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Determination of pyraclostrobin residue in wax gourd and its dietary risk assessment

Abstract

To study the residue behavior of pyraclostrobin in wax gourd, and evaluate the risk of dietary intake. Wax gourd samples were extracted with acetonitrile, purified with PSA and C18, detected by HPLC-MS/MS. In 2017, the supervised field trials of pyraclostrobin in wax gourd were carried out in Henan, Jiangsu, Sichuan, Beijing, Hunan, and Guangxi. The STMR of pyraclostrobin in wax gourd was acquired, and then the risk of dietary intake was evaluated. At the three spiked levels of 0.01, 0.1 and 1.0 mg kg-1, the average recoveries ranged from 95% to 104%, the RSDs ranged from 2.1% to 3.8%, and LOQ was 0.01 mg kg-1. The dissipation of pyraclostrobin in wax gourd in Sichuan fitted to the first order kinetics with the half-life of 4.1d. The 250 gL-1 emulsifiable concentrate of pyraclostrobin was sprayed at 150 and 225 g a.i./hm2 for 2-3 times on wax gourd. The final residue level of pyraclostrobin in wax gourd was between <LOQ and 0.11 mgkg-1. The national estimated daily intake (NEDI) of pyraclostrobin was 0.3722 mg, accounting for 19.69% of the acceptable daily intake (ADI) for general population, which means that it will not pose unacceptable risks to the health of the general population.

