Chemosphere 309 (2022) 136725

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Chemosphere
journal homepage: www.elsevier.com/locate/chemosphere

Simple and rapid method for 336 multiresidual pesticide analysis in saliva,
determination of their chemical stabilities, and biomonitoring of farmers
Eunyoung Park a, Jiho Lee a, Hye Suk Lee b, Jeong-Han Kim a, Yongho Shin c, *
a
Pesticide Chemistry and Toxicology Laboratory, Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National
University, Seoul, 08826, Republic of Korea
b
Drug Metabolism and Bioanalysis Laboratory, College of Pharmacy, The Catholic University of Korea, Bucheon, 14662, Republic of Korea
c
Department of Applied Biology, College of Natural Resources and Life Science, Dong-A University, Busan, 49315, Republic of Korea

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• A modified QuEChERS was developed

for multiresidual pesticide analysis in
saliva.
• Acidified acetonitrile exhibited excel­
lent extraction efficiency for the target
pesticides.
• Chemical stability of each pesticide was
evaluated under two different
conditions.
• It can be used for agricultural, forensic,
and clinical purposes.

A R T I C L E I N F O A B S T R A C T

Handling Editor: Keith Maruya Simultaneous multiresidual pesticide analysis of saliva samples was performed using scaled-down QuEChERS
extraction with LC-MS/MS and GC-MS/MS. The optimum extraction procedure using acidified acetonitrile was
Keywords: applicable to 336 pesticides (287 for LC-MS/MS and 49 for GC-MS/MS). To determine pesticide multiresidues in
Saliva saliva, 100 μL of the sample was extracted with 200 μL of 0.1% formic acid in acetonitrile, and the initial extract
Pesticide
was partitioned with 40 mg of MgSO4 and 10 mg of NaCl. The organic supernatants (120 μL) were then mixed
Multiresidues
with acetonitrile (30 μL) for matrix-matching (4:1, v/v), and the final extract solution was injected into the LC-
QuEChERS
Chemical stability MS/MS (4 μL) and GC-MS/MS (2 μL) systems. The established analytical method showed a good LOQs between 5
Exposure and 25 ng/mL with reliable accuracy/precision values and recovery results (50–140%) for the target pesticides.
Under the two different storage conditions, most of the analytes did not undergo chemical changes in the saliva
samples, whereas some pesticides were more stable in freeze-thaw processes than those left at room temperature.
Biomonitoring of farmers (ten mixers and ten sprayers) was successfully applied using the validated method, and
two carbamates (fenobucarb and propamocarb) were determined at trace concentrations (12.5–675.0 ng/mL
from 11 positively detected samples).

Abbreviations: CE, collision energy; CID, collision-induced dissociation; dSPE, dispersive solid-phase extraction; EI, electron ionization; ESI, electrospray ioni­
zation; GCB, graphitized carbon black; ME, matrix effect; PSA, primary secondary amine; QC, quality control; QuEChERS, quick, easy, cheap, effective, rugged, and
safe; S/N, signal-to-noise ratio; S/P, saliva/plasma.
* Corresponding author.
E-mail addresses: [email protected] (E. Park), [email protected] (Y. Shin).

https://doi.org/10.1016/j.chemosphere.2022.136725
Received 15 August 2022; Received in revised form 29 September 2022; Accepted 30 September 2022
Available online 5 October 2022
0045-6535/© 2022 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC license (http://creativecommons.org/licenses/by-
nc/4.0/).
E. Park et al. Chemosphere 309 (2022) 136725

1. Introduction was evaluated under two different conditions. To the best of our
knowledge, this is the first biological sample analysis for simultaneous
As pesticides are chemicals used to control and kill pests, they are pesticide multiresidue determination in saliva.
intrinsically toxic to living organisms, including humans (Maroni et al.,
2006). Agricultural workers (farmers) who use pesticides are more likely 2. Materials and methods
to be exposed to pesticides than other cohorts. Many studies have
investigated the various negative health effects during and after pesti­ 2.1. Reagents and materials
cide spraying (Hong et al., 2009; Nigatu et al., 2016; Sankoh et al.,
2016). In severe cases, acute toxicity that can lead to coma and even High-purity (>98%) pesticide standards or stock solutions (1000
death has been reported (Klatka et al., 2021; Zhao et al., 2021). Hence, it mg/L) were obtained from Sigma-Aldrich (St. Louis, MO, USA), Chem­
is important to quickly respond to any incidence of agricultural intoxi­ service (West Chester, PA, USA), Dr. Ehrenstorfer (Augsburg, Germany),
cation and monitor agricultural exposure by determining as many pes­ Wako (Osaka, Japan), and Ultra Scientific (North Kingstown, RI, USA).
ticides as possible from the biological samples of farmers. Acetone, acetonitrile, and methanol (HPLC grade) were purchased from
Biological fluids such as blood, urine, sweat, and saliva have been Merck (Darmstadt, Germany). Deionized water (18.2 MΩ/cm) was
used to estimate concentrations of pesticide multiresidues in the body prepared in-house using LaboStar TWF UV 7 (Siemens, MA, USA).
(Genuis et al., 2016; Lacassie et al., 2001; Nigg et al., 1993; Shin et al., Ammonium formate (LC-MS grade), formic acid (LC-MS grade), and
2019; Zuluaga et al., 2021). Among them, the use of saliva has some anhydrous magnesium sulfate (MgSO4 ≥99.5%) were obtained from
characteristics and is more advantageous. Saliva can be collected Sigma-Aldrich. NaCl (99.0%) was obtained from Samchun (Gyeonggi-
non-invasively even if the patient is unconscious. Saliva samples are do, South Korea). A ceramic homogenizer (2 mm) was purchased from
readily accessible and no medical expertise is required to collect them. Ultra Scientific.
They can also be easily transported to an analytical laboratory. Even if a
suspected person thoroughly rinses their mouth to remove evidence of 2.2. Preparation of standard solutions and quality control (QC) samples
intoxication, chemicals remaining in the bloodstream can be secreted
through the salivary glands, redissolve in the oral fluid, and be detect­ Individual stock solutions (100–1000 μg/mL) were prepared using
able in analytical levels (Haeckel, 1993; Timchalk et al., 2015). The ratio acetone, acetonitrile, and methanol. A mixed standard solution (2500
of pesticide concentration in saliva to that in plasma (saliva/plasma; ng/mL for each analyte) was prepared by combining each stock solution.
S/P) remains constant; therefore, pesticide levels in the body can be The mixed standard solution was further diluted with acetonitrile to
estimated from the S/P by determining them in the oral fluid (Lu et al., obtain working solutions (1,000, 750, 500, 250, 100, 50, and 25 ng/mL).
1997, 2006; Michalke et al., 2015; Timchalk et al., 2015). It also makes Calibration standards containing saliva matrices (100, 75, 50, 25, 10, 5,
it possible to verify recent pesticide exposure history related to inten­ and 2.5 ng/mL) were prepared according to the optimized sample
tional or unintentional acute poisoning. preparation procedures after adding 10 μL of each working solution into
Quick, easy, cheap, effective, rugged, and safe (QuEChERS) is the 100 μL of blank (pesticide-free) samples. QC samples of concentrations
most effective method for co-extracting pesticide multiresidues (Anas­ (5, 10, 25, and 75 ng/mL) were prepared using 100 μL of a blank saliva
tassiades et al., 2003; Rahman et al., 2017). The QuEChERS procedure sample by adding 10 μL of working solution. All working solutions were
for food or crop samples generally consists of three stages: acetonitrile stored at − 20 ◦ C when not in use, while QC samples were stored at − 80
extraction, partitioning by salting-out, and cleanup with dispersive ◦
C until analysis.
solid-phase extraction (dSPE). dSPE sorbents can effectively remove
various interferences such as sugars, amino acids, pigments, lipids, and 2.3. Collection of saliva from agricultural workers
fatty acids. However, they have a problem with co-elimination of spe­
cific pesticide classifications; Primary secondary amines (PSAs) remove Following previous studies (Dams et al., 2003; Haeckel, 1993; Hines
sulfonylureas, imidazolinones, and carboxylic acids, and graphitized et al., 2006; Höld et al., 1995; Lee et al., 2016; Lu et al., 2006; Pujadas
carbon black (GCB) eliminates planar pesticides (Kaczyński and Łoz­ et al., 2007), saliva was collected using a salivette (Sarstedt, Numbrecht,
owicka, 2017; Łozowicka et al., 2017; Shida et al., 2015). Fortunately, Germany). Briefly, a cotton roll was chewed for approximately 1 min to
saliva is composed of 99% water, so it is free from serious interference absorb saliva, placed in a collection tube, and then centrifuged to allow
issues (Carpenter, 2013). Therefore, a simpler but more efficient saliva to pass from the roll into the lower part of the tube. The subjects
QuEChERS method can be developed for saliva samples without the were not allowed to touch the roll to prevent exogenous contamination.
need for any dSPE step. Pesticide-free saliva for method validation was obtained from healthy
The stability of most of the pesticides depends on specific external six volunteers who were not related to agricultural occupation, and the
conditions such as pH, temperature, humidity, and media components samples were pooled to reduce differences in matrix effects between
(Turner, 2015). In general, saliva is a neutral aqueous medium con­ individuals. Samples for biomonitoring were collected from ten farmers
taining secreted salts and enzymes, which can make some pesticides after pesticide mixing/loading and from another ten farmers after
chemically unstable. Notably, quantitative loss or transformation due to pesticide spraying. Before pesticide treatments, the subjects were asked
degradation of pesticide ingredients or conversion to other pesticides to rinse their mouths with water twice to completely remove contami­
can lead to under- or overestimation of the actual concentration of nants. The samples were stored in a cooler with a frozen ice pack and dry
pesticides at the time of sampling. Therefore, it is important to know the ice until they were returned to the laboratory. All samples were stored at
pesticide stability in various scenarios. However, few studies have − 80 ◦ C prior to analysis. Saliva samples were collected after the
evaluated pesticide stability in biological samples. Understanding the approval of the Institutional Review Board (IRB) of Seoul National
stability patterns of pesticides under specific conditions will allow us to University, Seoul, Republic of Korea (IRB No. 1604/002–007).
establish guidelines for the appropriate transport, storage, and prepa­
ration of samples after collection. 2.4. LC-MS/MS and GC-MS/MS instruments
In this study, we established an LC-MS/MS and GC-MS/MS method
for 336 pesticides in human saliva and used the method for bio­ Multiresidual analysis was performed using a Nexera X2 UHPLC
monitoring 20 agricultural workers. A simple, convenient, and highly system (Shimadzu, Kyoto, Japan) coupled with an LCMS-8050 triple
effective QuEChERS extraction procedure without cleanup was newly quadrupole mass spectrometer (Shimadzu) and a GC-2010 Plus (Shi­
developed considering the characteristics of the oral fluid and target madzu) coupled with a GCMS-TQ8040 triple quadrupole mass spec­
pesticides. In addition, the chemical stability of each target pesticide trometer (Shimadzu).

