Journal of Chromatography B 1141 (2020) 122021

Contents lists available at ScienceDirect

Journal of Chromatography B
journal homepage: www.elsevier.com/locate/jchromb

Upgrading analytical methodology through comparative study for screening T

of 267 pesticides/metabolites in five representative matrices using UPLC-
MS/MS
Tae-Woong Naa,1, Md. Musfiqur Rahmanb,1, , Sung-Woo Kimc, M. Ershadul Haqued,
⁎

Jong-Bang Eune, Jae-Han Shimb,

⁎

a
National Agricultural Products Quality Management Service (NAQS), 141, Yongjeon-ro, Gimcheon-si, Gyeongsangbuk-do, Republic of Korea
b
Natural Products Chemistry Laboratory, Chonnam National University, Yongbong-ro 77, Buk-gu, Gwangju 500-757, Republic of Korea
c
Jeollanamdo Agricultural Research and Extension Services, Environment-Friendly Agricultural Research Institute, 1508, Senam-ro, Sanpo-myeon, Naju-si, Jeollanamdo
58213, Republic of Korea
d
Department of Statistics, University of Dhaka, Dhaka 1000, Bangladesh
e
Department of Food Science and Technology and BK 21 Plus Program, Graduate School of Chonnam National University. Yongbong-ro 77, Buk-gu, Gwangju 500-757,
Republic of Korea

ARTICLE INFO ABSTRACT

Keywords: A comparative study was conducted to replace the traditional screening method (MFDS#83) with the Quick,
Pesticides Easy, Cheap, Effective, Rugged and Safe (QuEChERS) EN method for the determination of 267 pesticides/me-
Multi-residue tabolites/plant activators/growth regulators in five representative crop matrices (mandarin, pepper, potato, rice,
QuEChERS and soybean). In the traditional method, samples were extracted with acetonitrile and salt, and purified with a
Traditional method
solid-phase extraction cartridge. In the QuEChERS method, the sample extraction was carried out using acet-
UPLC-MS/MS
onitrile and a mixture of salts, and purification was performed using dispersive solid phase extraction. The limit
of quantification (LOQ) for the MFDS#83 method was 0.0004 mg/kg, whereas for the QuEChERS EN method,
the LOQ varied from 0.002 to 0.006 mg/kg for all analytes in various matrices. A six-point matrix-matched
calibration curve was prepared for all analytes in five matrices for both methods. Both the MFDS#83 and
QuEChERS EN methods provided excellent linearity, with the coefficients of determination (R2) ≥ 0.99 for most
of the compounds. In both cases, the method was validated in terms of recovery and repeatability after the
fortification of two different concentrations with three replicates for each of the concentrations. The QuEChERS
EN method provided better recovery than the MFDS#83 method for all matrices except mandarin.

1. Introduction
The continued use of pesticides has the benefit of increasing pro-
duction in agro-food industries; however, it also negatively affects the
environment, crops, and livestock [1,2]. Primarily, the pesticide is ex-
posed by application to crops and resulting residues that are not ab-
sorbed by the crops but are washed in air or rainwater and flow into the
soil or river [3]. Therefore, this residue can be delivered to the crops via
soil or water, finally resulting in bioaccumulation in the livestock
through contaminated feed [4]. In addition, these pesticides can be
decomposed by light, moisture, or temperature according to their
properties, and some degraded metabolites can be formed as toxic
sulfoxide or sulfone. The toxic metabolites are absorbed and taken up
by the food chain and environmental circulation [5–7]. These processes
suggest that the management of pesticide is now an important issue for
both the local and international community [8]. To monitor pesticides
routinely, it is urgent to establish effective sample preparation and
detection methods, which include single-residue methods (SRM) for
analyzing individual compounds and multi-residue methods for si-
multaneous analysis of various compounds [9]. Although SRM have the
advantage of being more reliable in terms of precision and reproduci-
bility, the main drawback of these methods is their limited application
to specific pesticide and specific crops, [9,10]. In contrast, simultaneous
multi-residue methods have the advantage of analyzing various com-
pounds at the same time, although the precision and reproducibility of
specific compounds are slightly reduced. These methods are applicable

⁎
Corresponding authors.
E-mail address: jhshim@jnu.ac.kr (J.-H. Shim).
1
The first two authors contributed equally to this study.

https://doi.org/10.1016/j.jchromb.2020.122021
Received 22 October 2019; Received in revised form 2 February 2020; Accepted 3 February 2020
Available online 06 February 2020
1570-0232/ © 2020 Elsevier B.V. All rights reserved.
T.-W. Na, et al.
to various crop matrices over a wide range of pesticides [11–16].
Therefore, to monitor pesticide residue during the production and
distribution stages, and develop an approach that has versatility under
different international conditions, the multi-residue method is more
suitable than the SRM, and the development and verification of these
methods are essential [6,17,18]. In recent years, more rapid, simple,
and precise methods of analysis have become prominent. The most
popular analytical method in this regard is a method called the Quick,
Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method. The
QuEChERS method was first introduced by Anastassiades in 2003 to
effectively remove matrix co-extracts and successfully transfer polar
pesticides to the solvent layer in the shortest possible time using mag-
nesium sulfate (MgSO4). The purification was carried out by dispersive
solid-phase extraction (d-SPE) instead of time-consuming solid-phase
extraction [19]. The possible outcome of the method was the analysis of
a wide range of pesticides to various plant matrices [19–21]. The main
advantages of QuEChERS was its possibility of modification to suit the
characteristics of the analyte, matrices, and amount of sample or sol-
vent. Because of these advantages, two major modifications of QuE-
ChERS were observed for pH-dependent analytes: one is an acetate-
buffered version introduced by Lehotay in 2005, which is the official
AOAC method, and the other is a citrate-buffered version introduced in
2007 as a final modified version, which became the official EU method
called QuEChERS EN [20,22]. Furthermore, the method is still being
modified for analysis of a wide range of pesticides in various matrices
[23–27].
In the Republic of Korea, multi-class pesticides were analyzed using
the multi-residue methods named MFDS#83 (Ministry of Food and
Drug Safety#83) from the Korean Food Code. The method involved
acetonitrile and salts (sodium chloride,) for extraction, and purification
was carried out using an aminopropyl cartridge. Finally, analysis was
conducted via liquid chromatography (LC) and gas chromatography
(GC), coupled with a traditional detector. However, the method re-
quired mass spectrometric confirmation for avoiding false positive/
negative results. This was the only validation method before the
QuEChERS method was introduced [12]. The method has been used
since its initial introduction in 2000 as a representative liquid extrac-
tion method without any modification. The method was modeled by the
California Department Food and Agriculture (CDFA) and published by
the United States Food and Drug Administration (FDA). The CDFA
method was based on the Mills (1963) method, introduced as the first
multi-class pesticide multi-residue method, which was considered a
precise method established through long-term experiments and results
[28–31]. The method has been verified through modification and
supplementation over a period of time; thus, it can be applied to various
crops and is known to be suitable for quantification.
However, the method lacks research on the simultaneous analysis of
more than 200 pesticides. In addition, it is well-known that the cleanup
process is omitted or simplified, and is affected by the matrix.
Therefore, many studies on the applicability to various crops should be
conducted and verified. In this study, we compared the latest official
version of the QuEChERS EN method and traditional multi-residue
method (MFDS#83), which has been proven to be an effective method.
Thus, the purpose of this study was to investigate whether the
QuEChERS EN method can be a replacement of the existing traditional
method and determine the ability of both methods to simultaneously
analyze more than 200 pesticides.
2. Experimental
2.1. Chemicals and standards
A total of 267 pesticide/metabolite/plant activator/growth reg-
ulator standards were included in this study, from which 256 were
parent compounds, while the others were isomers and metabolites. The
pesticides were classified as insecticide (88), fungicide (76), herbicide
2
Journal of Chromatography B 1141 (2020) 122021
(72), acaricide (11), nematicide (2), plant activator (2), and plant
growth regulator (5). The pesticide standards were purchased from
AccuStandard, Inc. (New Haven, CT, USA), Wako Pure Chemical
Industries, Ltd. (Osaka, Japan), Dr. Ehrenstorfer GmbH (Augsburg,
Germany), Chem Service, Inc. (West Chester, USA), and LG Life
Sciences, Ltd. (Seoul, Korea). Analytical grade reagents and solvents
were used for extraction, purification, and detection. Acetonitrile, me-
thanol, and dichloromethane were purchased from Burdick & Jackson
(Muskegon, USA). Aminopropyl (NH2) SPE cartridge (1 g, 6 mL) was
obtained from Waters (Massachusetts, USA). An A-QTM EN QuEChERS
kits (4.0 g MgSO4, 1.0 g NaCl, 1.0 g Trisodium Citrate Dihydrate, 0.5 g
Disodium hydrogen citrates sesquihydrate, Lot No., 1505-13-E10) was
supplied by KRIAT (Daejeon, Republic of Korea). A syringe filter MN,
0.2 µm–15 mm was purchased from CHROMAFIL (Duren, Germany).
Sodium chloride and formic acid (less than98%) were purchased from
Junsei Chemical Co. Ltd. (Tokyo, Japan). Ammonium formate (greater
than99%) was supplied by Kanto Chemical Co. (Tokyo, Japan).
2.2. Preparation of standard stock solution and calibration
Standard stock solution for each of the analytes was individually
prepared at the level of 1000 mg/L. For the mixed standard solution,
200 μL of each standard solution was aliquoted into a 20-mL volumetric
flask and evaporated under nitrogen stream to prepare 20 mL of a
mixed standard solution at a level of 10 mg/L. All the prepared stan-
dard solutions were sealed in amber vials and stored at −20 °C for
freezing.
To prepare the calibration curve, the mixed standard solution was
diluted with acetonitrile at concentrations of 0.002, 0.005, 0.01, 0.02,
0.05, and 0.1 mg/kg. The calibration curve was constructed based on
the peak area of the chromatogram. Matrix-matched standards of the
QuEChERS method and the MFDS#83 method were prepared by using
a blank extract and diluted in a ratio of 4 : 1 (blank extract : standard
mixture).
2.3. UPLC-MS/Ms
A Waters AQUITY H-Class ultra-performance liquid chromato-
graphy (UPLC) (Waters, Hertfordshire, UK) system was coupled with a
triple quadrupole mass spectrometer (AB SCIEX QTRAP 4500, SCIEX,
Redwood, CA, USA) for instrumental analysis. The separation was
achieved using a CAPCELL CORE-C18 column (100 × 2.1 mm, 2.7 μm).
The mobile phase consisted of (A) water and (B) methanol, containing
0.1% (v/v) formic acid and 5-mM ammonium formate flowing in gra-
dient mode. The mobile phase gradient was started at 95% A for 1 min
with the flow rate of 0.3 mL/min, decreased to 45% A at 1.5 min and to
40% A at 5 min. The phase gradient continued to drop to 10% A with
the flow rate increasing to 0.4 mL/min and dropped to 2% A at
12.1 min, remaining the same until at 15 min. At that time, initial
condition was reinstated at 15.1 min, remaining constant until at
20 min. The injection volume was 2 μL with a flow rate of 0.3 mL/min,
and the total run time was 20 min for each injection. The column
temperature was set at 35 °C.
The conditions of mass spectrometry (MS) for some pesticides are
published; however, because the product and precursor ions are varied
owing to the instrument brand and analysis conditions, the MS condi-
tions were optimized and established for 267 target analytes. The in-
strument was operated using electrospray ionization in the positive and
negative modes. The curtain gas was 25 psi, ion spray voltages were
5500 V for positive and −4500 V for negative mode, and the tem-
perature was 500 °C. The Q1 mass scan of each component was per-
formed to confirm the precursor ion and intensity. Then, a scan for the
product ion was performed and the changes in the DP (declustering
potential), EP (entrance potential), CE (collision energy), and CXP
(collision cell exit potential) were noted. The most suitable MS/MS
product ion spectra were selected through the chromatogram. The
T.-W. Na, et al. Journal of Chromatography B 1141 (2020) 122021

