Enantioselective toxicity assessment of prothioconazole on earthworms (Eisenia foetida) in artificial soil environments.

The chiral fungicide prothioconazole (PTZ) is extensively employed in agricultural practices, prompting serious concern due to its environmental impact. PTZ is prone to undergo metabolism, leading to the formation of chiral prothioconazole-desthio (dPTZ) in the environment. However, limited knowledge exists regarding its enantioselective behavior and toxicity towards invertebrate organisms in soil ecosystems. In this study, R-(-)- and S-(+)- PTZ enantiomers were individually synthesized, and their stereoselective toxicity effects on earthworms (E. foetida) were studied in artificial soil under environmentally relevant concentration exposures. The results showed a significant accumulation of dPTZ in earthworms, surpassing the levels of PTZ. Moreover, the concentration of S-(-)- dPTZ in earthworms was notably higher than that of R-(+)- dPTZ after exposure, reaching peak levels on day 14. Concurrently, oxidative stress induced by S-(+)- PTZ enantiomers in earthworms exhibited a substantial increase compared to R-(-)- enantiomers on day 14, indicating a higher ecological risk associated with the former in non-target organisms. Transcriptome analysis unveiled distinct impacts on earthworm physiology. S-(+)-PTZ exposure significantly affected energy metabolism, immune responses and digestive systems. In contrast, R-(-)-PTZ exposure influenced the synthesis of carbohydrates, proteins, and lipids. These insights contribute to understanding the complex interactions between PTZ enantiomers and soil-dwelling organisms, providing a scientific foundation for advancing the application of high efficiency, low toxicity PTZ monomer pesticides.

Impacts of prothioconazole and prothioconazole-desthio on bile acid and glucolipid metabolism: Upregulation of CYP7A1 expression in HepG2 cells.

As an efficient triazole fungicide, prothioconazole (PTC) is widely used for the prevention and control of plant fungal pathogens. It was reported that the residues of PTC and prothioconazole-desthio (PTC-d) have been detected in the environment and crops, and the effects of PTC-d may be higher than that of PTC. Currently, PTC and PTC-d have been proven to induce hepatic metabolic disorders. However, their toxic effects on cellular bile acid (BA) and glucolipid metabolism remain unknown. In this study, HepG2 cells were exposed to 1-500 µM of PTC or PTC-d. High concentrations of PTC and PTC-d were found to induce cytotoxicity; thus, subsequent experimental exposure was conducted at concentrations of 10-50 µM. The expression levels of CYP7A1 and TG synthesis-related genes and levels of TG and total BA were observed to increase in HepG2 cells. Molecular docking analysis revealed direct interactions between PTC or PTC-d and CYP7A1 protein. To further investigate the underlying mechanisms, PTC and PTC-d were treated to HepG2 cells in which CYP7A1 expression was knocked down using siCYP7A1. It was observed that PTC and PTC-d affected the BA metabolism process and regulated the glycolipid metabolism process by promoting the expression of CYP7A1. In summary, we comprehensively analyzed the effects and mechanisms of PTC and PTC-d on cellular metabolism in HepG2 cells, providing theoretical data for evaluating the safety and potential risks associated with these substances.

Residue Behavior of Chiral Fungicide Prothioconazole and Its Major Chiral Metabolite in Flour Product Processing.

This study systematically investigates the stereoselective metabolism and residue behavior of chiral pesticide prothioconazole enantiomers during the steaming, baking, and frying of steamed buns, bread, and deep-fried dough sticks. The results show that steaming, baking, and frying can significantly promote the degradation of the prothioconazole enantiomers. In low- and high-concentration treatments, the degradation rates of prothioconazole enantiomers were over 96.0% and 45.4%, respectively, and the residual concentration of prothioconazole-desthio enantiomers was less than 32.7 µg/kg (excluding fried processing). During the processing of steamed buns, bread, and deep-fried dough sticks, the enantiomer fraction (EF) value of the prothioconazole enantiomer was close to 0.5, and the stereoselectivity was not significant. During the processing of steamed buns (low concentration), bread (low and high concentrations), and deep-fried dough sticks (low concentration), the stereoselectivity of prothioconazole-desthio was significant, and preferential enantiomer degradation occurred. Following the analysis of 120 flour product samples, the residual risk.