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E. Park et al. Chemosphere 309 (2022) 136725

For LC-MS/MS, pesticide residues were separated on a Kinetex C18 2.7. Method validation and matrix effect (ME)
column (100 × 2.1 mm, 2.6 μm, Phenomenex, Torrance, CA, USA)
coupled to a SecurityGuard Ultra guard column (Phenomenex). The The established analytical method was validated for limit of quan­
column temperature was 40 ◦ C, and the injection volume was 4 μL. The titation (LOQ), accuracy/precision, linearity of calibration, and recov­
eluent was composed of mobile phases A (0.1% formic acid and 5 mM ery. The LOQ was evaluated as the lowest concentration satisfying both
ammonium formate in water) and B (0.1% formic acid and 5 mM criteria: (i) the signal-to-noise ratio (S/N) of the peak was above 10 and
ammonium formate in methanol). The chromatographic separation was (ii) the percent relative error (% RE) of accuracy was between 80% and
initialized at 5% of mobile phase B with a constant flow rate of 0.2 mL/ 120% with percent relative standard deviation (% RSD) below 20%. The
min. After being maintained for 0.5 min, mobile phase B was linearly RE was defined as the ratio of the concentration of a QC measured on the
increased to 55% for 0.5 min and ramped to 95% for 7 min. This eluent instrument to the target (nominal) concentration of the corresponding
composition was held for 3 min, raised to 100% for 1 min, sharply QC.
dropped to the initial condition (5%) for 0.1 min, and then kept for 2.9 For intraday accuracy/precision, each single QC sample at the low
min. The total gradient time was 15 min. The MS/MS system was (LOQ; 5, 10, or 25 ng/mL) and high (75 ng/mL) concentrations were
operated using heated electrospray ionization (ESI) with a positive/ prepared and the concentrations were calculated by comparing peak
negative charge conversion system. The temperatures of the ion source responses (n = 5) to the saliva calibration curve. For the interday vali­
were set to 300 ◦ C (interface), 250 ◦ C (desolvation line), and 400 ◦ C dation, the same QC concentrations as those of intraday were evaluated
(heat block). The flow rates of the nebulizing (nitrogen), drying (nitro­ for five consecutive days. The linearity of the calibration ranging from
gen), and heating (air) gases were set to 3, 15, and 10 L/min, respec­ LOQ to 100 ng/mL for each analyte was evaluated (n = 5) using the
tively. Argon (≥99.999%) was used as a collision-induced dissociation correlation coefficient (r2). Recovery (n = 3) was studied by comparing
(CID) gas. The qualitative/quantitative data were processed using Lab­ pre-spiked and post-spiked saliva samples at two concentrations (low: 5
Solutions version 5.72 (Shimadzu). or 25 ng/mL; high: 75 ng/mL). The recovery values were calculated
For GC-MS/MS, the injection volume was 2 μL, the injection mode from the ratio of the response of the pre-spiking sample to that of the
was splitless, the inlet temperature was 280 ◦ C, and the pulsed injection post-spiking sample.
pressure was 250 kPa. Chromatographic separation was carried out The ME for each analyte was assessed by comparing the slope of
using an Rxi-5Sil MS capillary column (30 m × 0.25 mm i.d., 0.25 μm df, calibration (a) from the standards spiked into blank saliva extracts and
Restek, Bellefonte, PA, USA). Helium (≥99.999%) was used as the car­ that of calibration (b) from the corresponding non-matrix standard so­
rier gas at a constant flow rate of 1.50 mL/min. The oven temperature lutions. The ME was calculated using the following equation:
program was started at 90 ◦ C (held for 3 min), increased to 120 ◦ C at 20
(ME, %) = (a/b − 1) × 100
◦
C/min, and finally ramped to 300 ◦ C at 8 ◦ C/min, where it was held for
3 min. The total program duration was 30 min. The mass spectrometer
system was operated in the electron ionization (EI) mode at 70 eV. The
ion source and interface temperatures were 230 and 250 ◦ C, respec­ 2.8. Determination of chemical stabilities of pesticides in saliva samples
tively. Argon was used as a CID gas. The detector voltage was set at 1.4
kV. The qualitative/quantitative data were processed using GCMS so­ All of the high QC (75 ng/mL) samples were stored and prepared in
lution version 4.30 (Shimadzu). opaque tubes to minimize the effect of photolysis. For the stability test at
MRM optimization for LC- and GC-amenable compounds was con­ room temperature, the QC samples were stored at room temperature for
ducted. The precursor ion (parent ion) was selected for the full-scan over 8 h. In the freeze-thaw stability test, the QC samples were frozen at
mass spectrum. Two product ions (daughter ions) per compound were − 20 ◦ C for 12 h and thoroughly thawed at room temperature. This
selected as a quantifier ion and a qualifier ion with appropriate collision process was performed three times. All the treated stability samples
energies (CEs). The MRM conditions, CEs, and retention times of the were prepared as the final established method, and the % stability
target compounds are listed in Tables S1 and S2. (calculated from % RE) and % RSD (n = 3) of the target analytes were
evaluated. For comparative analysis of the accuracy/precision data, the
stability samples were analyzed with intraday QC samples in a single
2.5. Comparison of multiresidue extraction from saliva
batch.

Two scaled-down QuEChERS extraction procedures were used for

3. Results and discussion
100 μL of saliva: extraction with (i) acetonitrile (200 μL) and (ii) 0.1%
formic acid in acetonitrile (200 μL). Each extracted sample was parti­
3.1. Comparison of QuEChERS extraction solvent
tioned with 40 mg of MgSO4 and 10 mg of NaCl and then centrifuged.
The upper layer (120 μL) was matrix-matched with acetonitrile (30 μL)
QuEChERS methods without dSPE cleanup comprise two steps: sol­
for comparison with a post-spiked sample to evaluate the recoveries of
vent extraction and salt partitioning. The unbuffered salt combination,
336 target pesticides at 50 ng/mL (n = 3).
including MgSO4 and NaCl, has been reported to be optimal for multi­
residual extraction from biological samples such as serum and urine
2.6. Established sample preparation method (Shin et al., 2018, 2019). However, extraction solvent conditions for
biological fluids have not been compared. In the previous studies, we
The saliva sample (100 μL) in a 1.5-mL micro-tube was extracted confirmed that acidified acetonitrile including 0.1% formic acid was
with 200 μL of acetonitrile and two ceramic beads by vigorous shaking effective when extracting pesticides using QuEChERS procedures in
(1300 rpm, 1 min) using a Geno Grinder (1600 MiniG SPEX Sample crops and edible insects (mealworms) (Lee et al., 2018b; Shin et al.,
Prep, Metuchen, NJ, USA). The samples were treated with salts (40 mg 2020). In this study, two extraction solvents (acetonitrile and 0.1%
of MgSO4 and 10 mg of NaCl) and cooled in an ice bath to remove the formic acid in acetonitrile) were compared under the same unbuffered
heat generated by MgSO4 and water. The tube was shaken again and salt condition.
then centrifuged for 5 min at 13,000 rpm using a microcentrifuge (17 Each recovery value was verified (Fig. 1), and 321 of the 336 target
TR, Hanil Science, Seoul, Korea). The supernatants (120 μL) were mixed pesticides (black line box) showed excellent recoveries (70–120%) in
with acetonitrile (30 μL) for matrix matching (matrix/acetonitrile, 4:1 both solvents, according to the SANTE guidelines (European Commis­
v/v). The final extract solution was injected into the LC-MS/MS (4 μL) sion, 2019). When 0.1% formic acid in acetonitrile was used, 13 com­
and GC-MS/MS (2 μL) systems. pounds (A to M in Fig. 1) showed a superior recovery range (77–115%)

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E. Park et al. Chemosphere 309 (2022) 136725

