix interference in
environmental samples, is also diminished with NCI (4). Because of the added
selectivity of NCI, the coelution of other pesticides that are not electroncapturing is reduced and total run times also may be reduced. The most common
pesticides that exhibit greater sensitivity with NCI include organochlorines (OCs),
chlorinated cyclodiene insecticides, organophosphorus pesticides (OPs), and
pyrethroids (116). Other selected pesticides from a variety of chemical classes
that have nitro or halogen groups are also detected in NCI; these pesticides
include dinitroaniline herbicides and a range of halogenated fungicides (conazole,
imidazole, triazole, strobilurin, dicarboximide, and phthalimide fungicides), as
shown in Table I. For many OCs and OPs the greatest sensitivity is achieved with
GCNCI in selected-ion monitoring (SIM) mode (14), and GCNCI in selectedreaction monitoring (SRM) mode can provide additional confirmation ability or
reduce background interferences (1,2,4,15,16). The relative abundance of the
molecular and fragment ions formed in the ion source in NCI depends on the
ionization conditions, with the reagent gas influencing ion abundance levels (13).
Tandem Mass Spectrometry

Tandem MS with SRM can be used for GCamenable pesticides that are ionized
with NCI to provide even greater selectivity than GCNCI-SIM does. In general,
with the exception of the pyrethroids, lower detection limits are obtained with
NCI, SRM, AND LVI
Although isobutane, methane, ammonia in methane, and pure ammonia can be

used, methane is the most frequently used because of its reduced instrument
maintenance needs. However, for some fragment ions with a higher mass-tocharge ratio (m/z), the molecular ion abundance increases when ammonia or
ammonia with 5% methane is used (13). Detection limits will also vary depending
on separation conditions and the ions or SRM transitions that have been selected.
When isomers are present, chromatographic resolution is required, because
isomers are not isolated by MS. As shown in Table I, for the analysis of most
pesticides the molecular ion (quantitative or qualifier ion) is not monitored in
either EI or NCI because the abundance of molecular ions is too low at the
working ranges of trace analysis (1,13,24). In addition, the ion or SRM transition
selected may be dependent on the working calibration range, with more choices
available at higher concentrations. For a number of pesticides, only one abundant
fragment or product ion is observed and consequently isotopic masses or
transitions may be selected for confirmation. The relative standard deviation
(RSD) of ratios of response of the quantitative to the qualifier ion or SRM transition
may also vary and these values are more critical at trace levels; thus at trace levels
there are fewer possible confirmation ions that can provide the desired RSD of
<25% (1). When differences in mass selection exist in the literature they are often
a result of the fact that many pesticides are chlorinated or brominated, which
enables a choice of ions with similar abundances to be made between isotopic
masses or other unique fragment ions for confirmation as long as the criteria
for sensitivity, selectivity, and RSD of response ratio of the quantitative ion to
the qualifier ion can be obtained. In general, there is good agreement in the
literature about the ions selected for GCNCI-SIM for the quantitative ion (Table
I). Matrix interferences or coeluted peaks may also influence the choice; highermass ions (m/z > 100) of similar sensitivity generally are preferred because they
tend to suffer less from background interference in MS (1,4).
chromatographynegative chemical ionization-mass spectrometry (GCNCI-MS).
Pesticide
MW
NCI-SIM (m/z)
EI-SIM (m/z)
NCI-SRM m/z
(Collision Energy, eV)
EI-SRM m/z
Reference
Organochlorine Insecticides
-HCH
288
71, 73
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
-HCH
288
71, 73
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
-HCH (lindane)
288
71, 73
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
-HCH (lindane)
288
255, 257, 253
181, 183, 109
-HCH
288
71, 73
181, 111
HCH , , ,
88
71
Pentachloronitrobenzene
293
265, 249
237, 295
Hexachlorobenzene
285
284, 286, 282
284, 286, 282
o,p-DDE
316
318, 316
316, 318
316>246 (15),
246>176 (25)
p,p-DDE
316
318, 316
316, 318
316>246 (15),
246>176 (25)
o,p-DDD
306
71, 248
235, 165
235>165 (25),
235>200 (5)
p,p-DDD
306
71, 248
235, 165
235>165 (25),
235>200 (5)
p,p-DDT
352
71, 73
235, 165
235>165 (25),
235>199 (15)
Part 1 Aldrin
2
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
1
15
265>35 (35),
231>35 (25)
237>119 (15),
237>146 (25)
1
2,4
71>35 (5)
Cyclodiene Insecticides (Chlorinated) and Their Degradation Products

381
237
15
Chlordane
406
410, 408, 412
373, 375, 377
-Chlordane
406
266,232,71
373, 375
71>35 (5),
266>35 (15)
375>266 (25),
375>303 (10)
-Chlordane
406
266, 71, 232
375, 373
71>35 (5),
266>35 (15)
375>266 (25),
375>303 (10)
Dieldrin
381
237
-Endosulfan
404
372, 404,
406, 408, 410
15
241, 195
242>35 (15),
406>35 (10)
195>160 (5),
339>159 (15)
NCI, SRM, AND LVI
Table I: Selected-ion monitoring (SIM) or selected-reaction monitoring (SRM) for pesticides that can be analyzed by gas
316
318, 316
316, 318
316>246 (15),
246>176 (25)
235>165 (25),
235>200 (5)
235>165 (25),
235>200 (5)
306
71,ionization-mass
248
235, 165
chromatographynegative
chemical
spectrometry
(GCNCI-MS).
o,p-DDD
p,p-DDD
306
71, 248
235, 165
Pesticide
MW
NCI-SIM (m/z)
EI-SIM (m/z)
p,p-DDT
352
71, 73
235, 165
NCI-SRM m/z
(Collision
Energy,
eV)
71>35
(5)
EI-SRM m/z
235>165 (25),
(Collision
Energy, eV)
235>199 (15)
Reference
1
Cyclodiene Insecticides
(Chlorinated) Insecticides
and Their Degradation Products
Organochlorine
Aldrin
-HCH
381
288
237
71,
73
181, 111
Chlordane
-HCH
406
288
410, 408, 412

71, 73
373, 375, 377

181, 111
-Chlordane
-HCH (lindane)
406
288
266,232,71
71, 73
373, 375
181, 111
-Chlordane
-HCH (lindane)
406
288
266, 71, 232

255, 257, 253
375, 373
181, 183, 109
Dieldrin
-HCH
381
288
-Endosulfan
HCH , , ,
404
88
237
71, 73
372, 404,
406, 408,
71 410
-Endosulfan
404
293
-Endosulfan
Hexachlorobenzene
241, 195
241, 239, 195
237, 295
404
285
372, 406, 408

265, 249
372, 404,
406,
284, 408,
286, 411
282
-Endosulfan
o,p-DDE
404
316
372, 406, 408

318, 316
70, 239, 195

316, 318
Endosulfan sulfate
p,p-DDE
420
316
386, 352
318, 316
387, 272
316, 318
Endrin
o,p-DDD
Endrin
381
306
381
71, 248
237
235, 165
p,p-DDD
Endrin aldehyde
306
378
71, 248
272, 270
235, 165
67, 245
p,p-DDT
Endrin ketone
352
378
Heptachlor
Aldrin
370
381
Heptachlor
Chlordane
370
406
300, 266, 302

410, 408, 412
272, 100, 274

373, 375, 377
Heptachlor
-Chlordane
370
406
300
266,232,71
373, 375
Heptachlor epoxide
Dieldrin
Mirex
-Endosulfan
Mirex
386
406
386
381
540
404
540
282, 318
266, 71, 232
282
237
402, 368
372, 404,
406, 408, 410
404, 370, 439
epoxide
PartHeptachlor
2-Chlordane
181, 111
241, 196
284, 286, 282
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
15
1
71>35 (5),
73>37
71>35 (10)
(5),
266>35
(15)
71>35 (5),
73>37
71>35 (10)
(5),
181>145 (15),
181>109 (25),
(25)
375>266
2
1
375>303 (15),
(10)
181>145
181>109 (25),
(25)
375>266
266>35 (15)
375>303 (10)
71>35 (5),
73>37
242>35 (10)
(15),
406>35 (10)
181>145 (15),
181>109
195>160 (25)
(5),
265>35 (35),
231>35 (15),
(25)
242>35
237>119 (15),
237>146 (10),
(25)
195>160
406>35 (10)
339>159 (35)
386>97 (10),
352 >97 (15)
380>35 (5),
346>35 (5)
272>35 (20),
272>243 (15)
71, 73
235, 165
71>35 (35),
(5)
308>35
308, 272
317, 67
308>272 (15)
266>35 (30),
266, 232
100, 274
300>35 (25)
237
81, 353
375, 373
272, 274
241, 195
272, 274, 270
339>159 (15)
316>246 (15),
246>176 (25),
(25)
272>237
387>253 (15),
(10)
316>246
246>176 (25)
263>191 (35)
235>165 (25),
235>200 (5)
235>165 (25),
345>281
235>200(10),
(5)
345>245 (10)
235>165 (25),
317>281
235>199 (10),
(15)
317>245 (20)
100>65 (10),
272>237 (10)
1
1
1
2
15
1
1
15
2
1
1
2,4
2
1
1
1
6
1
15
1
1
1
1
1
15
4
2
71>35 (5),
266>35
(15)
71>35 (5),
375>266 (25),
375>303 (15),
(10)
353>263
282>35
(20)
71>35 (5),
266>35 (15)
353>317(25),
(5)
375>266
375>303 (10)
368>35 (20),
404>35 (15),
(15)
242>35
406>35 (10)
272>237 (10),
272>141
195>160 (30)
(5),
339>159 (15)
15
1
1
1
15
15
1
1
4
NCI, SRM, AND LVI
p,p-DDE
Table I:Heptachlor
Selected-ion monitoring
selected-reaction
monitoring
370 (SIM) or
300,
266, 302
272,
100, 274 (SRM) for pesticides that can be analyzed by gas
370
266, 232
100, 274
266>35 (30),
300>35 (25)
Heptachlor
370
300
Heptachlor epoxide
386
282, 318
81, 353
Pesticide
MW
NCI-SIM (m/z)
EI-SIM (m/z)
Mirex
540
402, 368
-HCH
Mirex
288
540
71, 73
404, 370, 439
Mirex
-HCH
540
288
368,71,
402,
73439
181, 111
181>145 (15),
181>109 (25)
8
1
Mirex
-HCH (lindane)
540
288
366, 368, 370,

402,71,
404,
73406
71>35 (5),
73>37 (10)
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
10
1
-HCH (lindane)
288
255, 257, 253
-HCH
Nitrofen
HCH , , ,
Nitrofen
288
283
88
283
293
71, 73
283, 138
71
283, 285, 284
283, 285, 202
265>35 (35),
265, 249
237, 295
231>35
(25)
Triazine Herbicide and Its Degradation
Products
Hexachlorobenzene
Atrazine
285
216
284, 286, 282
284, 286, 282

200, 215
o,p-DDE
DIA (desisopropyl-atrazine)
316
174
318, 316
316, 318
145, 158, 173
p,p-DDE
DEA (desethyl-atrazine)
316
188
318, 316
316,
318
172
o,p-DDD
306
71, 248
Acetochlor
p,p-DDD
Alachlor
270
306
270
71, 248
146, 162, 174

235, 165
160, 188
p,p-DDT
Alachlor
352
269
71, 73
235, 165
160, 188
Metolachlor
Aldrin
Propachlor
Chlordane
284
381
211
406
Part 3-Chlordane
DCPA
406
370
-Chlordane
406
Dieldrin
Ethalfluralin
381
333
-Endosulfan
Pendimethalin
404
281
Heptachlor epoxide
386
282
15
71>35 (5),
282>35 (20)
NCI-SRM
m/z
353>263 (15),
353>317m/z
(5)
EI-SRM
368>35 (20),
Organochlorine
Insecticides
272, 274
404>35 (15)
71>35 (5),
181, 111
272, 274, 270
73>37 (10)
Herbicides
181, 183, 109
Nitrophenyl Ether Herbicide
71>35 (5),
181, 111
73>37 (10)
283>138
(10),
283, 285
283>35 (35)
Chloroacetanilide
Herbicides
235, 165
71>35 (5)
276, 316
241, 195
252, 162, 281
333>46 (10),
333>303
(10)
242>35 (15),
406>35 (10)
15
1
1
4
2
181>145 (15),
181>109 (20),
(25)
283>162
283>253 (10)
237>119 (15),
237>146 (25)
Precursor>94,
122, 132
316>246 (15),
246>176 (25)
Precursor>68,
110

162, 238
237
120, 176
410, 408, 412
373, 375, 377
Phthalic Acid Herbicide 71>35 (5),
266,232,71
373, 375
266>35 (10),
(15)
330>35
332, 330
301, 332
332>302
(5)
71>35 (5),
266, 71, 232
375, 373
266>35
(15)
Dinitroaniline Herbicides
237
333,303
372, 404,
406,
281, 408,
251, 410
282
272>237 (10),
272>141 (30)
181>145 (15),
181>109 (25)
Reference
316>246 (15),
Precursor>79,
246>176
(25)
105, 130
235>165 (25),
235>200 (5)
Precursor>131
235>165 (25),
235>200 (5)
Precursor>160
235>165 (25),
188>160
235>199 (10),
(15)
160>130 (25)
Precursor>162
176>120 (10),
176>92 (15)
375>266 (25),
375>303
332>301 (10)
(5),
301>273 (25),
(15)
375>266
375>303 (10)
276>202 (15),
316>276 (5),
(5)
195>160
339>159 (15)
1
1
15
2
1
2,4
7
1
7
1
7
1
7
1
7
1
1
7
15
1
2
1
1
1
15
1
1
4
NCI, SRM, AND LVI
100>65 (10),
272>237 (10)
Heptachlor
270
146, 162, 174
Precursor>131
Alachlor
270
160, 188
Precursor>160
Table I: Selected-ion
monitoring (SIM) or selected-reaction monitoring (SRM) for pesticides that can be analyzed by gas

Alachlor
269
160, 188
Metolachlor
284
162, 238
Pesticide
MW
Propachlor
211
NCI-SIM (m/z)
Precursor>162
EI-SRM m/z
176>120 (10),
(Collision
Energy, eV)
176>92 (15)
Reference
330>35
(10),
71>35 (5),
332>302
(5)
73>37 (10)
332>301(15),
(5),
181>145
301>273 (25)
(15)
181>109
Dinitroaniline Herbicides 71>35 (5),

181, 111
73>37 (10)
333>46 (10),
276, 316
71>35 (5),
333>303
(10)
181, 111
73>37 (10)
252, 162, 281
181, 183, 109
335>305 (10),
306, 264
71>35 (5),
335>46
(10)
181, 111
73>37 (10)
306, 264, 307
181>145 (15),
181>109 (25)
276>202 (15),
181>145
316>276(15),
(5)
181>109 (25)
EI-SIM (m/z)
120, 176
NCI-SRM m/z
188>160 (10),
160>130 (25)
Phthalic Acid Herbicide

Organochlorine
Insecticides
DCPA
-HCH
370
288
332,
330
71, 73
-HCH
288
71, 73
Ethalfluralin
-HCH (lindane)
333
288
333,303
71, 73
Pendimethalin
-HCH (lindane)
281
288
281, 251, 282

255, 257, 253
Trifluralin
-HCH
335
288
335, 305
71, 73
Trifluralin
HCH , , ,
335
88
335, 305, 336