Table 1
Optimized MRM conditions of 256 (2 6 7) pesticides for multi-residue pesticide analysis using LC-MS/MS.
Compound Activity RT Q1 Q3 DP EP CE CXP

Abamectin B1a Insecticide 13.23 890.50 305.00 60 10 35 8

13.23 890.53 113.10 56 10 73 10
Acephate Insecticide 2.85 183.97 143.00 46 10 11 8
2.85 183.97 95.00 46 10 29 8
Acetamiprid Insecticide 3.63 223.10 126.00 86 10 29 11
3.63 223.10 99.10 86 10 49 11
Acibenzolar-S-methyl Plant activator 7.31 211.10 140.00 50 10 31 8
7.31 211.10 136.10 91 10 39 11
Alachlor Herbicide 8.52 270.12 238.00 41 10 13 10
8.52 270.12 162.10 41 10 27 6
Aldicarb Insecticide 4.09 212.98 116.00 56 10 15 4
4.09 208.10 116.00 41 10 11 8
Ametoctradin Fungicide 10.00 276.20 149.20 66 5 49 4
10.00 276.20 176.20 66 5 49 4
Amisulbrom Fungicide 10.87 465.83 227.00 76 10 29 10
10.87 465.83 148.10 76 10 63 10
Amitraz Insecticide 12.58 294.11 163.20 61 10 21 11
12.58 294.11 122.20 61 10 41 11
Atrazine Herbicide 5.61 216.12 174.00 36 10 23 8
5.61 216.12 103.90 36 10 39 8
Azimsulfuron Herbicide 5.77 425.00 182.10 71 10 23 11
5.77 425.00 156.10 71 10 45 11
Azinphos-methyl Insecticide 6.54 318.04 132.10 51 10 21 10
6.54 318.04 160.00 51 10 19 11
Azoxystrobin Fungicide 6.95 404.10 372.10 76 10 19 15
6.95 404.10 344.10 76 10 31 13
Bendiocarb Insecticide 4.57 224.20 167.20 71 10 13 11
4.57 224.20 109.20 71 10 23 11
Benfuracarb Insecticide 11.05 411.20 195.10 86 10 31 11
11.05 411.20 102.10 86 10 41 11
Benfuresate Herbicide 6.07 274.15 163.10 61 10 25 10
6.07 274.15 77.10 61 10 101 12
Bensulfuron-Methyl Herbicide 6.40 410.90 149.00 76 10 27 11
6.40 410.90 119.10 76 10 53 11
Benthiavalicarb-Isopropyl Fungicide 7.80 382.10 180.10 86 10 39 11
7.80 382.10 116.20 86 10 29 11
Benzobicyclon Herbicide 7.82 447.00 257.10 61 9 37 4
7.82 447.00 229.10 61 9 51 4
Benzoximate Acaricide 10.29 364.06 198.90 6 10 11 8
10.29 364.06 105.00 6 10 37 10
Compound Activity RT Q1 Q3 DP EP CE CXP
Bifenazate Acaricide 8.15 301.00 198.10 71 10 15 13
8.15 301.00 170.10 71 10 27 11
Bitertanol Fungicide 9.97 338.28 269.20 71 10 13 8
9.97 338.28 70.00 71 10 47 6
Boscalid Fungicide 7.34 343.00 307.00 111 10 27 17
7.34 343.00 140.00 116 10 27 13
Bromacil Herbicide 4.60 261.07 205.00 66 10 21 10
4.60 261.07 188.20 66 10 41 10
Buprofezin Insecticide 11.16 306.20 201.30 66 10 17 11
11.16 306.20 116.20 66 10 21 11
Cadusafos Insecticide 10.21 271.10 159.10 71 10 19 11
10.21 271.10 97.10 71 10 49 11
Cafenstrole Herbicide 7.95 351.20 100.20 71 10 19 11
7.95 351.20 72.10 71 10 39 11
Carbaryl Insecticide 4.87 202.10 145.10 71 10 16 10
4.87 202.10 127.10 71 10 39 10
Carbendazim Fungicide 3.35 192.10 160.10 91 10 27 13
3.35 192.10 132.00 91 10 41 11
Benomyl 3.35 192.01 160.10 60 10 25 6
3.35 192.01 132.10 60 10 41 12
Carbofuran Insecticide 4.58 222.10 165.10 70 10 17 9
4.58 222.10 123.00 70 10 29 9
Carboxin Fungicide 4.97 236.17 143.10 66 10 21 10
4.97 236.17 86.90 66 10 35 10
Carfentrazone-Ethyl Herbicide 9.32 412.00 345.80 91 10 27 13
9.32 412.00 365.90 91 10 21 15
Carpropamide Fungicide 9.44 334.10 139.10 76 10 29 11
9.44 334.10 103.10 76 10 57 11
Chlorantraniliprole Insecticide 6.37 482.04 283.90 6 10 21 10
6.37 482.04 450.90 6 10 25 18
Chlorfluazuron Insecticide 12.64 539.91 382.90 86 10 29 14
12.64 539.91 158.20 86 10 25 6
Chlorpyrifos Insecticide 11.88 349.93 97.00 71 10 53 8
(continued on next page)

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Table 1 (continued)