Determination of chiral prothioconazole and its chiral metabolite in water, juice, tea, and vinegar using emulsive liquid-liquid microextraction combined with ultra-high performance liquid chromatography.

Emulsive liquid-liquid microextraction (ELLME), a simple, rapid, and environmentally friendly technique, was established to identify chiral prothioconazole and its chiral metabolite in water, juice, tea, and vinegar using ultra-high-performance liquid chromatography (UPLC). Environmentally friendly extractant was mixed with pure water to prepare a high-concentration emulsion, which was added to samples to complete the emulsification and extraction in 1 s. Afterward, an electrolyte solution was added to complete the demulsification without centrifugation. ELLME did not use dispersants compared to the familiar dispersive liquid-liquid microextraction (DLLME), thus reducing the use of toxic solvents and avoiding the effect of dispersants on the partition coefficient. The linear range was from 0.01 to 1 mg/L. The limit of detection was 0.003 mg/L. The extraction recoveries ranged from 82.4 % to 101.6 %, with relative standard deviations of 0.7-5.2 %. The ELLME method developed has the potential to serve as an alternative to DLLME.

Enantioselective effects of chiral prothioconazole and its metabolites: Oxidative stress in HepG2 cells and lysozyme activity.

Chiral pesticides may exhibit enantioselectivity in terms of bioconcentration, environmental fate, and reproductive toxicity. Here, chiral prothioconazole and its metabolites were selected to thoroughly investigate their enantioselective toxicity and mechanisms at the molecular and cellular levels. Multispectral techniques revealed that the interaction between chiral PTC/PTCD and lysozyme resulted in the formation of a complex, leading to a change in the conformation of lysozyme. Meanwhile, the effect of different conformations of PTC/PTCD on the conformation of lysozyme differed, and its metabolites were able to exert a greater effect on lysozyme compared to prothioconazole. Moreover, the S-configuration of PTCD interacted most strongly with lysozyme. This conclusion was further verified by DFT calculations and molecular docking as well. Furthermore, the oxidative stress indicators within HepG2 cells were also affected by chiral prothioconazole and its metabolites. Specifically, S-PTCD induced more substantial perturbation of the normal oxidative stress processes in HepG2 cells, and the magnitude of the perturbation varied significantly among different configurations (P > 0.05). Overall, chiral prothioconazole and its metabolites exhibit enantioselective effects on lysozyme conformation and oxidative stress processes in HepG2 cells. This work provides a scientific basis for a more comprehensive risk assessment of the environmental behaviors and effects caused by chiral pesticides, as well as for the screening of highly efficient and less biotoxic enantiomeric monomers.

Exposure to the fungicide prothioconazole and its metabolite prothioconazole-desthio induced hepatic metabolism disorder and oxidative stress in mice.

Prothioconazole (PTC), as a popular triazole fungicide, with its main metabolite prothioconazole desthio (PTC-d), have attracted widespread concern due to their widely use and toxicological effects on non-target organisms. However, toxic effects of study analyzed PTC and PTC-d on the hepatic metabolism of mammalian still remains unclear. In this study, we conducted the study of the C57BL/6 mice which oral exposure to 30 mg/kg PTC and PTC-d via metabolomic analysis. In the liver, the metabolomics profile unveiled that exposure to 30 mg/kg PTC and PTC-d led to significantly altered 13 and 28 metabolites respectively, with 6 metabolites in common including significant decreased d-Fructose, Glutathione, showing the change of carbohydrate, lipid and amino acid metabolism. Via the further exploration of genes related to hepatic glycolipid metabolism and the biomarkers of oxidative stress, we found that liver was potentially damaged after exposure to 5 and 30 mg/kg PTC and PTC-d. Particularly, it was proved that PTC-d caused more adverse effect than its parent compound PTC on hepatotoxicity, and high concentration PTC or PTC-d exposure is more harmful than low concentration exposure.

Residue levels and dietary risk evaluation of prothioconazole-desthio and kresoxim-methyl in cucumbers after field application in twelve regions in China.