71
Triallate
293
303
Hexachlorobenzene
285
o,p-DDE
Acephate
316
183
318,
316
168
316, 318
Aspon
p,p-DDE
316
378
318, 316
316,
211, 318
253
Azinphos
methyl
o,p-DDD
317
306
157,
133
71, 248
Azinphos methyl
p,p-DDD
Azinphos ethyl
317
306
345
157
71, 248
185
p,p-DDT
Bromopropylate
352
428
Bromophos ethyl
Aldrin
Carbofenothion
Chlordane
Carbophenthion methyl
-Chlordane
Carbophenthion
394
381
342
406
215
406
343
-Chlordane
Carbophenthion
406
343
Chlorfenvinphos
Dieldrin
358
381
153
237
Chlorpyrifos
ethyl
-Endosulfan
349
404
313, 404,
315,
372,
214,
212
406,
408,
410
Part 4
10
301, 111
332
181,
Thiocarbamate Herbicide
265>35 (35),
237, 295
160>84
231>35 (15),
(25)
86, 268
160 >100(10)
284, 286, 282
284, 286, 282
Organophosphorus Pesticides (Insecticides)
265, 249
160, 161
306>264 (5),
181>145
264>206(15),
(5)
181>109 (25)
237>119 (15),
268>184
237>146 (15),
(25)
268>226 (10)
1
1
1
4
2
1
1
2,4
15
1
1
2,4
316>246 (15),
246>176 (25)
1
16
316>246
378>210 (15),
(10),
246>176
378>115 (25)
(30)
235, 165
235>165 (25),
235>200 (5)
13
1
235, 165
235>165 (25),
235>200 (5)
14,16
1
16
235>165 (25),
235>199 (15)
1
14
211>35 (20)
71, 81,
73
235, 165
71>35 (5)
79,
366, 368, 370
358
237
185>111 (20),
185, 145
157, 342
185>79 (35)
410, 408, 412
373, 375, 377
157
71>35 (5),
266,232,71
373, 375
266>35
(15)
185, 143
71>35 (5),
266,185
71, 232
375, 373
266>35 (15)
375>266 (25),
375>303 (10)
16
15
1
2
16
1
13
375>266 (25),
375>303 (10)
1
16
342>157 (10),
342>143 (15)
13,16
15
97, 197
241,
195
169>96 (15),
242>35
313>189
(10)
406>35 (10)
195>160 (5),
339>159 (15)
NCI, SRM, AND LVI
Acetochlor
317
157
14,16
ethyl monitoring
345 (SIM) or selected-reaction
185
Table Azinphos
I: Selected-ion
monitoring (SRM) for pesticides that can be analyzed by gas
chromatographynegative chemical
spectrometry (GCNCI-MS).
79,ionization-mass
81,
Bromopropylate
428
Bromophos ethyl
394
358
Pesticide
MW
NCI-SIM (m/z)
Carbofenothion
185, 145
EI-SIM (m/z)
157, 342
185>79 (35)
EI-SRM m/z
342>157
(10), eV)
(Collision
Energy,
342>143 (15)
16
Reference
1
Carbophenthion methyl
215
157
Carbophenthion
-HCH
343
288
185,
71, 143
73
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
13
1
Carbophenthion
-HCH
Chlorfenvinphos
343
288
358
185
71, 73
153
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
-HCH (lindane)
Chlorpyrifos ethyl
288
349
181, 111
97, 197
181>145 (15),
181>109 (25)
-HCH (lindane)
Chlorpyrifos ethyl
-HCH
Chlorpyrifos ethyl
HCH , , ,
Chlorpyrifos methyl
Chlorpyrifos methyl
Hexachlorobenzene
DEF
288
349
288
349
88
321
293
321
285
314
71, 315,
73
313,
214, 212
255, 257, 253
313, 314,
315, 212, 169
71, 73
313
71
214, 212
265, 249
212, 214, 285
284, 286, 282
257
71>35 (5),
169>96
(15),
73>37 (10)
313>189 (10)
16
1
13,16
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
141>126 (15),
141>96 (35),
(20)
265>35
231>35 (25)
321>268 (5),
321>208 (15),
(20)
237>119
237>146 (25)
o,p-DDE
Diazinon
316
304
318, 316
169, 171
316, 318
137, 179
p,p-DDE
Diazinon
316
304
318,
316
169,
303,
275
316, 318
Diazinon
o,p-DDD
304
306
167,
168,
71,
248169
287,
302,
288
235,
165
Dichlofenthion
p,p-DDD
314
306
278, 250
71, 248
223, 97
235, 165
Dimethoate
p,p-DDT
Dimethoate
229
352
229
Disulfoton
Aldrin
Dyfonate
Chlordane
274
381
246
406
EPN
Part 5-Chlordane
11
342
14
366, 368, 370
NCI-SRM m/z
185>111
(20), eV)
(Collision
Energy,
16
323
406
Ethion
-Chlordane
384
406
Ethion
Dieldrin
384
381
Ethyl-bromophos
-Endosulfan
394
404
16
181, 183, 109

181, 111
286, 125
237, 295
286, 288, 125
284, 286, 282
169>95 (20),
169>141 (10)
157, 159, 158

87, 93, 125
71, 73
235, 165
71>35 (5)
157
185
237
169>107 (20),
169, 109
109, 137
169>141 (5)
410, 408, 412
373, 375, 377
323, 138,
71>35 (5),
154,
307, 201
266,232,71
373, 375
266>35 (15)
185>111 (15),
185, 187
231, 97
71>35 (5),
185>157 (15)
266, 71, 232
375, 373
266>35 (15)
185
237
358, 257, 278,
372, 404,
242>35 (15),
79, 328, 169
241, 195
406, 408, 410
406>35 (10)
141>126 (15),
316>246 (15),
304>179 (25)
(5),
246>176
304>137 (30)
316>246 (15),
246>176 (25)
1
1
2
14
1
16
15
1
1
2,4
2,4
13,16
1
1
1
13
235>165 (25),
235>200 (5)
314>223 (25),
235>165 (25),
319>81 (25)
235>200 (5)
2,4
1
235>165 (25),
235>199 (15)
4
1
14,16
246>137 (5),
246>109 (15)
375>266 (25),
375>303 (10)
384>231 (5),
375>266 (25),
384>203 (15)
375>303 (10)
1
1
16
15
1
2
13
1
1
1
13,16
15
195>160 (5),
339>159 (15)
320>285 (5),
13
1
NCI, SRM, AND LVI
Azinphos methyl
Table I:Dimethoate
selected-reaction
monitoring
229 (SIM) or
157,
159, 158
87, 93, 125 (SRM) for pesticides that can be analyzed by gas
314
278, 250
223, 97
Dimethoate
229
157
Disulfoton
274
185
Pesticide
MW
NCI-SIM (m/z)
Dyfonate
169, 109
EI-SIM (m/z)
109, 137
EPN
323
-HCH
Ethion
288
384
323, 138,
154, 307, 201
71, 73
185, 187
-HCH
Ethion
288
384
71, 73
185
181, 111
-HCH (lindane)
Ethyl-bromophos
288
394
358,71,
257,
73278,
79, 328, 169
181, 111
-HCH (lindane)
Fenchlorphos
288
320
255, 257, 253

211, 213
181, 183, 109

125, 287
-HCH
Fenitrothion
288
277
71, 73
277, 168
181, 111
277, 125
HCH , , ,
Fenitrothion
88
277
71
277, 168, 278
277, 125, 109
Fenitrothion
293
277
265, 249
277
237, 295
Hexachlorobenzene
Fenthion
285
278
284, 286,
141 282
284, 286, 282
Fonofos
o,p-DDE
246
316
169, 316
109
318,
316, 318
Isocarbophos
p,p-DDE
316
318, 316
136, 121, 230, 289

316, 318
306
169>141 (5)
EI-SRM m/z
246>137 (5),
(Collision
Energy, eV)
246>109 (15)
181, 111
231, 97
16
Reference
1
13
71>35 (5),
185>111
(15),
73>37 (10)
185>157 (15)
71>35 (5),
73>37 (10)
181>145 (15),
384>231 (25)
(5),
181>109
384>203 (15)
181>145 (15),
181>109 (25)
1
13,16
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
1
13
141>126 (15),
211>35 (20)
71>35 (5),
73>37 (10)
320>285 (5),
320>204 (30)
181>145 (15),
277>260 (25)
(5),
181>109
277>109 (20)
265>35 (35),
231>35 (25)
237>119 (15),
237>146 (25)
1
1
2
1
1
1
15
2, 4
1
16
2,4
16
316>246 (15),
246>176 (25)
230>155,
316>246 (15),
198, 136, 212
246>176 (25)
199>121,
199
235>165167,
(25),
Isofenfos methyl
o,p-DDD
Isofenfos
p,p-DDD
Isofenfos
345
306
345
71, 248
380, 344, 182,
153, 244, 287, 302
71, 248
244
p,p-DDT
Leptophos
352
410
71, 73
241, 79
Malathion
Aldrin
330
381
Malathion
Chlordane
330
406
157, 159, 158

410, 408, 412
173, 125, 93
373, 375, 377
Malathion
-Chlordane
Malathion
330
406
330
157, 172, 315

266,232,71
157
373, 375
71>35 (5),
266>35 (15)
375>266 (25),
375>303 (10)
406
213
266,141
71, 232
375, 373
71>35 (5),
266>35 (15)
375>266 (25),
375>303 (10)
Dieldrin
Methidathion
381
302
237
157
-Endosulfan
Parathion
ethyl
404
291
372, 404,
291, 292, 97
406, 408, 410
Part 6-Chlordane
Omethoate
12
246
NCI-SRM m/z
169>107 (20),
(Collision
Energy, eV)
14,16
199, 121, 241, 231

235, 165
235>200 (5)
235, 165
235>165 (25),
235>200 (5)
235, 165
171, 377
235>165 (25),
235>199 (15)
71>35 (35),
(5)
241>81
257>81 (35)
157>142 (15),
172, 157
173, 125
172>84 (5)
237
13,16
1
17
1
17
1
13
1
16
1
1
1
15
2
2
13
1
16
1
16
15
13,16
241,
195
97, 291
242>35
291>154(15),
(5),
406>35 (10)
291>169
(10)
195>160 (5),
339>159 (15)
NCI, SRM, AND LVI
314>223 (25),
319>81 (25)
Dichlofenthion
345
244
16
241>81 (35),

241, 79
Table I:Leptophos
monitoring
257>81 (35)
Malathion
Malathion
330
330
172, 157
157, 159, 158
173, 125
173, 125, 93
NCI-SRM m/z
EI-SRM m/z
Pesticide
MW
NCI-SIM (m/z)
Malathion
330
157
Omethoate
-HCH
213
288
141
71,
73
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
16
1
Methidathion
-HCH
302
288
157
71, 73
181, 111
291
288
291
288
263
291, 292, 97
71, 73
291
255, 257, 253
263, 154, 141
181>145 (15),
181>109 (25)
13,16
1
Parathion ethyl
-HCH (lindane)
Parathion ethyl
-HCH (lindane)
Parathion methyl
71>35 (5),
73>37
291>154(10)
(5),
291>169
(10)
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
-HCH
Parathion
methyl
288
263
71, 263
73
154,
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
HCH , ,
,
Parathion
methyl
88
262
71
263
Profenophos
374
293
267
265,
249
237, 295
Phorate
Hexachlorobenzene
260
285
222,
224
284,
286,
282
121,
75282
284,
286,
Phorate
o,p-DDE
260
316
185
318,
316
316, 318
316>246 (15),
246>176 (25)
16
1
Phosalone
p,p-DDE
Phosalone
367
316
367
185
318, 316
185, 186, 187
316, 318
316>246 (15),
246>176 (25)
13
1
14
o,p-DDD
Pirimophos-ethyl
306
333
71,
248
141
235, 165
235>165 (25),
235>200 (5)
1
16
Profenofos
p,p-DDD
372
306
267, 308, 336,

71, 248
79, 186, 183
235, 165
235>165 (25),
235>200 (5)
13
1
Propetamphos
p,p-DDT
281
352
154
71,
73
235, 165
16
1
Ronnel
320
235>165 (25),
235>199 (15)
Ronnel
Aldrin
320
381
Sulfotep
Chlordane
322
406
410, 408, 412
322,
202
373,
375,
377
Sulprofos
Part 7-Chlordane
322
406
279, 247
266,232,71
140, 375
322
373,
Terbufos
-Chlordane
288
406
266,185
71, 232
375, 373
Tokuthion
Dieldrin
344
381
237,
301
237
113, 267
Tributylphosphorotrithioite
-Endosulfan
314
404
372, 404,
257
406, 408, 410
169,195
57
241,
Malathion
13
330
157, 172, 315
EI-SIM (m/z)
157>142 (15),
172>84 (5)
181, 183, 109

181, 111
13
16
97, 291
181, 111
Reference
1
1
16
2
13
1
14
15
16
265>35 (35),
231>35 (25)
185>111 (15),
185>157 (10)
71>35 (5)
211, 270, 141

211
237
237>119 (15),
237>146 (25)
260>75 (5),
263>231 (5)
322>202 (10),
322>146 (25)
279>139
(20),
71>35 (5),
279>124
(35)
266>35 (15)
322>156 (25),
(10),
375>266
322>97 (20)
375>303
(10)
71>35 (5),
266>35 (15)
269>161 (10),
237>79 (25)
375>266 (25),
375>303 (10)
344>328 (10),
344>73 (20)
257>79 (15),
242>35
257>89 (10)
(20)
406>35
314>115
195>160(20),
(5),
314>113 (15)
(20)
339>159
16
1
1
2,4
13
13
15
1
2
1
16
1
1
15
1
NCI, SRM, AND LVI
Isofenfos
372
267, 308, 336,

79, 186, 183
13
Table Propetamphos
I: Selected-ion monitoring
154
Ronnel
320
211, 270, 141
Ronnel
320
211
Pesticide
MW
NCI-SIM (m/z)
Sulfotep
EI-SIM (m/z)
322, 202
13
EI-SRM m/z
322>202
(10), eV)
(Collision
Energy,
322>146 (25)
13
Reference
1
279>139 (20),
Organochlorine
Insecticides
140, 322
279>124 (35)
71>35 (5),
181, 111
73>37 (10)
322>156 (10),
322>97 (20)
181>145 (15),
181>109 (25)
181,
113, 111
267
71>35 (5),
269>161
(10),
73>37 (10)
237>79
(25)
181>145
344>328 (15),
(10),
181>109
(25)
344>73 (20)
71,
73
257
181,
169,111
57
71>35 (15),
(5),
257>79
73>37 (10)
257>89
(20)
181>145 (20),
(15),
314>115
181>109 (20)
(25)
314>113
288
332
255, 257, 253

213, 296
181, 183, 109

109, 297
-HCH
288
71, 73
HCH , , ,
88
Captan
301
293
150,
265, 151
249
79,
107,
149
237,
295
Captan
Hexachlorobenzene
Captan
o,p-DDE
Captan
301
285
301
316
301
150, 149, 151

284, 286, 282
150, 182
318, 316
150
79, 151, 264

284, 286, 282
316, 318
316>246 (15),
246>176 (25)
4
2,4
8
1
15
p,p-DDE
Folpet
316
297
318,
316
146,
147,
148
316,
318
260,
104,
262
316>246 (15),
246>176 (25)
1
2
Folpet
o,p-DDD
Captafol
p,p-DDD
Captafol
297
306
146
71, 248
235, 165
8
1
349
306
349
150,151
71, 248
150, 217
79, 149, 137

235, 165
235>165 (25),
235>200 (5)
Captafol
p,p-DDT
352
349
71,
73
150
235, 165
Phthalimide
147
Tetrahydrophthalimide
Aldrin
151
381
149
237
Chlordane
406
410, 408, 412

250, 252, 251
266,232,71
250, 262
139, 251, 253

373, 375
Dicofol
370
406
370
71>35 (5),
266>35 (15)
375>266 (25),
375>303 (10)
-Chlordane
Dicofol
406
370
266, 71, 232

250, 252, 251
375, 373
71>35 (5),
266>35 (15)
375>266 (25),
375>303 (10)
Dieldrin
Dicofol
381
370
237
250
-Endosulfan
404
372, 404,
406, 408, 410
Sulprofos
322
279, 247
-HCH
Terbufos
288
288
71, 73
185
-HCH
Tokuthion
288
344
71, 73
237,
301
-HCH (lindane)
Tributylphosphorotrithioite
288
314
-HCH (lindane)
Trichloronate
Part 8-Chlordane
Dicofol
14
322
NCI-SRM m/z
16
181,Fungicides
111
296>108 (10),
296>79 (20)
71>35 (5),
73>37 (10)
71>35 (5)
1
16
2
1
181>145 (15),
181>109 (25)
Phthalimide
Fungicides and Their Degradation Products
71
265>35 (35),
231>35 (25)
1
15
237>119 (15),
237>146 (25)
3
1
235>165 (25),
235>200 (5)
3
1
8
235>165 (25),
235>199 (15)
1
15
147 Insecticides (Chlorinated) and Their Degradation Products

Cyclodiene
8
8
15
Bridged
Diphenyl
Acaricide
373, 375,
377
2
2
1
8
1
12
15
Anilide
241, 195Fungicides
242>35 (15),
406>35 (10)
307>307 (5),
195>160 (5),
339>159 (15)
NCI, SRM, AND LVI
Profenofos
297
146
Captafol
349
150,151
8
79, 149, 137
Captafol
349
150,
217
chromatographynegative
chemical
ionization-mass
spectrometry (GCNCI-MS).
Captafol
15
Phthalimide
Pesticide
Tetrahydrophthalimide
349
147
MW
151
150
147
NCI-SIM (m/z)
149
EI-SIM (m/z)
NCI-SRM m/z
EI-SRM m/z
Reference
2
1
Bridged Diphenyl
Acaricide
Organochlorine
Insecticides
Dicofol
-HCH
370
288
250,71,
252,
73251
139,
251,
253
181,
111
Dicofol
-HCH
Dicofol
370
288
370
250, 262
71, 73
250, 252, 251
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
8
1
12
-HCH
(lindane)
Dicofol
288
370
71,
73
250
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
1
15
-HCH (lindane)
288
255, 257, 253
Anilide
Fungicides
181,
183, 109
2
307>307
(5),
71>35 (5),
307>195.2
(5)
73>37 (10)
Boscalid
-HCH
343
288
HCH
, , ,
Fenhexamid
88
302
293
Fipronil
Hexachlorobenzene
437
285
Fipronil
o,p-DDE
437
316
Fluquinconazole
p,p-DDE
376
316
318, 316
316, 318
348>203 (5),
348>320 (15)
Hexaconazole
o,p-DDD
314
306
71, 248
235, 165
257>221 (5),
257>69 (15)
Myclobutanil
p,p-DDD
289
306
288, 289, 290