Compound Activity RT Q1 Q3 DP EP CE CXP

11.88 349.93 198.00 71 10 25 8

Chlorsulfuron Herbicide 4.85 358.00 141.00 81 10 23 11
4.85 358.00 167.10 81 10 25 11
Chromafenozide Insecticide 8.32 395.20 175.10 71 10 21 11
8.32 395.20 147.20 71 10 61 11
Cinosulfuron Herbicide 4.18 414.16 182.90 111 10 21 8
4.18 414.16 83.00 111 10 59 8
Compound Activity RT Q1 Q3 DP EP CE CXP
Clethodim Herbicide 10.87 360.10 164.00 76 10 27 13
10.87 360.10 77.00 76 10 99 11
Clofentezine Acaricide 10.38 303.00 138.10 71 10 19 11
10.38 303.00 102.10 71 10 55 11
Clomazone Herbicide 6.49 240.10 125.10 81 10 25 11
6.49 240.10 89.00 81 10 61 11
Clothianidin Insecticide 3.54 250.10 169.10 66 10 17 21
3.54 250.10 132.10 66 10 17 21
Cyazofamid Fungicide 8.64 325.10 107.90 71 10 19 11
8.64 325.10 261.00 71 10 15 13
Cyclosulfamuron Herbicide 8.26 422.00 260.80 71 10 23 13
8.26 422.00 217.90 71 10 33 13
Cyflufenamid Fungicide 10.15 413.08 295.10 71 10 21 12
10.15 413.08 203.00 71 10 49 8
Cyhalofop-butyl Herbicide 10.58 358.11 256.00 76 10 17 10
10.58 358.11 120.00 76 10 35 10
Cymoxanil Fungicide 3.82 199.02 128.00 51 10 13 10
3.82 199.02 83.00 51 10 35 8
a
Cyproconazole (I,II) Fungicide 8.01,8.68 292.10 70.14 81 10 33 11
8.01,8.68 292.10 125.17 81 10 41 11
Cyprodinil Fungicide 8.79 226.10 93.00 96 10 41 8
8.79 226.10 77.10 96 10 63 8
Cyromazine Insecticide 1.58 167.03 85.00 56 10 23 8
1.58 167.03 125.00 56 10 23 10
Demeton-S-methyl Insecticide 4.66 231.03 89.00 16 10 13 8
4.66 231.03 61.00 16 10 41 6
Diafenthiuron Insecticide 12.41 385.00 329.00 46 10 33 14
12.41 385.00 278.00 46 10 43 12
Diazinon Insecticide 9.72 305.20 169.20 86 10 27 11
9.72 305.20 153.20 86 10 29 11
Dichlofluanid Fungicide 7.95 350.00 332.90 31 10 9 12
7.95 350.00 127.00 31 10 21 12
Dichlorvos Insecticide 4.51 220.87 109.00 66 10 23 8
4.51 220.87 79.10 66 10 50 8
Diethofencarb Fungicide 6.86 268.00 226.00 71 10 15 15
6.85 268.00 180.00 66 10 23 14
Difenoconazole Fungicide 10.25 406.00 251.00 81 10 23 19
10.25 406.00 337.00 126 10 23 19
Diflubenzuron Insecticide 8.86 311.00 158.10 76 10 19 11
8.86 311.00 141.10 76 10 45 11
Compound Activity RT Q1 Q3 DP EP CE CXP
Dimepiperate Herbicide 10.49 264.13 146.10 6 10 13 6
10.49 264.13 119.10 6 10 23 10
Dimethametryn Herbicide 8.38 256.20 186.10 91 10 25 11
8.38 256.20 71.00 91 10 43 11
Dimethenamide Herbicide 7.10 276.10 244.10 76 10 19 13
7.10 276.10 168.30 76 10 31 11
Dimethoate Insecticide 3.68 229.98 199.00 46 10 13 8
3.68 229.98 125.00 46 10 27 10
Dimethomorph (E) Fungicide 7.04 388.10 301.10 101 10 25 11
7.04 388.10 165.20 101 10 45 11
Dimethomorph (Z) 7.54 388.11 301.10 101 10 25 11
7.54 388.11 165.20 101 10 45 11
Dimethylvinphos Insecticide 7.95 330.96 127.00 66 10 15 10
7.94 330.96 169.90 66 10 47 8
Diniconazole Fungicide 10.04 326.10 70.00 21 10 61 6
10.04 326.10 158.70 21 10 37 18
Diphenamid Herbicide 6.13 240.20 134.20 91 10 27 11
6.13 240.20 165.10 91 10 55 11
Dithiopyr Herbicide 10.95 402.10 354.00 96 10 23 13
10.95 402.10 271.90 96 10 39 13
Diuron Herbicide 5.84 233.10 72.10 86 10 35 11
5.84 233.10 160.10 86 10 35 11
Dymron Herbicide 7.74 269.10 151.10 86 10 17 11
7.74 269.10 119.10 86 10 25 11
Edifenphos Fungicide 9.39 311.10 111.10 86 10 29 11
9.39 311.10 109.10 86 10 41 11
(continued on next page)

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T.-W. Na, et al. Journal of Chromatography B 1141 (2020) 122021

Table 1 (continued)

Compound Activity RT Q1 Q3 DP EP CE CXP

Emamectine B1a Insecticide 10.89 886.90 82.20 131 10 119 11

10.89 886.90 158.30 131 10 49 11
Emamectine B1b 10.48 872.80 82.20 126 10 119 11
10.48 872.80 158.30 126 10 51 11
EPN Insecticide 10.62 324.10 296.00 16 10 19 12
10.62 324.10 156.90 16 10 29 14
Esprocarb Herbicide 11.26 266.20 91.10 81 10 33 11
11.26 266.20 65.10 81 10 79 11
Ethaboxam Fungicide 5.16 321.02 183.10 86 10 31 8
5.16 321.02 200.20 86 10 35 8
Ethiofencarb Insecticide 5.09 226.10 106.90 61 10 21 9
5.09 226.10 164.10 61 10 11 10
Ethofenprox Insecticide 13.68 394.30 177.20 71 10 21 11
13.68 394.30 135.20 71 10 33 11
Compound Activity RT Q1 Q3 DP EP CE CXP
Ethoprophos Insecticide 8.35 243.00 130.90 66 10 27 10
8.35 243.00 96.90 66 10 43 10
Ethoxyquin Fungicide 6.14 218.21 160.20 81 10 43 10
6.14 218.21 174.20 81 10 39 10
Ethoxysulfuron Herbicide 7.79 399.00 260.90 76 10 21 13
7.79 399.00 217.90 76 10 33 13
Etoxazole Acaricide 12.15 360.20 141.10 96 10 43 11
12.15 360.20 63.00 96 10 129 11
Etrimfos Insecticide 9.58 293.10 265.00 86 10 21 13
9.58 293.10 125.10 86 10 33 11
Famoxadone Fungicide 9.83 392.16 331.20 36 10 13 10
9.83 392.16 238.10 36 10 25 10
Fenamidone Fungicide 7.24 312.10 236.00 61 10 19 10
7.24 312.10 92.00 61 10 37 8
Fenamiphos Nematicide 8.83 304.20 217.00 56 10 29 8
8.83 304.20 202.00 56 10 45 8
Fenarimol Fungicide 8.28 331.00 268.00 101 10 27 13
8.28 331.00 81.00 106 10 27 13
Fenazaquin Acaricide 12.67 307.20 161.20 91 10 19 11
12.67 307.20 147.10 91 10 59 11
Fenbuconazole Fungicide 8.67 337.20 125.10 96 10 41 11
8.67 337.20 70.00 96 10 35 11
Fenobucarb Insecticide 6.66 208.10 95.20 67 10 19 10
6.66 208.10 152.00 67 10 11 10
Fenothiocarb Acaricide 9.06 254.08 72.00 56 10 33 8
9.06 254.08 160.00 56 10 13 8
Fenoxanil Fungicide 6.95 329.10 156.10 156 10 43 13
6.95 329.10 172.20 156 10 49 11
Fenoxaprop-P-Ethyl Herbicide 11.00 362.00 288.10 86 10 23 13
11.00 362.00 121.00 86 10 37 11
Fenoxycarb Insecticide 9.06 302.20 116.20 76 10 15 11
9.06 302.20 256.20 76 10 17 13
Fenpyroximate Acaricide 12.42 422.00 366.00 66 10 23 21
12.42 422.00 135.00 56 10 41 13
Fenthion Insecticide 9.66 279.10 247.10 86 10 17 13
9.66 279.10 169.20 86 10 23 11
Fentrazamide Herbicide 9.48 350.19 154.10 61 10 17 10
9.48 350.19 83.00 61 10 35 10
Ferimzone (z) Fungicide 6.26 255.20 132.00 86 10 27 17
6.26 255.20 124.20 86 10 27 17
Compound Activity RT Q1 Q3 DP EP CE CXP
Flonicamid Insecticide 3.34 229.95 203.10 86 10 23 13
3.34 229.95 148.00 86 10 39 11
TFNA 3.21 192.02 148.00 21 10 31 6
3.21 192.02 98.10 21 10 45 11
TFNG 3.27 248.98 203.10 86 10 31 13
3.27 248.98 148.10 86 10 39 13
Fluacrypyrim Acaricide 10.55 427.00 145.10 71 10 33 11
10.55 427.00 205.10 71 10 17 13
Flubendiamide Insecticide 9.39 408.10 274.10 45 10 23 10
9.39 683.03 408.00 45 10 15 10
Flucetosulfuron (Er) Herbicide 6.59 488.16 156.00 56 10 23 6
6.59 488.16 273.10 56 10 35 10
Flucetosulfuron (Th) 6.79 488.16 156.00 56 10 23 6
6.79 488.16 273.10 56 10 35 10
Fludioxonil Fungicide 7.27 266.09 229.00 1 10 15 8
7.27 266.09 158.00 1 10 45 8
Flufenacet Herbicide 8.46 364.10 152.20 71 10 25 11
8.46 364.10 194.30 71 10 17 11
Flufenoxuron Insecticide 12.18 489.00 158.10 91 10 27 11
(continued on next page)