had LOQs of 10 ng/mL (23.5%) and 25 ng/mL (6.8%), respectively. The

LOQ of each pesticide (5–25 ng/mL) was verified to be sufficiently
sensitive for forensic studies based on a case report where pesticide
concentrations of ≥600 ng/mL were determined in patients attempting
suicide (Lanaro et al., 2011). Several studies on occupational pesticide
exposure detected an average of 5.4–4130 ng/mL of pesticides in saliva
at representative sampling periods (Denovan et al., 2000; Hines et al.,
2006; Nigg et al., 1993), except for one report (≤0.061 ng/mL) (Lu et al.,
2006). To verify the carryover, blank samples were injected after in­
jection of the highest calibration point. There were negligible carryover
effects for the analytes (LOQ <20%). Therefore, the sensitivity of our
analytical method can be used for screening pesticide multiresidues in
occupational exposure studies, unintentional acute intoxication cases,
and self-poisoning cases.
The accuracy and precision of the analytical method were deter­
mined at both low (LOQ) and high (75 ng/mL) QC concentrations (Ta­
bles 1 and 2). All 336 pesticides at each LOQ had quantitative properties
satisfying acceptable accuracy (% RE; 80–120%) and precision (% RSD;
≤20%) in both intra- and interday tests, according to the Food and Drug
Administration (FDA) guidelines (US FDA, 2018). The intra- and inter­
day % RE at the high QC were determined to be acceptable between 85%
and 115% with % RSD ≤15% based on the FDA guidelines (US FDA,
2018). Therefore, the established scaled-down QuEChERS with LC- and
GC-MS/MS achieved sufficient sensitivity, reliability, and ruggedness.
Fig. 1. Comparison of two QuEChERS extraction solvents (acetonitrile vs 0.1% Linearity was evaluated by the correlation coefficient (r2) of cali­
formic acid (FA) in acetonitrile) through recovery rates for 336 pesticides (n = bration (Tables 1 and 2), and the values between 0.9884 and 0.9999
3) at 50 ng/mL. Single letters (A to O) indicate pesticide names: A, imazapic; B, were found to be excellent for simultaneously detecting pesticide mul­
imazamox; C, imazethapyr; D, quinmerac; E, asulam; F, imazaquin; G, half­ tiresidues. More accurate results at lower concentrations, particularly
enprox; H, permethrin; I, bifenthrin; J, chlorothalonil; K, pyridalyl; L, milbe­
LOQ, were obtained using a regression factor (1/x) for all target analyte
mectin A4; M, etofenprox; N, cyromazine; O, cinmethylin.
calibrations. The established analytical method demonstrated a
reasonable quantitative relationship between the concentration and
compared to those in non-acidified acetonitrile (12–65%). Among them, signal. Chromatograms of LC- and GC-MS/MS for the matrix-matched
imidazolinone herbicides (imazamox, imazapic, imazaquin, and ima­ standard at 100 ng/mL were shown in Fig. S1.
zethapyr) and quinmerac contain carboxylic acid moieties. Therefore, The recovery ranged from 73% to 120% with % RSD ≤20% for 333 of
ion suppression of carboxylate was required to increase the recovery 336 pesticides (Tables 1 and 2), showing excellent results according to
rates by lowering the pH of the extraction solvent. In various QuEChERS the SANTE guidelines (European Commission, 2019). Among the three
methods for imidazolinones or multiresidual pesticides, the pH condi­ compounds with out of the recovery ranges, cyromazine and dicofol
tions are controlled during sample preparation (Martins et al., 2014; showed a slightly lower recovery range (50–66%, Table 1). Cyromazine
Shin et al., 2020). Pyrethroids (bifenthrin, etofenprox, halfenprox, and is classified as a weak base; thus, water-soluble ionic forms (e.g.,
permethrin) also require solvent acidification to improve their recovery ammonium) are activated by the low pH of the extraction solvent
rates (54–65% to 81–96%). Hou et al. reported a slight increase in the (Turner, 2015). Chlorothalonil, the GC-MS/MS analyte, showed exces­
recovery of pyrethroids when acidified acetonitrile (1% acetic acid) was sive recovery (140%) at low QC levels. This phenomenon occurred due
used during QuEChERS extraction (Hou et al., 2019). Only the re­ to blockage of active sites in the GC injector liner at the initial analysis,
coveries of cinmethylin and cyromazine slightly decreased from 79% to as reported elsewhere too (Lehmann et al., 2018; Park et al., 2021). The
61% and from 83% to 69%, respectively, when 0.1% formic acid was recovery rates of these three analytes were also acceptable for multi­
added (N and O in Fig. 1). Lowering the pH of an extraction solvent has residue analysis based on a wider range of recovery criteria (30–140%)
been reported to be more effective for pesticide multiresidues (Lee et al., with consistent % RSD (≤20%) (European Commission, 2019). As re­
2018b; Park et al., 2021). Based on the overall results, 0.1% formic acid covery data are generally considered as accuracies in crop matrices, they
in acetonitrile was selected as the optimal extraction solvent. can be used as an alternative trueness parameter for pesticides in bio­
Except for the 336 target pesticides, captafol, captan, and folpet logical samples (Hiemstra and de Kok, 2007).
which are the phthalimide fungicides were further verified and not
recovered in either solvent. These pesticides have been reported to be 3.3. ME
rapidly hydrolyzed under aqueous conditions and degrade into phtha­
limides in the body (Gordon et al., 2001; Turner, 2015). Therefore, these In LC- and GC-MS/MS, signal suppression/enhancement is a com­
compounds were excluded from the target pesticides. mon phenomenon caused by sample matrices (Peters and Remane,
2012; Rahman et al., 2013). We divided % ME into three regions
3.2. Method validation (Table 3) according to the magnitude of the absolute values: negligible
ME (− 20%–20%), moderate ME (− 50% to − 20% or 20%–50%), and
Four validation factors, LOQ, accuracy/precision, linearity of cali­ strong ME (<− 50% and >50%).
bration, and recovery, were evaluated (Table 1). The validation results In LC-MS/MS, 280 (98%) of 287 target analytes showed negligible
are summarized in Table 2. The LOQ was defined as the lowest amount MEs. Among the five compounds included in the moderate to strong
(ng) in saliva (mL) which not only satisfy S/N 10 or higher but also show suppression area (Table 3), three pyrethroids (bifenthrin, deltamethrin,
constant responses on the chromatogram. Among the 336 target pesti­ and etofenprox) were found to be most affected (ME; − 51% to − 37%,
cides (287 for LC-MS/MS and 49 for GC-MS/MS), 234 (69.6% of the Table 1). While LC-MS/MS generally shows signal suppression for ana­
total: 190 for LC-MS/MS and 44 for GC-MS/MS) showed an LOQ of 5 ng/ lytes in biological samples (Park et al., 2021; Shin et al., 2018), most of
mL, with the S/N of greater than 10 (Table 2). The remaining pesticides the pesticides used in this study showed no ME. In GC-MS/MS, the MEs

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E. Park et al. Chemosphere 309 (2022) 136725

Table 1
Limit of quantitation (LOQ), linearity of calibration (r2), recovery, and matrix effect (ME) results for 336 target pesticides.
No. Compound Name LOQ ng/mL r2 Accuracy and Precision RE, % (RSD, %) Recovery ME
% %
Intraday Interday

Lowa Highb Lowa High2 Level 1c Level 2b

LC-MS/MS analytes (No. 1–287)

1 Acephate 5 0.9980 98 (12) 110 (1) 98 (5) 101 (9) 89 (17) 82 (8) − 9.6
2 Acetamiprid 5 0.9978 102 (4) 98 (3) 96 (4) 95 (5) 97 (6) 96 (2) 4.9
3 Acibenzolar-S-methyl 25 0.9995 113 (11) 101 (4) 100 (18) 101 (7) 96 (11) 94 (5) 8.4
4 Alachlor 10 0.9975 91 (14) 94 (4) 109 (12) 93 (13) 102 (10) 83 (7) 4.9
5 Allidochlor 5 0.9971 107 (15) 94 (7) 95 (8) 94 (9) 88 (15) 98 (9) − 0.7
6 Ametoctradin 5 0.9973 89 (8) 97 (5) 98 (12) 99 (3) 100 (9) 95 (4) 6.3
7 Ametryn 5 0.9984 85 (11) 101 (12) 95 (13) 98 (4) 94 (8) 97 (13) 3.8
8 Anilofos 5 0.9978 81 (14) 101 (8) 99 (9) 98 (4) 104 (1) 100 (6) − 3.4
9 Asulam 5 0.9978 92 (11) 99 (4) 106 (10) 100 (4) 107 (5) 94 (4) − 5.9
10 Atrazine 5 0.9972 88 (8) 96 (3) 96 (9) 100 (10) 105 (9) 92 (5) − 1.8
11 Azaconazole 10 0.9967 104 (6) 97 (4) 111 (9) 99 (6) 102 (5) 94 (3) 4.8
12 Azamethiphos 5 0.9990 100 (10) 92 (6) 101 (6) 98 (5) 97 (13) 97 (8) − 5.3
13 Azimsulfuron 5 0.9975 81 (9) 94 (10) 83 (20) 92 (3) 88 (19) 103 (2) − 2.4
14 Azinphos-methyl 5 0.9984 93 (7) 93 (3) 103 (13) 97 (6) 104 (5) 93 (2) 9.7
15 Azoxystrobin 5 0.9988 89 (11) 94 (8) 105 (3) 98 (4) 96 (12) 96 (7) − 1.0
16 Benfuresate 25 0.9967 99 (9) 107 (7) 105 (10) 101 (10) 85 (6) 86 (11) 8.2
17 Bensulfuron-methyl 10 0.9946 100 (11) 101 (5) 111 (13) 97 (6) 99 (14) 91 (13) 1.3
18 Bensulide 25 0.9969 97 (7) 98 (4) 95 (6) 101 (8) 100 (13) 92 (5) 4.9
19 Benthiavalicarb-isopropyl 5 0.9986 81 (13) 100 (10) 107 (14) 99 (4) 98 (12) 99 (12) 5.0
20 Benzobicyclon 5 0.9967 108 (8) 95 (8) 91 (20) 100 (7) 96 (13) 100 (11) − 1.4
21 Bifenthrin 5 0.9971 108 (14) 93 (7) 110 (6) 94 (6) 87 (5) 96 (8) − 60.9
22 Bitertanol 25 0.9982 101 (9) 101 (10) 101 (15) 100 (10) 92 (4) 98 (13) 18.2
23 Bromacil 5 0.9972 94 (16) 96 (4) 104 (11) 99 (6) 100 (2) 96 (5) − 3.1
24 Bromobutide 5 0.9984 106 (4) 102 (6) 103 (8) 99 (6) 89 (9) 105 (10) 2.4
25 Bromoxynil 25 0.9953 107 (17) 99 (10) 104 (11) 97 (6) 91 (12) 102 (3) − 1.4
26 Bupirimate 25 0.9961 102 (6) 103 (8) 102 (12) 104 (8) 96 (9) 92 (4) 13.4
27 Butachlor 5 0.9991 97 (8) 95 (5) 111 (11) 98 (7) 102 (9) 89 (10) 3.5
28 Butafenacil 5 0.9989 97 (16) 94 (4) 113 (10) 94 (4) 95 (11) 96 (10) − 1.4
29 Butocarboxim 5 0.9953 83 (14) 94 (2) 85 (4) 96 (3) 98 (10) 94 (2) − 8.1
30 Cadusafos 5 0.9981 104 (5) 97 (5) 104 (8) 97 (4) 85 (14) 99 (4) − 0.7
31 Cafenstrole 10 0.9942 108 (12) 101 (10) 104 (12) 94 (14) 90 (6) 94 (7) 2.7
32 Carbaryl 5 0.9991 102 (4) 99 (5) 99 (8) 100 (5) 95 (9) 92 (6) 2.1
33 Carbendazim 5 0.9978 112 (7) 97 (1) 103 (7) 97 (1) 80 (8) 102 (1) − 11.7
34 Carboxin 5 0.9975 100 (4) 99 (4) 101 (3) 97 (3) 97 (7) 96 (1) 1.0
35 Carfentrazone-ethyl 5 0.9986 92 (10) 96 (4) 101 (13) 99 (6) 94 (16) 95 (4) 9.6
36 Carpropamid 10 0.9976 96 (19) 109 (8) 116 (8) 102 (6) 99 (15) 97 (10) 9.1
37 Chinomethionat 25 0.9989 98 (5) 96 (7) 102 (5) 102 (6) 91 (4) 91 (12) 4.8
38 Chlorantraniliprole 10 0.9965 84 (7) 100 (6) 108 (18) 104 (6) 89 (5) 97 (7) 4.3
39 Chlorfluazuron 5 0.9988 99 (15) 104 (6) 100 (6) 99 (10) 87 (17) 78 (10) − 5.9
40 Chloridazon 5 0.9958 97 (8) 102 (10) 107 (13) 98 (6) 102 (10) 91 (10) 6.4
41 Chlorimuron-ethyl 10 0.9968 102 (9) 91 (10) 98 (14) 92 (6) 98 (2) 95 (5) − 0.6
42 Chlorotoluron 5 0.9993 93 (6) 98 (5) 101 (2) 99 (7) 100 (4) 97 (3) 7.0
43 Chlorpyrifos 10 0.9970 97 (11) 98 (11) 111 (10) 96 (8) 98 (5) 94 (14) 8.8
44 Chlorpyrifos-methyl 5 0.9991 111 (12) 98 (6) 110 (17) 97 (7) 112 (9) 90 (6) 17.4
45 Chlorsulfuron 5 0.9988 96 (11) 98 (3) 101 (12) 101 (11) 93 (9) 96 (7) − 2.0
46 Cinmethylin 10 0.9951 113 (19) 98 (4) 104 (13) 97 (10) 94 (15) 94 (15) − 5.6
47 Clethodim 5 0.9972 85 (18) 95 (11) 84 (19) 98 (9) 107 (9) 95 (12) − 3.2
48 Clofentezine 5 0.9978 87 (6) 98 (4) 102 (6) 100 (3) 106 (4) 99 (3) − 3.6
49 Clomazone 5 0.9987 91 (3) 93 (2) 102 (6) 99 (5) 96 (6) 96 (4) 3.3
50 Clothianidin 5 0.9982 113 (6) 97 (6) 105 (3) 100 (4) 106 (8) 100 (3) 2.1
51 Cyanazine 10 0.9987 103 (8) 88 (3) 104 (5) 94 (8) 103 (6) 98 (6) 0.3
52 Cyazofamid 5 0.9958 90 (9) 99 (4) 93 (7) 98 (3) 97 (9) 96 (2) 0.8
53 Cycloate 10 0.9992 98 (9) 97 (3) 102 (11) 99 (1) 95 (2) 92 (5) 6.4
54 Cyclosulfamuron 10 0.9959 106 (5) 95 (10) 101 (6) 90 (8) 105 (6) 94 (10) 0.2
55 Cyflufenamid 25 0.9963 114 (13) 99 (6) 103 (7) 96 (5) 109 (3) 93 (6) − 2.5
56 Cyhalofop-butyl 10 0.9947 110 (9) 94 (6) 114 (17) 101 (9) 102 (9) 99 (6) − 10.9
57 Cymoxanil 5 0.9983 108 (4) 92 (11) 99 (7) 95 (12) 98 (4) 95 (0) 3.1
58 Cyproconazole 5 0.9979 114 (9) 103 (4) 109 (13) 100 (5) 101 (12) 93 (4) 6.9
59 Cyprodinil 25 0.9935 88 (19) 100 (4) 99 (13) 103 (9) 94 (15) 96 (8) − 1.5
60 Cyromazine 5 0.9988 103 (2) 93 (7) 105 (4) 97 (5) 65 (9) 57 (5) − 4.1
61 Daimuron 5 0.9970 95 (4) 97 (4) 101 (4) 99 (5) 94 (10) 103 (6) 0.3
62 Deltamethrin 25 0.9924 86 (17) 95 (10) 97 (12) 102 (6) 90 (10) 91 (3) − 37.3
63 Demeton-S-Methyl 10 0.9923 112 (7) 101 (8) 115 (10) 107 (14) 114 (15) 97 (14) − 0.2
64 Di-allate 5 0.9995 81 (8) 87 (11) 97 (19) 97 (9) 101 (10) 91 (6) 4.0
65 Diazinon 5 0.9989 95 (8) 101 (6) 101 (10) 98 (3) 98 (6) 99 (6) 5.0
66 Dichlorvos 10 0.9992 102 (7) 99 (6) 98 (7) 99 (12) 95 (3) 97 (6) − 5.9
67 Diclofop-methyl 10 0.9970 109 (12) 97 (6) 114 (9) 103 (11) 106 (5) 99 (8) 0.4
68 Dicrotophos 5 0.9994 98 (3) 101 (3) 103 (5) 98 (3) 97 (2) 96 (4) − 3.3
69 Diethofencarb 5 0.9989 90 (5) 99 (2) 101 (10) 101 (7) 95 (1) 89 (4) 5.7
70 Difenoconazole 5 0.9997 96 (6) 100 (3) 100 (9) 99 (2) 105 (1) 95 (5) 3.8
(continued on next page)