71, 248
179, 245, 288

235, 165
Propiconazole
p,p-DDT
Prochloraz
342
352
377
Tetraconazole
Aldrin
372
381
Chlordane
406
Metribuzin
-Chlordane
214
406
410, 408, 412

198, 199,
200, 184
266,232,71
Part 9-Chlordane
Linuron
406
249
266, 71, 232

248, 250, 217
Dieldrin
381
237
Iprodione
-Endosulfan
330
404
15
71, 73
181, 111
265>222 (5),
265>168 (5)
265>35 (35),
Conazole,
IImidazole, Pyrazole,
265, 249
237, 295Triazinone, and Triazole Fungicides
231>35 (25)
366, 384, 331
367, 369, 213
284, 286, 282
284, 286, 282
366>318 (5),
318, 316
316, 318
366>250 (5)
181>145 (15),
181>109 (25)
71
256>220 (5),
256>109 (20)
71>35 (5)
71, 73
235, 165
377, 375, 379
308, 310, 266
117>97.5 (5),
117>117 (5)
237
15
5
237>119 (15),
237>146 (25)
1
4
2,4
316>246 (15),
246>176 (25)
5
1
316>246 (15),
246>176 (25)
5
1
235>165 (25),
235>200 (5)
5
1
235>165 (25),
235>200 (5)
2
1
235>165 (25),
235>199 (15)
5
1
2
5
15
Triazinone Herbicide
373, 375, 377
198, 41, 57
373, 375
5
1
2
71>35 (5),
266>35 (15)
Phenylurea Herbicide
71>35 (5),
375, 373
266>35
(15)
61, 248, 160
Dichlorophenyl Dicarboximide Fungicides

372, 404,
242>35 (15),
329, 330, 331
314,
316,
187
241,
195
406, 408, 410
406>35 (10)
375>266 (25),
375>303 (10)
2,4
1
375>266 (25),
375>303 (10)
1
2
15
195>160 (5),
339>159 (15)
2,4
1
NCI, SRM, AND LVI
Folpet
256>220 (5),
256>109 (20)
342
377 (SIM) or
377,
375, 379
308,
Table I:Prochloraz
selected-reaction
monitoring

117>97.5 (5),
Tetraconazole
372
Pesticide
Metribuzin
MW
214
117>117 (5)
Triazinone Herbicide
NCI-SIM
(m/z)
198, 199,
200, 184
EI-SIM (m/z)
198, 41, 57
NCI-SRM m/z
EI-SRM m/z
Reference
181>145 (15),
181>109 (25)
2
1
181>145 (15),
181>109 (25)
1
2,4
181>145 (15),
181>109 (25)
1
2
2,4
Phenylurea Herbicide
Organochlorine
Insecticides
Linuron
-HCH
249
288
-HCH
Iprodione
288
330
-HCH
(lindane)
Procymidone
288
284
71>35 (5),
73>37 (10)
Dichlorophenyl Dicarboximide Fungicides
71>35 (5),
71, 73
181, 111
73>37
(10)
329, 330, 331
314, 316, 187
71>35 (5),
73284
181,
111
283,71,
285,
96,
283,
285
73>37 (10)
Vinclozolin
-HCH
(lindane)
286
288
241, 257,
243, 253
245
255,
-HCH
288
71, 73
Azoxystrobin
HCH , , ,
403
88
71
Azoxystrobin
403
293
371, 356, 301

265, 249
237, 295
Trifloxystrobin
Hexachlorobenzene
408
285
284, 286, 282
284, 286, 282
o,p-DDE
Acrinathrin
316
541
318, 316
167, 305, 333
p,p-DDE
Acrinathrin
316
541
o,p-DDD
Allethrin
306
302
318, 305,
316
333,
334, 167
71, 248
235, 165
p,p-DDD
Allethrin
306
302
71,
248168
167,
134,
235, 165
-Cyfl
uthrin
p,p-DDT
352
434
71, 73
235, 165
Bifenthrin
423
Aldrin
Bifenthrin
Chlordane
Bifenthrin
-Chlordane
Bifenthrin
381
423
406
423
406
423
Part 10
-Chlordane
Bifenthrin
16
406
423
248,71,
250,
73217
61,
248,
160
181,
111
212, 183,
187, 109
124
181,
Strobilurin Fungicides
181, 111
2,4
2
71>35 (5),
73>37 (10)
371>356 (10),
371>371 (5)
181>145 (15),
181>109 (25)
265>35 (35),
231>35 (25)
202>184 (10),
202>164 (10)
237>119 (15),
237>146 (25)
Pyrethroid Insecticides
316, 318
316>246 (15),
246>176 (25)
316>246 (15),
246>176 (25)
316, 318
167>41 (30),
167>111 (20)
406>257
(15),
71>35 (5)
207>35 (10)
(5), Products
Cyclodiene Insecticides (Chlorinated) and Their 386>205
Degradation
386>141 (20)
237
205, 241, 386
410, 408, 412
373, 375, 377
386, 241,
387, 205
71>35 (5),
266,232,71
373, 375
266>35 (15)
205, 241, 206
71>35 (5),
266,387,
71, 232
375,
373
386,
388
181,
165,
166
266>35 (15)
Dieldrin I-II
Cyfenothrin
381
375
237
167,
198
Cyfl
uthrin I-IV
-Endosulfan
434
404
372,
404,
171,
207,
209
406, 408, 410
207, 209,
1
5
15
12
1
5
2,4
1
8
1
11
235>165 (25),
235>200
(5)
123>81 (6)
235>165 (25),
235>200 (5)
1
11
235>165 (25),
163>127(15)
(5)
235>199
1
6
1
6
375>266 (25),
375>303 (10)
375>266 (25),
375>303 (10)
15
8
2
11
1
12
1
2,4
15
8
241, 195
242>35 (15),
406>35 (10)
195>160 (5),
339>159 (15)
8
1
NCI, SRM, AND LVI
Propiconazole
Table
302
167, 134, 168
11
406>257 (15),
163>127 (5)
207>35 (10)
I: Selected-ion monitoring (SIM) or selected-reaction monitoring (SRM) for pesticides
that can be analyzed by gas
-Cyfluthrin
434

386>205 (5),
Bifenthrin
423
Bifenthrin
423
205, 241, 386
Pesticide
MW
NCI-SIM
(m/z)
386, 241,
Bifenthrin
EI-SIM (m/z)
387, 205
EI-SRM m/z
Reference
11
Bifenthrin
423
205, 241, 206
Bifenthrin
-HCH
423
288
386,71,
387,
73388
181,
165,
166
181,
111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
2,4
1
Cyfenothrin I-II
-HCH
Cyfluthrin I-IV
375
288
434
167, 198
71, 73
171, 207, 209
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
8
1
8
-HCH (lindane)
Cyfluthrin I-IV
288
434
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
1
9
-HCH (lindane)
Cyfluthrin I-IV
-HCH
Cyfluthrin I-IV
HCH , , ,
Cyfluthrin I-IV
288
434
288
434
88
434
71, 209,
73
207,
171, 173, 211
255, 257, 253
171, 173,
207, 209, 211
71, 73
207, 171, 209
71
207
12
-Cyhalothrin
293
450
Hexachlorobenzene
-Cyhalothrin
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
2
10
1
11
15
15
265, 249
237, 295
265>35 (35),
241>205
(5),
231>35 (25)
241>121 (20)
237>119 (15),
237>146 (25)
1
5
285
450
284, 286, 282

205, 241, 243
284, 286, 282
o,p-DDE
-Cyhalothrin
316
450
318,
316
241,
205,
243
316, 318
316>246 (15),
246>176 (25)
1
11
-Cyhalothrin
p,p-DDE
450
316
205,
241,
243
318,
316
316, 318
12
1
Cypermethrin
o,p-DDD
416
306
207, 209, 171

71, 248
163, 165, 181

235, 165
316>246 (15),
246>176 (25)
Cypermethrin
p,p-DDD
416
306
71, 248
235, 165
171>127 (5),
171>91 (5),
207>207 (5)
Cypermethrin
p,p-DDT
416
352
171, 173,
73211
207,71,
209,
235, 165
71>35 (5)
Cypermethrin
416
Cypermethrin
Aldrin
416
381
207, 209,
237 171
Cypermethrin
Chlordane
416
406
410, 207
408, 412
373, 375, 377
-Cypermethrin
-Chlordane
416
406
266,232,71
373, 375
-Cypermethrin
416
406
171, 207, 209

266, 71, 232
375, 373
Deltamethrin
Dieldrin
505
381
237
-Endosulfan
Deltamethrin
404
505
372, 404,
406, 408, 410
-Chlordane
Part 11
17
423
386>141 (20)
NCI-SRM m/z
181, 183, 109
2,4
8
235>165 (25),
235>200 (5)
2
1
235>165 (25),
235>200 (5)
5
1
235>165 (25),
235>199 (15)
10
1
207, 171, Insecticides

209
Cyclodiene
(Chlorinated) and Their Degradation Products
11
12
15
241, 195
15
2
388>239
(15),
71>35 (5),
388>211
(25)
266>35 (15)
375>266
181>152 (25),
(30)
375>303 (10)
6
1
71>35 (5),
266>35 (15)
297>81 (15),
297>297 (15),
297>79 (15)
242>35 (15),
297>79
406>35 (15),
(10)
297>81 (15)
375>266 (25),
375>303 (10)
8
1
5
15
195>160 (5),
339>159
181>125 (15)
(25)
1
6
NCI, SRM, AND LVI
Allethrin
416
171, 173,
207, 209, 211
10
Table Cypermethrin
I: Selected-ion monitoring
selected-reaction
416 (SIM) or
207,
171, 209
Cypermethrin
416
207, 209, 171
Cypermethrin
416
207
Pesticide
MW
NCI-SIM (m/z)
-Cypermethrin
416
EI-SIM (m/z)
NCI-SRM m/z
388>239
(15), eV)
(Collision
Energy,
388>211 (25)
11
12
EI-SRM m/z
(Collision
Energy,
181>152
(30) eV)
15
Reference
6
-Cypermethrin
416
171, 207, 209
-HCH
Deltamethrin
288
505
71, 73
181, 111
-HCH
Deltamethrin
288
505
71, 73
181, 111
-HCH (lindane)
Deltamethrin
288
505
71, 73
79,81, 297
181, 111
-HCH (lindane)
Deltamethrin
288
505
255, 257, 253

79, 81, 137
181, 183, 109
-HCH
Deltamethrin
288
505
181, 111
HCH , , ,
Deltamethrin
88
505
71, 295,
73
297,
505, 217
71
79, 81, 137, 139
Deltamethrin
293
505
265,
79249
237, 295
Deltamethrin
Hexachlorobenzene
505
285
297,286,
299,282
79
284,
181, 286,
253, 282
251
284,
Fenfluthrin
o,p-DDE
389
316
207,
209,
171
318,
316
316, 318
Fenpropathrin
p,p-DDE
349
316
318, 316
316, 318
Fenpropathrin
o,p-DDD
Fenpropathrin
349
306
349
141
71, 248
141, 142, 143
235, 165
235>165 (25),
235>200 (5)
8
1
11
p,p-DDD I-II
Fenvalerate
306
420
71, 248
211,
213
235, 165
235>165 (25),
235>200 (5)
1
8
Fenvalerate
p,p-DDT I-II
420
352
167, 169,
71, 73
211,
213
235, 165
235>165 (25),
235>199 (15)
10
1
Fenvalerate I-II
420
Fenvalerate
Aldrin I-II
420
381
211
237
Chlordane
Flucythrinate
406
451
410, 408, 412
373, 375, 377
-Chlordane
Flucythrinate
406
451
266,232,71
243,
244, 245
373, 375
Flucythrinate
-Chlordane
Part 12
Flumethrin
451
406
243
266, 71, 232
375, 373
510
381
503
404
503
237
294, 296, 502
372, 404,
406, 408,
294 410
18
Dieldrin
Fluvalinate
-Endosulfan
Fluvalinate
71>35 (5),
297>81
(15),
73>37 (10)
297>297
(15),
297>79
(15)
71>35 (5),
73>37 (10)
297>79
(15),
297>81
(15)
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
181>145 (15),
181>109 (25)
181>125 (25)
181>145 (15),
181>109 (25)
1
5
1
6
1
8
2
10
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
1
11
15
12
265>35 (35),
231>35 (25)
237>119 (15),
237>146 (25)
1
15
2,4
141>141 (5),
141>97.4 (10)
71>35 (5)
316>246 (15),
246>176 (25)
11
1
316>246 (15),
246>176 (25)
5
1

212
Cyclodiene
11
15
15
241, 195
243>199 (5),
243>133.5 (15)
71>35 (5),
266>35 (15)
2
5
375>266 (25),
375>303 (10)
1
11
71>35 (5),
266>35 (15)
207>91 (25)
375>266 (25),
375>303 (10)
15
1
242>35 (15),
406>35 (10)
195>160 (5),
339>159 (15)
6
15
11
1
15
NCI, SRM, AND LVI
Cypermethrin
420
167, 169,
211, 213
10
Table Fenvalerate
I: Selected-ion
selected-reaction
I-II monitoring
420 (SIM) or
211,
213, 212
Fenvalerate I-II
420
Flucythrinate
451
Pesticide
MW
NCI-SIM (m/z)
Flucythrinate
451
243
Flumethrin
-HCH
510
288
71, 73
181, 111
71>35 (5),
207>91
(25)
73>37 (10)
181>145 (15),
181>109 (25)
6
1
Fluvalinate
-HCH
Fluvalinate
503
288
503
294, 296, 502

71, 73
294
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
-HCH
(lindane)
Fluvalinate-I-II
288
503
71, 296
73
294,
181, 111
71>35 (5),
73>37 (10)
181>145 (15),
181>109 (25)
11
1
15
-HCH (lindane)
288
255, 257, 253
181, 183, 109
tau-Fluvalinate
-HCH
503
288
71, 73
181, 111
HCH , , ,
cis/trans-Permethrin
88
391
71 209
171, 207,
391
293
207, 209,
265, 171,
249 173
237, 295
Hexachlorobenzene
391
285
171, 173,
284, 286, 282
207, 209, 211
284, 286, 282
o,p-DDE
316
391
207,
318, 390,
316
209, 171
316, 318
316>246 (15),
246>176 (25)
1
11
p,p-DDE
391
316
207,
209,
171
318,
316
316, 318
316>246 (15),
246>176 (25)
12
1
o,p-DDD
Phenothrin
391
306
350
207
71, 248
167
235, 165
235>165 (25),
235>200 (5)
p,p-DDD
Prallethrin
306
300
71, 248
167, 132, 168
235, 165
235>165 (25),
235>200 (5)
15
1
8
1
11
p,p-DDT
Tefl
uthrin
352
419
241,
71, 382,
73
205, 243
235, 165
235>165 (25),
235>199 (15)
1
11
Tetradifon
356
Aldrin
Tetradifon
381
356
320, 237
318, 245
Chlordane
Tetramethrin
406
331
410,
165, 408,
167, 412
331
373, 375, 377
Tetramethrin
-Chlordane
331
406
331,
165, 332
266,232,71
373, 375
71>35 (5),
266>35 (15)
375>266 (25),
375>303 (10)
11
1
Transfluthrin
-Chlordane
371
406
207, 209, 171

266, 71, 232
375, 373
71>35 (5),
266>35 (15)
375>266 (25),
375>303 (10)
11
1
381
237
404
372, 404,
406, 408, 410
Flucythrinate
Dieldrin
Part
13
19
-Endosulfan
451
211
11
15
243>199 (5),
243, 244, 245
EI-SIM (m/z)
243>133.5 m/z
(15)
NCI-SRM
EI-SRM m/z
11
15
294>179 (10),
294>250 (10),
71>35 (5),
294>194
(15),
73>37
(10)
294>294
(5)
Reference
1
8
2
181>145 (15),
181>109 (25)
5
1
15
8
265>35 (35),
231>35 (25)
237>119 (15),
237>146 (25)
9
1
10
2,4
71>35 (5)
Cyclodiene
356
8
15
12
2
8
15
241, 195
242>35 (15),
406>35 (10)
195>160 (5),
339>159 (15)
NCI, SRM, AND LVI
Fenvalerate I-II
NCI, SRM, AND LVI
SPONSORED
QuEChERS GCMS Analysis of
Pesticides in Cucumber
QuEChERS Dispersive Solid Phase
Extraction for the GC-MS Analysis of
Pesticides in Cucumber
Anila I Khan, Thermo Fisher Scientific, Runcorn, Cheshire, UK
A p p li c a ti o n N o te 20549
Click to
view PDF
GCNCI-SIM than with GCNCI-SRM (1,5,6). However, GCNCI-SRM can provide

additional confirmation, particularly when qualifier ions in GCNCI-SIM are too
low in abundance. Literature data are more limited for GCNCI-SRM, however
(Table I).
Key Words
QuEChERS, pesticide residue analysis, cucumber, food safety
Abstract
QuEChERS dispersive SPE is a simple, fast and quantitative sample
preparation method. This application demonstrates the effectiveness of this
technique in the GC/MS analysis of pesticides in cucumber, using a Thermo
Scientific TraceGOLD TG-5MS GC column for analysis.
The recoveries for the spiked pesticides in cucumber matrix at 50 ng/g were
between 75.2 to 119.6% with relative standard deviations ranging from
2.1 to 8.9% using the QuEChERS method described in EN15662.
Introduction
QuEChERS (Quick, Easy, Cheap, Effective, Rugged and
Safe) is a dispersive Solid Phase Extraction (SPE)
technique for extracting multi-residue pesticides from
fruits and vegetables. The advantages of this methodology
are speed, ease of execution, minimal solvent requirement
and cost. The QuEChERS methodology was developed by
Anastassiades et al1 and has become widely used in food
safety analyses.
The method is:
Quick high sample throughput, typically 8 samples
can be prepared in under 30 min
Easy it requires less handling of extracts than other
techniques i.e. fewer steps are required
Cheap less sorbent material is needed and less time is
required to process samples compared to other techniques
Effective the simple technique gives high and accurate
recovery levels for a range of different compound types.
Rugged the method can detect a large number of
pesticides including charged and polar pesticides
Safe unlike other techniques, it does not require the
use of chlorinated solvents. Extraction is typically carried
out using acetonitrile, which is both GC and LC
compatible.
The sample preparation approach described in the