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Table 1 (continued)

Compound Activity RT Q1 Q3 DP EP CE CXP

12.18 489.00 141.10 91 10 71 11

Flumioxazin Herbicide 6.34 355.00 327.10 45 10 30 8
6.34 355.00 299.10 45 10 36 8
Fluopicolide Fungicide 7.68 383.00 173.00 91 10 35 10
7.68 383.00 109.00 91 10 91 12
Fluopyram Fungicide 8.20 397.02 208.00 81 10 29 8
8.20 397.02 173.00 81 10 35 8
Fluquinconazole Fungicide 8.17 375.90 307.10 91 10 33 13
8.17 375.90 108.00 91 10 69 11
Flusilazole Fungicide 8.84 316.10 247.10 101 10 23 13
8.84 316.10 165.20 101 10 39 11
Flutolanil Fungicide 7.62 324.00 262.00 81 10 23 11
7.62 324.00 242.00 86 10 23 11
Forchlorfenuron Plant growth regulator 5.77 248.10 129.00 81 10 21 11
5.77 248.10 155.10 81 10 19 11
Fosthiazate Insecticide 5.14 284.10 228.10 76 10 15 11
5.14 284.10 104.20 76 10 29 11
Furathiocarb Insecticide 11.16 383.20 195.20 81 10 23 11
11.16 383.20 252.10 81 10 17 11
Gibberellic acid Plant growth regulator 3.59 364.15 239.10 56 10 21 8
3.59 364.15 311.00 56 10 19 14
Compound Activity RT Q1 Q3 DP EP CE CXP
Halosulfuron-Methyl Herbicide 8.37 435.00 182.10 76 10 31 11
8.37 435.00 139.00 76 10 59 11
Haloxyfop Herbicide 9.32 362.08 316.00 76 10 23 10
9.32 362.08 287.90 76 10 35 12
Haloxyfop-P-Methyl 10.52 376.00 315.90 96 10 21 13
10.52 376.00 287.90 96 10 29 13
Hexaconazole Fungicide 9.63 314.10 70.00 96 10 39 11
9.63 314.10 159.10 96 10 43 11
Hexaflumuron Insecticide 10.82 461.00 158.20 91 10 25 11
10.82 461.00 141.10 91 10 59 11
Hexazinone Herbicide 4.59 253.23 171.10 71 10 23 10
4.59 253.23 71.10 71 10 45 10
Hexythiazox Acaricide 11.76 353.10 228.00 76 10 23 15
11.76 353.10 168.10 76 10 37 13
Imazalil Fungicide 4.58 297.05 158.90 41 10 27 8
4.58 297.05 255.10 41 10 23 10
Imazosulfuron Herbicide 7.43 412.90 153.10 76 10 19 11
7.43 412.90 257.90 76 10 33 13
Imibenconazole Fungicide 11.35 411.00 125.00 88 10 50 11
11.35 411.00 171.00 88 10 28 11
Imicyafos Nematicide 4.18 305.10 201.10 51 9 31 4
4.18 305.10 235.20 51 9 25 4
Imidacloprid Insecticide 3.50 256.00 209.00 65 10 21 14
3.50 256.00 175.00 65 10 25 14
Inabenfide Plant growth regulator 6.92 339.14 320.90 81 10 23 12
6.92 339.14 80.00 81 10 67 6
Iprobenfos Fungicide 9.10 288.97 205.00 61 10 15 8
9.10 288.97 91.00 61 10 12 8
Iprovalicarb Fungicide 8.24 321.20 119.10 66 10 35 11
8.24 321.20 203.20 66 10 15 13
Isoprocarb Insecticide 5.49 194.20 95.20 71 10 19 11
5.49 194.20 137.10 71 10 13 11
Isoprothiolane Fungicide 7.71 291.10 231.10 71 10 15 13
7.71 291.10 189.10 71 10 27 11
Isopyrazam Fungicide 10.30 360.20 244.20 56 10 33 4
10.30 360.20 320.30 56 10 29 4
Kresoxim-methyl Fungicide 9.38 314.16 116.00 56 10 23 10
9.38 314.16 131.10 56 10 33 12
Linuron Herbicide 6.91 249.00 160.00 75 10 23 11
6.91 249.00 182.10 75 10 19 11
Compound Activity RT Q1 Q3 DP EP CE CXP
Lufenuron Insecticide 11.70 510.90 158.20 60 10 27 8
11.70 510.90 141.20 60 10 67 8
Malathion Insecticide 7.75 331.00 127.00 71 10 17 11
7.75 331.00 99.10 71 10 31 11
Mandipropamid Fungicide 7.57 411.80 328.10 76 10 21 13
7.57 411.80 125.00 76 10 47 11
Mefenacet Herbicide 7.98 299.10 148.20 81 10 19 11
7.98 299.10 120.20 81 10 35 11
Mepanipyrim Fungicide 8.35 224.00 106.00 96 10 35 11
8.35 224.00 77.00 96 10 35 11
Mepronil Fungicide 7.67 270.20 119.10 91 10 31 11
7.67 270.20 91.20 91 10 53 11
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6
T.-W. Na, et al. Journal of Chromatography B 1141 (2020) 122021

Table 1 (continued)

Compound Activity RT Q1 Q3 DP EP CE CXP

Metalaxyl Fungicide 5.67 280.20 220.20 76 10 17 11

5.67 280.20 160.20 76 10 31 11
Metamifop Herbicide 11.07 441.10 288.00 101 10 23 13
11.07 441.10 180.20 101 10 27 11
Metazosulfuron Herbicide 7.25 476.10 182.20 41 9 31 4
7.25 476.10 295.10 41 9 25 4
Metconazole Fungicide 9.73 320.20 70.00 91 10 43 11
9.73 320.20 125.00 91 10 61 11
Methabenzthiazuron Herbicide 5.55 222.10 165.20 76 10 23 11
5.55 222.10 150.10 76 10 45 11
Methidathion Insecticide 6.29 303.10 85.10 76 10 27 11
6.29 303.10 145.10 76 10 15 13
Methiocarb Insecticide 7.02 226.20 169.20 71 10 13 11
7.02 226.20 121.00 71 10 23 11
Methomyl Insecticide 3.33 163.10 88.10 61 10 13 10
3.33 163.10 106.20 61 10 13 10
Methoxyfenozide Insecticide 7.83 369.30 149.20 66 10 21 11
7.83 369.30 133.10 66 10 31 11
Metobromuron Herbicide 5.48 259.00 170.00 66 10 23 8
5.48 259.00 148.10 66 10 21 8
Metolachlor Herbicide 8.59 284.16 252.00 31 10 19 10
8.59 284.16 176.20 31 10 33 8
Metolcarb Insecticide 4.30 166.03 109.00 46 10 15 8
4.30 166.03 94.00 46 10 41 8
Metrafenone Fungicide 10.23 409.04 209.10 66 10 23 10
10.23 409.04 227.00 66 10 29 10
Metribuzin Herbicide 4.64 215.06 187.10 61 10 23 8
4.64 215.06 59.80 61 10 71 6
Compound Activity RT Q1 Q3 DP EP CE CXP
Mevinphos Insecticide 3.86 224.99 192.90 56 10 9 8
3.86 224.99 127.00 56 10 19 10
Milbemectin A3 Acaricide 12.93 511.29 95.10 86 10 47 10
12.93 511.29 105.10 86 10 89 12
Milbemectin A4 13.37 525.36 91.00 86 10 109 12
13.37 525.36 55.10 86 10 93 12
Molinate Herbicide 7.69 188.20 126.20 71 10 17 11
7.69 188.20 55.10 71 10 35 11
Monocrotophos Insecticide 3.37 224.10 127.10 66 10 21 11
3.37 224.10 109.10 66 10 41 11
Myclobutanil Fungicide 7.71 289.20 70.00 86 10 27 11
7.71 289.20 125.10 86 10 41 11
Napropamide Herbicide 8.46 272.20 171.20 86 10 23 13
8.46 272.20 129.20 86 10 21 11
Nereistoxin Insecticide 0.82 150.00 105.00 41 10 21 10
0.82 150.00 61.00 41 10 33 6
Nicosulfuron Herbicide 10.23 411.08 209.10 71 10 19 8
10.23 411.08 228.90 71 10 23 10
Novaluron Insecticide 10.97 493.10 158.10 86 10 27 11
10.97 493.10 141.10 86 10 69 11
Nuarimol Fungicide 6.90 315.00 252.00 101 10 31 13
6.90 315.00 81.00 101 10 31 13
Ofurace Fungicide 4.55 282.10 254.10 86 10 17 13
4.55 282.10 160.20 86 10 31 11
Omethoate Insecticide 3.08 214.14 125.00 66 10 31 10
3.08 214.14 109.10 66 10 39 10
Oxadiazon Herbicide 11.48 345.10 303.00 61 10 20 11
11.48 362.10 220.00 61 10 31 11
Oxadixyl Fungicide 4.09 279.20 219.20 76 10 15 13
4.09 279.20 133.20 76 10 27 11
OxamyI Insecticide 3.23 237.03 72.00 6 10 27 6
3.23 237.03 90.10 6 10 11 8
Oxaziclomefon Herbicide 11.02 376.00 190.10 86 10 21 11
11.02 376.00 133.10 86 10 47 11
Paclobutrazole Plant growth regulator 7.44 294.18 70.00 46 10 47 6
7.44 294.18 125.10 46 10 45 11
Penconazole Fungicide 9.12 284.10 159.10 81 10 39 11
9.12 284.10 70.10 81 10 27 11
Pencycuron Fungicide 10.16 329.00 125.00 86 10 33 11
10.16 329.00 99.00 86 10 83 15
Compound Activity RT Q1 Q3 DP EP CE CXP
Pendimethalin Herbicide 11.97 282.16 212.10 31 10 15 8
11.97 282.16 194.10 31 10 23 8
Penoxsulam Herbicide 4.94 484.05 195.10 56 10 37 8
4.94 484.05 194.50 56 10 41 8
Penthiopyrad Fungicide 9.33 360.13 276.00 76 10 19 10
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7
T.-W. Na, et al. Journal of Chromatography B 1141 (2020) 122021