Keywords:
pyraclostrobin; wax gourd; residue; dietary risk

1 Introduction

Wax gourd (Benincasa hispida), also known as ash gourd, white pumpkin, and white gourd, belonging to the Cucurbitaceae family, genus Benincasa, is long season vegetable widely planted in Asia and other semi-tropical countries (Xie et al., 2019Xie, D., Xu, Y. C., Wang, J. P., Liu, W. R., Zhou, Q., Luo, S. B., Huang, W., He, X. M., Li, Q., Peng, Q. W., Yang, X. Y., Yuan, J. Q., Yu, J. G., Wang, X. Y., Lucas, W. J., Huang, S. W., Jiang, B., & Zhang, Z. H. (2019). The wax gourd genomes offer insights into the genetic diversity and ancestral cucurbit karyotype. Nature Communications, 10(1), 5158. http://dx.doi.org/10.1038/s41467-019-13185-3. PMid:31727887.
http://dx.doi.org/10.1038/s41467-019-131...
). It is popular and healthy food because of containing rich nutrients, polysaccharides, dietary fiber, low sodium and high potassium (Huang et al., 2011Huang, G., Tan, J. T., Tan, X. C., & Peng, D. Q. (2011). Preparation of polysaccharides from wax gourd. International Journal of Food Sciences and Nutrition, 62(5), 480-483. http://dx.doi.org/10.3109/09637486.2010.550275. PMid:21355748.
http://dx.doi.org/10.3109/09637486.2010....
; Shakya et al., 2019Shakya, A., Gogoi, N., Chaudhary, S. K., Bhat, H. R., & Ghosh, S. K. (2019). Development and validation of a high-performance thin-layer chromatography method for the quantification of rutin in the fruit pulp of Benincasa hispida (Thunb.) cogniaux. Jpc-Journal of Planar Chromatography-Modern Tlc, 32(5), 371-377. http://dx.doi.org/10.1556/1006.2019.32.5.4.
http://dx.doi.org/10.1556/1006.2019.32.5...
). Wax gourd has also the medical effects of anti-inflammation, diuresis and detumescence (Dobre et al., 2014Dobre, C., Elena, T., Ionita, L., & Berca, L. M. (2014). Benincasa hispida (wax gourd) technological characteristics and biological proprieties. Romanian Biotechnological Letters, 19(4), 9504-9509.; Varghese & Raj, 2021Varghese, H. S., & Raj, G. G. (2021). Protective role of Benincasa hispida (Thunb.) Cogn. fruit extract against gentamicin induced nephrotoxicity in rats. Indian Journal of Experimental Biology, 59(9), 633-639.). However, it is vulnerable to some diseases of early blight, gray mold, powdery mildew, and which seriously affect the yield and quality of wax gourd (Jiang et al., 2018Jiang, D., Duan, L. F., Li, R. N., Pan, X. L., Tao, Y., Wu, X. H., Xu, J., Liu, X. G., Dong, F. S., & Zheng, Y. Q. (2018). Evaluation of the safe application of copper nonylphenolsulfonate and hexaconazole in wax gourd under field conditions. Ecotoxicology and Environmental Safety, 159, 71-76. http://dx.doi.org/10.1016/j.ecoenv.2018.04.037. PMid:29730411.
http://dx.doi.org/10.1016/j.ecoenv.2018....
). Pyraclostrobin has a good control effect on powdery mildew, gray mildew, downy mildew and other diseases (Bowness et al., 2016Bowness, R., Gossen, B. D., Chang, K. F., Goswani, R., Willenborg, C. J., Holtz, M., & Strelkov, S. E. (2016). Sensitivity of mycosphaerella pinodes to pyraclostrobin fungicide. Plant Disease, 100(1), 192-199. http://dx.doi.org/10.1094/PDIS-03-15-0350-RE. PMid:30688575.
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; Chen et al., 2012Chen, Y., Zhang, A. F., Gao, T. C., Zhang, Y., Wang, W. X., Ding, K. J., Chen, L., Sun, Z., Fang, X. Z., & Zhou, M. G. (2012). Integrated use of pyraclostrobin and epoxiconazole for the control of fusarium head blight of wheat in Anhui Province of China. Plant Disease, 96(10), 1495-1500. http://dx.doi.org/10.1094/PDIS-01-12-0099-RE. PMid:30727306.
http://dx.doi.org/10.1094/PDIS-01-12-009...
; Fan et al., 2019Fan, K., Wang, J., Fu, L., Zhang, G. F., Wu, H. B., Feng, C. C., & Qu, J. L. (2019). Baseline sensitivity and control efficacy of pyraclostrobin against botryosphaeria dothidea isolates in China. Plant Disease, 103(7), 1458-1463. http://dx.doi.org/10.1094/PDIS-07-18-1214-RE. PMid:31025906.
http://dx.doi.org/10.1094/PDIS-07-18-121...
; Skandalis et al., 2016Skandalis, N., Dimopoulou, A., Beri, D., Tzima, A., Malandraki, I., Theologidis, I., Bitivanos, S., Varveri, C., Klitsinaris, T., & Vassilakos, N. (2016). Effect of pyraclostrobin application on viral and bacterial diseases of tomato. Plant Disease, 100(7), 1321-1330. http://dx.doi.org/10.1094/PDIS-10-15-1216-RE. PMid:30686190.
http://dx.doi.org/10.1094/PDIS-10-15-121...
).