European EN15662 QuEChERS procedure2 was used for
extracting pesticides from cucumber. This is a two stage
process: sample extraction, followed by dispersive SPE.
In the sample extraction stage, the food sample is
homogenized to increase the available surface area of the
sample to provide optimal extraction efficiencies. The
homogenized sample is placed in the extraction tube
containing magnesium sulfate and salts (sodium chloride,
sodium citrate tribasic dihydrate, sodium citrate dibasic
sesquihydrate). Magnesium sulfate ensures that, upon
addition of acetonitrile, a phase separation is induced
between water and organic solvent with the pesticides of
interest being extracted into the organic phase. When
acetonitrile is poured into the extraction tube containing
the homogenized sample,
In both EI and NCI, pesticides generally fragment easily in the ion source. NCI
is a softer ionization process than EI so there is a tendency to observe highermass ions with NCI than with EI. However, even with these higher-mass ions
sometimes only one small product ion of significant abundance is formed
from collision-induced dissociation (CID), such as Cl for a number of the OCs,
chlorinated cyclodiene insecticides, and a few OPs: In Table I, see SRMs for
hexachlorocyclohexanes (HCHs), dichloro-diphenyl-trichloroethane (DDT),
chlordane, endosulfan, heptachlor epoxide, and mirex. For other chemical
classes such as most OPs, strobilurin fungicides, and pyrethroids, highermass product ions from CID are generally produced. GCNCI-SRM has been
applied successfully to the quantitative and confirmation analysis of a number of
pyrethroids (5,6). Pyrethroids generally have larger-mass fragment ions present
in the ion source that can be selected as the parent ion for CID. The added
selectivity of NCI coupled with SRM provides the opportunity for reduced
analysis times because matrix interferences and the occurrence of coeluted
pesticides are often reduced.
EI frequently is used for multiresidue pesticide analysis with GCtandem MS
(1722). GCEI-SRM is applicable to a large number of target compounds, and
information is more readily available from MS libraries for parent ions formed in
the EI ion source. This approach allows for the analysis of a larger range of GC
amenable pesticides together with the chloroacetanilides and triazines. GCEISRM also can be used for pesticide classes that are amenable to NCI; however,
in general detection limits are higher in GCEI-SRM. Some OPs have similar
sensitivity for GCEI-SIM and GCEI-SRM (17). GCEI-SRM does not suffer from
the background and matrix interferences observed with GCEI-SIM and in some
cases can shorten the total analysis time by reducing both sample preparation
and GC run time with fewer problems of coeluted compounds (18). In our work
on atmospheric samples, however, we found that GCEI-SRM was not suitable at
the low picogram-per-microliter level for OCs, cyclodiene insecticides, or most
20
NCI, SRM, AND LVI

organophosphorus pesticides (1). The response ratios of quantitative to qualifier
SRM transitions at these trace levels often exceeded an RSD of 25%, thus another
approach was required for confirmation (1). For other sample types, such as food
commodities, where sample concentration or size can be adjusted to provide
preconcentration, the use of GCEI-SRM analysis has been sufficient to meet
regulatory needs (1722).
Large-Volume Injections Using Cold On-Column

Injection or a Programmable-Temperature
Vaporizer
For standard pesticide analysis, a splitsplitless injector in splitless mode
frequently is used with a variety of liners ranging from those containing glass
wool to prepacked inlet liner material (such as CarboFrit, Restek) to improve the
efficiency of vaporization. With splitless injections sample volume is limited to
~12 L depending upon solvent selection. Phthalimide fungicides are prone to
degradation in the injector port and must be analyzed with a different injector.
Phthalimide fungicides, including both trichlorothio-phthalimides (captan
and folpet) and tetrachlorothio-phthalimides (captafol), exhibit significant
improvements in sensitivity with NCI and tend to form higher-mass ions in NCI
(4,23). To minimize degradation in the injector port and improve detection limits,
analysis can be completed with a large-volume cold on-column (COC) injection
approach where a volume of up to 100 L is injected (4). Another option is to
use a PTV. The choice between using COC injection and a PTV often depends
on the boiling point of the analyte and solvent used as well as the method used
to clean up the extracts before analysis. Phthalamide fungicides have low boiling
points and thus are less suitable to the use of a PTV inlet because certain types
of PTV liners may increase sample degradation and analyte loss may occur during
the solvent venting stage (23). However, large-volume (up to 20 L) dirty matrix
injection (DMI), where the sample extract is held in a microvial in a glass liner of
a PTV inlet, has been shown to be successful for captan, captafol, and folpet,
even with ethyl acetate as the solvent (24). Some degradation products of these
fungicides have been monitored by GCEI-SIM but sensitivity generally is too low,
21
NCI, SRM, AND LVI

and as a result liquid chromatography (LC) methods generally are preferred (23).
Stabilizers or solvent acidification can be used in sample preparation to reduce
degradation, however (23).
Although PTV and COC inlets are commonly used for more thermally labile
pesticides, another advantage of these approaches lies in their adaptation
for large-volume injections for any GCamenable pesticide. In both cases the
chromatography system is adapted to include a guard column and retaining
column or retention gap and the speed of the injection is controlled to 1 L/s
to accommodate larger injection volumes. In some cases shorter analytical
columns are also selected to reduce the potential for thermal degradation on
column. The temperature of an NCI ion source is also lower than that of an EI
source. PTV can be used for splitless pulsed injection with a total volume up to
120 L or more by multiple pulsed injections of 20 L each when the temperature
is low (75 C) and the split vent is open with hexane as solvent (25). Solvent
evaporation speed depends on the injector (liner) temperature, pressure, and
split flow rate. After the sample injection and solvent venting step are completed,
the PTV temperature can be ramped to transfer the analytes to the analytical
column (24,25). Some researchers have used ambient pressure during the solvent
vent stage for PTV injections (26). These PTV approaches have a setup during
the venting step similar to that of large volume injectiondirty matrix injection
(LVIDMI) techniques but these PTV approaches allow for multiple injections, to
increase the total volume to a range that is comparable to LVICOC injections.
With DMI injections the GC liner is replaced after each injection with a robotic
injection system, thus removing the potential for buildup of nonvolatiles in the
injection liner over time. Concurrent solvent recondensation is another approach
that has been used for splitless large-volume injections of 5 L with solvents such
as toluene, isooctane, and acetonitrile (27).
Conclusions
With the exception of the chloracetanilides and triazines, negative chemical
ionization provides added selectivity and sensitivity for many electron-capturing
pesticides (with halogenated, NO2, or P ester groups) that are GCamenable.
22
References
1. R. Raina and P. Hall, Anal. Chem. Insights 3, 111125 (2008).
2. R. Huskova, E. Matisova, S. Hrouzkova, and L. Svorc, J. Chromatogr. A 1216, 63266334 (2009).
3. R. Huskova, E.Matisova, L. Svorc, J. Mocak, and M. Kirchner, J. Chromatogr. A 1216, 49274932
(2009).
4. R. Bailey and W. Belzer, J. Agric. Food Chem. 55, 11501155 (2007).
5. R. Montes, I. Rodriguez, M. Ramil, E. Rubi, and R. Cela, J. Chromatogr. A 1216, 54595466
(2009).
6. P. Deme, T. Azmeera, B.L.A. P. Devi, P.R. Jonnalagadda, R.B.N. Prasad, and U.V.R.V. Sarathi, Food
Chem. 142, 144151 (2014).
7. T. Dagnac, S. Bristeau, R. Jeannot, C. Mouvet, and N. Baran, J. Chromatogr. A 1067, 225233
(2005).
8. I.R. Pizzutti, A. de Kok, C.D. Cardoso, B. Reichert, M. de Kroon, W. Wind, L. W. Righi and R. C.
da Silva, J. Chromatogr. A 1251, 1626 (2012).
9. G.A. Bonwick, C. Sun, P. Abdul-Latif, P.J. Baugh, C.J. Smith, R. Armitage, and D.H. Davies, J.
Chromatogr. A 707, 293-302 (1995).
10. M. Yasin, P.J. Baugh, G.A. Bonwick, D.H. Davies, P. Hancock, and M. Leinoudi, J. Chromatogr.
A 754, 235243 (1996).
11. C.-Y. Shen, X.-W. Cao, W.-J. Shen, Y. Jiang, Z.-Y. Zhao, B. Wu, K.-Y. Yu, H. Liu, and H.-Z. Lian,
Talanta 84, 141147, (2011).
12. D.I. Kolberg, O.D. Prestes, M.B. Adaime, and R. Zanella, Food Chem. 125, 14361442 (2011).
23
NCI, SRM, AND LVI
Although GCNCI-SRM has not been used as frequently as GCNCI-SIM because

of generally slightly higher detection limits, it can provide added selectivity and
confirmation ability for difficult sample matrices. GCEI-SRM also can reduce the
occurrence of background and matrix interferences observed with GCEI-SIM
methods. When lower detection limits are necessary and not obtainable with
preconcentration of samples or when pesticides are more thermally labile, LVI
COC or LVIPTV options typically can provide a 10- to 100-fold improvement in
detection limits, and conditions during injection can be optimized to minimize
thermal degradation and loss of analytes of interest. A wide range of pesticides in
different classes can be analyzed with GCNCI methods.
NCI, SRM, AND LVI

13. L. Amendola, F. Botre, A.S. Carollo, D. Longo, and L. Zoccolillo, Anal. Chim. Acta 461, 97108
(2002).
14. K.S. Liapis, P. Aplada-Sarlis, and N.V. Kyriakidis, J. Chromatogr. A 996, 181187 (2003).
15. M. Okihashi, Y. Kitagawa, K. Akutsu, H. Obana, and Y. Tanaka, J. Pestic. Sci. 30, 368377 (2005).
16. M.V. Russo, L. Campanella, and P. Avino, J. Chromatogr. B 780, 431441 (2002).
17. M. Mezcua, C. Ferrer, J.F. Garcia-Reyes, M. J. Martinez-Bueno, M. Sigrist, and A.R. FernandezAlba, Food Chem. 112, 221225 (2009).
18. Y. Sapozhnikova and S.J. Lehotay, Anal. Chim. Acta 758, 8092 (2013).
19. A. Garrido Frenich, J.L. Martinez Vidal, J.L. Fernandez Moreno, and R. Romero-Gonzalez, J. Chromatogr. A 1216, 47984808 (2009).
20. J.L. Martinez Vidal, F.J. Arrebola Liebanas, M.J, Gonzalez Rodriguez, A. Garrido Frenich, and J.L.
Fernandez Moreno, Rapid Commun. Mass. Spectrom. 20, 365375 (2006).
21. T. Cajka, C. Sandy, V. Bachanova, L. Drabova, K. Kalachova, J. Pulkrabova, and J. Hajslova, Anal.
Chim. Acta 743, 5160 (2012).
22. K. Mastovska and P.L. Wylie, J. Chromatogr. A 1265, 155164 (2012).
23. R. Raina-Fulton, J AOAC Int., in press (2014).
24. R. Raina, in Pesticides Strategies for Pesticide Analysis, M. Stoytcheva, Ed. (InTech, Rijeka, Croatia, 2011), pp. 105130.
25. Y. Zhao, L. Yang, and Q. Wang, J. Am. Soc. Mass Spectrom. 18, 13751386 (2007).
26. R.J.C.A. Steen, I.L. Freriks, W.P. Cofino, and U.A. Th. Brinkman, Anal. Chim. Acta 353, 153163
(1997).
27. S. Walorczyk, J. Chromatogr. A 1222, 98108 (2012).
Renata Raina-Fulton, PhD, is an associate professor and department head in the

Department of Chemistry and Biochemistry and at the Trace Analysis Facility at the
University of Regina, in Regina, Saskatchewan, Canada.
24
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in Fruits and Vegetables by Gas Chromatography

Time-of-Flight Mass Spectrometry
By N
oelia Belmonte,
Samanta Ucls, Miguel
Gamn, Carmen Ferrer,
Milagros Mezcua, and
Amadeo R. Fernndez
Alba
The main objective of this study was to evaluate the capabilities of gas
chromatography (GC) with timeofflight mass spectrometry (MS) for screening
pesticides in fruits and vegetables using a purpose-built accurate-mass database.
Analytical performance was tested on four matrices: potato, tomato, spring onion,
and orange. Two resolution modes, 7000 FWHM and 12,000 FWHM, were tested
to establish the concentration range within which automatic identification was
possible, considering the retention time window of 0.2 min and at least two ions
with a mass error lower than 10 ppm as the identification criteria. The effects of the
matrix on identification and quantification were also studied for the four selected
matrices. The developed method was applied to real samples and the qualitative
and quantitative results were compared with those obtained using GC with triplequadrupole MS.
The use of agrochemicals at various stages of crop cultivation has an important impact on food protection
and quality preservation. For this reason, there are different regulations in each country on pesticide
use but the lists of approved pesticides are not harmonized worldwide. This situation is becoming more
important because the number of pesticides authorized in Europe (1) has been reduced to around 50%
of the total number of compounds manufactured. The practical target analysis approach applied in
many routine laboratories for pesticide residue testing consists of selecting a list of compounds that are
amenable for analysis by gas chromatography (GC) and liquid chromatography (LC) using an established
combined priority list. This means that the majority of low-frequency or misused compounds are not
studied.
GC coupled to mass spectrometry (MS) is the technique of choice for the analysis of many classes of
pesticides, particularly volatile, semivolatile, and low-polarity compounds. The large number of GCMS
26
ACCURATE-MASS DATABASE
AN ACCURATE-MASS
DATABASE FOR SCREENING
PESTICIDE RESIDUES
SPONSORED
Ultra-Selective Detection
of Analytes in Complex
Matrices
U-SRM Ultra Selective Detection of
Analytes in Complex Matrix Samples on
the TSQ Quantum XLS Ultra GC-MS/MS
Hans-Joachim Huebschmann, Paul Silcock, Inge de Dobbeleer, Thermo Fisher Scientific, Bremen and Dreieich,
Germany
W hi te Pape r 52283
Click to
view PDF
Key Words
GC-MS/MS, U-SRM, Mass Resolution, Matrix, Selectivity,
TSQ Quantum XLS Ultra
Introduction
The Thermo Scientific TSQ Quantum XLS Ultra triple
quadrupole mass analyzer breaks the barrier of low level
matrix interferences for a reliable compound quantitation.
Where single quadrupole analyzers are limited to isobaric
interferences on the same selected ion monitoring mass
(SIM), a triple quadrupole mass spectrometer offers
increased matrix selectivity. With this, triple quadrupole
detection has provided a large step forward in our ability
to detect at very low concentrations in complex matrix.
But, the selectivity of this nominal mass analyzer can be
challenged when some matrix/target combinations are
considered. When analyzing at extremely low concentrations, the overwhelming intensity of matrix compounds
can still provide interferences in some cases, as chemical
noise, on the product ion mass traces. It is not surprising
in this context that different matrices interfere with
different impact. So, what is the technical solution the
TSQ Quantum XLS Ultra triple quadrupole GC-MS/
MS system is using to break these barriers and overcome
these typical matrix limitations?
Of course, high performance quadrupoles are a prerequisite for a good GC triple quad system as the mass
analyzers lie at the heart of the performance delivery. In a
triple quadrupole analyzer, the first quadrupole (Q1) that
sits directly behind the ion source and pre-filter, has a
large influence on the target analyte selectivity of the mass
spectrometer. The desired precursor ions, after being
generated in the ion source, are selected by Q1 and are
transmitted to the collision cell (Q2) for fragmentation.
The third quadrupole (Q3) is set to transmit to the
detector only preselected products ions generated in Q2 as
a result of collision induced fragmentation (CID).
Without high-performing quadrupoles, mass selection
capabilities become compromised and tend towards a
lower resolution to transmit the same number of ions.
Systems that do not employ HyperQuad technology have
to open the mass transmission window (above unit mass
resolution) to transmit the same number of ions as a
HyperQuad analyzer in unit mass mode. This decreased
selectivity for target ions can give rise to chemical
Delivering High Performance Triple