Table 1 (continued)

Compound Activity RT Q1 Q3 DP EP CE CXP

9.33 360.13 255.90 76 10 27 10

Pentoxazone Herbicide 11.16 354.05 286.00 86 10 17 10
11.16 354.05 186.00 86 10 35 8
Phenthoate Insecticide 9.35 321.03 79.00 66 10 57 6
9.35 321.03 135.00 66 10 27 10
Phorate Insecticide 10.08 261.04 75.00 46 10 13 8
10.08 261.04 46.90 46 10 45 6
Phosalone Insecticide 10.02 368.10 182.10 81 10 21 13
10.02 368.10 75.20 81 10 95 11
Phosphamidone Insecticide 4.10 300.10 127.10 86 10 25 11
4.10 300.10 174.20 86 10 19 11
Phoxim Insecticide 10.11 299.10 129.00 71 10 17 11
10.11 299.10 76.90 71 10 45 11
Picoxystrobin Fungicide 9.22 368.09 205.10 56 10 13 8
9.22 368.09 145.00 56 10 29 6
Piperophos Herbicide 10.44 354.10 255.00 91 10 19 15
10.44 354.10 171.00 91 10 29 13
Pirimicarb Insecticide 4.47 239.20 72.10 73 10 34 10
4.47 239.20 182.20 73 10 21 9
Pirimiphos-methyl Insecticide 10.04 306.20 164.20 96 10 29 11
10.04 306.20 108.20 96 10 39 11
Probenazole Plant activator 4.33 224.01 41.10 61 10 27 4
4.33 224.01 39.00 61 10 57 4
Prochloraz Fungicide 9.51 376.05 308.00 36 10 17 12
9.51 376.05 265.90 36 10 23 10
Profenofos Insecticide 10.89 373.00 302.70 86 10 23 13
10.89 373.00 128.00 86 10 59 11
Prometryn Herbicide 7.26 242.14 158.00 56 10 31 8
7.26 242.14 200.10 56 10 25 8
Propamocarb Fungicide 2.98 189.20 102.20 76 10 23 11
2.98 189.20 74.00 76 10 35 11
Propanil Herbicide 6.89 218.10 162.10 86 10 19 11
6.89 218.10 127.10 86 10 37 11
Propaquizafop Herbicide 11.32 444.12 100.10 81 10 33 10
11.32 444.12 56.00 81 10 51 10
Compound Activity RT Q1 Q3 DP EP CE CXP
Propiconazole Fungicide 9.48 342.07 158.90 66 10 33 14
9.48 342.07 69.00 66 10 39 8
Propoxur Insecticide 4.53 210.20 111.10 66 10 19 11
4.53 210.20 93.10 66 10 33 11
Pymetrozin Insecticide 3.05 218.20 105.10 81 10 27 11
3.05 218.20 79.20 81 10 55 11
Pyraclofos Insecticide 9.95 361.10 111.10 101 10 85 11
9.95 361.10 138.10 101 10 55 11
Pyraclostrobin Fungicide 9.96 388.00 194.00 46 10 19 14
9.96 388.00 163.00 51 10 29 14
Pyrazolynate Herbicide 10.19 439.10 173.10 91 10 25 11
10.19 439.10 155.00 91 10 25 11
Pyrazophos Fungicide 10.16 374.00 222.20 96 10 27 11
10.16 374.00 194.20 96 10 43 11
Pyrethrin Insecticide 12.27 329.28 161.20 66 10 17 10
12.27 329.28 105.20 66 10 43 10
Pyribenzoxim Herbicide 11.50 610.20 413.10 61 10 23 15
11.50 610.20 180.20 61 10 47 11
Pyributicarb Herbicide 11.76 331.10 181.10 70 10 21 10
11.76 331.10 108.00 70 10 40 10
Pyridaben Insecticide 12.76 365.00 147.00 96 10 31 11
12.76 365.00 309.00 91 10 31 11
Pyridaphenthion Insecticide 7.94 341.04 189.10 61 10 29 8
7.94 341.04 205.10 61 10 29 8
Pyrifluquinazon Insecticide 7.75 465.10 423.10 61 11 27 8
7.75 465.10 107.20 61 11 45 8
Pyriftalid Herbicide 6.65 319.17 139.20 96 10 41 10
6.65 319.17 83.10 96 10 65 10
Pyrimethanil Fungicide 6.56 200.20 107.20 96 10 31 11
6.56 200.20 82.10 96 10 35 11
Pyrimidifen Insecticide 11.05 378.10 184.20 96 10 31 11
11.05 378.10 150.20 96 10 45 11
Pyriminobac-methyl (E) Herbicide 6.63 362.10 330.10 76 10 19 13
6.63 362.10 75.10 66 10 109 12
Pyriminobac-methyl (Z) 7.54 362.13 330.20 66 10 21 10
7.54 362.13 284.10 76 10 33 13
Pyrimisulfan Herbicide 6.11 420.10 370.00 36 8 23 6
6.11 420.10 388.10 36 8 21 6
Pyriproxyfen Insecticide 11.84 322.00 96.00 56 10 21 13
(continued on next page)

8
T.-W. Na, et al. Journal of Chromatography B 1141 (2020) 122021

Table 1 (continued)