Pyraclostrobin, N-[2-[[1-(4-chlorophenyl)pyrazol-3-yl]oxymethyl]phenyl]-N-methoxycarbamate is a methoxyacrylate fungicide developed by BASF, which inhibits mitochondrial respiration by blocking electron transfer between cytochromes b and c1, so that the recipient cell cannot produce and supply the energy for normal metabolism, resulting in cell death (Bowness et al., 2016Bowness, R., Gossen, B. D., Chang, K. F., Goswani, R., Willenborg, C. J., Holtz, M., & Strelkov, S. E. (2016). Sensitivity of mycosphaerella pinodes to pyraclostrobin fungicide. Plant Disease, 100(1), 192-199. http://dx.doi.org/10.1094/PDIS-03-15-0350-RE. PMid:30688575.
http://dx.doi.org/10.1094/PDIS-03-15-035...
). Pyraclostrobin has the functions of protection, treatment and internal absorption in the control of crop diseases. It has the characteristics of broad spectrum and high efficiency and been used in more than 150 crops in more than 60 countries around the world. According to the pesticide registration data of China Pesticide Information Network (http://www.chinapesticide.org.cnhysj/index.jhtml), pyraclostrobin has been registered in 79 crops, such as rice, wheat, peanut, cucumber, pepper, potato, onions, watermelon, apple, honeysuckle, tea and so on. Besides, it can improve crop physiological function and enhance crop stress resistance (Li et al., 2020Li, P., Sun, P. Y., Li, D., Li, D. L., Li, B. H., & Dong, X. L. (2020). Evaluation of pyraclostrobin as an ingredient for soybean seed treatment by analyzing its accumulation-dissipation kinetics, plant-growth activation, and protection against phytophthora sojae. Journal of Agricultural and Food Chemistry, 68(43), 11928-11938. http://dx.doi.org/10.1021/acs.jafc.0c04376. PMid:33078613.
http://dx.doi.org/10.1021/acs.jafc.0c043...
; Xiong et al., 2020Xiong, H., Liu, X. F., Fan, Y. H., Luan, S. R., & Huang, Q. C. (2020). Effect of membrane integrity on survival competition ofBotrytis cinereaupon QoI fungicide pyraclostrobin. Journal of Phytopathology, 168(10), 601-608. http://dx.doi.org/10.1111/jph.12940.
http://dx.doi.org/10.1111/jph.12940...
). However, pyraclostrobin is highly toxic to fish, invertebrates and amphibians in aquatic ecosystems (Mao et al., 2020Mao, L., Jia, W., Zhang, L., Zhang, Y. N., Zhu, L. Z., Sial, M. U., & Jiang, H. Y. (2020). Embryonic development and oxidative stress effects in the larvae and adult fish livers of zebrafish (Danio rerio) exposed to the strobilurin fungicides, kresoxim-methyl and pyraclostrobin. The Science of the Total Environment, 729, 139031. http://dx.doi.org/10.1016/j.scitotenv.2020.139031. PMid:32387777.
http://dx.doi.org/10.1016/j.scitotenv.20...
; Skanes et al., 2021Skanes, B., Warriner, K., & Prosser, R. S. (2021). Hazard assessment using an in-silico toxicity assessment of the transformation products of boscalid, pyraclostrobin, fenbuconazole and glyphosate generated by exposure to an advanced oxidative process. Toxicology In Vitro, 70, 105049. http://dx.doi.org/10.1016/j.tiv.2020.105049. PMid:33171224.
http://dx.doi.org/10.1016/j.tiv.2020.105...
; Zhang et al., 2017Zhang, C., Wang, J., Zhang, S., Zhu, L. S., Du, Z. K., & Wang, J. H. (2017). Acute and subchronic toxicity of pyraclostrobin in zebrafish (Danio rerio). Chemosphere, 188, 510-516. http://dx.doi.org/10.1016/j.chemosphere.2017.09.025. PMid:28910725.
http://dx.doi.org/10.1016/j.chemosphere....
). Therefore, it is necessary to investigate the dissipation fate in agricultural products and environmental samples to ensure food and environment safety.