Quadrupole Experiments
The TSQ Quantum XLS Ultra, as shown in Figure 1,
provides enhanced mass resolution capabilities that are
unique to GC-triple quadrupole systems. The specially
designed high precision hyperbolic quadrupoles (Thermo
Scientific HyperQuad quadrupoles), as pictured in
Figure 2, provide enhanced mass resolution and ion
transmission. This increased analyzer performances
allows an extremely low background signal which
simplifies quantification of trace compounds even when
shadowed by hugely intense matrix components, of which
a significant portion exhibit isobaric ions interfering with
the selected reaction monitoring (SRM) process. For trace
analysis in the ppt range, the increased radius of the
HyperQuad quadrupole assembly used a 50% larger
diameter between the rods for increased ion transmission
and hence sensitivity.
applications reported for pesticides analysis in fruits and vegetables (27) is

the result of efficient GC separation, together with spectral information and
satisfactory sensitivity provided by MS. Until recently, low-resolution singlequadrupole MS detectors working in selected-ion monitoring (SIM) mode
were the instruments most commonly used because of their relatively low cost,
compactness, and simplicity (8).
Single-quadrupole analyzers show limitations in analyzing complex matrices,
however ion-trap detection and, more recently, triple-quadrupole analyzers
allow operation in tandem MS (MS2) mode, which achieves high sensitivity and
selectivity, as demonstrated in an increasing number of applications reported in
various fields (9,10). Recent progress in instrumentation design (mainly optics), the
use of fast recording electronics, and signal processing improvements also have
led to a renaissance for time-of-flight mass spectrometry (TOF-MS) in the study
of organic compounds in complex matrices, particularly in LC-based applications.
TOF-MS provides greater sensitivity in full-spectrum acquisition mode compared
to conventional scanning instruments, principally because of its high massanalyzer efficiency: GCTOF-MS can screen hundreds of compounds at high
sensitivity in a single run.
Figure 1. TSQ Quantum XLS Ultra enhanced mass resolution

GC-MS/MS system
In addition, GCTOF-MS data can be acquired and reprocessed without prior

knowledge of the presence of certain compounds (that is, no analyte-specific
information is required). Equally important is that the presence of other
compounds of interest can be investigated at any time, simply by reprocessing
the data (known as retrospective analysis). The high mass-resolving power and
mass accuracy provided by GCTOF-MS make it feasible to obtain extracted ion
chromatograms using narrow mass windows thus excluding a large proportion
of the chemical background and isobaric interferences, which significantly
improves signal-to-noise ratios. Under these conditions, pesticide identification is
improved in comparison to other conventional analyzers.
Over recent years, TOF-MS has been widely used for the development of
screening methods for pesticide determination in fruits and vegetables, although
LC-based methods are still more widely used (11,12). Nonetheless, compounds
that exhibit a very low ionization yield with electrospray sources and that have
27
AN ACCURATE-MASS
PESTICIDE RESIDUES
100
100
90
90
80
80
70
60
50
40
30
20
70
60
50
40
30
Experimental
20
Chemicals and Reagents
10
10
0
10 g/kg
20 g/kg
50 g/kg
10 g/kg
100 g/kg
Potato
20 g/kg
50 g/kg
100 g/kg
Orange
100
100
Compounds (%)
90
Compounds (%)
Nowadays, GCTOF-MS is becoming a

technique of interest within the scientific
community for the determination of organic
compounds (13,14). The main objective of this
study was to develop a purpose-built database
that includes the retention time and at least
two ions for a total of 110 pesticides and to
use this database in the identification of these
compounds in fruit and vegetable matrices at
different resolving powers.
Spring Onion
Compounds (%)
Compounds (%)
Tomato
low LCMS amenability typically are outside the

scope of those LCMS screening methods.
80
70
60
50
40
30
20
10
90
80
70
60
50
40
30
20
Sample Treatment
10
0
10 g/kg
20 g/kg
50 g/kg
100 g/kg
0
10 g/kg
20 g/kg
50 g/kg
100 g/kg
Figure 1: Percentage of compounds included in the databases identified automatically with more than two ions
(green), one ion (orange), and not identified (red), working at low resolution.
28
All pesticide analytical standards used in this

study were purchased from Dr. Ehrenstorfer
(Ausburg, Germany) and Sigma Aldrich
(Steinheim, Germany) at analytical grade (purity
> 95%). A mixture standard solution contained
all the pesticides studied and was prepared at
10 g/mL in ethyl acetate and stored at 20 C.
Ethyl acetate was obtained from Fluka Analytical
Pestanal (Steinheim, Germany).
The tomatoes, spring onions (called scallions in

the United States), potatoes, and oranges used
in this study were obtained from an organic
farm in Almeria, Spain. The fruit and vegetable
samples used for the application of the
developed method were purchased from local
markets.
AN ACCURATE-MASS
PESTICIDE RESIDUES
Tomato
Spring Onion
90
Compounds (%)
100
90
Compounds (%)
100
80
70
60
50
40
30
80
70
60
50
40
30
20
20
10
10
0
10 g/kg
20 g/kg
50 g/kg
Gas Chromatography
10 g/kg
100 g/kg
20 g/kg
50 g/kg
100 g/kg
g
Orange
Potato
100
100
90
90
Compounds (%)
Compounds (%)
The extraction method employed (15) is as

follows: A representative 10-g portion of
previously homogenized sample was weighed
in a 200-mL PTFE centrifuge tube. Then 10 mL
of ethyl acetate was added, and the tube was
shaken vigorously for 3 s by hand. Next, 1.5 g of
sodium chloride and 8 g of magnesium sulfate
were added, and the tube was shaken in an
automatic axial extractor (Agytax, Cirta Lab)
for 15 min. The extract was then centrifuged at
3700 rpm for 5 min. Then the extract containing
the equivalent of 1 g of sample per mL in 100%
ethyl acetate was injected directly into the GC.
80
70
60
50
40
30
80
70
60
50
40
30
20
20
10
10
0
0
10 g/kg
20 g/kg
50 g/kg
100 g/kg
10 g/kg
20 g/kg
50 g/kg
100 g/kg
Figure 2: Percentage of compounds included in the databases identified automatically with more than two ions
(green), one ion (orange), and not identified (red), working at high resolution.
29
The separation of the pesticides from the

whole fruit or vegetable extract was carried out
using an Agilent 7890 GC system with two 15
m X 0.25 mm, 0.25-m df Agilent HP5MS UI
Ultra Inert GC columns connected through an
auxiliary programmable control module (PCM).
The samples were injected in splitless mode
through an ultra-inert liner with a glass wool frit
(Agilent). The injection volume was 2 L. The
injector temperature was held at 280 C during
the entire run time. Helium (99.999% purity) was
used as the carrier gas. The oven temperature
program was as follows: 60 C for 1 min,
then 60120 C at 40 C/min, and 120310
C at 5 C/min. The analytical separation
was performed under retention time locking
conditions, using clorpyrifos methyl as the
AN ACCURATE-MASS
PESTICIDE RESIDUES
locking compound at a retention time

of 18.11 min. The flow rates in column 1
and column 2 were 1.225 mL/min and
1.425 mL/min, respectively. The total
run time was 40.5 min with 3 additional
minutes for backflushing.
x106
Backflushing was carried out to

eliminate unwanted heavy materials
from column 1, thereby shortening
the analysis time, reducing system
maintenance, and prolonging column
life. The backflush conditions were set
as follows: the oven temperature was
set at 310 C, the PCM pressure was
50 psi, and the inlet pressure was 1 psi.
These conditions allow a negative flow
on the column 1.
1.4
1.3
1.2
1.1
Counts (%)
1
0.9
0.8
0.7
0.6
0.5
0.4
Mass Spectrometry
0.3
0.2
0.1
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
Acquisition time (min)
Figure 3: Total ion chromatogram for orange (red), tomato (blue), potato (green), and spring onion (black)
obtained by GCTOF-MS.
30
31
32
33
37
The GC system was connected to an

Agilent Technologies model 7200
TOF-MS instrument equipped with
an electron ionization source. The
ion source and quadrupole analyzer
temperatures were set at 280 C
and 150 C, respectively. The TOF
analyzer was operated at two different
acquisition rates, 2 GHz and 4 GHz,
acquiring data in the m/z 45550 mass
range. Perfluorotributylamine was used
for daily MS calibration. The accuracy
of the generated ions was controlled
through an internal mass calibration
performed before each injection.
AN ACCURATE-MASS
PESTICIDE RESIDUES
x101
2.6
-34.07 ppm
2.4
7000 FWHM
2.2
-23.7 ppm
Counts (%)
2
1.8
1.6
1.4
1.2
2.1 ppm
5 ppm
0.8
0.4
0.2
0
160
170
180
190
200
210
220
230
240
250
260
270
280
290
Mass-to-charge ratio (m/z)

1
x10
2.8
2.6
1.13 ppm
2.4
12,000 FWHM
2.2
1.8
1.6
The information obtained from this study has been discussed considering two
different goals. The first was to perform screening intended to produce a yesor-no answer while considering only two identification criteria: the retention time
and a single ion with a mass accuracy of less than 10 ppm mass error. The second
goal was to obtain confirmation of results and semiquantification considering
two ions with a mass accuracy of less than 10 ppm mass error and the correct
retention time.
6.4 ppm
1.4
1.2
1
1.8 ppm
0.8
0.6
0.4
0.2
0
160
170
180
190
200
210
220
The selected matrices were spiked with all the pesticides at four concentration
levels, 10, 20, 50, and 100 g/kg. Automatic identification was performed at two
acquisition rates, 2 GHz and 4 GHz, which allowed respective mass resolutions of
7000 and 12,000 full width at half maximum (FWHM). The search parameters were
0.2 min for the retention time window and 10 ppm for the mass error tolerance.
Results and Discussion
0 ppm
Counts (%)
The experimental conditions described above were applied to create an

accurate-mass database of 110 pesticides. The selected pesticides were run in
a tomato matrix at a concentration of 100 g/kg. The full-scan spectrum was
studied and at least two characteristic ions (with a relative abundance higher
than 30% with respect to the base peak) were selected. In most cases, the
molecular ion of the pesticides was not present in the mass spectrum or its
relative abundance was too low to be considered. After the ions were selected, a
molecular formula was assigned, and their exact masses were calculated and used
to create the database together with their retention times. All the information
was collected in an Excel file, which was converted into a CSV format to be
used as the library and linked to Mass Hunter data analysis software (Agilent
Technologies).
Automatic Identification Using the Pesticide Database
0.6
230
240
250
260
270
280
290
Figure 4: Mass spectra for benalaxyl in tomato at 20 g/kg obtained

at low and high resolution using GCTOF-MS.
31
Creation of the Database
AN ACCURATE-MASS
PESTICIDE RESIDUES
x10 4
7000 FWHM
7
6.5
Goal 1: Screening and Automatic

Identification
The four selected extracts spiked at 10,
20, 50, and 100 g/kg were analyzed at
low resolution (7000 FWHM) and at high
resolution (12,000 FWHM). The analysis was
processed with data analysis software using
the purpose-built database as the library.
6
5.5
Counts (%)
The pesticides, identified automatically with

the correct retention time and one ion with
a mass accuracy of less than 10 ppm mass
error, were counted and compared with the
total number of pesticides included in the
database to calculate the percentage of
pesticides identified.
139.0005
235.0324
-43.0 ppm
4.5
4
-1.5 ppm
-7.8 ppm
203.0631
99.1161
3.5
179.0452
3
2.5
107.0697
2
226.2169
1.5
279.0957
314.0592
1
0.5
0
50
60
70
80
90
100
110
120
130
140
150
160
170
180
190
200
210
220
230
240
250
260
270
280
290
300
310
320
x10 4
3.6
12,000 FWHM
3.4
3.2
3
2.8
138.9942
Counts (%)
2.6
107.0242
2.2
235.0314
2.3 ppm
2.4
98.0700
2.8 ppm
184.1690
1.8
203.0633
1.6
1.4
-8.8 ppm
151.0382
1.2
1
314.0587
0.8
281.0950
0.6
0.4
0.2
0
50
60
70
80
90
100
110
120
130
140
150
160
170
180
190
200
210
220
230
240
250
260
270
280
290
300
310
320
The results obtained operating at 7000

FWHM are shown in Figure 1. The green
columns for each concentration level are
the pesticide percentages with at least two
ions having an error lower than 10 ppm
and a retention time difference of less
than 0.2 min. The orange columns are the
pesticide percentages with at least one
ion with a mass error lower than 10 ppm
and a retention time error of less than 0.2
min. The columns in red are the compound
percentages not detected or detected with
a mass error higher than 10 ppm.
Figure 5: Mass spectra for nuarimol in orange at 20 g/kg obtained at

low and high resolution using GCTOF-MS.
32
At the 10-g/kg level, the percentages

of pesticides identified with at least one
fragment and the correct retention time
AN ACCURATE-MASS
PESTICIDE RESIDUES
Sample
GCTOF-MS
GCTriple-Quadrupole MS
Comments
Pepper
Boscalid (0.25 mg/kg)
Iprodione (0.20 mg/kg)
Pyriproxifen (0.054 mg/kg)
Pyriproxifen (0.062 mg/kg)
Tomato 2
Bifenthrin 1F (0.12 mg/kg)
Bifenthrin (0.13 mg/kg)
Ion 166.099 error

> 10 ppm
Melon 1
Bupirimate (0.15 mg/kg)
Buprofezin (0.49 mg/kg)
Buprofezin (0.16 mg/kg)
Metalaxyl (0.060 mg/kg)
Metalaxyl (0.068 mg/kg)
Pirimicarb (0.33 mg/kg)
Pirimicarb (0.24 mg/kg)
Penconazole (0.091 mg/kg)
Penconazole (0.074 mg/kg)
Brussels sprouts
Tomato 4
Ion 166.099 error

> 10 ppm
Bean 1*
Iprodione 1F (0.830 mg/kg)
After dilution
Azoxystrobin ND
Azoxystrobin (0.337 mg/kg)
Coelution
Azoxystrobin ND
Coelution
Azoxystrobin ND
Coelution
Bean 4*
After dilution
Bean 5*
After dilution
Myclobutanil (0.041 mg/kg)
Myclobutanil (0.041 mg/kg)
Tomato 1
Tomato 3
Zucchini
Bean 2
Bean 3
Melon 2
Grape
* These samples were injected after being diluted 1:2 to achieve correct identication and quantication.
1F = one fragment detected
ND = not detected
33
were as follows: 80% in tomato, 50%

in potato, 76% in spring onion, and
70% in orange. As expected, at higher
concentration levels, the percentage of
pesticides found was higher.
Figure 2 shows the results of the studied
matrix at the different concentration
levels obtained when operating the
system at high resolution (12,000 FWHM).
At this acquisition rate, the percentage
of positives at all concentration levels,
and in all matrices, is higher than when
operating at low resolution. At 10 g/kg,
the percentages of pesticides identified
with at least one fragment and the correct
retention time were as follows: 88% in
tomato, 91% in potato, 88% in spring
onion, and 82% in orange. At 20 g/kg,
100% of the pesticides included in the
database for tomato were identified with at
least two ions and a mass error < 10 ppm
along with a retention time < 0.2 min. The
pesticide identification rates for the spring
onion, potato, and orange were 95%,
97%, and 85%, respectively. Operating at
high resolution, the number of pesticides
found with a mass error below the level
established as the mass tolerance was
higher than at low resolution.
An important influence on mass
assignments is the ability of a mass
spectrometer to resolve two peaks on the
m/z scale when they are close together.
Table I: Real samples identified and quantified with the proposed method and compared to GCtriple-quadrupole
MS. Values listed in parentheses are the concentrations that were found in the samples.
SPONSORED
Analysis of OCPs, PAHs,
and PCBs in Environmental
Samples
Consolidated GC-MS/MS Analysis of OCPs,
PAHs, and PCBs in Environmental Samples
Inge de Dobbeleer, Joachim Gummersbach, Hans-Joachim Huebschmann, Anton Mayer, Paul Silcock
Thermo Fisher Scientific, Dreieich, Germany
Key Words
Organochlorine pesticides, polychlorinated biphenyls, polyaromatic
hydrocarbons, soil samples, water samples, building material, repeatability,
linearity
Goal
To describe the analysis of semivolatile compounds in various environmental
matrices, showing the productivity and high quality results of the GC-MS/
MS system.
This application note describes a high performance, highly