Compound Activity RT Q1 Q3 DP EP CE CXP

11.84 322.00 185.00 51 10 29 14

Compound Activity RT Q1 Q3 DP EP CE CXP
Pyroquilon Fungicide 4.42 174.10 132.00 101 10 31 11
4.42 174.10 117.10 101 10 41 11
Quinalphos Insecticide 9.31 299.14 97.10 76 10 51 10
9.31 299.14 163.10 76 10 33 10
Quinmerac Herbicide 3.83 222.10 204.00 71 10 19 11
3.83 222.10 141.10 71 10 43 11
Quinoclamine Herbicide 4.40 208.10 105.10 101 10 33 11
4.40 208.10 77.10 101 10 51 11
Quizalofop-ethyl Herbicide 11.01 373.13 299.20 101 10 25 10
11.01 373.13 163.10 101 10 59 10
Salflufenacil Herbicide 6.65 501.10 349.00 46 10 35 6
6.65 501.10 198.00 46 10 59 6
Sethoxydim Herbicide 11.28 328.20 282.30 81 10 17 11
11.28 328.20 178.10 81 10 25 13
Silafluofen Insecticide 14.33 426.15 287.00 30 10 15 10
14.33 426.15 168.00 30 10 49 8
Simeconazole Fungicide 8.31 294.20 70.00 41 10 53 8
8.31 294.20 72.90 41 10 49 6
Simetryn Herbicide 4.84 214.06 124.10 71 10 25 12
4.84 214.06 68.00 71 10 49 6
Spinetoram (J) Insecticide 9.95 748.52 98.10 101 10 101 12
9.95 748.52 142.20 101 10 45 10
Spinetoram (L) 10.55 760.49 98.20 106 10 91 12
10.55 760.49 142.20 106 10 43 10
SpinosynA Insecticide 9.24 732.50 142.30 116 10 39 11
9.24 732.50 99.10 116 10 67 11
SpinosynD 9.85 746.50 142.20 116 10 39 11
9.85 746.50 99.20 116 10 65 11
Spirodiclofen Insecticide 12.38 411.20 71.10 55 10 25 4
12.38 411.20 313.10 55 10 17 8
Spiromesifen Insecticide 11.98 371.20 273.20 65 10 19 6
11.98 371.20 255.20 65 10 29 6
Sulfoxaflor Insecticide 3.73 278.08 174.00 61 10 11 8
3.73 278.08 153.90 61 10 37 12
Tebuconazole Fungicide 9.28 308.00 70.00 96 10 41 11
9.28 308.00 125.00 96 10 41 11
Tebufenozide Insecticide 8.98 353.10 133.10 71 10 25 11
8.98 353.10 297.10 66 10 25 11
Tebufenpyrad Acaricide 11.18 334.20 145.20 116 10 37 11
11.18 334.20 117.20 116 10 51 11
Compound Activity RT Q1 Q3 DP EP CE CXP
Tebupirimfos Insecticide 11.44 319.12 277.00 56 10 19 10
11.44 319.12 153.10 56 10 37 8
Teflubenzuron Insecticide 11.40 381.10 158.20 66 10 23 13
11.40 381.10 141.10 66 10 57 11
Terbuthylazine Herbicide 7.15 230.10 174.20 86 10 21 13
7.15 230.10 104.00 86 10 43 11
Terbutryn Herbicide 7.40 242.13 186.10 36 10 25 8
7.40 242.13 91.00 36 10 35 8
Tetraconazole Fungicide 8.42 372.00 159.00 76 10 47 11
8.42 372.00 70.00 81 10 37 15
Thenylchlor Herbicide 8.42 324.09 127.10 46 10 19 6
8.42 324.09 97.00 46 10 59 8
Thiabendazole Fungicide 3.50 202.10 175.10 96 10 33 13
3.50 202.10 131.10 96 10 43 11
Thiacloprid Insecticide 3.77 253.10 126.10 81 10 27 17
3.77 253.10 186.10 121 10 19 16
Thiamethoxam Insecticide 3.34 292.00 211.20 66 10 17 11
3.34 292.00 181.20 66 10 27 11
Thiazopyr Herbicide 9.53 397.06 377.00 56 10 31 12
9.53 397.06 335.00 56 10 39 14
Thidiazuron Plant growth regulator 4.53 221.10 102.00 81 10 19 11
4.53 221.10 128.00 81 10 25 11
Thifensufuron-Methyl Herbicide 4.34 388.00 167.10 81 10 21 11
4.34 388.00 204.90 81 10 33 13
Thiobencarb Herbicide 10.11 258.10 125.00 81 10 27 11
10.11 258.10 89.30 81 10 67 11
Thiodicarb Insecticide 5.02 355.10 88.10 71 10 23 11
5.02 355.10 108.10 71 10 21 11
Thiophanate-methyl Fungicide 4.47 343.10 151.10 81 10 25 11
4.47 343.10 226.20 81 10 15 11
Tiadinil Fungicide 7.83 268.10 101.10 61 10 27 8
7.83 270.10 101.10 61 10 27 8
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T.-W. Na, et al. Journal of Chromatography B 1141 (2020) 122021

Table 1 (continued)

Compound Activity RT Q1 Q3 DP EP CE CXP

Tolclofos-methyl Fungicide 10.21 301.02 124.90 81 10 21 12

10.21 301.02 175.00 81 10 33 14
Tolyfluanid Fungicide 9.44 363.97 238.00 51 10 19 8
9.44 363.97 137.00 51 10 37 12
Triadimefon Fungicide 7.70 294.10 197.20 91 10 19 11
7.70 294.10 225.00 91 10 19 11
Triazophos Insecticide 8.15 314.10 162.20 81 10 27 11
8.15 314.10 119.20 81 10 47 11
Compound Activity RT Q1 Q3 DP EP CE CXP
Tricyclazole Fungicide 3.94 189.99 163.10 46 10 31 6
3.94 189.99 136.00 46 10 39 6
Trifloxystrobin Fungicide 10.72 409.00 186.00 51 10 23 14
10.72 409.00 206.00 46 10 21 14
Triflumizole Fungicide 10.44 346.07 278.00 26 10 15 8
10.44 346.07 73.00 26 10 23 11
Triflumuron Insecticide 9.98 359.10 156.10 91 10 23 13
9.98 359.10 139.10 91 10 45 11
Uniconazole Fungicide 7.62 292.11 70.00 46 5 41 4
7.62 292.11 125.10 46 5 39 4
Vamidothion Insecticide 3.59 287.91 146.10 66 10 19 11
3.59 287.91 118.00 66 10 35 11
Bentazone (-) Herbicide 4.44 238.90 196.90 −5 −10 −26 −13
4.44 238.90 174.90 −5 −10 −26 −13

a
Cyproconazole including two peaks.

intensity, selectivity, and sensitivity of the ion were measured for the
obtained spectrum, and the quantifier and qualifier were determined.
The optimized MRM conditions are shown in Table 1.
2.4. Selection of experimental samples
Five types of crops, which can represent the characteristics of each
taxon, were selected in this study. Rice represents high starch; soybean
represents oil seeds,; potato represents roots/tubers, which contain
much water; pepper represents fruiting vegetables, which also contain
large amounts of water; and mandarin represents fruits, with high acid
and water content. All samples were purchased from an organic agri-
cultural farm and stored after being crushed separately with dry ice
at −40 °C (soybean and rice were crushed without dry ice).
2.5. Sample preparation
2.5.1. MFDS#83 method protocol
A 50-g representative chopped sample was homogenized with
100 mL of acetonitrile for 5 min and filtered with Whatman 6 filter
paper. The filtrate was transferred to a 500-mL separatory funnel, and
15 g of NaCl was added. The sample mixture was then shaken for 5 min
with a mechanical shaker and kept 1 h for separation. From the upper
layer, 20 mL was transferred to a 100-mL separatory funnel and par-
titioned with 20 mL of n-hexane saturated with acetonitrile. The lower
acetonitrile layer was collected, and the upper n-hexane layer was re-
extracted with 20 mL of acetonitrile saturated with n-hexane. Both the
acetonitrile layers were combined and evaporated for purification.
The extract was re-dissolved in a 4-mL mixture of MeOH:DCM (1:99
v/v) and loaded to a 500-mg amino cartridge, which was previously
conditioned with 6 mL of DCM. The cartridge was then eluted with
6 mL of MeOH:DCM (1:99 v/v) mixture and evaporated. The purified
extract was then reconstituted with 2 mL of acetonitrile and filtered
with a membrane filter.
2.5.2. QuEChERS EN method protocol
From the homogenized sample, 10 g (5 g for rice and soybean) was
added to a 50-mL Teflon centrifuge tube to which 10 mL of acetonitrile
(15 mL for soybean) was added and shaken for 1 min. Then, 4.0 g of
MgSO4, 1.0 g of NaCl, 1.0 g of trisodium citrate dihydrate, and 0.5 g of
disodium hydrogen citrate sesquihydrate was added and shaken again
for 1 min. The tube was thereafter centrifuged for 5 min at 4000 rpm.
Afterwards, 1.0 mL of the supernatant was transferred to a 2 mL micro-
centrifuge tube to which 150 mg of MgSO4, and 25 mg of PSA (primary
secondary amine) were added as purification sorbent and vortexed for
20 s. The micro-centrifuge tube was then centrifuged for 5 min at
4000 rpm. The purified supernatant was filtered through a 0.2-µm
syringe filter and transferred to a 2-mL vial for UPLC-MS/MS analysis.
2.6. Method validation
Method was validated through the standard parameters of the
SANTE guidelines, such as limit of quantification, linearity, selectivity,
specificity, accuracy, and precision [32]. Limit of quantification (LOQ)
represents the lowest concentration or mass of the analyte that pro-
vided signal 10 times higher than the noise level. Linearity was assessed
from the coefficient of determination (R2) after constructing a six-point
calibration curve using concentration vs area. Selectivity is the identical
retention time of the standard peak, and the peak recovered after for-
tification of the same standard to the blank sample. Specificity is the
absence of interference or interference ≤ 30% of the LOQ at the
standard retention time in the blank samples. Accuracy is the average
recovery of the fortified standard from the blank samples with three
replicates. Precision was measured from the relative standard deviation
derived from the three-replicate analysis.
2.7. Statistical analysis
The recovery and RSD value of 267 pesticides extracted from two
different methods (MFDS#83 and QuEChERS EN) in five matrices
(mandarin, pepper, potato, rice, and soybean) at two different con-
centration level (10 × LOQ, 50 × LOQ) were analyzed and compared.
The acceptable range of the recovery values and the %RSD values were
considered as 70–130% and 0–30 respectively. Data analysis and entry
were accomplished by the use of statistical package SPSS. The data
were analyzed in two sections. The first section of the analysis was
made up of a non-parametric binomial test for comparing two methods
(MFDS#83 and QuEChERS EN) and also for comparing two con-
centration levels (10 × LOQ, 50 × LOQ), the second section was made
up of a non-parametric chi-square test for comparing the matrices
(Mandarin, pepper, Potato, Rice, and Soybean). The null hypotheses of
interest were (i) MFDS#83 and QuEChERS EN methods were equally