According to the research reported, a variety of detection methods have been used to detect pyraclostrobin residues in different plant substrates. Pyraclostrobin residue analysis is mainly conducted through gas chromatography(GC) (Lagunas-Allue et al., 2015Lagunas-Allue, L., Sanz-Asensio, J., & Martinez-Soria, M. T. (2015). Mobility and distribution of eight fungicides in surface, skin and pulp in grapes. An application to pyraclostrobin and boscalid. Food Control, 51, 85-93. http://dx.doi.org/10.1016/j.foodcont.2014.10.028.
http://dx.doi.org/10.1016/j.foodcont.201...
; Podbielska et al., 2018Podbielska, M., Szpyrka, E., Piechowicz, B., Sadlo, S., & Sudol, M. (2018). Assessment of boscalid and pyraclostrobin disappearance and behavior following application of effective microorganisms on apples. Journal of Environmental Science and Health. Part B, Pesticides, Food Contaminants, and Agricultural Wastes, 53(10), 652-660. http://dx.doi.org/10.1080/03601234.2018.1474554. PMid:30024824.
http://dx.doi.org/10.1080/03601234.2018....
), GC coupled with mass spectrometry (GC-MS) (Menezes et al., 2010Menezes, A. Fo., Santos, P. N., & Pereira, P. A. D. (2010). Development, validation and application of a method based on DI-SPME and GC-MS for determination of pesticides of different chemical groups in surface and groundwater samples. Microchemical Journal, 96(1), 139-145. http://dx.doi.org/10.1016/j.microc.2010.02.018.
http://dx.doi.org/10.1016/j.microc.2010....
), GC coupled with tandem mass spectrometry (GC-MS/MS) (González-Rodríguez et al., 2009González-Rodríguez, R. M., Cancho-Grande, B., & Simal-Gandara, J. (2009). Multiresidue determination of 11 new fungicides in grapes and wines by liquid-liquid extraction/clean-up and programmable temperature vaporization injection with analyte protectants/gas chromatography/ion trap mass spectrometry. Journal of Chromatography. A, 1216(32), 6033-6042. http://dx.doi.org/10.1016/j.chroma.2009.06.046. PMid:19576591.
http://dx.doi.org/10.1016/j.chroma.2009....
), high-performance liquid chromatography (HPLC) (Lin et al., 2020Lin, S. K., Yang, L. P., Zheng, Q., Wang, Y. Q., Cheng, D. M., & Zhang, Z. X. (2020). Dissipation and distribution of pyraclostrobin in bananas at different temperature and a risk assessment of dietary intake. International Journal of Environmental Analytical Chemistry, 102(17), 1-13.), HPLC coupled with tandem mass spectrometry (HPLC-MS/MS) (Zhao et al., 2020Zhao, H., Zhao, Y., & Hu, J. Y. (2020). Dissipation, residues and risk assessment of pyraclostrobin and picoxystrobin in cucumber under field conditions. Journal of the Science of Food and Agriculture, 100(14), 5145-5151. http://dx.doi.org/10.1002/jsfa.10563. PMid:32519755.
http://dx.doi.org/10.1002/jsfa.10563...
, 2021Zhao, Z., Sun, R. X., Su, Y., Hu, J. Y., & Liu, X. L. (2021). Fate, residues and dietary risk assessment of the fungicides epoxiconazole and pyraclostrobin in wheat in twelve different regions, China. Ecotoxicology and Environmental Safety, 207, 111236. http://dx.doi.org/10.1016/j.ecoenv.2020.111236. PMid:32911182.
http://dx.doi.org/10.1016/j.ecoenv.2020....
), ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) (Gao et al., 2019Gao, Y., Yang, S., Li, X. X., He, L. F., Zhu, J. M., Mu, W., & Liu, F. (2019). Residue determination of pyraclostrobin, picoxystrobin and its metabolite in pepper fruit via UPLC-MS/MS under open field conditions. Ecotoxicology and Environmental Safety, 182, 109445. http://dx.doi.org/10.1016/j.ecoenv.2019.109445. PMid:31330408.
http://dx.doi.org/10.1016/j.ecoenv.2019....
; He et al., 2021He, L., He, F. L., Yang, S., Gao, Y. Y., Li, B. X., Liu, F., & Mu, W. (2021). Dissipation kinetics and safety evaluation of pyraclostrobin and its desmethoxy metabolite BF 500-3 in a cucumber greenhouse agroecosystem. Environmental Science and Pollution Research International, 28(14), 17712-17723. http://dx.doi.org/10.1007/s11356-020-11798-6. PMid:33400109.
http://dx.doi.org/10.1007/s11356-020-117...
), and so on. However, no studies have been reported on any environmental residual behavior of pyraclostrobin in wax gourd at present. QuEChERS (quick, easy, cheap, effective, rugged, and safe) is a sample pretreatment technology based on dispersive solid phase extraction, and it has been widely used in recent years (Fazal et al., 2022Fazal, A., Ismail, A., Naeem, I., Oliveira, C. A. F., Shaukat, S., Saleem, M. U., Saima, S., Nasir, U., Alam, A., Aslam, Z., & Aslam, R. (2022). Exposure assessment of selected pesticide residues using occurrence data in foods and serum samples in Pakistan. Food Science and Technology (Campinas), 42, e01222. http://dx.doi.org/10.1590/fst.01222.
http://dx.doi.org/10.1590/fst.01222...
; Wang et al., 2022Wang, Y., Xing, L. J., Luo, R. F., Li, X. Y., Zhang, F., & Lu, S. L. (2022). Modified QuEChERS combined with UPLC-MS/MS to determine eight biogenic amines in Xinjiang smoked horsemeat sausages. Food Science and Technology (Campinas), 42, e93521. http://dx.doi.org/10.1590/fst.93521.
http://dx.doi.org/10.1590/fst.93521...
). This research develops a modified QuEChERS method coupled with HPLC-MS/MS and determine the residual pyraclostrobin in wax gourd.