5
E. Park et al. Chemosphere 309 (2022) 136725

Table 1 (continued )
No. Compound Name LOQ ng/mL r2 Accuracy and Precision RE, % (RSD, %) Recovery ME
% %
Intraday Interday

Lowa Highb Lowa High2 Level 1c Level 2b

71 Diflubenzuron 5 0.9984 86 (16) 96 (7) 88 (15) 98 (7) 101 (8) 100 (5) 3.6
72 Diflufenican 5 0.9989 100 (5) 96 (1) 109 (10) 99 (4) 92 (6) 96 (7) 0.8
73 Dimethachlor 5 0.9987 100 (6) 98 (4) 103 (8) 97 (2) 96 (7) 94 (2) 4.7
74 Dimethametryn 5 0.9993 95 (8) 98 (6) 98 (4) 99 (4) 103 (5) 97 (4) 5.2
75 Dimethenamid 5 0.9988 88 (7) 99 (3) 98 (12) 101 (5) 106 (8) 93 (4) 4.2
76 Dimethoate 5 0.9986 106 (4) 97 (3) 102 (4) 97 (9) 98 (8) 96 (1) 1.7
77 Dimethomorph 5 0.9994 97 (8) 94 (3) 98 (9) 92 (4) 98 (10) 93 (4) 2.8
78 Diniconazole 5 0.9984 115 (12) 98 (4) 104 (14) 99 (5) 95 (14) 94 (7) 7.0
79 Dinotefuran 5 0.9976 93 (7) 99 (4) 99 (6) 97 (1) 88 (8) 92 (5) − 1.3
80 Diphenamid 5 0.9983 92 (2) 98 (3) 96 (7) 99 (4) 96 (5) 100 (5) 0.5
81 Diuron 5 0.9969 95 (17) 99 (3) 102 (6) 100 (4) 93 (11) 99 (3) 1.2
82 Edifenphos 5 0.9985 85 (8) 106 (6) 93 (10) 99 (5) 97 (10) 96 (0) 6.0
83 Emamectin B1a 5 0.9965 103 (4) 94 (14) 99 (5) 97 (6) 104 (3) 93 (12) − 0.9
84 EPN 5 0.9984 84 (9) 97 (2) 94 (13) 99 (7) 110 (5) 97 (11) − 3.4
85 Epoxiconazole 5 0.9985 90 (8) 98 (5) 102 (5) 99 (4) 90 (6) 93 (4) 6.2
86 Esprocarb 5 0.9949 102 (8) 93 (15) 92 (8) 103 (4) 90 (3) 92 (1) 0.4
87 Ethaboxam (EBX) 10 0.9959 106 (8) 93 (7) 105 (9) 94 (10) 113 (9) 98 (8) − 2.9
88 Ethametsulfuron-methyl 5 0.9977 86 (16) 95 (6) 96 (11) 94 (4) 103 (10) 97 (9) − 2.5
89 Ethiofencarb 5 0.9980 91 (9) 93 (4) 96 (6) 92 (6) 92 (3) 95 (5) 6.3
90 Ethion 10 0.9995 100 (12) 104 (4) 113 (4) 101 (8) 90 (5) 82 (12) 11.3
91 Ethoprophos 5 0.9985 103 (9) 92 (5) 103 (10) 97 (5) 91 (20) 99 (7) − 3.6
92 Ethoxysulfuron 10 0.9884 108 (6) 89 (8) 119 (10) 92 (5) 95 (8) 87 (5) − 1.8
93 Etofenprox 5 0.9943 103 (3) 99 (5) 108 (5) 99 (7) 91 (11) 100 (4) − 51.3
94 Etoxazole 10 0.9985 88 (18) 95 (8) 120 (8) 103 (12) 99 (19) 80 (12) 17.3
95 Etrimfos 5 0.9983 91 (15) 106 (6) 104 (18) 99 (4) 101 (3) 102 (8) − 1.7
96 Fenamidone 5 0.9985 99 (11) 93 (3) 112 (17) 103 (11) 101 (20) 88 (7) 5.2
97 Fenamiphos 5 0.9980 92 (2) 99 (5) 102 (6) 99 (6) 101 (6) 99 (2) 3.7
98 Fenarimol 10 0.9988 105 (12) 96 (6) 102 (19) 102 (9) 98 (7) 97 (5) 3.3
99 Fenazaquin 5 0.9988 87 (8) 94 (7) 88 (10) 96 (13) 94 (12) 97 (7) − 18.9
100 Fenbuconazole 5 0.9995 88 (11) 97 (4) 97 (9) 100 (4) 95 (17) 95 (4) 2.6
101 Fenhexamid 5 0.9956 91 (16) 97 (5) 87 (20) 100 (5) 119 (5) 98 (7) 2.2
102 Fenobucarb (BPMC) 5 0.9985 96 (6) 98 (3) 100 (7) 102 (9) 98 (5) 97 (5) 10.4
103 Fenoxanil 5 0.9989 96 (7) 97 (3) 103 (6) 99 (4) 98 (8) 97 (5) 4.9
104 Fenoxaprop-p-ethyl 5 0.9984 90 (7) 95 (10) 99 (13) 99 (11) 120 (17) 103 (7) − 3.9
105 Fenoxycarb 5 0.9984 96 (8) 98 (4) 104 (13) 99 (6) 102 (11) 99 (2) 2.2
106 Fenthion 25 0.9962 115 (7) 111 (8) 106 (7) 98 (4) 102 (3) 96 (5) 1.9
107 Fipronil 10 0.9975 109 (14) 94 (3) 116 (7) 100 (6) 101 (8) 94 (3) 1.9
108 Fluacrypyrim 5 0.9947 92 (7) 96 (7) 96 (14) 99 (9) 104 (10) 94 (12) 6.8
109 Fluazinam 5 0.9961 92 (20) 101 (3) 103 (16) 93 (10) 111 (14) 96 (3) − 1.6
110 Flucetosulfuron 10 0.9936 98 (5) 94 (3) 113 (9) 95 (3) 92 (3) 93 (5) 3.4
111 Fludioxonil 25 0.9941 95 (10) 94 (5) 103 (12) 94 (8) 94 (8) 92 (5) 11.0
112 Flufenacet 10 0.9945 88 (14) 98 (9) 116 (10) 104 (7) 99 (4) 93 (16) 8.4
113 Flumioxazin 5 0.9958 81 (14) 104 (4) 109 (18) 97 (6) 116 (9) 97 (1) 1.5
114 Fluopicolide 10 0.9978 92 (7) 102 (8) 104 (9) 96 (8) 98 (5) 89 (7) 4.6
115 Fluopyram 10 0.9965 107 (5) 93 (5) 99 (12) 94 (4) 98 (3) 97 (7) − 3.9
116 Fluquinconazole 5 0.9988 91 (16) 92 (2) 89 (18) 95 (11) 105 (8) 102 (9) − 1.5
117 Flusilazole 5 0.9960 90 (19) 90 (6) 98 (1) 96 (4) 104 (8) 91 (3) 3.9
118 Flusulfamide 25 0.9932 113 (9) 99 (5) 111 (12) 98 (5) 112 (15) 96 (3) − 5.2
119 Flutolanil 10 0.9951 98 (11) 102 (5) 99 (10) 98 (3) 103 (9) 93 (8) 2.6
120 Forchlorfenuron 5 0.9979 104 (10) 98 (3) 98 (12) 101 (8) 105 (15) 96 (6) 0.7
121 Fosthiazate 5 0.9989 99 (4) 96 (5) 102 (4) 95 (4) 102 (6) 97 (7) 3.0
122 Furathiocarb 5 0.9995 99 (14) 92 (15) 101 (15) 105 (5) 112 (14) 104 (7) 2.8
123 Halfenprox 5 0.9993 102 (2) 96 (4) 102 (4) 99 (4) 97 (1) 98 (9) − 3.2
124 Halosulfuron-methyl 10 0.9989 95 (9) 100 (4) 96 (6) 97 (4) 94 (2) 100 (8) 7.3
125 Haloxyfop-R-Methyl 10 0.9968 94 (12) 107 (5) 117 (6) 106 (5) 98 (19) 105 (5) 7.7
126 Hexaconazole 5 0.9993 89 (9) 102 (2) 100 (10) 98 (7) 91 (14) 97 (6) 3.4
127 Hexazinone 5 0.9973 96 (9) 93 (10) 97 (9) 94 (10) 94 (3) 92 (7) 2.1
128 Hexythiazox 5 0.9964 93 (6) 102 (5) 100 (7) 95 (11) 101 (10) 84 (18) 18.7
129 Imazalil 5 0.9976 107 (6) 98 (6) 102 (8) 99 (5) 108 (1) 96 (4) 3.7
130 Imazamox 25 0.9991 91 (19) 89 (10) 104 (20) 103 (10) 84 (9) 91 (5) 8.7
131 Imazapic 10 0.9982 98 (11) 100 (11) 112 (13) 101 (6) 98 (10) 94 (10) 11.3
132 Imazaquin 10 0.9963 105 (4) 100 (8) 114 (9) 94 (5) 112 (8) 98 (4) 1.4
133 Imazethapyr 10 0.9970 101 (12) 98 (2) 114 (7) 98 (4) 103 (17) 105 (6) − 5.0
134 Imazosulfuron 10 0.9965 102 (15) 91 (4) 91 (18) 96 (6) 102 (9) 90 (3) 4.5
135 Imidacloprid 5 0.9972 96 (10) 96 (6) 103 (7) 99 (1) 100 (13) 99 (2) 5.2
136 Inabenfide 10 0.9931 96 (17) 102 (5) 100 (20) 91 (8) 90 (14) 103 (4) 9.7
137 Indanofan 10 0.9954 98 (16) 97 (7) 107 (16) 94 (5) 102 (9) 100 (5) 0.7
138 Indoxacarb 10 0.9986 100 (14) 99 (4) 112 (16) 102 (5) 93 (0) 91 (2) 9.0
139 Iprobenfos 5 0.9984 94 (9) 99 (6) 94 (9) 97 (4) 101 (9) 100 (2) − 1.2
140 Iprodione 25 0.9983 104 (10) 93 (9) 104 (3) 104 (6) 97 (10) 99 (8) 1.1
141 Iprovalicarb 5 0.9968 91 (9) 100 (6) 92 (11) 99 (8) 92 (10) 99 (15) 6.1
142 Isazofos 10 0.9970 101 (2) 100 (6) 106 (5) 97 (2) 105 (2) 100 (7) 3.2
(continued on next page)