A commercial formulation called prothioconazole·kresoxim-methyl 50% water dispersible granule (WG), used to control the powdery mildew, is under registration for use on cucumbers. Therefore it is urgent to validate the reliability of the recommended good agricultural practices (GAP) conditions (187.5 g a.i. ha-1, three sprays with a 7-d interval, and a pre-harvest interval of 3 d) by conducting field trials in 12 regions in China, as required by national regulations, with a subsequent risk assessment. The residues of prothioconazole-desthio and kresoxim-methyl in field samples were determined using QuEChERS coupled with high-performance liquid chromatography - tandem mass spectrometry (HPLC-MS/MS). On the suggested pre-harvest interval (PHI, 3 d), the residual concentrations of prothioconazole-desthio (no maximum residue limit set in China) and kresoxim-methyl (maximum residue limit = 0.5 mg/kg) in cucumbers were 0.01-0.020 and 0.01-0.050 mg/kg, respectively. The acute risk quotients of prothioconazole-desthio in cucumbers were no higher than 0.079% for Chinese consumers. The chronic dietary risk quotient of kresoxim-methyl and of prothioconazole-desthio for different groups of consumers in China ranged from 2.3% to 5.3% and from 1.6% to 4.6%, respectively. Thus, prothioconazole·kresoxim-methyl 50% WG sprayed on cucumbers under the recommended GAP information can pose a negligible risk for Chinese consumers.

Widening the Lens on Prothioconazole and Its Metabolite Prothioconazole-Desthio: Aryl Hydrocarbon Receptor-Mediated Reproductive Disorders through in Vivo, in Vitro, and in Silico Studies.

Reproductive disorders are a serious public health problem worldwide. Epidemiological data suggest that exposure to environmental pollutants is associated with the onset of reproductive disorders. However, the effects in reproductive health and exact mechanism of action of representative agricultural compounds prothioconazole (PTC) and its metabolite prothioconazole-desthio (dPTC) on mammals remain unclear. Here, we studied the physiological effects of the exposure to environmentally relevant doses of PTC and dPTC in mice reproductive systems. Combining in vivo, in vitro, and in silico studies, we observed that PTC and dPTC disrupt reproductive health by inducing metabolic perturbation, induction of apoptosis, and inflammation in gonadal tissue, which are achieved via activation of the aryl hydrocarbon receptor (AhR). Convincingly, the addition of alternate-day injections of CH223191 (an AhR inhibitor) to the 30-day exposure regimen ameliorated ovarian tissue damage, as evidenced by decreased TUNEL-positive cells and partially restored the inflammation and apoptotic factor levels. This study comprehensively reports the toxic effects of low-dose PTC and dPTC in the reproductive system in vivo and identifies AhR as a potential therapeutic target for the amelioration of reproductive disorders caused by similar endocrine-disrupting chemicals.

The fungicide prothioconazole and its metabolite prothioconazole-desthio disturbed the liver-gut axis in mice.

The triazole fungicide prothioconazole (PTC) can cause adverse effects in animals, and its main metabolite prothioconazole-desthio (PTC-d) is even much more harmful. However, the toxic effects of PTC and PTC-d on the liver-gut axis of mice are still unknown. In the present experiment, we found that oral exposure to PTC and PTC-d increased total bile acids (TBAs) levels in the serum, liver, and feces. Correspondingly, the transcription of genes involved in bile acids (BAs) disposition was significantly influenced by PTC or PTC-d exposure. Furthermore, the BAs composition of serum BAs was analyzed by LC-MS, and the results indicated that PTC and PTC-d exposure changed the BAs composition, lowered the ratio of conjugated/unconjugated BAs, elevated the ratio of CA/b-MCA, and enhanced the hydrophobicity of BAs pool. 16s RNA gene sequencing of the DNA from colonic contents uncovered that PTC and PTC-d exposure altered the relative abundance and constitution of intestinal microbiota, increasing the relative level of Lactobacillus with bile salt hydrolase (BSH) activity. Furthermore, PTC and PTC-d exposure impaired the gut barrier function, causing an increase in mucus secretion. In particular, the effects of PTC-d on some endpoints in the BAs metabolism and gut barrier function had been proven to be more significant than the parent compound PTC. All these findings draw attention to the health risk of PTC and PTC-d exposure in regulating BAs metabolism, which might lead to some metabolic disorders and occur of related diseases in animals.

