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I. INTRODUCTION
Pesticides have been widely used throughout the world since the
middle of the 20th century. Based on the compilation of the
British Crop Protection Council, approximately 860 active
substances are formulated in pesticide products currently
(Tomlin, 2003). These substances belong to more than 100
substance classes. Benzoylureas, carbamates, organophosphorous compounds, pyrethroids, sulfonylureas, or triazines are the
most important groups. The chemical and physical properties
of pesticides may differ considerably. There are several
acidic pesticides; others are neutral or basic. Some compounds
contain halogens, others phosphorous, sulfur, or nitrogen. These
heteroatoms may have relevance for the detection of pesticides. A
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TABLE 1. Typical ions selected for GC-EI-MS or transitions used in LC-ESI-MS/MS and the sensitivity obtained with
both techniques
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a
EU regulation can be found on the websites: http://europe.eu.int/eur-lex/en/search/search_lif.html or http://europe.eu.int/eurlex/lex/en/repert/035020.htm
b
[M NH4] used as quasimolecular ion.
c
Regulated metabolite in the EU.
d
Analyte requires special HPLC conditions for detection with ESIMS/MS.
e
Quasimolecular ion was [MOH].
f
Reference 2 does not report the product ion.
g
Quasimolecular ion was [(MO)/2].
h
Quantitative degradation of the pesticide occurs in the GC injector.
i
Reference 1 does not report the product ion.
A. GCMS
Ionization of pesticides in GCMS can be done by EI, and
positive or negative chemical ionization (PCI, NCI). For ion
separation, single quad instruments are used most frequently.
Additionally, GCMS systems with quadrupole ion traps, timeof-flight (TOF) mass spectrometers or tandem mass spectrometers are available.
Most of the published studies on residue analysis by GC
MS report on results obtained by single quadrupole instruments
and EI ionization. Advantages of EI ionization are a low influence
of molecular structure on response, and a large number of
characteristic fragments. Extensive studies describe the simultaneous determination of 245400 pesticides by GCEIMS with
single quadrupole mass filters (Cairns et al., 1993; Fillion, Sauve,
& Selwyn, 2000; Stan, 2000; Chu, Hu, & Yao, 2005). The use of
ion traps in scan mode is more simple because no selection of
characteristic ions is necessary during data acquisition. In full
scan mode these instruments are quite sensitive, and confirmation
by library search is possible at lower concentrations. But,
Mass Spectrometry Reviews DOI 10.1002/mas
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B. LCMS
If pesticides are not amenable to GC, the application of LC is the
best alternative. Likewise, LC may be combined with single
quadrupole instruments, quadrupole ion traps, triple quadrupole
(tandem) mass spectrometers, TOF spectrometers, or hybrid
quadrupole TOF instruments.
In contrast to GCMS, single quadrupole mass spectrometers
are not used in the majority of recent studies dealing with LCMS.
A disadvantage of single quadrupole instruments (and ion traps
operated in the SIM mode) is the high intensity of background
signals produced from sample matrix and HPLC solvent clusters.
Due to this chemical noise in real samples very low limits of
quantification cannot be achieved, even if the sensitivity of these
instruments is high (Hernandez, Sancho, & Pozo, 2005).
The chemical background can be reduced significantly if
tandem MS in combination with selected reaction monitoring
(SRM) is applied. Even if a co-extracted matrix component has the
molecular mass of a pesticide, usually both isobaric ions can be
separated in SRM experiments, because their fragmentation in the
collision cell most often results in different product ions. Therefore,
tandem mass spectrometers offer excellent sensitivity and unsurpassed selectivity. For this reason, triple quadrupole mass analyzers
have been the most often applied MS detectors until now (Pico,
Blasco, & Font, 2004). Quadrupole ion traps may also be operated in
the MS/MS mode, which reduces the background to a level known
from tandem mass spectrometers. However, ion collection,
fragmentation, and mass analysis of fragments is a step by step
process in traps and requires much more time than in triple
quadrupole instruments, which do this in parallel. Furthermore, ion
traps suffer from a limited dynamic range, a smaller potential to
fragment very stable ions and the inefficiency to trap low mass
fragments (Pico, Blasco, & Font, 2004).
Time-of-flight mass spectrometers in combination with LC
are more often used in high-resolution mode (typical mass
error <2 mDa), which provides better discrimination of background (Hogenboom et al., 1999; Ferrer et al., 2005). The main
advantage of this type of instrument is the identification of
unknown peaks in a sample even if analytical standards are
not available (Garcia-Reyes et al., 2005; Thurman, Ferrer, &
Fernandez-Alba, 2005). But, this advantage is usually not needed
in the enforcement of maximum residue levels. Furthermore,
identification of pesticides in samples is less certain by LC
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C. Final Decision
Any of the instruments discussed above have special merits, but
none of them can detect the full range of all pesticides. However,
if the selection of the most appropriate techniques is focused on
the enforcement of maximum residue levels, simultaneous
identification, and quantification of a very large number of
target analytes will be more important than the detection,
identification, and quantification of non-regulated (non-target)
pesticides and/or metabolites. Under these conditions, EI
ionization and single quadrupole MS was identified as the
preferred GC detection system. If LC is used, most benefits
should be obtained from tandem mass spectrometers operating in
the electrospray mode. Therefore, in the next section scope and
sensitivity of GC-EIMS will be compared to pesticide detection
by LCESItandem MS.