6
E. Park et al. Chemosphere 309 (2022) 136725

Table 1 (continued )
No. Compound Name LOQ ng/mL r2 Accuracy and Precision RE, % (RSD, %) Recovery ME
% %
Intraday Interday

Lowa Highb Lowa High2 Level 1c Level 2b

143 Isoprocarb 5 0.9994 94 (7) 98 (9) 102 (8) 99 (2) 100 (6) 101 (4) 2.2
144 Isoprothiolane 5 0.9962 98 (6) 95 (5) 100 (5) 98 (3) 94 (3) 96 (9) 0.6
145 Isoproturon 5 0.9983 88 (8) 99 (4) 96 (7) 98 (5) 100 (4) 95 (2) − 2.4
146 Isopyrazam 5 0.9983 102 (13) 104 (3) 100 (14) 99 (9) 96 (9) 91 (12) − 5.5
147 Isoxathion 5 0.9985 100 (7) 107 (8) 103 (7) 101 (7) 100 (5) 97 (10) 4.2
148 Kresoxim-methyl 5 0.9972 92 (11) 97 (7) 89 (12) 99 (4) 94 (4) 96 (3) 1.8
149 Linuron 5 0.9996 105 (10) 98 (4) 109 (8) 99 (4) 100 (5) 89 (16) 1.9
150 Malathion 5 0.9975 111 (11) 97 (7) 102 (6) 96 (8) 74 (8) 97 (12) − 1.8
151 Mandipropamid 5 0.9991 93 (9) 96 (4) 99 (8) 100 (7) 99 (3) 98 (5) 4.0
152 Mecarbam 10 0.9983 107 (3) 94 (5) 111 (3) 96 (3) 93 (11) 97 (9) 5.6
153 Mefenacet 5 0.9978 93 (5) 96 (3) 97 (5) 94 (14) 98 (5) 94 (3) 3.4
154 Mefenpyr-diethyl 5 0.9986 94 (9) 102 (5) 103 (8) 100 (4) 100 (3) 97 (4) 3.4
155 Mepanipyrim 25 0.9969 89 (10) 90 (4) 100 (17) 103 (8) 95 (2) 97 (9) 5.3
156 Metalaxyl 5 0.9988 93 (4) 97 (4) 103 (12) 99 (4) 99 (9) 92 (4) 0.7
157 Metamifop 5 0.9986 93 (19) 100 (3) 97 (8) 102 (3) 98 (6) 97 (2) 6.2
158 Metazosulfuron 5 0.9983 86 (8) 94 (2) 98 (20) 95 (8) 99 (5) 90 (1) 5.9
159 Metconazole 5 0.9989 103 (4) 99 (4) 105 (5) 98 (3) 100 (1) 98 (6) 6.2
160 Methabenzthiazuron 10 0.9981 93 (8) 101 (8) 113 (16) 101 (10) 95 (15) 90 (12) 7.9
161 Methamidophos 5 0.9991 102 (10) 92 (4) 100 (11) 96 (6) 78 (5) 80 (7) − 21.4
162 Methidathion 5 0.9954 95 (13) 93 (9) 108 (10) 96 (7) 98 (11) 98 (3) 1.5
163 Methiocarb 5 0.9987 92 (5) 100 (4) 98 (8) 101 (7) 91 (5) 98 (7) − 1.1
164 Methomyl 5 0.9964 103 (7) 95 (4) 103 (3) 100 (7) 109 (8) 103 (4) 0.3
165 Methoxyfenozide 10 0.9963 109 (10) 90 (11) 112 (10) 92 (11) 98 (6) 99 (9) 9.9
166 Metobromuron 10 0.9988 114 (8) 99 (4) 102 (17) 95 (6) 103 (7) 110 (3) − 5.0
167 Metolachlor 5 0.9989 90 (6) 92 (4) 93 (7) 96 (5) 99 (2) 90 (2) 9.3
168 Metolcarb 5 0.9976 109 (6) 93 (4) 102 (12) 95 (3) 99 (8) 100 (8) − 0.2
169 Metominostrobin 5 0.9978 100 (10) 100 (3) 104 (8) 98 (6) 99 (8) 95 (2) 9.3
170 Metribuzin 5 0.9986 116 (4) 103 (2) 98 (20) 98 (5) 90 (18) 95 (12) − 1.9
171 Mevinphos 5 0.9992 109 (3) 98 (7) 100 (7) 96 (6) 98 (1) 95 (2) 4.7
172 Milbemectin A4 10 0.9985 100 (14) 94 (9) 108 (12) 101 (11) 94 (9) 98 (5) − 3.6
173 Molinate 5 0.9983 91 (11) 105 (9) 103 (4) 98 (4) 108 (6) 93 (8) 4.0
174 Monocrotophos 5 0.9973 100 (5) 97 (4) 100 (4) 96 (4) 100 (5) 98 (2) − 5.0
175 Myclobutanil 5 0.9986 104 (4) 96 (7) 106 (18) 100 (3) 80 (10) 94 (6) 7.1
176 Napropamide 10 0.9980 115 (9) 99 (2) 103 (6) 100 (6) 91 (7) 93 (3) 2.4
177 Nicosulfuron 5 0.9972 93 (10) 90 (5) 96 (8) 89 (3) 97 (10) 96 (9) 3.3
178 Novaluron 5 0.9987 102 (16) 92 (7) 92 (11) 99 (3) 108 (8) 97 (8) − 1.3
179 Nuarimol 5 0.9992 93 (10) 96 (6) 107 (8) 98 (4) 108 (16) 99 (4) 2.1
180 Ofurace 5 0.9982 112 (8) 92 (10) 107 (7) 91 (5) 109 (4) 89 (2) − 2.4
181 Orysastrobin 5 0.9995 101 (6) 95 (5) 97 (16) 100 (4) 99 (7) 94 (3) 3.2
182 Oxadiazon 5 0.9972 113 (14) 106 (4) 105 (11) 97 (8) 99 (9) 92 (10) − 0.8
183 Oxadixyl 5 0.9982 109 (9) 92 (6) 102 (11) 93 (5) 100 (9) 96 (11) 8.3
184 Oxamyl 5 0.9982 95 (10) 100 (5) 100 (8) 97 (4) 101 (4) 93 (7) − 0.3
185 Oxaziclomefone 10 0.9986 107 (2) 98 (5) 105 (6) 104 (2) 96 (4) 99 (4) 2.4
186 Oxydemeton-methyl 5 0.9989 98 (3) 99 (3) 101 (9) 98 (1) 98 (9) 92 (4) − 4.2
187 Paclobutrazol 5 0.9983 107 (5) 99 (5) 94 (12) 98 (5) 94 (5) 99 (1) 4.5
188 Parathion 5 0.9990 91 (11) 97 (11) 99 (9) 97 (5) 92 (5) 103 (3) − 0.4
189 Pebulate 10 0.9983 98 (16) 104 (3) 107 (9) 100 (8) 98 (7) 93 (9) 3.5
190 Penconazole 5 0.9982 91 (8) 100 (5) 99 (18) 98 (4) 103 (4) 98 (6) 3.6
191 Pendimethalin 10 0.9978 95 (7) 91 (9) 97 (5) 97 (3) 95 (17) 92 (15) − 2.0
192 Penoxsulam 10 0.9992 89 (7) 98 (2) 102 (8) 99 (3) 110 (1) 90 (6) 3.4
193 Penthiopyrad 5 0.9976 92 (14) 94 (5) 102 (5) 97 (6) 90 (13) 99 (8) 9.2
194 Permethrin 10 0.9977 111 (7) 90 (4) 101 (12) 96 (11) 100 (7) 95 (8) − 14.2
195 Phenmedipham 10 0.9985 104 (4) 97 (4) 107 (6) 98 (3) 101 (1) 99 (2) − 2.5
196 Phenothrin 5 0.9976 82 (18) 92 (2) 100 (17) 100 (8) 93 (6) 94 (7) − 4.7
197 Phenthoate 5 0.9985 97 (11) 97 (5) 99 (5) 98 (4) 100 (8) 98 (3) 5.0
198 Phorate 10 0.9990 112 (11) 100 (8) 120 (10) 102 (5) 92 (3) 100 (6) 3.4
199 Phosalone 5 0.9987 95 (8) 105 (3) 95 (6) 102 (4) 86 (6) 98 (3) 6.2
200 Phosmet 5 0.9971 93 (4) 98 (2) 98 (14) 99 (5) 94 (3) 99 (5) 4.7
201 Phosphamidon 5 0.9985 103 (6) 96 (2) 102 (5) 98 (4) 101 (5) 92 (6) 5.8
202 Phoxim 10 0.9985 100 (11) 105 (8) 105 (12) 100 (8) 100 (9) 85 (9) 5.4
203 Picolinafen 10 0.9971 102 (12) 93 (8) 107 (10) 100 (5) 104 (2) 93 (14) 3.4
204 Picoxystrobin 5 0.9979 96 (9) 103 (9) 103 (11) 96 (5) 107 (2) 97 (8) 3.6
205 Piperophos 25 0.9984 98 (13) 101 (8) 108 (10) 107 (8) 97 (20) 94 (6) 7.0
206 Pirimicarb 5 0.9991 98 (10) 97 (1) 93 (14) 102 (7) 99 (1) 98 (9) − 0.4
207 Pirimiphos-ethyl 5 0.9974 102 (4) 96 (8) 96 (9) 101 (9) 97 (7) 85 (15) 4.3
208 Pirimiphos-methyl 5 0.9987 98 (9) 96 (6) 95 (6) 103 (3) 99 (7) 100 (6) 0.3
209 Pretilachlor 5 0.9988 97 (7) 94 (5) 97 (11) 97 (3) 101 (5) 97 (1) − 2.1
210 Probenazole 5 0.9970 104 (12) 90 (10) 102 (13) 92 (9) 104 (9) 100 (7) − 0.1
211 Prochloraz 5 0.9982 104 (10) 100 (6) 100 (9) 100 (2) 100 (8) 96 (4) 5.1
212 Profenofos 25 0.9982 99 (4) 97 (4) 97 (18) 99 (5) 108 (3) 96 (8) 4.2
213 Promecarb 5 0.9984 100 (7) 98 (5) 106 (4) 99 (7) 96 (3) 96 (7) 1.0
214 Prometryn 5 0.9992 80 (13) 93 (5) 92 (11) 96 (3) 96 (16) 100 (8) 3.7
(continued on next page)