Acute and chronic toxicities of prothioconazole and its metabolite prothioconazole-desthio in Daphnia magna.

Current research on prothioconazole (PTC), a broad-spectrum triazole fungicide, mainly focuses on its efficacy and residues; only a few studies have been assessing its toxicological effects. Using acute and chronic toxicity tests, we assessed the effects of PTC and its metabolite prothioconazole-desthio (PTCd) on the inhibition of the activity, growth, and reproduction of Daphnia magna. A dose-response relationship was established to determine sensitive biological indicators. In the acute and chronic toxicity tests, the 48-h EC50 (concentration for 50% of the maximal effect) of PTC and PTCd for D. magna were 2.82 and 5.19 mg/L and 0.0807 and 0.132 mg/L, respectively; in the latter test, PTC was 1.64 times more toxic than PTCd. Acute-to-chronic toxicity ratios were calculated using chronic toxicity data; the ratios were 227 and 27.5 for PTC and PTCd, respectively. Our results indicate that both PTC and PTCd affect the growth and reproduction of D. magna and that the toxicity of PTC is greater than that of PTCd. In conclusion, the metabolites of PTD are toxic to D. magna at certain concentrations, and their environmental risks should not be neglected.

Toxification metabolism and treatment strategy of the chiral triazole fungicide prothioconazole in water.

Toxification metabolism of the chiral triazole fungicide prothioconazole in the environment has attracted an increasing amount of attention. To better understand the fate of prothioconazole in aquatic ecosystems and develop a treatment strategy, the stereoselective toxicity, degradation and bioconcentration of prothioconazole were investigated in water with algae at the enantiomer level. There was remarkable enantioselectivity against Chlorella pyrenoidosa, and the highly toxic S-prothioconazole was preferentially degraded with enantiomer fraction values ranging from 0.5 to 0.74. Metabolism experiment results showed that the parent compound was quickly eliminated driven by biodegradation and abiotic degradation (hydrolysis, photolysis). Fourteen phase I and two phase II metabolites involved in the reactions of hydroxylation, methylation, dechlorinating, desulfuration, dehydration and conjugation were identified, where prothioconazole-desthio was the major metabolite. The highly toxic metabolite prothioconazole-desthio persisted in water and hardly degraded with or without C. pyrenoidosa. Furthermore, the reaction system including 1 mg of cobalt coated in nitrogen doped carbon nanotubes and 0.156 g of peroxymonosulfate was used to eliminate prothioconazole-desthio. Approximately 96% prothioconazole-desthio was eliminated and transformed to low toxicity metabolites. This work provides a strategy for the risk evaluation of prothioconazole in aquatic ecosystems and proposes a workable plan for the elimination of pesticide residues in water.

Toxicological effects of acute prothioconazole and prothioconazole-desthio administration on liver in male Chinese lizards (Eremias argus).

Prothioconazole (PTC) is a high effective systemic fungicide, and one of its major metabolites is prothioconazole-desthio (PTC-d). Because of its wildly use in the farmland of China, the local eco-toxicological effects of PTC as well as PTC-d are needed to be concerned. This study investigated hepatoxicity of Chinese lizards (Eremias argus), a local non-target organism, after single dose oral treated (100 mg kg-1 BW) through pathological, enzyme activity and gene expression analysis. PTC treatment caused ballooning and PTC-d treatment led to macrovesicular steatosis of hepatocyte. The elevation of serum indexes, including the activities of aspartate aminotransferase (AST), alkaline phosphatase (ALP) and alanine aminotransferase (ALT), further confirmed the hepatic injury. PTC and PTC-d treatments altered oxidative status reflected by the inhibition of superoxide dismutase (SOD) activity , meanwhile, the stimulation of catalase (CAT) activity, glutathione peroxidase (GPx) activity and malondialdehyde (MDA) content. The mRNA expression changes of apoptosis-related factors and cytokines genes, including Bax, Bcl-2, TNF-α, NF-κB, Caspase-3 and Nrf2, deeply uncovered the potential mechanism of hepatotoxicity caused by PTC and PTC-d. In brief, the results indicated that both of these two compounds altered oxidative status, then were likely to trigger caspase-3 by affecting the ratio of pro- and anti-apoptotic factors which belong to intrinsic apoptosis pathway. Specifically, more serious impacts were induced by PTC-d than its parent compound. This study is the first to provide specific insight into potential hepatotoxicity resulted from PTC and PTC-d in male Chinese lizards.