10
T.-W. Na, et al.
Fig. 1. Recovery and repeatability percentage of 267 pesticides in five representative matrices (a) MFDS#83 method at 10 × LOQ level; (b) MFDS#83 method at
50 × LOQ level; (c) QuEChERS EN at 10 × LOQ level; and (d) QuEChERS EN at 50 × LOQ level.
likely to fulfill the requirement (value at acceptable range) (ii)
10 × LOQ and 50 × LOQ concentrations levels were equally likely to
fulfill the requirement, and (iii) All five matrices (mandarin, pepper,
potato, rice, and soybean) were equally likely to fulfill the requirement.
In hypothesis testing, the null hypothesis will be rejected at 5% level of
significance if p-value of that test is less than 0.05. Rejection of null
hypothesis in (i), (ii) and (iii) indicates significant differences in the
proportions fulfill the requirement.
3. Results and discussion
3.1. Method validation
Linearity at the level of 0.002–0.1 mg/kg was verified with the es-
tablished analytical conditions of UPLC-MS/MS for both methods.
Excellent linearity was observed for the matrix-matched calibration
curve constructed with the extract of QuEChERS EN, showing coeffi-
cients of determination (R2) ≥ 0.99 for all compounds except three.
Meanwhile, good linearity was also observed from the matrix-matched
calibration extracted from MFDS#83 with coefficients of determination
(R2) ≥ 0.99 for most of the compounds except ten. In both methods, a
lower linearity was observed for a few compounds with a determination
of coefficient (R2) ≥ 0.97.
The limit of quantification was calculated using a S/N ratio based on
the detection limit. This value was dependent on the dilution ratio of
the dilution method used. The limit of quantification for all the analytes
in the five representative matrices following the MFDS#83 method was
11
Journal of Chromatography B 1141 (2020) 122021
0.0004 mg/kg. On the other hand, the quantitative limits of the
QuEChERS EN method were set to 0.002 mg/kg for potato, citrus and
pepper, 0.004 mg/kg for brown rice, and 0.006 mg/kg for soybean. The
LOQ of the MFDS#83 method was approximately ten times lower than
that of the QuEChERS EN method because of the concentration process
(evaporation step) involved in the traditional method.
For the recovery test, five representative agricultural products were
prepared and spikes at 10 times LOQ and 50 times LOQ. According to
the project guideline the recovery and RSD tolerances were estimated at
the 70%–130% and ≤30% levels, respectively. A table with the re-
coveries at 10 × LOQ is provided as supplementary information.
3.2. Statistical interpretation
The MFDS#83 and QuEChERS EN method were compared in terms
of recovery and the null hypothesis is rejected as the observed p-value
was 0.029. This result indicates the proportion of two methods differs
significantly and QuEChERS EN method predominant (51.7%) in terms
of pesticides at an acceptable range. The p-value for the concentration
levels 10 × LOQ and 50 × LOQ was observed as 0.795 in the
MFDS#83 method and was 0.737 in the QuEChERS EN method con-
sequently the null hypothesis of equality of proportion couldn’t be re-
jected in both the methods. Therefore the two concentration levels were
equally likely to fulfill the requirement within the MFDS#83 and
QuEChERS EN methods. The p-value for testing the equality of pro-
portion among the five matrices was 0.535 in the MFDS#83 method but
it was 0.001 in the QuEChERS EN method. Therefore five matrices were
T.-W. Na, et al.
equally likely to fulfill the requirement within the MFDS#83 methods.
In contrast, five matrices differ significantly in fulfilling the require-
ment (value at an acceptable range) within the QuEChERS EN method.
In QuEChERS EN method Rice was predominant (21.6%) in terms of the
acceptable range.
In the case of RSD, the MFDS#83 and QuEChERS EN method were
compared and the null hypothesis is rejected as the observed p-value
was 0.000. This result indicates the proportion of two methods differs
significantly and QuEChERS EN method predominant (52.5%) in terms
of pesticides at an acceptable range. The p-value for the concentration
levels 10 × LOQ and 50 × LOQ was observed as 0.934 in the
MFDS#83 method and was 0.829 in the QuEChERS EN method con-
sequently the null hypothesis of equality of proportion couldn’t be re-
jected in both the methods. Therefore the two concentration levels were
equally likely to fulfill the requirement within the MFDS#83 and
QuEChERS EN methods. Similarly, the p-value for testing the equality
of proportion among the five matrices was 0.295 in the MFDS#83
method and was 0.986 in the QuEChERS EN method consequently the
null hypothesis of equality of proportion couldn’t be rejected in both
the methods. Therefore the five matrices were equally likely to fulfill
the requirement within the MFDS#83 and QuEChERS EN methods.
3.3. Comparative study of MFDS#83 and QuEChERS EN methods
The recovery and repeatability percentages of 267 compounds in
the MFDS#83 and QuEChERS EN methods are shown in Fig. 1.
In the MFDS#83 method, the number of compounds that satisfy the
guidelines of 70–130% recovery and RSD of 30% or less at the
10 × LOQ level were 191–216, where the highest number of com-
pounds was found in the potato matrix, followed by pepper, mandarin
rice, and soybean. At 50 × LOQ, 191–220 compounds were recovered
in the order of potato, mandarin, pepper, rice, and soybean.
On the other hand, in QuEChERS EN method, at the 10 × LOQ
level, 189–247 compounds satisfied the guidelines (recovery 70–130%
and RSD 30% or less). The highest recovery rate was found for rice,
followed by soybean, potato, pepper, and mandarin. Only 189 com-
pounds were recovered for mandarin, which were somewhat lower than
those for other crops. At 50 × LOQ, 179–241 compounds were re-
covered, in which the highest number of compounds were recovered
from potato, followed by soybean, rice, pepper, and mandarin.
3.4. Comparison in terms of recovery percentage
The 267 compounds and the compounds that satisfy the guidelines
in terms of recovery rate and RSD at the levels of 10 × LOQ and
50 × LOQ in both methods are shown in Fig. 2. To observe the
Fig. 2. Average percent recovery of 267 compounds at the (A) 10 × LOQ level; (B) 50 × LOQ level.
12
Journal of Chromatography B 1141 (2020) 122021
difference based on the fortified concentrations, the recovery percent
for each concentration was compared. As shown in the figure, at the
10 × LOQ level, the QuEChERS EN method showed better results than
the MFDS#83 method by 4.87% for pepper, 5.62% for potato, 13.86%
for rice, and 16.48% for soybean. However, MFDS#83 showed better
results by 7.86% for mandarin than the QuEChERS EN method at the
same concentration level. On the other hand, at 50 × LOQ level, the
QuEChERS EN method showed 1.12% better results for pepper, 7.87%
for potato, 10.11% for rice, and 17.98% for soybean, but for mandarin,
the extraction efficiency in terms of number of compounds was 13.86%
lower than that of the MFDS#83 method. The results at the 10 × LOQ
and 50 × LOQ levels were slightly different because of the different
spiking concentrations. Therefore, the number of compounds that sa-
tisfy the guidelines at both concentration levels were compared. The
results showed that the QuEChERS EN method was 3.00% better for
potato, 11.60% for rice, and 11.60% for soybean. For mandarin, the
MFDS#83 method showed 14.4% better results, and the results for
pepper were similar in both methods. As opposed to these two matrices,
potato, rice, and soybean showed good results in the QuEChERS EN
method for all comparison results.
3.5. Traditional to modern methodology
Although the QuEChERS EN method showed no significant differ-
ence with the MFDS#83 method in the recovery test, it showed com-
paratively better results in the method validation test. The recovery
results showed that none of the compounds met the guidelines. At the
low level (10 × LOQ level), the QuEChERS EN method showed better
results compared with the MFDS#83 method except mandarin, which
might be due to the low pH of mandarin influencing recovery during
the QuEChERS EN method. Five representative matrices were selected
in this study with regard to pH, fat, pigment, and starch with high
consumption on a regular basis. Monitoring methods are primarily used
as qualitative method and fastness is important in analyzing the field
sample. The QuEChERS EN method involves a lower amount of toxic
solvent consumption, no use of a glass apparatus, and no time-con-
suming cartridge purification, like the traditional MFDS#83 method
does. Therefore, it is expected that QuEChERS EN method can be
practically applied for multi-residue analysis.
4. Conclusions