productive analysis of OCPs, PAHs and PCBs in
environmental samples through a consolidated GC-MS/MS
method using the Thermo Scientific TRACE 1310 GC
and the TSQ 8000 triple quadrupole GC-MS/MS. Also
described is the use of smart software tools that are
integrated into the method development and analysis
workflow to minimize the time needed to implement
and maintain the methodology in routine.
Experimental Conditions
Introduction
Organochlorine pesticides (OCP), polyaromatic
hydrocarbons (PAH), and polychlorinated biphenyls
(PCB) are compound classes that are highly familiar to
routine environmental or contract testing laboratories.
Various approaches are taken to address these compound
classes in the diverse matrix environment experienced by
these laboratories.
Gas chromatography-mass spectrometry (GC-MS) is well
suited for the analysis of OCPs, PAHs, and PCBs. Single
quadrupole GC-MS has offered the opportunity for the
environmental laboratory to increase selectivity for these
analytes over that of classical detectors, such as UV and
fluorescence detectors in HPLC and ECD and FID
detectors in GC. This has allowed for limited optimization
of sample preparation procedures to increase time to
result and ultimately reduce laboratory costs.
Triple quadrupole GC-MS/MS provides a significant
increase in selectivity when compared to single quadrupole
GC-MS. This selectivity has a profound effect on the
ability to cut through chemical background (interference),
which enhances the capability and productivity of an
environmental testing laboratory. This technique is being
increasingly utilized by many environmental laboratories,
especially those looking for a competitive edge.
Unfortunately, for laboratories newer to GC-MS/MS, the
adoption of this technique presents a challenge in realizing
the productivity advantages offered without significantly
impacting continued laboratory operations.1,2
A ppl ica tion N ote 52389
Click to
view PDF
Sample Preparation
Water samples
To 1 L of sample, n-hexane was added and the mixture
was shaken. After the separation of water and organic
phases, the organic phase was removed and dried with
anhydrous Na2SO4. An aliquot of the organic extract was
evaporated to a volume of 34 mL and then evaporated
under a gentle nitrogen stream to the final volume.
Solid samples
Into a glass jar, 10 g of the sample (soil, sediment, or
building material) was weighed, then anhydrous Na2SO4
and 40 mL of extraction solvent mixture (hexane and
acetone) were added. The glass jar was sealed with a
Teflon seal and sonicated for 20 min. An aliquot of the
sample extract was placed into a Kuderna Danish
apparatus, and another 40 mL of extraction solvent
mixture was added to the sample and the extraction was
repeated. An aliquot of second extraction was added to
the first extraction aliquot. The extract was evaporated to
a volume of 34 mL and then evaporated under a gentle
nitrogen stream to the final volume.
When peaks are not (fully) resolved, the resulting measured mass profile will be
the sum of the two individual mass profiles, and the maximum of the combined
profile will lie somewhere between the exact masses of the two individual
peaks. As a consequence, the mass assignment, which is based on a centroiding
algorithm of the detected profile, will result in an incorrect analyte mass. As
sample complexity becomes greater (a larger number of matrix peaks at high
levels relative to the analytes), the mass resolution (defined as an MS instruments
ability to distinguish between two ions with similar m/z values) of a mass
spectrometer can become a key parameter in the correct assignment of analyte
masses.
Figure 3 shows the full-scan chromatogram of the four matrices selected for this
study. The total ion chromatogram in red corresponds to the orange matrix;
the chromatogram in black corresponds to spring onion; green corresponds to
potato; and blue corresponds to tomato. Orange shows the largest number of
matrix interferences.
Figure 4 is an example showing the analysis of benalxyl at 20 ppb in tomato
analyzed at both low and high resolution. If we set the mass tolerance to 10 ppm,
only two ions are identified when operating at 7000 FWHM, whereas all the ions
are identified at 12,000 FWHM.
Figure 5 is an example of an analysis in the orange matrix. In this case the mass
spectrum of the pesticide nuarimol is shown at low and high resolution. At low
resolution only one ion is correctly identified, whereas at high resolution two ions
were identified with a mass error lower than that established.
Goal 2: Confirmation and Semiquantification

With the purpose of using the developed method for confirmation and
semiquantification, we considered the identification criteria to be the
identification of at least two ions with a mass error lower than 10 ppm and a
retention time error lower than 0.2 min. The pesticide percentages that meet
these criteria are as follows: Operating at high resolution, more than 70% of the
pesticides were detected at the 20 g/kg level in tomato, potato, and spring
34
AN ACCURATE-MASS
PESTICIDE RESIDUES
onion (Figure 2) with a mass error lower than 10 ppm, and with a retention time
error lower than 0.2. In the orange, 60% of the pesticides were detected.
After it was known that identification and further confirmation worked far better
at high resolution, the linearity of the response was studied between 10 and 100
g/kg operating at high resolution, and very good linearity was observed in this
range, showing correlation coefficients better than 0.999, except in these specific
cases: benalaxyl, tetraconazole, and flutolanil in orange; butralin, etoxyquine, and
p,pDDT in tomato; and pirimifos methyl, pyriproxyfen, tetraconazole, tetradifon,
and tolylfluanide in spring onion.
Given that screening methods are intended to be used with a large number of
matrices, a semiquantification study of samples was performed in the tomato
matrix-matched solution at two concentration levels, 10 and 100 g/kg. The
results were compared with those obtained using a GCtriple-quadrupole
MS system in which the concentration of the samples was calculated with
a calibration curve in matrix-matched solution performed with different
commodities, depending on the analyzed samples.
The selection of tomato as the quantification matrix was based on the results
obtained from a matrix effect study. The slope of all the matrices obtained from
the linear curve in the range between 10 and 100 g/kg was compared with the
slope obtained in solvent. All matrices showed a marked signal enhancement
effect when compared with solvent; however, the differences in the slopes
between matrices were very small.
Real samples were analyzed with the developed method, and some of the
pesticides found are listed in Table I. The results were compared with those
obtained by GCtriple-quadrupole MS; the samples were also semiquantified
as described above, and the results obtained are also shown in Table I. The
quantification differences for both systems are mostly within 50%. In some cases
the pesticide concentration was high, which caused detector saturation; such
samples were injected after dilution for better identification and quantification.
This was the case of iprodione in the bean samples. Another special case was
bifenthrin in tomato; this compound was identified with only one ion because
other characteristic ions showed errors higher than those established.
35
AN ACCURATE-MASS
PESTICIDE RESIDUES
Conclusions
The created database of 110 pesticides, which included the retention time and
at least two ions, was applied to automatic pesticide identification in tomato,
potato, spring onion, and orange. This automatic identification was made and
compared at two resolution powers, showing better results in high-resolution
mode. The identification of nearly 100% of pesticides included in the database
was demonstrated at low concentration levels (10 and 20 g/kg) in all the studied
matrices except in orange, where the pesticide identification percentage was
85%. In addition, the method was applied for the analysis of real samples, and
the obtained results were compared with those using GCtriple-quadrupole MS,
showing differences in the quantification results of less than 50%.
References
1. The European Parliament and the European Council, Regulation (EC) No. 396/2005 of the European Parliament and of the Council of 23rd February, 2005 on maximum residue levels of pesticides in or on food and feed of plant and animal origin and amending Council Directive 91/414/
ECC (Brussels, Belgium, 2005).
2. K. Banerjee, S. Mujawar, S.C Utture, S. Dasgupta, and P.G. Adsule, Food Chem. 138(1), 600607
(2013).
3. M.L. de Oliveira, F.D. Madureira, F. Aurlio, A.P. Pontelo, G. Silva, R. Oliveira, and C. Paes, Food
Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess. 29(4), 657664 (2012).
4. V.C. Fernandes, J.L. Vera, V.F. Domingues, L.M.S. Silva, N. Mateus, and C. DelerueMatos, J. Am.
Soc. Mass Spectrom. 23(12), 21872197 (2012).
5. C.J. Anagnostopoulos, G. Balagiannis, and G.E. Miliadis, Spectroscopy Letters 45(3), 202218 (2012).
6. L. Cherta, J. Beltran, T. Portols, and F. Hernndez, Anal. Bioanal. Chem. 402(7), 23012314
(2012)
7. M.A., MartnezUroz, M. Mezcua, N.Belmonte Valles, and A.R FernndezAlba. Anal. Bioanal.
Chem. 402(3), 13651372 (2012).
8. M. Mezcua, M.A. MartnezUroz, P.L. Wylie, and A.R. FernndezAlba, J. AOAC Int. 92(6),
17901806 (2009).
9. M. Zhiling, Z. Wen, L. Lingyun, Z. Shuning, L. Huan, Z. Yanguo, G. Qingzhen, and L. Su, Chin.
J. Chromatogr. 31(3), 228239 (2013).
10. U. Koesukwiwat, S.J. Lehotay, and N.J. Leepipatpiboon, J. Chromatogr. A 1218(39), 70397050
(2011).
11. M. Mezcua, O. Malato, M.A. MartinezUroz, A. Lozano, A. Agera, and A.R. FernndezAlba, J.
AOAC Int. 94(6), 16741684 (2011).
12. C. Ferrer, O. Malato, A. Agera, and A.R. FernandezAlba, Comprehensive Anal. Chem. 58, 160
(2012)
13. F. Hernandez, T. Portoles, E. Pitarch, and F.J. Lopez, Trends Anal. Chem. 30(2), 388400 (2011).
36
AN ACCURATE-MASS
PESTICIDE RESIDUES
14. F. Zhang, C. Yu, W. Wang, R. Fan, Z. Zhang, and Y. Guo. Anal. Chim. Acta 757, 3937 (2012).
15. S. Ucls, N. Belmonte, M. Mezcua, A.B. Martnez, M.J. MartnezBueno, M. Gamn, and A.R.
FernndezAlba, J. Envir. Sci. Health, Part B. (submitted).
Noelia Belmonte, Samanta Ucls,

Carmen Ferrer, Milagros Mezcua, PhD, and
Professor Amadeo R. FernndezAlba, PhD, are all at the Agrifood Campus
of International Excellence (ceiA3) and the European Union Reference Laboratory

for Pesticide Residues in Fruit & Vegetables, at the University of Almeria, in
Almeria, Spain. Miguel Gamn, PhD, is with the Agro-Food Analysis Service of
the Generalitat Valenciana and the European Union Reference Laboratory. Direct
correspondence to Milagros Mezcua
at mmezcua@ual.es
37
AN ACCURATE-MASS
PESTICIDE RESIDUES
Pesticide Analysis
Center of Excellence
Productivity simplified
Community reference
Scientific exchange
Leading technologies
Productive solutions
thermoscientific.com/pesticideanalysis
Featured Webinar: Validation of a Routine Multi-Residue Pesticide

Method Using QuEChERS and GC-MS/MS Workflow Solutions
2014 Thermo Fisher Scientific. All trademarks are the property

of Thermo Fisher Scientific Inc. and its subsidiaries.
for the Analysis of Pesticides and

Environmental Contaminants in Fish
By Yelena
Sapozhnikova
A rapid, high-throughput analytical method was developed and evaluated for the
simultaneous determination of pesticides and environmental contaminants in fish.
The compounds included polycyclic aromatic hydrocarbons (PAHs), polychlorinated
biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and flame retardants.
The method was based on a QuEChERS (quick, easy, cheap, effective, rugged, and
safe) technique with acetonitrile extraction, and a dispersive solid-phase extraction
(dSPE) cleanup. Three sorbent combinations were compared for cleanup efficiency
and recoveries of the contaminants: C18+PSA, traditionally used for lipid removal
in dSPE, and two novel sorbents, based on silica coated with zirconium dioxide
(ZrO2) and ZrO2/C18, designed for phospholipid removal. The dSPE cleanup with
ZrO2 sorbent provided the highest efficiency with the lowest baseline, as well as
satisfactory recoveries (70120% calculated based on isotope-labeled internal
standards) for the most analytes. The method allows for quick sample preparation of
fish samples for the analysis of almost 200 targeted contaminants using fast, lowpressure gas chromatography with tandem mass spectrometry (GCMS-MS), thus
providing a wide scope of analysis.
Advances in pesticide residue analysis in recent years led to development of methods for the analysis of
multiple pesticides of different classes with a single sample preparation technique and preferably one
chromatographic run. Other classes of contaminants, which were previously analyzed by separate methods,
requiring either a different sample preparation technique or an additional chromatographic run, are now
integrated into multiclass, multiresidue methods, allowing for faster, less expensive, high-throughput analysis.
Thus, we recently reported a method for the simultaneous determination of >140 pesticides (1) and 50
environmental contaminants (2) in fish using fast sample preparation based on QuEChERS (quick, easy, cheap,
effective, rugged, and safe) extraction and dispersive solid-phase extraction (dSPE) and low-pressure vacuumoutlet gas chromatography with tandem mass spectrometry (low-pressure GCMS-MS). The reported method
38
RAPID SAMPLE PREP
RAPID SAMPLE PREPARATION