7
E. Park et al. Chemosphere 309 (2022) 136725

Table 1 (continued )
No. Compound Name LOQ ng/mL r2 Accuracy and Precision RE, % (RSD, %) Recovery ME
% %
Intraday Interday

Lowa Highb Lowa High2 Level 1c Level 2b

215 Propachlor 5 0.9986 97 (10) 101 (4) 104 (9) 100 (7) 89 (7) 92 (5) 3.4
216 Propamocarb 5 0.9984 106 (5) 92 (3) 101 (10) 103 (11) 86 (15) 86 (3) 23.7
217 Propanil 10 0.9997 94 (12) 97 (4) 102 (9) 97 (7) 97 (14) 93 (7) 8.6
218 Propaquizafop 5 0.9982 93 (16) 103 (3) 99 (15) 105 (6) 111 (2) 96 (4) 6.9
219 Propazine 5 0.9977 98 (9) 99 (5) 97 (6) 104 (8) 103 (1) 95 (9) 0.5
220 Propiconazole 5 0.9980 101 (6) 102 (11) 108 (11) 102 (4) 95 (8) 97 (3) 3.4
221 Propisochlor 25 0.9986 105 (8) 101 (5) 99 (12) 98 (5) 86 (5) 94 (6) 5.8
222 Propyzamide 5 0.9977 88 (15) 99 (7) 104 (19) 96 (1) 110 (2) 97 (6) 0.8
223 Pyraclofos 10 0.9956 92 (11) 101 (12) 105 (6) 102 (11) 87 (15) 93 (17) 21.0
224 Pyraclostrobin 5 0.9990 94 (7) 100 (3) 102 (3) 101 (4) 104 (7) 99 (4) 0.9
225 Pyrazoxyfen 10 0.9986 106 (5) 96 (5) 108 (6) 97 (4) 98 (5) 96 (3) − 1.4
226 Pyribenzoxim 5 0.9972 107 (6) 103 (5) 100 (11) 97 (12) 104 (3) 99 (5) 7.0
227 Pyridaben 10 0.9967 108 (7) 93 (10) 98 (13) 92 (8) 92 (5) 92 (5) − 27.4
228 Pyridalyl 5 0.9985 103 (3) 96 (4) 99 (4) 98 (5) 101 (5) 96 (8) − 1.3
229 Pyrifenox 5 0.9977 99 (8) 99 (4) 104 (9) 98 (4) 90 (15) 90 (7) 3.7
230 Pyrimethanil 10 0.9999 97 (9) 89 (5) 98 (10) 96 (9) 97 (8) 95 (10) 1.3
231 Pyriminobac-methyl E 10 0.9981 105 (8) 95 (7) 99 (6) 97 (5) 103 (14) 100 (14) 0.1
232 Pyriminobac-methyl Z 5 0.9972 95 (6) 95 (4) 101 (4) 95 (6) 98 (2) 96 (4) − 4.4
233 Pyroquilon 5 0.9986 108 (8) 95 (6) 109 (9) 97 (7) 101 (9) 91 (4) 0.7
234 Quinalphos 10 0.9988 89 (11) 100 (8) 113 (11) 95 (13) 97 (12) 98 (8) 12.7
235 Quinmerac 10 0.9925 118 (15) 97 (5) 106 (7) 95 (8) 106 (7) 100 (4) 12.6
236 Quinoclamine 5 0.9967 108 (10) 98 (1) 109 (13) 97 (10) 83 (17) 92 (4) 1.8
237 Quizalofop-ethyl 10 0.9973 86 (13) 93 (14) 110 (9) 105 (4) 95 (9) 96 (5) 14.9
238 Rimsulfuron 5 0.9982 94 (11) 98 (5) 95 (15) 93 (5) 82 (13) 89 (2) − 3.5
239 Saflufenacil 10 0.9979 85 (13) 95 (2) 100 (7) 94 (13) 107 (7) 103 (9) − 2.8
240 Simazine 10 0.9985 106 (12) 102 (2) 104 (11) 100 (6) 104 (2) 94 (1) − 2.2
241 Simeconazole 5 0.9960 109 (9) 99 (3) 104 (10) 97 (3) 109 (10) 97 (5) 3.1
242 Simetryn 5 0.9989 99 (5) 96 (1) 105 (6) 96 (4) 95 (4) 93 (1) 3.1
243 Spinetoram (XDE-175-J) 5 0.9993 87 (7) 97 (4) 89 (13) 103 (4) 100 (15) 97 (5) 4.4
244 Spinetoram (XDE-175-L) 5 0.9991 91 (7) 99 (5) 92 (4) 104 (4) 96 (3) 95 (4) 8.2
245 Spinosyn A 5 0.9989 89 (11) 100 (9) 98 (11) 101 (5) 95 (15) 100 (8) 8.4
246 Spinosyn D 5 0.9995 102 (6) 98 (3) 96 (9) 100 (2) 102 (6) 97 (3) 9.8
247 Spirodiclofen 5 0.9966 80 (8) 89 (9) 96 (14) 104 (7) 97 (6) 81 (5) − 7.8
248 Sulfoxaflor 5 0.9927 105 (7) 100 (2) 96 (6) 95 (9) 100 (12) 98 (6) 1.8
249 Sulprofos 5 0.9991 92 (8) 86 (11) 103 (15) 93 (11) 100 (13) 82 (15) 1.7
250 Tebuconazole 5 0.9991 93 (9) 98 (4) 104 (7) 98 (4) 107 (2) 99 (5) 7.0
251 Tebufenpyrad 10 0.9986 92 (11) 100 (8) 111 (13) 106 (5) 98 (8) 101 (9) − 2.5
252 Tebupirimfos 5 0.9983 93 (7) 90 (7) 104 (4) 95 (4) 94 (4) 90 (9) − 1.7
253 Terbuthylazine 5 0.9975 94 (8) 95 (6) 104 (14) 98 (8) 98 (10) 99 (1) 0.4
254 Terbutryn 5 0.9990 89 (9) 97 (6) 96 (7) 94 (5) 109 (6) 94 (6) 11.6
255 Tetraconazole 5 0.9971 107 (8) 96 (4) 111 (7) 98 (6) 105 (11) 100 (6) 3.3
256 Thenylchlor 5 0.9964 86 (15) 100 (4) 101 (12) 99 (4) 92 (8) 96 (4) 2.7
257 Thiabendazole 5 0.9994 91 (14) 99 (5) 95 (11) 99 (10) 94 (6) 93 (8) − 0.5
258 Thiacloprid 10 0.9977 98 (3) 97 (7) 110 (6) 96 (6) 100 (3) 98 (10) 1.1
259 Thiamethoxam 5 0.9941 95 (11) 97 (3) 101 (7) 101 (3) 112 (3) 100 (5) 0.4
260 Thiazopyr 10 0.9970 117 (18) 115 (7) 113 (12) 100 (8) 106 (14) 93 (8) 1.4
261 Thidiazuron 10 0.9976 96 (7) 99 (3) 100 (8) 100 (5) 110 (6) 97 (8) 3.5
262 Thifensulfuron-methyl 10 0.9987 102 (6) 96 (5) 104 (6) 98 (5) 98 (5) 99 (2) − 3.0
263 Thifluzamide 5 0.9938 90 (20) 100 (11) 103 (18) 101 (5) 81 (11) 98 (6) 6.5
264 Thiobencarb 5 0.9975 90 (13) 95 (3) 100 (14) 99 (3) 102 (6) 91 (14) 0.0
265 Thiocyclam 10 0.9982 109 (19) 108 (5) 100 (14) 103 (12) 73 (8) 78 (9) − 12.7
266 Thiodicarb 5 0.9987 95 (2) 97 (2) 100 (9) 102 (4) 98 (2) 93 (3) 4.7
267 Tolfenpyrad 25 0.9954 95 (9) 108 (7) 102 (13) 100 (14) 102 (16) 93 (11) 3.4
268 Tolylfluanid 25 0.9964 94 (8) 96 (10) 102 (8) 98 (7) 102 (4) 101 (4) 3.4
269 Triadimefon 10 0.9994 90 (11) 96 (7) 97 (7) 99 (5) 96 (4) 97 (6) 1.0
270 Triadimenol 25 0.9983 105 (8) 96 (4) 113 (10) 98 (8) 91 (11) 112 (7) 2.0
271 Tri-allate 5 0.9992 94 (13) 97 (11) 101 (4) 100 (7) 105 (15) 92 (9) 15.4
272 Triazophos 10 0.9952 81 (7) 109 (9) 112 (17) 104 (7) 100 (12) 102 (12) 5.0
273 Tribenuron-methyl 5 0.9993 83 (8) 96 (5) 88 (4) 90 (8) 97 (12) 92 (6) − 1.0
274 Tribufos 10 0.9981 96 (9) 103 (10) 109 (6) 97 (9) 94 (7) 93 (3) − 19.3
275 Trichlorfon 10 0.9967 103 (4) 95 (2) 111 (4) 97 (7) 106 (2) 93 (4) 0.6
276 Triclopyr 5 0.9986 93 (9) 95 (7) 89 (13) 101 (5) 118 (11) 100 (1) 2.0
277 Tricyclazole 5 0.9977 95 (14) 94 (6) 98 (12) 97 (10) 109 (5) 89 (8) 5.0
278 Trifloxystrobin 5 0.9988 104 (7) 94 (4) 95 (13) 95 (5) 94 (5) 96 (5) − 0.4
279 Triflumizole 5 0.9992 100 (3) 99 (2) 102 (5) 100 (4) 99 (8) 97 (4) − 1.0
280 Triflumuron 5 0.9971 107 (7) 98 (4) 107 (13) 101 (5) 93 (14) 96 (4) − 1.2
281 Trimethacarb 10 0.9959 80 (7) 102 (9) 108 (15) 105 (9) 94 (7) 94 (13) − 1.5
282 Triticonazole 5 0.9994 91 (10) 98 (4) 99 (8) 100 (3) 92 (15) 92 (5) 4.3
283 Uniconazole 5 0.9976 109 (6) 101 (6) 103 (11) 97 (3) 98 (11) 96 (3) 2.2
284 Vamidothion 5 0.9992 107 (6) 102 (4) 99 (6) 99 (5) 100 (5) 94 (3) 1.5
285 Vernolate 10 0.9980 92 (11) 96 (5) 101 (13) 99 (3) 99 (3) 90 (7) 8.4
286 XMC 5 0.9989 104 (5) 103 (3) 100 (5) 99 (4) 93 (2) 97 (5) 1.3
(continued on next page)