To upgrade the multi-residue analytical method, the QuEChERS EN
method and traditional MFDS#83 method were compared for the
screening of 267 pesticides/metabolites/plant activators/growth reg-
ulators in five representative matrices using UPLC-MS/MS. The
T.-W. Na, et al.
matrices (mandarin, pepper, potato, rice, and soybean) were selected in
terms of pH, pigments, starch, and fat along with high consumption
rate. Both methods provided good linearity and low LOQ for most of the
analytes. The QuEChERS EN method provided better results in terms of
recovery and repeatability than the MFDS#83 method for all matrices
except mandarin. As a quick screening method, the QuEChERS EN
method can be an upgraded replacement of the MFDS#83 method for
pepper, potato, rice, and soybean, or similar types of matrices. Further
modification of the QuEChERS EN version is needed for acidic/low pH
matrices, such as mandarin.
Conflict of interest
The authors declare no conflict of interest.
CRediT authorship contribution statement
Tae-Woong Na: Conceptualization, Methodology, Software, Data
curation, Writing - original draft. Md. Musfiqur Rahman:
Conceptualization, Methodology, Software, Data curation, Writing -
original draft. Sung-Woo Kim: Visualization, Investigation. M.
Ershadul Haque: . Jong-Bang Eun: Writing - review & editing. Jae-
Han Shim: Supervision.
Acknowledgments
This study was supported by a grant of Ministry of Food and Drug
Safety (MFDS) (Project no. 15162MFDS071), Republic of Korea.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.jchromb.2020.122021.
References
[1] B.M. Keikothaile, P. Spanoghe, W. Steurbaut, Effects of food processing on pesticide
residues in fruits and vegetables, Food Chem. Toxicol. 48 (2010) 1–6.
[2] A. Hercegova, M. Domotorova, E. Matisova, M. Kirchner, R. Otrekal, V. Stefuca,
Fast gas chromatography with solid phase extraction clean-up for ultratrace ana-
lysis of pesticide residues in baby food, J. Chromatogr. A 1084 (2005) 46–53.
[3] O. Tiryaki, C. Temur, The Fate of Pesticide in the Environment, J. Biol. Environ. Sci.
4 (2010) 29–38.
[4] W.G. Fong, H.A. Moye, J.N. Seiber, J.P. Toth, Pesticide residues in foods: Methods,
Techniques, and Regulations, John wiley & Sons Inc, 1999.
[5] A. Hercegova, M. Domotorova, E. Matisova, Sample preparation methods in the
analysis of pesticide residues in baby food with subsequent chromatographic de-
termination, J. Chromatogr. A 1153 (2007) 54–73.
[6] Pesticide analysis method practical guide of Food code (4th edition), National
Institute of Food and Drug Safety Evaluation, 2013.
[7] A. Yang, A.M. Abd El-Aty, J.H. Park, A. Goudah, M.M. Rahman, J.A. Do, O.J. Choi,
J.H. Shim, Analysis of 10 systemic pesticide residues in various baby foods using
liquid chromatography, Biomed. Chromatogr. 28 (2014) 735–741.
[8] WHO, International Code of Conduct on the Distribution and Use of Pesticides:
Guidelines for the Registration of Pesticides. World Health Organization; Rome,
Italy: 2010.
[9] E. Bempelou, K.S. Liapis, G.E. Miliadis, Validation of single residue methods for the
determination of dithiocarbamates and inorganic bromide residues in plant pro-
ducts, Hell. Plant Prot. J. 5 (2012) 57–64.
[10] S. Grimalt, P. Dehouck, Review of analytical methods for the determination of
pesticide residues in grapes, J. Chromatogr. A 1433 (2016) 1–23.
[11] S. Walorczyk, Development of a multi-residue method for the determination of
pesticides in cereals and dry animal feed using gas chromatography–tandem
quadrupole mass spectrometryII. Improvement and extension to new analytes, J.
Chromatogr. A 1208 (2008) 202–214.
13
Journal of Chromatography B 1141 (2020) 122021
[12] T.D. Nguyen, E.M. Han, M.S. Seo, S.R. Kim, M.Y. Yun, D.M. Lee, G.H. Lee, A multi-
residue method for the determination of 203 pesticides in rice paddies using gas
chromatography/mass spectrometry, Anal. Chim. Acta 619 (2008) 67–74.
[13] J.M. Lee, J.W. Park, G.C. Jang, K.J. Hwang, Comparative study of pesticide multi-
residue extraction in tobacco for gas chromatography–triple quadrupole mass
spectrometry, J. Chromatogr. A 1187 (2008) 25–33.
[14] T. Cajka, J. Hajslova, O. Lacina, K. Mastovska, S.J. Lehotay, Rapid analysis of
multiple pesticide residues in fruit-based baby food using programmed temperature
vaporiser injection–low-pressure gas chromatography-high-resolution time-of-flight
mass spectrometry, J. Chromatogr. A 1186 (2008) 281–294.
[15] K. Zhang, J.W. Wong, D.G. Hayward, P. Sheladia, A.J. Krynitsky, F.J. Schenck,
M.G. Webster, J.A. Ammann, S.E. Ebeler, Multiresidue Pesticide Analysis of Wines
by Dispersive Solid-Phase Extraction and Ultrahigh-Performance Liquid
Chromatography-Tandem Mass Spectrometry, J. Agric. Food Chem. 57 (2009)
4019–4029.
[16] C. Diez, W.A. Traag, P. Zommer, P. Marinero, J. Atienza, Comparison of an acet-
onitrile extraction/partitioning and “dispersive solid-phase extraction“ method
with classical multi-residue methods for the extraction of herbicide residues in
barley samples, J. Chromatogr. A 1131 (2006) 11–23.
[17] W.J. Choe, Simultaneous analysis of pesticide multi-residues in agricultural pro-
ducts using GC-MS/MS, Gyeongsang National University, doctoral thesis, 2013.
[18] National Agricultural Products Quality Management Service, hazardous substances
analysis method for agricultural products: Korea Food and Drug Safety notice 2013-
138 (amendment, 2013.4.5).
[19] M. Anastassiades, S.J. Lehotay, D. Štajnbaher, F.J. Schenck, Fast and Easy
Multiresidue Method Employing Acetonitrile Extraction/Partitioning and
“Dispersive Solid-Phase Extraction” for the Determination of Pesticide Residues in
Produce, J. AOAC Int. 86 (2003) 412–431.
[20] S.J. Lehotay, M. Hiemstra, P. van Bodegraven, Validation of Fast and Easy Method
for the Determination of Residues from 229 Pesticides in Fruits and Vegetables
Using Gas and Liquid Chromatography and Mass Spectrometric Detection, J. AOAC
Int. 88 (2005) 595–614.
[21] P. Payá, M. Anastassiades, D. Mark, I. Sigalova, B. Tasdelen, J. Oliva, A. Barba,
Analysis of pesticide residues using the Quick Easy Cheap Effective Rugged and Safe
(QuEChERS) pesticide multiresidue method in combination with gas and liquid
chromatography and tandem mass spectrometric detection, Anal. Bioanal. Chem.
389 (2007) 1697–1714.
[22] QuEChERS: A Mini-multiresidue method for the analysis of pesticide residues in
low-fat products.(www.Quechers.com, 2009).
[23] S.J. Lehotay, K.A. Son, H. Kwon, U. Koesukwiwat, W. Fu, K. Mastovska, E. Hoh,
N. Leepipatpiboon, Comparison of QuEChERS sample preparation methods for the
analysis of pesticide residues in fruits and vegetables, J. Chromatogr. A 1217 (2010)
2548–2560.
[24] K. Mastovska, K.J. Dorweiler, S.J. Lehotay, W.J. Segscheid, K.A. Szpylka, Pesticide
Multiresidue Analysis in Cereal Grains Using Modified QuEChERS Method
Combined with Automated Direct Sample Introduction GC-TOFMS and UPLC-MS/
MS Techniques, J. Agric. Food Chem. 58 (2010) 5959–5972.
[25] U. Koesukwiwat, S.J. Lehotay, K. Mastovska, K.J. Dorweiler, N. Leepipatpiboon,
Extension of the QuEChERS Method for Pesticide Residues in Cereals to Flaxseeds,
Peanuts, and Doughs, J. Agric. Food Chem. 58 (2010) 5950–5958.
[26] F.J. Schenck, J.E. Hobbs, Evaluation of the Quick, Easy, Cheap, Effective, Rugged,
and Safe (QuEChERS) Approach to Pesticide Residue Analysis, Bull. Environ.
Contam. Toxicol. 73 (2004) 24–30.
[27] U. Koesukwiwat, K. Sanguankaew, N. Leepipatpiboon, Rapid determination of
phenoxy acid residues in rice by modified QuEChERS extraction and liquid chro-
matography–.tandem mass spectrometry, Anal. Chim. Acta 626 (2008) 10–20.
[28] Multi-residue pesticide screens, California Department of Food and Agriculture,
Division of Inspection Services, Chemistry Laboratory Services Branch, Pesticide
Residue Program. Joe T, 1988.
[29] M. Luke, J.E. Froberg, H.T. Masumoto, Extraction and cleanup of organochlorine,
organophophate, organonitrogen and hydrocarbon pesticides in produce for de-
termination by gas-liquid chromatography, J. Assoc. Off. Anal. Chem. 58 (1975)
1020–1026.
[30] P.A. Mills, J.H. Omley, R.A. Guither, Rapid method for chlorinated pesticide re-
sidues in nonfatty foods, J. Assoc. Off. Anal. Chem. 46 (1963) 186–191.
[31] S.M. Lee, M.L. Papathakis, H.M.C. Feng, G.F. Hunter, J.E. Carr, Multipesticide re-
sidue method for fruits and vegetables: California Department of Food and
Agriculture, Fresenjus Journal of Analytical Chemists 339 (1991) 376–383.
[32] Directorate General for Health and Food Safety, Guidance document on analytical
quality control and method validation procedures for pesticide residues and ana-
lysis in food and feed, Doc. no. SANTE/11813/2017. European Commission,
Brussels, 2007.