In this study, we evaluated the residual dissipation dynamics of pyraclostrobin in wax gourd, hoping to provide a reference for the research and application of prevention and control of wax gourd diseases. In addition, we also evaluated the potential risk of pyraclostrobin exposure through the dietary intake of general populations.

2 Materials and methods

2.1 Materials

Standard of pyraclostrobin (99.0%, purity) was purchased from Dr. Ehrenstorfer GmbH (Augsburg, Germany). Chromatography-grade acetonitrile (ACN) and methanol were obtained from Sigma-Aldrich (Steinheim, Germany). Chromatography-grade formic acid was supplied by Anaqua Global International Inc, Ltd.(Cleveland, the United States of America). Sodium chloride (NaCl) and anhydrous magnesium sulfate (MgSO4) were of analytical-grade and obtained from Sinopharm Chemical Reagent Co., Ltd (Beijing, China). The sorbents of primary secondary amine (PSA), octadecylsilyl (C18) and 0.22 μm nylon syringe filters were provided by Agela Technologies (Tianjin, China).

2.2 Field experimental design

The experiment was carried out according to NY/T 788-2004 (Guideline on Pesticide Residue Trials) issued by the Institute of the Control of Agrochemicals, Ministry and Agriculture, People's Republic of China. The field trials were conducted during 2017-2018 at six different provinces of China, including Henan, Jiangsu, Sichuan, Beijing, Hunan, and Guangxi. The area of each experimental plot was 15 m2 and each treatment had three replications. No pesticide was used during the whole period of wax gourd growth in the control treatment. A buffer area of 10 m2 was used to separate the plots of different treatments. The 250 gL-1 emulsifiable concentrate of pyraclostrobin was used as tested pesticide.

The dissipation dynamics trials were carried out in Henan province and Sichuan province, and 250 gL-1emulsifiable concentrate of pyraclostrobin was dissolved in water and sprayed at 300 g a.i. ha-1 (2 times of the recommended high dosage) when wax gourd grows to half the size of mature individuals with one time. The samples were collected at 2 h (calculated as the initial residue), 1, 3, 5, 7, 14, 21, 28,35,45 days after spraying. According to the Standard operating procedures for Field trial of Pesticide Registration residues, wax gourd fruits (about 2 kg) were randomly collected in different directions and different parts of the each experimental plot, chopped and mixed, and the samples were prepared by quadruple method and stored at -20 °C.

To investigate the residue levels of the pyraclostrobin in wax gourd, the terminal residue experiment was carried out at the dosage level of 150 g a.i. ha−1 (recommended high dosage) and 225 g a.i. ha−1 (1.5 times of the recommended high dosage) for 2 and 3 times with 5 days interval, respectively. The experiment locations were in six different provinces of Henan, Jiangsu, Sichuan, Beijing, Hunan, and Guangxi. The wax gourd samples were collected from each plot at preharvest intervals of 5, 7, and 10 days. All samples were stored at −20 °C until analysis.