8
E. Park et al. Chemosphere 309 (2022) 136725

Table 1 (continued )
No. Compound Name LOQ ng/mL r2 Accuracy and Precision RE, % (RSD, %) Recovery ME
% %
Intraday Interday

Lowa Highb Lowa High2 Level 1c Level 2b

287 Zoxamide 5 0.9986 97 (13) 100 (7) 98 (9) 98 (6) 106 (7) 97 (9) 7.1
GC-MS/MS analytes (No. 288–336)
288 Aldrin 5 0.9986 96 (15) 98 (6) 102 (8) 99 (8) 88 (15) 93 (5) 11.3
289 BHC-alpha 5 0.9995 98 (9) 96 (6) 97 (9) 98 (5) 94 (1) 89 (8) 19.7
290 BHC-beta 5 0.9990 101 (4) 101 (3) 100 (8) 100 (5) 98 (4) 89 (5) 19.3
291 BHC-delta 5 0.9987 97 (4) 100 (3) 96 (3) 99 (4) 89 (9) 92 (4) 13.6
292 BHC-gamma 5 0.9991 99 (7) 100 (2) 96 (5) 100 (4) 89 (6) 92 (4) 14.3
293 Bromophos-methyl 5 0.9949 115 (17) 94 (3) 95 (11) 97 (7) 86 (8) 91 (3) 20.0
294 Bromopropylate 5 0.9984 107 (3) 94 (1) 95 (9) 95 (3) 97 (6) 93 (3) 194.7
295 Chlordane-cis 5 0.9952 91 (14) 100 (3) 93 (13) 99 (4) 76 (11) 97 (3) 4.9
296 Chlordane-trans 5 0.9989 83 (7) 97 (4) 95 (13) 97 (4) 85 (8) 96 (5) 10.8
297 Chlorfenapyr 10 0.9912 115 (14) 100 (5) 109 (12) 103 (7) 88 (10) 87 (1) 26.1
298 Chlorobenzilate 5 0.9983 109 (8) 96 (2) 98 (9) 96 (4) 94 (4) 92 (4) 107.7
299 Chlorothalonil 5 0.9905 108 (6) 101 (3) 100 (6) 99 (5) 140 (3) 95 (4) − 0.2
300 Chlorpropham 5 0.9987 99 (4) 95 (5) 100 (3) 98 (4) 93 (1) 95 (7) 40.8
301 Chlorthal-dimethyl 5 0.9988 94 (8) 99 (3) 97 (11) 98 (3) 97 (11) 94 (4) 11.4
302 Cyanophos 5 0.9979 106 (4) 98 (3) 99 (6) 99 (3) 94 (2) 91 (4) 23.9
303 Cyfluthrin 5 0.9978 112 (7) 96 (2) 95 (6) 96 (4) 91 (4) 92 (7) 75.9
304 DDD-o,p’ 5 0.9988 97 (2) 100 (2) 97 (3) 99 (5) 95 (2) 94 (3) 9.0
305 DDD-p,p’ 5 0.9970 110 (5) 90 (1) 96 (8) 94 (5) 91 (3) 85 (6) 69.6
306 DDE-o,p’ 5 0.9988 101 (2) 99 (2) 100 (4) 98 (4) 90 (2) 92 (4) 9.1
307 DDE-p,p’ 5 0.9989 100 (4) 99 (3) 98 (3) 98 (5) 95 (2) 93 (4) 12.5
308 DDT-o,p’ 5 0.9985 103 (5) 100 (2) 97 (3) 98 (5) 92 (6) 93 (3) 15.0
309 DDT-p,p’ 5 0.9960 114 (4) 85 (0) 96 (11) 92 (5) 89 (3) 85 (4) 111.1
310 Dichlobenil 5 0.9992 93 (8) 99 (9) 97 (11) 98 (9) 99 (1) 90 (8) 8.7
311 Dicloran 5 0.9977 108 (4) 97 (4) 95 (9) 98 (4) 102 (2) 88 (4) 39.4
312 Dicofol 25 0.9936 105 (3) 96 (7) 100 (9) 96 (5) 50 (2) 66 (5) − 51.1
313 Dieldrin 5 0.9979 92 (8) 101 (3) 102 (16) 100 (5) 87 (14) 94 (2) 6.2
314 Endosulfan-alpha 5 0.9991 107 (7) 100 (5) 91 (16) 99 (6) 102 (5) 91 (5) 11.8
315 Endosulfan-beta 10 0.9992 115 (17) 96 (6) 115 (10) 96 (9) 74 (16) 94 (2) 9.5
316 Endosulfan-sulfate 5 0.9992 107 (8) 94 (4) 96 (18) 98 (3) 96 (5) 94 (4) 22.0
317 Ethalfluralin 5 0.9987 112 (17) 101 (7) 96 (11) 98 (8) 104 (6) 88 (10) 38.4
318 Etridiazole 5 0.9990 105 (10) 86 (9) 97 (15) 92 (13) 95 (5) 86 (11) 103.7
319 Fenclorim 5 0.9995 97 (9) 98 (6) 98 (6) 98 (5) 97 (4) 90 (8) 14.1
320 Fenitrothion 10 0.9948 107 (2) 92 (2) 96 (10) 95 (5) 88 (6) 90 (5) 48.5
321 Fthalide 5 0.9984 107 (9) 99 (3) 101 (6) 98 (4) 100 (3) 95 (3) 16.7
322 Heptachlor 5 0.9989 99 (2) 94 (5) 91 (6) 95 (4) 94 (0) 89 (6) 53.7
323 Heptachlor epoxide 5 0.9989 111 (8) 97 (3) 100 (12) 96 (8) 90 (15) 81 (5) 15.4
324 Isofenphos 5 0.9983 108 (4) 98 (3) 96 (7) 96 (3) 96 (4) 91 (3) 26.1
325 Isofenphos-methyl 5 0.9981 106 (7) 97 (2) 96 (6) 96 (4) 96 (2) 92 (3) 29.4
326 Nitrothal-isopropyl 5 0.9966 116 (14) 97 (3) 100 (4) 96 (5) 99 (8) 92 (4) 53.1
327 Oxyfluorfen 10 0.9951 101 (8) 94 (4) 90 (11) 95 (5) 86 (3) 92 (2) 70.2
328 Parathion-methyl 5 0.9949 119 (11) 93 (1) 100 (8) 95 (3) 100 (3) 90 (5) 54.7
329 Pentachloroaniline 5 0.9992 91 (8) 100 (4) 97 (9) 99 (5) 93 (1) 93 (2) 11.4
330 Pentachlorothioanisole 5 0.9992 99 (6) 99 (4) 94 (13) 99 (5) 97 (5) 92 (3) 13.7
331 Procymidone 5 0.9985 99 (6) 100 (4) 102 (6) 99 (3) 101 (1) 93 (5) 13.5
332 Quintozene 5 0.9984 104 (16) 94 (4) 95 (14) 96 (3) 100 (6) 89 (8) 25.4
333 Silafluofen 5 0.9992 111 (7) 92 (1) 96 (8) 93 (4) 98 (3) 86 (4) 19.0
334 Tefluthrin 5 0.9989 97 (9) 100 (4) 97 (8) 98 (5) 96 (1) 91 (6) 21.3
335 Tetradifon 5 0.9985 106 (5) 96 (3) 97 (6) 96 (6) 96 (3) 88 (4) 19.0
336 Vinclozolin 5 0.9979 92 (5) 98 (3) 101 (6) 98 (3) 89 (17) 93 (3) 14.8
a
LOQ (5, 10, or 25 ng/mL).
b
75 ng/mL.
c
5 ng/mL for analytes with LOQ 5 ng/mL; 25 ng/mL for analytes with LOQ 10 or 25 ng/mL.