AND FAST GCMS-MS
Multi-Residue Pesticide
Analysis of Ayurvedic Churna
1 1
2
3
3
1
Adi
Purwanto
, Alex
Chen
, Kuok1,Shien
Hans-Joachim
Huebschmann
Manoj
Surwade
, Sunil
T Kumar
Aarti ,Karkhanis
, Manish
Kumar1, Soma Dasgupta1, Hans-Joachim Huebschmann2,
2 2
3
PT
AlphaFisher
Analytical
Indonesia,
Jakarta,
Alpha
Analytical
Singapore,
Thermo Fisher Scientific,
Thermo
Scientific,
Mumbai,
India;
Thermo
FisherPte.,
Scientific,
Singapore
Singapore
Click to
view PDF
pnpilci caal ti
tSu
i o np pNo
No
t eNo
TeA
c
Ap
h
o rte
1100te
332-60910 0
Detection,
and
Quantitation
of in
Analysis ofIdentification,
Multi-Residue
Pesticides
Present
Azo
Dyes inChurna
Leatherby
and
Textiles by GC/MS
Ayurvedic
GC-MS/MS
1
Traditional
herbal medicine,
liquid/liquid
dyes, cancerogenic
amines, fast
textiles,
leather, ENextraction,
ISO
Keywords:
Keywords: azo
QuEChERS,
timed-SRM,
time
MRM, ion
standard method,
ISQ Seriesretention
GC/MS, fast
fullsynchronization,
scan detection, quantitation,
ratio
confirmation,
data processing
confirmation,
library TraceFinder
search.
Introduction
Introduction
Azo
dyes are
characterized
their
vivid
Ayurveda
is acompounds
Sanskrit term,
made up ofbythe
words
colors
They
"ayus"and
andprovide
"veda. excellent
"meaningcoloring
life and properties.
science; together
are
important
and widely
usedAasblend
coloring
agentsherbs
in the
translating
to 'science
of life'.
of several
textile
and make
leatherupindustries.
The risk
in theknown
use of as
azo
and spices
the powdered
mixture
dyes
arises Depending
mainly fromonthe
products
that can
"churna".
itsbreakdown
intended use
for medicinal,
be
created
in vivo by
reductive
of theAvipittakara
azo group
beauty,
or culinary
purpose,
thecleavage
recipe varies.
into
aromatic
DueAyurvedic
to the toxicity,
carcinogenicity
"churna"
is a amines.
traditional
formula
used
and
potential
mutagenicity
thus formed
widely
and almost
daily to of
control
vitiated aromatic
pitta dosha,
amines,
of certain
azosystem,
dyes ascontrol
textile indigestion,
and leather
remove the
heatuse
in the
digestive
colorants,
andvomiting
the exposure
of consumers
usinganalytical
the textile
constipation,
and anorexia.
A major
and
leatherfor
colored
with azo
compounds
causes
a serious
challenge
these types
of samples
is mainly
addition
[1]
health
concern
main
of consumer
of multiple
herbs. The
withtwo
sugar
androutes
the natural
color of
exposure
herbs[1]. are the skin absorption of the azo compounds
from the dyed clothes worn, and potential oral ingestion,
mainly
referring
the sucking
textiles by
babiesfrom
and
The dried
leaves to
result
in highlyofcomplex
extracts
young
children.
The manufacturing
workers
canofalso
be
the sample
preparation
due to the rich
content
active
exposed
via the
inhalation
route.
ingredients,
essential
oils and
the typical high boiling
natural polymer compounds. Due to the use of pesticides
in the
herbs, theclassified
"churna"
contain
residual
The
EUfresh
Commission
22may
amines
as proven
pesticides.
Analysis
pesticide residues
is thus
important
or
suspected
human of
carcinogens.
Azo dyes
which,
by
[2]
and governmentally
. Strict
quality may
parameters
reductive
cleavage of regulated
one or more
azo groups,
release
haveorbeen
mented
to aromatic
preserve the
quality
and efficacy of
one
more
of these
amines
in detectable
these "churnas".
concentrations,
i.e. above 30 ppm in the finished articles or
in the dyed parts thereof may not be used in textile and
leather
articles
which may come into direct and prolonged
Sample
Preparation
contact with the human skin or oral cavity [2]. The EU
In brief, the QuEChERS sample preparation (see Figure
Directive 2002/61/EC has banned the use of dangerous
1) involved the extraction of 15 g of a powder sample
azo colorants, placing textiles and leather articles colored
of Avipittakara "churna" with 15 mL acetonitrile
with such substances on the market, and requested the
(containing 1% acetic acid) in the presence of 3 g
development of a validated analytical methodology for
magnesium sulfate, 1.5 g sodium acetate and 1 g NaCl.
control. Since the azo dyes are one of the longest known
The supernatant (1 mL) was collected after centrifugation,
synthetic dyes, simple and inexpensive in preparation,
and dispersive cleanup was performed using 200 mg
available easily in bulk and in great variety, and rarely
PSA and 10 mg GCB. The extract was centrifuged at
cause acute symptoms, the textile manufactures can be
10 000 rpm for 5 min, and 3 L of supernatant was
persuaded to use them despite the regulations if the
strict and reliable analytical control is not imposed.
Experimental Conditions
Weigh 15 g of Churna
Sample Preparation
The sample preparation for the analysis of textile samples
depends on the nature of the textile. The textiles made of
15 mL acetonitrile (with 1% acetic acid) was added, shaken
cellulose
and protein
fibers,
for example cotton, viscose,
well.Further
3 g MgSO
4+1.5 g NaOAc+1 g NaCl was added
[3]
wool, or silkand
make
the azousing
dyesa accessible
homogenized
Vortex mixerto a reducing
agent without prior extraction. The EN ISO 17234-1
standard method for the analysis of such textiles is based
on theAfter
chemical
reduction
azo5 dyes
followed
centrifugation
(5000ofrpm,
min), cleanup
of 1bymLsolid
phase
extraction
(SPE) with
ethyl acetate
providing
a
supernatant
performed
by dispersive
SPE using
200 mg PSA
10 solvent
mg GCB concentration.
ready-to-inject extractand
after
The analysis of synthetic fibers like polyester, polyamide,
Centrifuge acrylic
at 1000 rpm
for 5 min, 3 L ofmaterials
supernatant
polypropylene,
or polyurethane
requires
prior extraction of injected
the azotodyes
and is described in the
GC-MS/MS
EN 14362-2 standard method. The analysis of leather
Figure 1. follows
Sample preparation
for 17234
extraction
of pesticides
from
samples
the EN ISO
standard
method.
ayurvedic churnas
The azo group of most azo dyes can be reduced in the
presence of sodium dithionite (Na2S2O4) under mild
conditions (pH = 6, T = 70 C), resulting in the cleavage
of the diazo group and formation of two aromatic amines
as the reaction products. The amines are extracted by
liquid-liquid extraction with t-butyl methyl ether (MTBE),
concentrated, adjusted to a certain volume with MTBE,
then analyzed by GC/MS. The quantitation is performed
covers a wide range of pesticides along with legacy and novel environmental
contaminants: polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons
(PAHs), polybrominated diphenyl ethers (PBDEs), and flame retardants,
providing wide analytical scope. The method allows for rapid and simple sample
preparation (10 homogenized fish samples can be prepared in 1 h), and lowpressure GCMS-MS affords a fast (9-min) separation of more than 200 analytes,
achieving high throughput. The method can easily be extended to include
additional GC-amenable emerging contaminants as the need arises.
During the extraction of complex matrices, multiple unwanted components
from the matrix are extracted along with the compounds of interest. These
coextractive matrix compounds may fill the active sites of the GC inlet and
column causing matrix-induced ion enhancement (3), resulting in overestimating
of the calculated concentrations. After continuous accumulation of matrix
compounds on the inlet liner and the front part of the GC column, opposite
effects can be observed a matrix-induced diminishing effect (4), or a
reduced ion signal, leading to underestimating of the calculated concentrations.
Matrix compounds may also cause retention time shifts and analyte peak
distortion, thus compromising quality of the analysis.
To remove interfering matrix compounds from extracts, various cleanup
procedures such as gel permeation chromatography, solid-phase extraction,
column chromatography, and others are commonly used. These techniques
are laborious and time consuming, and require large amounts of organic
solvents. Sample cleanup by dSPE is fast, inexpensive, and removes coextractive
interfering matrix components while requiring no solvents for conditioning
sorbents or eluting compounds of interest. Sorbents such as C18 and primary
secondary amine (PSA) are traditionally used for dSPE cleanup of fish extracts.
Recently, two novel zirconium dioxidebased (ZrO2) sorbents, specifically
designed to remove phospholipids, were introduced. A sorbent based on
silica coated with ZrO2 is recommended for samples with <15% fat, and a
sorbent based on silica coated with ZrO2 and incorporated C18 (ZrO2/C18) is
recommended for samples with >15% fat.
In this article, we aim to evaluate and compare the C18+PSA, ZrO2, and ZrO2/C18
39
RAPID SAMPLE PREP
SPONSORED
Figure 1: Coextractive materials removal efficiency.
Experimental
Sample Preparation
Sample preparation was as follows:
First, 10 g of homogenized fish sample
was placed into a 50-mL polypropylene
centrifuge tube, and internal standards
and analytes were added for recovery
experiments. After 15 min, 10 mL of
acetonitrile was added and the tube was
vigorously shaken by hand for 30 s. Then
the entire extract, including the tissue,
was poured into another 50-mL centrifuge
tube containing 4 g of anhydrous
magnesium sulfate and 1 g of sodium
chloride and was vigorously shaken
by hand for 1 min. The tube was then
centrifuged for 2 min at 3250 rcf. Then, 1
mL of the extract was placed into a 2-mL
centrifuge tube for dSPE cleanup with 150
mg of anhydrous magnesium sulfate and
either 50 mg of C18 plus 50 mg of PSA; 50
mg of ZrO2; or 50 mg of ZrO2/C18. The
40
RAPID SAMPLE PREP
sorbents for dSPE in terms of coextractive

removal efficiencies and target analyte
and internal standard recoveries, evaluate
matrix effects and identify matrix
compounds causing them, and investigate
how these matrix compounds alter the
chromatographic behavior of coeluted
analytes. Lastly, we sought to examine
the relationship of analyte recoveries on
different sorbents and their log Kow values.
(a)
4500
4000
Peak area (counts)
C18+PSA
3500
3000
ZrO2
2500
ZrO2/C18
Sorbents for dSPE
2000
1500
1000
500
0
0
50
100
150
200
250
Low-Pressure GCMS-MS
Added concentration (ng/mL)
Calculated concentration normalized to

ISTD
(b)
Solvent only
250
C18+PSA
200
ZrO2
ZrO2/C18
150
100
50
0
0
50
100
150
200
250
Figure 2: Solvent-only and matrix-matched calibration curves for

triphenyl phosphate prepared using dSPE with C18+PSA, ZrO2,
and ZrO2 /C18 based on (a) external calibration and (b) normalized to an internal standard.
41
J.T.Baker Bakerbond C18 sorbent (40 m) was purchased from Thomas Scientific
(Swedesboro, New Jersey). Primary secondary amine (PSA) was purchased from
UCT (Bristol, Pennsylvania), and zirconium dioxide-based Supel QuE Z-Sep
(ZrO2) and QuE Z-Sep Plus (ZrO2/C18) sorbents were purchased from Supelco
(Bellefonte, Pennsylvania).
An Agilent Technologies 7000B triple-quadrupole mass spectrometer with

electron ionization (EI) interfaced to an Agilent 7890A gas chromatograph was
used. The GC instrument had a 220-V fast oven heating upgrade, which allowed
for fast heating rates. A 15 m 0.53 mm, 1-m df Rti-5ms Restek GC column
was used, and it was connected to a 5 m 0.18 mm non-coated guard column
(Restek) at the injector, and to a 17 cm 0.53 mm phenyl-methyl deactivated
guard column (Restek) at the transfer line with Ultimate union column connectors
(Agilent) as previously described (2). The carrier gas was ultrahigh purity helium at
2 mL/min constant flow rate. The GC oven temperature program was as follows:
70 C for 1.5 min, 70180 C at 80 C/min, 180250 C at 40 C/min, 250320
C at 70 C/min, and then 320 C until a total run time of 9 min was reached.
The transfer line and ion source were set at 250 C and 320 C, respectively.
The triple-quadrupole collision gas was nitrogen at 1.5 mL/min and the quench
gas was helium at 2.25 mL/min. The EI energy was 70 eV, the quadrupole
temperatures were set at 150 C, and the solvent delay was at 2 min. A multimode
inlet (MMI, Agilent) was operated as a programmable temperature vaporizer
(PTV) with solvent vent mode of He flow at 50 mL/min, and the injection volume
was 5 L. The inlet temperature program was as follows: 80 C for 0.31 min (at
which point the vent was closed), 80420 C at 320 C/min, and 420 C for the
rest of the GC run.
RAPID SAMPLE PREP
tubes were then shaken vigorously for 30 s and centrifuged for 2 min at 3250 rcf.
Lastly, 0.5 mL of the treated extract was transferred into an autosampler vial and
subjected to low-pressure GCMS-MS analysis.
Solvent only
(a)
Peak area (counts)
C18+PSA
20,000
ZrO2
ZrO2/C18
15,000
10,000
5000
0
0
100
200
300
400
(b)
Calculated concentration normalized to ISTD

400
Solvent only
350
C18+PSA
300
ZrO2
250
ZrO2/C18
Three sets of matrix-matched calibration curves were prepared in catfish extracts

after each dSPE cleanup (C18+PSA, ZrO2, and ZrO2/C18) as described above.
Analyte and internal standard solutions were added to the treated extracts to
yield needed concentrations for six calibration points. The solvent-only calibration
curve was prepared in acetonitrile.
200
150
100
50
0
0
100
200
300
400
Figure 3: Solvent-only and matrix-matched calibration

curves for o,p-DDE prepared using dSPE with C18+PSA, ZrO2,
and ZrO2 /C18 based on (a) external calibration and (b) normalized to an internal standard.
42
A sample of 10 g of catfish was extracted with 10 mL of acetonitrile by shaking

for 1 min, then the entire extract, including the tissue, was poured into another
50-mL centrifuge tube with 4 g of anhydrous magnesium sulfate and 1 g of sodium
chloride, vigorously shaken for 1 min, and centrifuged as described above. An
aliquot of the extract (2.5 mL) was placed into a previously weighed glass tube,
evaporated to dryness with nitrogen, and dried in the oven at 100 C for 20 min;
at that point the tube with extractable residue was weighed again. To compare
the coextractive removal efficiency, portions of 5 mL of the extract were placed
into three previously weighed glass tubes and were treated with C18+PSA, ZrO2,
and ZrO2/C18, 250 mg in each case, all in combination with 750 mg of anhydrous
magnesium sulfate. After the tubes were centrifuged, 2.5 mL of the treated
extracts was evaporated to dryness and dried in the oven at 100 C for 20 min,
and the tube was weighed again. The experiment was conducted in triplicate.
Results and Discussion
Coextractive Compounds and Matrix Effects

Coextractive matrix compounds may alter chromatographic behavior of analytes
and cause matrix effects, leading to inaccurate quantification. Therefore, it
is important to develop a sample preparation procedure that yields as few
coextractive components as possible. In this method, we selected acetonitrile
as an extraction solvent because of its ability to extract compounds with a wide
range of polarities without coextracting a significant amount of fat or lipids
compared to nonpolar solvents (such as hexane or ethyl acetate). For example,
the amount of total lipid and fat in catfish is approximately 6% (5), and using
acetonitrile for extraction, the total measured amount of coextractives was 8
mg/g, or 0.8%.
RAPID SAMPLE PREP
Coextractives Determination
Solvent only
25,000
Octadecanoic acid
C18+PSA
0.95
ZrO2
0.9
0.85
ZrO2 /C18
0.8
Coextractive material removal efficiency was estimated gravimetrically

by measuring the difference in weight before and after each dSPE
treatment and calculated using the following formula:
0.75
0.7
Oleic acid
0.65
removal efficiency % =
0.55
before dSPE coextractive weight after dSPE

( coextractive weight
coextractive weight before dSPE
0.5
0.45
X 100% [1]
0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
-0.05
-0.1
3.8 3.9
4.1 4.2
4.3 4.4 4.5 4.6
4.7 4.8 4.9
5.1 5.2 5.3 5.4 5.5 5.6 5.7 5.8 5.9
6.1 6.2 6.3
6.4 6.5 6.6
Figure 4: Overlaid total ion chromatograms for C18+PSA, ZrO2, and ZrO2 /C18
dSPE treatments.
43
Counts (%)
0.6
6.7 6.8
Figure 1 shows the glass tubes from our experiments with coextractive
materials before and after dSPE with C18+PSA, ZrO2, and ZrO2/C18.
As it can be seen, the tube after dSPE with ZrO2/C18 still had an
appreciable amount of coextractive material that was not removed.
The calculated removal efficiency was 80% for C18+PSA, 66% for ZrO2,
and 56% for ZrO2/C18 dSPE. This means that 1 mL of extract after dSPE
cleanup contained 1.6 mg (C18+PSA), 2.72 mg (ZrO2), or 3.52 mg (ZrO2/
C18) of coextractive materials, and with an injection volume of 5 L,
8 g (C18+PSA), 13.2 g (ZrO2), or 17.6 g (ZrO2/C18) of coextractives
were introduced in the GC injector with each injection. Although the
amounts of matrix compounds left in the extract seem very small, they
still may affect chromatographic behavior of coeluted analytes and
cause matrix effects. Matrix-matched calibrations were used to account
for matrix effects in each dSPE treatment. Difference in adsorption of
analytes on active sites in neat solvent and sample extract containing
matrix compounds can be seen as a difference between slopes of
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Dispersive SPE is commonly used to remove coextractive materials that

interfere with the analysis of target analytes. In this work, three different
sets of sorbents in dSPE were compared: C18+PSA, ZrO2, and ZrO2/C18,
each in the amount of 50 mg per 1 mL of extract, all in combination with
150 mg of anhydrous magnesium sulfate.
x102
5.2
5
4.8
4.6
4.4
4.2
4
3.8
3.6
ME, % =
C18+PSA
3.4
ZrO2 / C18
Counts (%)
3.2
3
2.8
2.4
2.2
2
1.8
ZrO2
1.4
X 100% [2]
Calculated matrix effects for triphenyl phosphate were 4% (C18+PSA),

24% (ZrO2), and 25% (ZrO2/C18) (Figure 2a), based on peak areas (not
normalized to an internal standard), illustrating signal enhancements in
matrix-matched calibration by up to 25%. However, when normalized
to the internal standard, 13C18-triphenyl phosphate, calculated matrix
effects were 4% (C18+PSA), 2% (ZrO2), and 1% (ZrO2/C18) (Figure 2b).
2.6
1.6
calibration curve - slope of SO calibration

( slope of MMcurve
slope of SO calibration curve
5.4
1.2
1
0.8
0.6
0.4
0.2
0
-0.2
-0.4
4.76
4.78
4.8
4.82
4.84
4.86
4.88
4.9
4.92
4.94
4.96
4.98
5.02
5.04
5.06
5.08
5.1
Figure 5: Extracted ion chromatogram for ion 176 representing o,p-DDE at

tR = 4.854 min for extracts treated with C18+PSA, ZrO2, and ZrO2 /C18.
5.2
In another example, matrix effects calculated for o,p-DDE based on

peak areas were 3% (C18+PSA), 19% (ZrO2), and 51% (ZrO2/C18)
(Figure 3a), revealing a less common example of signal diminishing
effect (4) by up to 51%. However, when using the internal standard
fenthion-d 6, calculated matrix effects were (3%) for C18+PSA, (0%) for
ZrO2, and (3%) for ZrO2/C18 (Figure 3b). These examples demonstrate
the value of internal standards to compensate for matrix effects and
eliminate the need for matrix-matched calibration in some cases.
Similar findings were reported for matrix effects of multiple pesticides
in various food crops (6). Indeed, it is difficult, if not impossible, to
have clean, contaminant-free materials to prepare matrix-matched
calibrations for many matrices. In case of megamethods with
multiple analytes of different types, it is rare to find such an
analyte-free matrix.
Interestingly, calculated matrix effect values for triphenyl phosphate
and o,p-DDE gave Pearson correlation coefficients of 0.93 and 0.96,
respectively, when linked to the amounts of coextractive materials for
each dSPE treatment.
44
RAPID SAMPLE PREP
solvent-only and matrix-matched calibration curves. For example,