Prothioconazole and prothioconazole-desthio induced different hepatotoxicities via interfering with glycolipid metabolism in mice.

Prothioconazole (PTA), a new triazole fungicide, has been widely used worldwide. A recent study has confirmed that PTA and its main metabolite prothioconazole-desthio (dPTA) interfere with the liver metabolism in reptiles. However, little is known about liver toxicity of these two pollutants in mammals. Here, female mice were orally exposed to PTA (1.5 mg/kg body weight/day) and dPTA (1.5 mg/kg body weight/day) for 30 days. Additionally, growth phenotype and indexes related to serum and liver function were examined. Using metabolomics and gene expression analysis, PTA- and dPTA-induced hepatotoxicity was studied to clarify its potential underlying mechanism of action. Together, the results indicated that PTA and dPTA exposure caused changes in growth phenotypes, including elevated blood glucose levels, triglyceride accumulation, and damage of liver function. Additionally, exposure to PTA and dPTA caused changes in genes and metabolites related to glycolipid metabolism in female mice, thereby interfering with the pyruvate metabolism and glycolysis/gluconeogenesis pathways, ultimately leading to hepatic metabolism disorders. In particular, the effect of dPTA on hepatotoxicity has been proven to be more significant than that of PTA. Thus, these findings help us understand the underlying mechanism of action of PTA and dPTA exposure-induced hepatotoxicity in mammals and possibly humans.

Enantiomeric separation of prothioconazole and prothioconazole-desthio by Capillary Electrophoresis. Degradation studies in environmental samples.

In this work, two analytical methodologies by Capillary Electrophoresis were developed. The first one enabled the rapid and cost-effective enantioseparation of prothioconazole and was applied to the analysis of prothioconazole-based commercial agrochemical formulations. The second methodology enabled the simultaneous enantioseparation of prothioconazole and its metabolite prothioconazole-desthio and was applied to degradation studies of both compounds in soil and sand samples. The influence of several experimental variables was investigated to develop both methodologies. The separation of prothioconazole enantiomers was achieved in 4.5 min with a resolution of 2.8 employing a neutral cyclodextrin (heptakis(2,3,6-tri-O-methyl)-ß-cyclodextrin). Given the nature of prothioconazole-desthio, a neutral cyclodextrin cannot be used for its chiral separation. For this reason, the simultaneous enantioseparation of prothioconazole and prothioconazole-desthio was achieved in 5.5 min with resolution values of 1.9 and 8.2, respectively, using a negatively charged cyclodextrin (sulfated-γ-cyclodextrin). The analytical characteristics of the developed methodologies were evaluated and both methods showed good performance to be applied to the quantitation of the enantiomers of prothioconazole in commercial agrochemical formulations (LOD 0.7 mg L-1) and to carry out degradation studies for both compounds in environmental matrices (LODs lower than 0.9 and 1.3 mg L-1 for prothioconazole and prothioconazole-desthio enantiomers, respectively). The recovery values obtained were in the range between 94-104 % for the agrochemical formulations, between 96-99 % for the sand samples and between 97-100 % for the soil samples.

Terminal residue and dietary intake risk assessment of prothioconazole-desthio and fluoxastrobin in wheat field ecosystem.

BACKGROUND: Wheat is one of the most important cereal crops worldwide, and use of fungicides is an essential part of wheat production. Both prothioconazole and fluoxastrobin give excellent control of important seed and soilborne pathogens. The combination of these two fungicides shows a complementary mode of action and has a wide usage around the world. But the residue levels of these fungicides in the wheat matrix are still unknown. RESULTS: In the current study, a simple, low-cost and highly sensitive method using modified QuECHERS procedure combined with high-performance liquid chromatography-tandem mass spectrometry was developed to simultaneously quantify E- and Z-fluoxastrobin and the main metabolite prothioconazole-desthio of prothioconazole in the wheat matrix. The recoveries of prothioconazole-desthio, E-fluoxastrobin and Z-fluoxastrobin ranged from 84% to 101%, with relative standard deviation of less than 13.2%. The terminal residues of prothioconazole-desthio and E- and Z-fluoxastrobin were studied in wheat grain and straw under field conditions. The results showed that the terminal residue of the target compounds ranged from <0.01 to 0.029 mg kg-1 and <0.05 to 7.6 mg kg-1 in wheat grain and straw (expressed as dry weight), respectively. The risk quotients of prothioconazole-desthio and fluoxastrobin were 0.2% and 3.2%. CONCLUSIONS: The residue levels of the target analytes in wheat grain were lower than the maximum residue limits recommended by the Chinese Ministry of Agriculture. And the calculated risk quotient values were far below 100%, indicating a low dietary intake health risk to consumers. © 2021 Society of Chemical Industry.