2.3 Sample extraction and purification

10.0 g wax gourd samples (accurate to 0.01 g) were weighed into a 50 mL centrifuge tube, then 10 mL acetonitrile and 2.5 g sodium chloride were added. The mixtures were vortexed vigorously for 10 minutes. The supernatant was centrifuged at 4000 r/min for 5 min, and the 1.5 mL of supernatant was loaded into a centrifuge tube containing 0.15 g anhydrous magnesium sulfate, 0.05 g C18 and 0.05 g PSA. The tubes were vortexed for 1 min and then centrifuged at 4000 r/min for 5 min. The upper layer filtered by 0.22 μm Nylon syringe filter was transferred to an auto-sampler vial for HPLC-MS/MS analysis.

2.4 HPLC-MS/MS detection

The chromatographic separation of pyraclostrobin was performed on a LC-30AD HPLC system connected to a triple-quadrupole LCMS-8050 equipped with an electrospray ionization source (ESI). The analyte was separated on a Agilent ZORBAX SB-Aq column (100 mm × 3.0 mm, 3.5 μm particle size). The mobile phase consisted of 0.1% formic acid aqueous solution (solvent A) and chromatography grade acetonitrile (solvent B), and VA:VB = 20:80, with a flow rate of 0.3 mL/min. The column oven temperature was maintained at 35 °C in order to decrease viscosity. The injection volume was set at 10 µL.

The MS/MS detection was performed in positive mode for pyraclostrobin and monitoring conditions were optimized for the target compound. The typical conditions were as follows: the capillary voltage was set at 4.0 kV, the desolvation temperature was held at 350 °C. The cone and desolvation gas flows were 3.0 and 10 L min-1, respectively. Multi-reaction monitoring (MRM) mode was selected as the scan mode and the MS/MS parameters are shown in Table 1.

Table 1
MS/MS parameters of pyraclostrobin.

2.5 Recovery experiments

The standard solutions of pyraclostrobin were spiked to the blank sample of wax gourd at appropriate concentrations of 0.01 mg/kg, 0.1 mg/kg, 1.0 mg/kg. Five parallel treatments for each spiked-level were carried out. These samples were pretreated and detected following the procedure as mentioned previously above.

2.6 Data analysis

The dissipation curves of pyraclostrobin in wax gourd were plotted according to the data of pesticide concentrations, and the degradation rate constant and half-life of degradation (DT50) were calculated using the first-order Equation 1 followed:

C t = C 0 e k t (1)

where C0 and Ct were represented sample initial concentration (mg kg-1) and residue concentration (mg kg-1) at time t (d). And the DT50 was calculated using the k value for each site separately (DT50 = ln 2/k).

The national estimated daily intake (NEDI) of pyraclostrobin and the risk quotient (RQ) were calculated using the following Formulas (2 and 3) (Xing et al., 2022Xing, L. J., Wang, Y., Luo, R. F., Li, X. Y., & Zou, L. J. (2022). Determination of 31 pesticide residues in wolfberry by LC-MS/MS and dietary risk assessment of wolfberry consumption. Food Science and Technology (Campinas), 42, e61921. http://dx.doi.org/10.1590/fst.61921.
http://dx.doi.org/10.1590/fst.61921...
).

N E D I = S T M R i × F i (2)
R Q = N E D I / A D I × b . w . × 100 % (3)

where STMRi (mg/kg) represented supervised trials median residue of pyraclostrobin in wax gourd in China, Fi referred to the daily intake of a certain agricultural product or food in China (kg), acceptable daily intake (ADI) represented the acceptable daily intake, b.w. was the average body weight of Chinese adult (63 kg). The ADI of pyraclostrobin formulated by China was 0.03 mg/kg b.w.

3 Results and discussion

3.1 Method validation

In this study, the external standard method accompanied by matrix matched calibration standards was used for quantitation to reduce matrix effect. A certain amount of pyraclostrobin was added to the wax gourd blank extracts to get the final working standard solution. And satisfactory linearity (y = 182083285.44x + 1847149.19) and correlation coefficient of determination (R2 = 09974, R2 > 0.99) were achieved over the concentration range of 0.01 -1.0 mg kg−1.