Figure 2a shows solvent-only and matrix-matched calibration curves for
triphenyl phosphate, a flame retardant. Matrix effects (ME in equation
2) were calculated as a difference between the slope of the matrixmatched (MM) calibration curve for each dSPE treatment and solventonly (SO) calibration curve.
x10-2
C18+PSA
120
ZrO2
100
ZrO2/C18
80
60
40
20
Figure 6: Average recoveries (%) of PBDEs, PCBs, PAHs, flame retardants, and pesticides and representative internal
standards. Error bars represent standard deviations (%).
45
Fe
nt
hi
on
-d
Pe
st
ic
id
es
-T
rip
he
ny
lp
ho
sp
ha
te
13
18
re
ta
rd
an
ts
Fl
am
e
PA
H
s
A
ce
na
ph
th
yl
en
ed
13
12
-P
CB
15
3
PC
Bs
12
6
FB
D
E
PB
D
Es
Furthermore, we looked into what

particular coextractive compounds may
cause matrix effects. Extracts treated
with the three dSPE sorbents were run in
scan mode, and total ion chromatograms
(TICs) for these extracts are shown
in Figure 4. Deconvolution software
(Agilent Mass Hunter) and the NIST
2011 library identified 24 compounds
in the range of 37 min on the TIC after
dSPE with C18+PSA, 29 compounds in
extracts treated with ZrO2 dSPE, and
17 compounds in extracts treated with
ZrO2/C18. Most of these compounds
were lipids, fatty acids, and their
derivatives. Although fewer coextracted
compounds were identified in the ZrO2/
C18 chromatogram, their intensities were
greater than those treated with C18+PSA
or ZrO2 (Figure 4). Two of the largest
peaks, identified on the TICs, were oleic
acid (tR = 4.4 min) and octadecanoic acid
(tR = 4.9 min), fatty acids from animal
and vegetable fat and oil. Oleic acid
amounts, calculated from peak areas,
which remained in the extracts after dSPE
cleanup with C18+PSA and ZrO2, were
20% and 13%, respectively, of the amount
of oleic acid in the extract after dSPE
with ZrO2/C18 (100%), suggesting that
ZrO2/C18 was not effective in removing
this fatty acid. Likewise, octadecanoic
acid amounts were 13% (C18+PSA) and
2% (ZrO2) of the amount of this acid in
ZrO2/C18 (100%). Pearson correlation
RAPID SAMPLE PREP
140
140
130
120
110
100
90
80
70
60
50
40
p= 0.81
ZrO2
p= 0.69
70
ZrO2/C18
p= 0.53
65
60
C18+PSA
p= 0.44
ZrO2
p= 0.28
ZrO2/C18
p= 0.90
coefficients for peak areas of oleic and octadecanoic acids

and amounts of coextractive materials in these extracts
were 0.89 and 0.88, respectively, proposing their significant
contributions to the total amount of coextractive materials.
The compound o,p-DDE is coeluted with octadecanoic
acid, and therefore may be affected by this coextractive
interference. In fact, a good correlation was observed for
matrix effects calculated for o,p-DDE and the peak areas
of octadecanoic acid for the three dSPE treatments, with r 2
= 0.97.
55
50
45
40
35
30
25
20
Log Kow
Log Kow
Flame Retardants
70
p= 0.76
ZrO2
p= 0.68
ZrO2/C18
p= 0.82
50
40
30
20
PBDEs
50
45
Recovery (%)
Recovery (%)
60
C18+PSA
C18+PSA
p= 0.95
ZrO2
p= 0.95
ZrO2/C18
p= 0.99
40
35
30
25
10
20
0
0
10
12
5.5
14
6.5
PCBs
50
45
Recovery (%)
7.5
Log Kow
Log Kow
C18+PSA
p= 0.70
ZrO2
p= 0.85
ZrO2/C18
p= 0.88
40
35
30
25
20
6.2
6.4
6.6
6.8
7.0
7.2
7.4
7.6
7.8
Log Kow
Figure 7: Correlation between recoveries (%) and log Kow values for pesticides, PAHs, flame
retardants, PBDEs, and PCBs.
46
8.5
Figure 5 shows an extracted ion chromatogram for ion 176

representing o,p-DDE at tR = 4.854 min for the three dSPE
treatments and corresponding o,p-DDE multiple reaction
monitoring (MRM) chromatograms of the same extracts.
MRM peak widths for o,p-DDE were 2.98 0.19 s for
C18+PSA treatment, 2.97 0.33 s for ZrO2, and 3.90 0.39
s for ZrO2/C18 for n = 10 injections. The peak on the ZrO2/
C18 MRM chromatogram was wide and asymmetrical with
visible tailing, perhaps showing an example of interference
by coeluted matrix compounds.
ZrO2 treatment removed the highest amounts of both
oleic and octadecanoic acids in our experiment, and
thus provided the cleanest chromatograms with lowest
background levels. Several publications to date have
described successful applications of ZrO2 in dSPE for
cleanup of matrices with high amounts of lipids, and
agreed that compared to C18+PSA and ZrO2/C18, ZrO2
provided the cleanest extracts, lowest background noise
on the chromatograms, lowest amounts of coextractive
matrix compounds (7), and highest amounts of analyzed
contaminants showing acceptable recoveries and lowest
standard deviations.
RAPID SAMPLE PREP
PAHs
C18+PSA
Recovery (%)
Recovery (%)
Pesticides
Extraction of Organochlorine
Pesticides from Oysters
Pranathi Perati and SM Rahmat Ullah
Thermo Fisher Scientific, Sunnyvale, CA, USA
Click to
view PDF
A ppl i cati on Bri ef 1 5 2
Extraction of Organochlorine Pesticides

from Oyster Tissue Using Accelerated
Solvent Extraction
Key Words
Persistent organic pollutants, moisture absorbing polymer, wet samples,
accelerated solvent extraction, sample preparation
Introduction
Organochlorine pesticides (OCPs) are a class of chemicals
that were used to control insect pests since the 1940s.
The use of OCPs was banned in the later part of the last
century due to their longevity, a trait that made them
effective for long term pest control, but also increased
concerns of potential health outcomes such as cancer in
humans and ecosystem disruption. Pesticides are regulated
in the U.S. by the Environmental Protection Agency (EPA)
under the Federal Insecticide, Fungicide, and Rodenticide
Act (FIFRA). Some states also regulate pesticides under
FIFRA, in a more restrictive manner than the EPA. In the
European Union, water intended for human consumption
must meet a maximum level of 0.1 g/L for each pesticide
and a maximum of 0.5 g/L for total pesticides, except for
aldrin, dieldrin, heptachlor, and heptachlor epoxide, which
are each limited to maximum levels of 0.03 g/L. Maximum contaminant levels have been established for OCPs
by the United States Environmental Protection Agency
ranging from 0.2 g/L for Lindane to 2 g/L for Endrin.
Many OCPs are endocrine disrupting chemicals, meaning
they have subtle toxic effects on the bodys hormonal
systems. Endocrine disrupting chemicals often mimic the
bodys natural hormones, disrupting normal functions
contributing to adverse health effects. OCPs are persistent
organic pollutants (POPs), a class of chemicals that are
ubiquitous environmental contaminants because they
break down very slowly in the environment and accumulate in lipid rich tissue such as body fat. According to the
Centers for Disease Control and Prevention (CDC), most
people have OCPs present in their bodies. Exposure to
low concentrations of organochlorine chemicals over a
long period may eventually lead to a substantial body
burden of toxic chemicals. Organochlorine compounds
have long been recognized as the most deleterious
contaminants to biota in the world's marine and estuarine
waters. Various biomonitoring strategies have therefore
been developed to monitor and evaluate the adverse
impact of these compounds on the marine ecosystems.
Analyses of OCPs are becoming increasingly important,
and often with the need to isolate and analyze trace levels
of compounds from a variety of matrices such as soil,
sediment, animal tissue, fruits, and vegetables. Sample
pretreatment constitutes an important step prior to
analysis. The purpose of the sample pretreatment step
is to selectively isolate the analytes of interest from
matrix components and present a sample suited for
routine analysis by an established analytical techniques
such as gas chromatography or high-pressure liquid
chromatography. Accelerated solvent extraction is an
established technique for extracting analytes of interest
from a solid, semisolid or an adsorbed liquid sample
using an organic solvent at an elevated temperature and
pressure. The elevated pressure elevates the boiling
temperature of the solvent thereby allowing faster
extractions to be conducted at relatively high temperatures. Thus the extraction process is significantly faster
than traditional methods such as Soxhlet extraction.
This Application Brief discusses the use of Thermo
Scientific Dionex ASE Prep MAP, a proprietary polymer
designed to remove moisture and increase extraction
efficiencies from wet samples including soils, tissues and
food products. This polymer is useful for in-cell extraction
of trace level organics from a variety of moisture containing
samples with no additional pre or post extraction steps.
The Dionex ASE Prep MAP polymer has a high-capacity
for water removal and does not suffer from some of the
limitations of clumping or precipitation observed in some
of the traditional drying methods.
Recovery Study
As the need for multiresidue, multiclass analysis of contaminants in foods and
environmental samples continues to increase, megamethods covering multiple
classes of contaminants in one uncomplicated technique are being introduced.
As we have demonstrated, the use of internal standards helps to account for
matrix effects and enables quite accurate quantification. Because it is impractical
to match every single analyte with the isotope-labeled internal standard,
efforts were made in our method development to use one representative
internal standard labeled with an isotope or an alternate label for each class
of contaminants. Thus, all organophosphate flame retardants were quantified
with 13C18-triphenyl phosphate, all PCB congeners with 13C12-PCB 153, all PBDE
congeners and brominated flame retardants with fluoro-BDE 126 (FBDE 126),
and all pesticides with fenthion-d 6. PAHs were quantified with various isotopelabeled PAHs because surrogate PAH cocktails are readily available and relatively
inexpensive.
The recoveries of nearly all analytes were within the acceptable range of 70120%
while using calculations normalized to the internal standards (1,2). In the present
study, we attempted to estimate real recoveries of the analytes without
internal standard correction and recoveries of representative internal standards.
This information gives a valuable insight into what quantity of analytes and
internal standards were lost during the sample extraction and cleanup process.
Recoveries were calculated as an average for all representative compounds of
one class: PCBs, PBDEs, PAHs, flame retardants, and pesticides. For the three
dSPE treatments, the average recoveries of representative compounds in a class
and corresponding internal standards were as follows: PBDEs: 3238%, internal
standard FBDE 126: 3446%; PCBs: 3138%, internal standard 13C12-PCB 153:
3138%; PAHs: 3946%, internal standard acenaphthylene-d 8: 3364%; flame
retardants: 3645%, internal standard 13C18-triphenyl phosphate 5058%;
and pesticides: 71108%, internal standard fenthion-d 6: 104121% (Figure 6).
For all classes of contaminants, except for pesticides, the recoveries were 23
times lower when not normalized to an internal standard, reflecting incomplete
recovery during the sample preparation process. For pesticides, however, the
recoveries were comparable with and without normalizing to an internal standard.
47
RAPID SAMPLE PREP
SPONSORED
Correlation Between Recoveries and Log Kow Values

As is well known, the octanolwater partition coefficient, log Kow, is one of the
factors affecting sorption of organic compounds. During a dSPE step, organic
compounds in the extract undergo a sorptiondesorption process on the
sorbent, with their recoveries representing their retention on the sorbent. We
explored a correlation between recoveries of targeted analytes from various
classes and their log Kow values. For all classes of contaminants, except for PAHs,
Pearson correlation coefficients were negative, indicating that increasing log Kow
values lead to decreased recoveries. Chemicals with higher log Kow values are
more lipophilic, and therefore tend to have a higher affinity for lipids, adsorbed
by the sorbent during the SPE cleanup process. Similar to our study, Rajski and
colleagues (7) examined the relationship of 92 pesticide recoveries from fatty
matrices and log Kow, and concluded that the majority of pesticides with higher
log Kow values exhibited lower recoveries.
The correlations were more pronounced for PBDE and PCB congeners, which are
homologues with similar structures, unlike other classes of chemicals that include
compounds with rather diverse chemical structures. Thus, for PBDE congeners,
the calculated correlation coefficients were as follows: 0.95 for C18+PSA, 0.99
for ZrO2, and 0.99 for ZrO2/C18 (Figure 7). For PCB congeners, the correlation
coefficients were 0.70 for C18+PSA, 0.85 for ZrO2, and 0.88 for ZrO2/C18
(Figure 7). In contrast to the negative correlation observed for all pesticides,
PCBs, PBDEs and flame retardants on all sorbents, the correlation coefficient for
PAHs was negative only for ZrO2/C18 treatment (0.90), and weakly positive for
C18+PSA (0.44) and for ZrO2 (0.28).
48
RAPID SAMPLE PREP
For all classes, except for PAHs, the recoveries using dSPE with ZrO2 were greater
compared to the use of dSPE with C18+PSA or ZrO2/C18 (Figure 6). Also, standard
deviations for dSPE with ZrO2/C18 were greatest compared to the other two
treatments, compromising the repeatability of the method. Some of the matrix
effects and coextractive compounds discussed above may be responsible for
higher standard deviations in extracts treated with ZrO2/C18.
Conclusions
In this study we compared the efficiency of the three sorbents for dSPE cleanup
of fish extracts for the analysis of pesticides and environmental contaminants:
C18+PSA, ZrO2, and ZrO2/C18. C18+PSA removed the highest amounts of
coextractive materials by weight (80%), followed by ZrO2 (66%) and ZrO2/C18
(55%). In terms of target analyte recovery, ZrO2 provided the highest recoveries
for most contaminants, and ZrO2/C18 showed the highest standard deviations,
possibly caused by matrix interferences. Recoveries of analytes not normalized
to internal standards were 23 times lower for all classes, except for pesticides,
reflecting losses during the sample preparation.
Two major coextractive compounds identified in the extracts that contributed to
matrix effects were oleic and octadecanoic acid, and dSPE with ZrO2 removed
these compounds most effectively. Peak shapes and widths were altered by
coeluted matrix interferences as we observed in the case of o,p-DDE.
Calculated matrix effects for triphenyl phosphate and o,p-DDE based on peak areas
were lowest for dSPE with C18+PSA, followed by ZrO2 and ZrO2/C18; however,
they were 4% for all the treatments when normalized to internal standards,
indicating the need to use an internal standard to compensate for matrix effects.
Finally, Pearson correlations were reported for compound recoveries and tR and
log Kow values.
Acknowledgments
Technical assistance of Tawana Simons of the USDA/ Agricultural Research
Service is greatly appreciated. Dr. Olga Shimelis of Supelco is acknowledged for
providing Z-Sep and Z-sep Plus sorbents.
49
RAPID SAMPLE PREP
Another interesting relationship was observed between tR and log Kow. Pearson
correlation coefficients were 0.99 for PAHs, 0.95 for PBDEs, 0.98 for PCBs, 0.92 for
flame retardants, and 0.55 for pesticides.
Mention of trade names or commercial products in this publication is

solely for the purpose of providing specific information and does not imply
recommendation or endorsement by the U.S. Department of Agriculture. USDA is
an equal opportunity employer.
References
1.
2.
3.
4.
Y. Sapozhnikova, J. Agric. Food Chem. http://dx.doi.org/10.1021/jf404389e. (2014).

Y. Sapozhnikova and S.J. Lehotay, Anal. Chim. Acta 758, 8092. (2013).
D.R. Erney, A.M. Gillespie, D.M. Gilvydis, and C.F. Poole. J. Chromatogr. 638, 5763. (1993).
J. Hajslova and T. Cajka, in Food Toxicants Analysis, Y. Pico, Ed. (Elsevier, Oxford, UK, 2006),
pp. 419473.
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Yelena Sapozhnikova, PhD, is a research chemist at the Agricultural Research

Service in the Eastern Regional Research Center, at the United States Department of
Agriculture in Wyndmoor, Pennsylvania.
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Table of contents

NCI, SRM, and LVI

Improving Sensitivity and Selectivity in

An Accurate-Mass Database for Screening

Rapid Sample Preparation and Fast GCMS-MS

NCI, SRM, AND LVI

The selection of analytical methods for gas chromatography (GC)-amenable

NCI, SRM, AND LVI

A ppli cati o n N o te 52291

Pesticides, Tea, Herbal products, ASE, SRM, MRM, Multi-residue analysis,

A routine screening method for more than 200 pesticide

Negative Chemical Ionization

Tandem Mass Spectrometry

NCI, SRM, AND LVI

Although isobutane, methane, ammonia in methane, and pure ammonia can be

chromatographynegative chemical ionization-mass spectrometry (GCNCI-MS).

255, 257, 253

181, 183, 109

284, 286, 282

284, 286, 282

Cyclodiene Insecticides (Chlorinated) and Their Degradation Products

410, 408, 412

373, 375, 377

266, 71, 232

NCI, SRM, AND LVI

410, 408, 412

373, 375, 377

266, 71, 232

372, 406, 408

372, 406, 408

70, 239, 195

300, 266, 302

272, 100, 274

NCI, SRM, AND LVI

chromatographynegative chemical ionization-mass spectrometry (GCNCI-MS).

366, 368, 370,

255, 257, 253

284, 286, 282

284, 286, 282

146, 162, 174

(Collision Energy, eV)

Cyclodiene Insecticides (Chlorinated) and Their Degradation Products

NCI, SRM, AND LVI

146, 162, 174

chromatographynegative chemical ionization-mass spectrometry (GCNCI-MS).

Dinitroaniline Herbicides 71>35 (5),

Phthalic Acid Herbicide

281, 251, 282

335, 305, 336

NCI, SRM, AND LVI

366, 368, 370

181, 183, 109

157, 159, 158

NCI, SRM, AND LVI

chromatographynegative chemical ionization-mass spectrometry (GCNCI-MS).

255, 257, 253

181, 183, 109

277, 125, 109

284, 286, 282

136, 121, 230, 289

157, 159, 158

157, 172, 315

199, 121, 241, 231

NCI, SRM, AND LVI