A. GCMS Data
Gas chromatographymass spectrometry (GCMS) has been
practiced in analyzing pesticides for several decades and
most characteristic ions are available from pesticide analytical
manuals, applications of instrument producers, or spectral
databases supplied by producers of analytical standards. In
addition, some excellent articles covering GCMS analysis of a
broad range of pesticides are published (Fillion, Sauve, &
Selwyn, 2000; Wong et al., 2003; Chu, Hu, & Yao, 2005).
Only for some pesticides, the characteristic ions in Table 1 are
obtained from recent articles or studies conducted by the pesticide
industry.
For estimation of sensitivity, several mixed standards in
solvent containing all pesticides were analyzed. A state-of-theart GCMS system (Agilent 6890N GC and 5975 inert MSD)
using a pulsed pressure injection of 1 mL onto a HP-5 MS column
(30 m 0.25 mm 0.25 mm), EI ionization, and a dwell time
of 40 msec were applied for each characteristic ion. The same
GC conditions were used for all injections. Obviously, a longer
dwell time would result in better sensitivity, butat the same
timeit would reduce the number of pesticides analyzed in
one run.
Standard solutions containing 10,000, 1,000, 100, 10, and
1 ng/mL were injected. If a pesticide was not detected at the
highest concentration in the SIM mode, no data were added to
Table 1. In all other cases, characteristic ions are presented. The
LOQ was set to the lowest concentration, which gave a signal-tonoise ratio of 10 for the most intense peak of the analyte.
B. LCMS/MS Data
Typical transitions from precursor to product ions are taken from
recent publications, since analogous data collections do not
exist for LCMS/MS. SRM transitions from studies conducted
with triple quadrupole or quadrupole ion trap instruments
are preferred. If such studies were not found, data on
those product ions are cited, which are produced in single
quadrupole instruments by increasing the potential between
the entrance capillary and the first skimmer (fragmentor or cone
voltage). Often, transitions for a selected pesticide are published
by more than two authors. In such cases, the citation in Table 1
prefers the first or at least the earlier publications. However, no
transitions were found in published studies for approximately
one-third of the pesticides listed in Table 1. In such cases, typical
transitions are taken from unpublished studies of pesticide
producers or from the world wide web (BfR, 2005).
The sensitivity of LCMS/MS instruments was assessed
using a triple quadrupole mass spectrometer (API 4000,
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B. Comparison of Sensitivity
Both, GCMS- and LCMS-based methods, reveal a significant
variation of sensitivity, covering at least a range of 34 orders of
magnitude, depending on the pesticide. However, a comparison
of the median of the limits of quantification clearly shows much
higher sensitivity if determinations are based on LC and tandem
MS. Most analytes may be quantified reliably by LCMS/MS
(at least in standard solutions) at concentrations between 0.1 and
1 ng/mL. In contrast, the median of the limits of quantification
observed by GCMS is distinctly higher, that is, at 100 ng/mL.
The distribution of LOQ data from Table 1 is summarized
separately for both techniques in Figure 1. Nearly the same
distribution is found for organophosphorus pesticides, which are
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*This calculation/list does not contain four organotin compounds, four quaternary ammonium salts,
glyphosate, and picloram, which require special LC conditions for ESIMS/MS detection.
cides.
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ACKNOWLEDGMENTS
We thank Volker Happel and Marilyn Menden for their support
needed for the determination of limits of quantification. Natasa
Markovic, Birgit Mueller, and Annamaria Melcher provided
important technical assistance throughout this work.
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Lutz Alder studied chemistry in Berlin/Germany with a focus on organic and analytical
chemistry. After his Ph.D. in 1978, he managed the mass spectrometry laboratory at
Humboldt University for several years. He has been employed by the Federal Health
Office, which is now the Federal Institute for Risk Assessment (BfR) since 1991. Most of
his research has been concerned with residue analytical methods and its standardization
for official use. He has published 50 scientific journal articles and book chapters.
Baerbel Vieth studied chemistry at the Humboldt University in Berlin. After finishing the
Ph.D. in 1981, her research has been focused on the development and application of HPLC
methods in trace analysis of drugs, enzyme activities, and of pesticides. In 1991, she joined
the former Federal Health Office of Germany, which is now the Federal Institute for Risk
Assessment (BfR). Besides residue analytical methods for pesticides, she has been
responsible for evaluation of residues in human milk with special focus on POPs. She has
published about 30 articles in scientific journals.
At present, Lutz Alder, Kerstin Greulich, and Baerbel Vieth are responsible for the
evaluation of residue analytical methods provided for registration of new pesticides in
Germany and contribute to authorization of plant protection products in Europe.
Guenther Kempe studied food chemistry and analytical chemistry in Dresden/Germany.
He started his career in the Hygiene Institute Chemnitz, which became the State Institute
for Food and Health Protection of Saxony/Germany in 1991. Most of his research has been
concerned with pesticide residue analysis in food. Today he is the head of the residue
laboratory of the State Institute and chairmen of the Working Group Pesticides of the
Gesellschaft Deutscher Chemiker (Chemical Society of Germany). His publication list
contains 20 scientific articles and book chapters.
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