The accuracy of this method was evaluated by recoveries with five replications (n = 5) at three different fortifications. The relative standard deviation values (RSD, %) between tested concentrations were used to reveal the precision of the method. At the three spiked levels of 0.01, 0.1 and 1.0 mg kg−1, the average recoveries ranged from 95% to 104%, the relative standard deviations (RSDs) ranged from 2.1% to 3.8% (Table 2). The LOQ was defined as the minimum concentration of the analyte that has been validated with acceptable accuracy by applying the complete analytical method. In this paper, the LOQ for pyraclostrobin in wax gourd was 0.01 mg kg−1.

Table 2
Recoveries and RSDs of pyraclostrobin in wax gourd samples (n = 5).

3.2 Dissipation of pyraclostrobin in wax gourd

The initial concentrations of pyraclostrobin in wax gourds were 0.14 mg kg-1 in Sichuan and lower than LOQ in Henan. Because of the rapid descent of concentration, pyraclostrobin could not be detected in Henan and only one digestion curve in Sichuan was obtained (Figure 1). The dissipation equation in the representative location (Sichuan province) was Ct = 0.0754e-0.1690t. It demonstrated the residues of pyraclostrobin in wax gourd declined significantly with time and the reduction prominently followed first-order kinetic equation. According to the degrade curve, the degradation half-life of pyraclostrobin in Sichuan province was calculated through “DT50 = ln2/k” as 4.1 d. The dissipation behavior was influenced by the sunlight, temperature, humidity and other climatic factors. In a previous research, the half-lives of pyraclostrobin were 7.6 days on wheat in Shanxi province of China (Zhao et al., 2021Zhao, Z., Sun, R. X., Su, Y., Hu, J. Y., & Liu, X. L. (2021). Fate, residues and dietary risk assessment of the fungicides epoxiconazole and pyraclostrobin in wheat in twelve different regions, China. Ecotoxicology and Environmental Safety, 207, 111236. http://dx.doi.org/10.1016/j.ecoenv.2020.111236. PMid:32911182.
http://dx.doi.org/10.1016/j.ecoenv.2020....
), 8.2 days on cucumbers in southern China (Zhao et al., 2020Zhao, H., Zhao, Y., & Hu, J. Y. (2020). Dissipation, residues and risk assessment of pyraclostrobin and picoxystrobin in cucumber under field conditions. Journal of the Science of Food and Agriculture, 100(14), 5145-5151. http://dx.doi.org/10.1002/jsfa.10563. PMid:32519755.
http://dx.doi.org/10.1002/jsfa.10563...
), 6.20~7.79 days on Rosa roxburghii in China (Han et al., 2022aHan, L., Wu, Q., & Wu, X. M. (2022a). Dissipation and residues of pyraclostrobin in rosa roxburghii and soil under filed conditions. Foods, 11(5), 11. http://dx.doi.org/10.3390/foods11050669. PMid:35267302.
http://dx.doi.org/10.3390/foods11050669...
), 1.5-2.3 days on cowpea in China (Han et al., 2022bHan, S., Bi, Y. Y., Han, L. J., Song, S. Y., Ye, Z. Y., Qin, F. Y., & Lv, X. R. (2022b). Residue behavior and risk assessment of pyraclostrobin and thifluzamide in Cowpea. Bulletin of Environmental Contamination and Toxicology, 108(4), 786-790. http://dx.doi.org/10.1007/s00128-021-03421-2. PMid:35067727.
http://dx.doi.org/10.1007/s00128-021-034...
), 9.2-13 days in pomegranate fruits and 13.5-17 days in the leaves (Mohapatra et al., 2021Mohapatra, S., Siddamallaiah, L., & Matadha, N. Y. (2021). Behavior of acetamiprid, azoxystrobin, pyraclostrobin, and lambda-cyhalothrin in/on pomegranate tissues. Environmental Science and Pollution Research International, 28(22), 27481-27492. http://dx.doi.org/10.1007/s11356-021-12490-z. PMid:33506422.
http://dx.doi.org/10.1007/s11356-021-124...
). The degradation rate in the present study showed that pyraclostrobin was easily degradable pesticide.

Figure 1
Dissipation curve of of pyraclostrobin in wax gourd in Sichuan Province.