of 26 (53%) pesticides were negligible, whereas the rest (22 analytes), temperature and freeze-thaw stability) tests expressed as % stability and
except for dicofol, showed moderate to strong enhancement (Tables 1 % RSD (n = 3) are shown in Fig. 2. The dots outside the boxes in the plots
and 3). Large matrix effects are inevitable in GC-based instruments indicate that the corresponding pesticides degraded during the stability
(Rahman et al., 2013), and the relatively weak ME patterns observed in tests. The details of the stability results for all pesticides are presented in
saliva samples were remarkable. As saliva is mostly composed of 99% Supporting Table S3.
water (Carpenter, 2013), its matrix is very simple. Therefore, it can be For the samples left at room temperature for over 8 h, approximately
used in simple QuEChERS extraction without cleanup. For pesticides 20% of the 336 pesticides decomposed and did not satisfy the strict FDA
with negligible ME and excellent recovery range (70–120%), solvent guidelines (out of red box, Fig. 2 a). Among them, 13 compounds
standard calibration could be an alternative to matrix-matched cali­ (probenazole, chinomethionate, spirodiclofen, benzobicyclon, phen­
bration as it reduces the preparation time. medipham, phosmet, cymoxanil, flumioxazin, trichlorfon, tolylfluanid,
azinphos-methyl, iprodione, and triallate) were found to be most
decomposed (% Stability ≤38%). Except for probenazole, for which
3.4. Chemical stabilities in saliva samples stability data are not available, they were found to be susceptible to
hydrolysis in neutral to alkaline media (Turner, 2015). The pH of saliva
The summarized results of two types of stability (short-term room

9
E. Park et al. Chemosphere 309 (2022) 136725

Table 2
Summary of method validation results; The numbers of pesticides and their
percentages for 336 target pesticides (287 for LC-MS/MS and 49 for GC-MS/MS)
satisfying the criteria of limit of quantitation (LOQ), linearity of calibration (r2),
and recovery.
Range No. of Pesticides (%)

Total LC-MS/MS GC-MS/MS

LOQ
5 ng/mL 234 (69.6%) 190 (66.2%) 44 (89.8%)
10 ng/mL 79 (23.5%) 75 (26.1%) 4 (8.2%)
25 ng/mL 23 (6.8%) 22 (7.7%) 1 (2.0%)
Sum 336 (100%) 287 (100%) 49 (100%)
r2
>0.990 335 (99.7%) 286 (99.7%) 49 (100.0%)
0.988–0.990 1 (0.3%) 1 (0.3%) 0 (0.0%)
Sum 336 (100%) 287 (100%) 49 (100%)
RE of accuracy
Lowa (RSD ≤20%)
within 80–120% (intraday) 336 (100%) 287 (100%) 49 (100%)
within 80–120% (intraday) 336 (100%) 287 (100%) 49 (100%)
Highb (RSD ≤15%)
within 85–115% (intraday) 336 (100%) 287 (100%) 49 (100%)
within 85–115% (intraday) 336 (100%) 287 (100%) 49 (100%)
Recovery
Level 1c (RSD ≤20%)
50%–70% 2 (0.6%) 1 (0.3%) 1 (2.0%)
70%–120% 333 (99.1%) 286 (99.7%) 47 (95.9%)
120%–140% 1 (0.3%) 0 (0.0%) 1 (2.0%)
Sum 336 (100%) 287 (100%) 49 (100%)
Level 2b (RSD ≤20%)
50%–70% 2 (0.6%) 1 (0.3%) 1 (2.0%)
70%–120% 334 (99.4%) 286 (99.7%) 48 (98.0%)
120%–140% 0 (0.0%) 0 (0.0%) 0 (0.0%)
Sum 336 (100%) 287 (100%) 49 (100%)
a
LOQ (5, 10, or 25 ng/mL).
b
75 ng/mL.
c
5 ng/mL for analytes with LOQ 5 ng/mL; 25 ng/mL for analytes with LOQ 10
or 25 ng/mL.

Table 3
Summary of matrix effect (ME) results for 336 target pesticides (287 for LC-MS/
MS and 49 for GC-MS/MS).
ME Range Magnitude LC-MS/MS GC-MS/MS

<− 50% Strong suppression 2 1

− 50% to − 20% Moderate suppression 3 0
− 20%–20% Negligible 280 26
20%–50% Moderate enhancement 2 12
>50% Strong enhancement 0 10
Sum 287 49

is almost neutral, and salivary components (e.g., salts and enzymes) can Fig. 2. Chemical stabilities for 336 target pesticides in saliva samples under (a)
accelerate hydrolysis of pesticides. In fact, the half-life of chinome­ room temperature and (b) freeze-thaw condition at a high QC level of 75 ng/mL
thionine in pH 7 media is 80 h at room temperature (Turner, 2015), (n = 3). The insides of red and blue boxes in the plots showed the pesticides
satisfying % RE within 85–115% (% RSD ≤15%) and % RE within 70–130% (%
which decreased by more than 90% to only 8 h in saliva.
RSD ≤30%), respectively. Single letters (A to O) in both plots indicate pesticide
Chemical interactions between pesticides in the QC samples cannot
names: A, fenobucarb (BPMC); B, chlorothalonil; C, probenazole (nd; not
be ignored. Interestingly, the % stability of dichlorvos at room temper­ detectable); D, chinomethionat; E, spirodiclofen; F, benzobicyclon; G, phen­
ature was 148%, far exceeding the 100% criterion (Fig. 2 a and medipham; H, phosmet; I, cymoxanil; J, flumioxazin; K, trichlorfon; L, tolyl­
Table S3). This is because trichlorfon, which showed <5% stability, fluanid; M, azinphos-methyl; N, iprodione; O, tri-allate. (For interpretation of
converted to dichlorvos. In a further stability study in which trichlorfon the references to colour in this figure legend, the reader is referred to the Web
was the sole compound in saliva samples, we verified that 99.3% of 75 version of this article.)
ng/mL of trichlorfon degraded, and 40.1 ng/mL of dichlorvos was newly
produced. Conversion of trichlorfon into dichlorvos has also been re­ satisfied the strict FDA criteria (red box in Figs. 2 b), and 331 (98.5%)
ported in various environmental samples such as soil, seawater, and pesticides met the wider criteria for multiresidue screening in biological
cabbage (Li et al., 2011; Samuelsen, 1987). samples (blue box) (Gray et al., 2008; Lehmann et al., 2018; Schummer
Although it was confirmed that more than 80% of the target pesti­ et al., 2012). Only five compounds, fenobucarb (BPMC), chlorothalonil,
cides were stable at room temperature (Fig. 2 a), alternative sample probenazole, chinomethionate, and spirodiclofen, showed poor % sta­
storage and usage strategies are required for some unstable pesticides. In bility (11–68%, Fig. 2 b and Table S3). Interestingly, fenobucarb and
the other stability test, in which three cycles of freezing (− 20 ◦ C) and chlorothalonil were less stable after repeated freeze-thaw cycles than
thawing procedures were performed, 311 (92.6%) of the 336 analytes those left at room temperature.

10
E. Park et al. Chemosphere 309 (2022) 136725

Table 4 precision. Most of the pesticides were chemically stable in saliva left for
Unintentional pesticide exposures verified through saliva samples from 20 several hours at room temperature; very few pesticides lost their sta­
agricultural workers during mixing/loading and spraying. bility even after several freezing and thawing processes. The established
Pesticide Name (ng/mL) method was successfully used for biomonitoring farmers (ten mixers and
Sample No. Fenobucarb (BPMC) Propamocarb
ten sprayers). Two carbamates (fenobucarb and propamocarb) were
determined at trace concentrations (12.5–675.0 ng/mL from 11 posi­
Mixing/loading
tively detected samples). This bioanalysis method can be a powerful tool
#1 38.3 -a
#3 25.3 – for detecting various pesticides in agriculture as well as for forensic and
#4 38.5 – clinical purposes.
#5 34.8 –
#8 31.0
Author contributions statement
–
#9 44.0 –
#2, #6, #7, #10 – –
Spraying Eunyoung Park: Formal analysis, Validation, Writing – original
#2 54.3 – draft, Jiho Lee: Investigation, Resources, Hye Suk Lee: Conceptuali­
#3 320.0
–
zation, Funding acquisition, Jeong-Han Kim: Project administration,
#5 108.5 –
#8 – 12.5
Visualization, Yongho Shin: Data curation, Methodology, Software,
#9 – 675.0 Supervision, Writing – review & editing.
#1, #4, #6, #7, #10 – –
a
Not detected. Declaration of competing interest

In further experiments where saliva was thawed only once in a frozen The authors declare that they have no known competing financial
state at − 20 ◦ C or − 80 ◦ C, followed by immediate sample preparation, interests or personal relationships that could have appeared to influence
all the target pesticides were stable (data not shown). Because of this the work reported in this paper.
stability issue, we maintained the ice box temperature below − 20 ◦ C
after real sample collection and immediately conducted preparation Data availability
without refreezing to prevent the decomposition of pesticides in saliva.
Data will be made available on request.
3.5. Application
Acknowledgement
In general, use of pesticides on farmlands involves two tasks: mixing/
loading and spraying (Lee et al., 2018a). Saliva samples collected from This research was supported by the Bio and Medical Technology
the agricultural workers (ten mixers and ten sprayers) were extracted Development Program of the National Research Foundation (NRF) and
using the established method, and the pesticide multiresidues were funded by the Korean government (MSIT) (grant number NRF-
determined (Table 4). 2015M3A9E1028325), Republic of Korea.
Two carbamates, fenobucarb (BPMC) and propamocarb, were
detected. Fenobucarb was found in the mixers, from which it was Appendix A. Supplementary data
detected in six out of ten operations, and the detection frequency in the
mixers was higher than that in the sprayers (two out of ten operations). Supplementary data to this article can be found online at https://doi.
However, the concentration range (25.3− 44.0 ng/mL) among the org/10.1016/j.chemosphere.2022.136725.
detected samples was lower than that of the sprayer (54.3− 108.5 ng/
mL). Propamocarb was detected in only three sprayer saliva samples, References
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