3.3 Terminal residues of pyraclostrobin in wax gourd

The data for the final residues of pyraclostrobin in wax gourd with a longer harvest interval was shown in Table 3. As the data shown, after 5 days of last application of pyraclostrobin, the residue of pyraclostrobin in wax gourd is < 0.01-0.092 mg kg-1, STMR is 0.01 mg kg-1, the highest residue (HR) is 0.092 mg kg-1; after 7 days of the last application, the residue of pyraclostrobin in wax gourd is < 0.01-0.11 mg kg-1, STMR is 0.01 mg kg-1, HR is 0.11 mg kg-1; after 10 days of the last application, the residual amount of pyraclostrobin in wax gourd was < 0.01-0.055 mg kg-1, STMR was 0.01 mg kg-1, HR was 0.055 mg kg-1. Pesticide residues in agricultural products are affected by the nature of pesticides, dosage, times of application, environmental and climatic conditions and other factors. After 3 days from the last treatment, the residues of copper nonylphenolsulfonate were below 0.38 mg kg−1 in wax gourd, and the residues of hexaconazole ranged from< 0.01 to 0.19 mg kg−1 in wax gourd (Jiang et al., 2018Jiang, D., Duan, L. F., Li, R. N., Pan, X. L., Tao, Y., Wu, X. H., Xu, J., Liu, X. G., Dong, F. S., & Zheng, Y. Q. (2018). Evaluation of the safe application of copper nonylphenolsulfonate and hexaconazole in wax gourd under field conditions. Ecotoxicology and Environmental Safety, 159, 71-76. http://dx.doi.org/10.1016/j.ecoenv.2018.04.037. PMid:29730411.
http://dx.doi.org/10.1016/j.ecoenv.2018....
). In this study, there are some differences in pyraclostrobin residues in wax gourd in different areas. It may be related to the differences in climatic conditions, wax gourd varieties and sample size in different experimental areas.

Table 3
Final residues of in wax gourd samples of pyraclostrobin.

3.4 Dietary risk assessment

The dietary risk probability was assessed by RQ, which was calculated by comparing the value of NEDI of pyraclostrobin with ADI. The ADI of pyraclostrobin is 0.03 mg kg−1 b.w. and the average Chinese adult body weight is 63 kg. Therefore, the ADI of pyraclostrobin for each person is 1.89 mg (0.03 mg kg−1 bw × 63 kg bw). According to the above calculation method, the NEDI values were calculated from the reference residue limit for a maximum dietary risk (Table 4). The supervised trials median residue (STMR) of pyraclostrobin concluded from the field trials were 0.01 mg kg−1 and as the reference residue limit of evaluated wax gourd. As shown in Table 4, the RQ of pyraclostrobin was 19.69%. Hence, the above results indicated that the application of pyraclostrobin in wax gourd with recommended dosage will not bring potential dietary risk for Chinese consumers.

Table 4
Dietary risk assessment form of pyraclostrobin.

4 Conclusion

In summary, this study established an HPLC-MS/MS method based on QuEChERS (quick, easy, cheap, effective, rugged, and safe)to detect the residues of pyraclostrobin in wax gourd samples through field trials in six locations of China. Samples were extracted with the acetonitrile solution and purified with dispersive solid phase extraction. The results of field trials showed that the terminal residues pyraclostrobin were all below 0.11 mg kg-1, which was far below the official MRLs recommended by China. In addition, the RQ value pyraclostrobin was 19.69%. In conclusion, the application of the commercial 250 gL-1 pyraclostrobin emulsifiable concentrate(EC) at the recommended dosage is safe.

  • Practical Application: It is urgent to evaluate the dietary intake risk of pyraclostrobin to general populations and monitor its residual dissipation in wax gourd.
  • Funding

    This work was supported by the Natural Science Foundation of Henan Province (No.182300410080) and the Key Scientific and Technological Projects of Henan Province (212102110456).

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Publication Dates

  • Publication in this collection
    16 Jan 2023
  • Date of issue
    2023

History

  • Received
    31 Oct 2022
  • Accepted
    21 Dec 2022
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