Popping candy-assisted dispersive liquid-liquid microextraction for enantioselective determination of prothioconazole and its chiral metabolite in water, beer, Baijiu, and vinegar samples by HPLC.

To simultaneously determine the enantiomers of prothioconazole and its chiral metabolite prothioconazole-desthio in water, beer, Baijiu, and vinegar samples by HPLC, a simple, fast, environmentally-friendly popping candy-assisted dispersive liquid-liquid microextraction technique was developed. A green medium-chain fatty acid (decanoic acid) and popping candy could be used as the extractant and solid dispersant respectively to avoid the use of toxic organic solvents. Decanoic acid was collected after extraction by solidification at room temperature. The linear range of this technique was from 27.1 to 1000 µg L-1. The limits of detection and quantification were within the ranges of 8.1-11.2 µg L-1 and 27.1-37.3 µg L-1, respectively. The extraction recovery was 80.8% to 102.5% with the relative standard deviation ranged from 1.1 to 7.1%. This technique has been successfully applied to enantioselectively determine the residues of prothioconazole and prothioconazole-desthio in water, beer, Baijiu, and vinegar samples.

Effects of exposure to prothioconazole and its metabolite prothioconazole-desthio on oxidative stress and metabolic profiles of liver and kidney tissues in male mice.

Prothioconazole (PTC), a popular agricultural fungicide, and its main metabolite prothioconazole-desthio (PTCd) are receiving great attention due to their toxicological effects in the non-target organisms. This study investigated their dosage-dependent (1 and 5 mg/kg BW/day) toxicological effects on oxidative stress and metabolic profiles of liver and kidney tissues using male mice. PTC and PTCd significantly inhibited the growth phenotype including body weights gain, liver and kidney indices. Furthermore, these effects were deeply investigated using the biomarkers of oxidative stress, and metabolomics. Notably, these effects were dose and tissue-dependent. Specifically, the more serious impacts involving oxidative stress and metabolic disorders were observed in the high concentration treatment groups. Also, the liver tissue was more severely affected than the kidney tissue. Lastly, the change in oxidative stress biomarkers and metabolomics profile revealed that PTCd induced more severe toxic effects than the parent compound PTC. In brief, these results indicate that exposure to PTC and PTCd could cause potential health risks in mammals.

Enantioselective Separation and Dissipation of Prothioconazole and Its Major Metabolite Prothioconazole-desthio Enantiomers in Tomato, Cucumber, and Pepper.

In this study, a simple and effective chiral analytical method was developed to monitor prothioconazole and prothioconazole-desthio at the enantiomeric level using supercritical fluid chromatography-tandem triple quadrupole mass spectrometry. The baseline enantioseparation for prothioconazole and prothioconazole-desthio was achieved within 2 min on a Chiralcel OD-3 column with CO2/0.2% acetic acid-5 mmol/L ammonium acetate 2-propanol (85:15, v/v) as the mobile phase at a flow rate of 1.5 mL/min and column temperature of 25 °C. The limit of quantitation for each enantiomer was 5 µg/kg, with a baseline resolution of >3.0. The results of enantioselective dissipation showed that R-(-)-prothioconazole was preferentially degraded in tomato, cucumber, and pepper under greenhouse conditions. S-(-)-prothioconazole-desthio was preferentially degraded in tomato and cucumber; however, R-(+)-prothioconazole-desthio was preferentially degraded in pepper. Results of this study may help to facilitate more accurate risk assessment of prothioconazole and its major metabolite in agricultural products.

Dissipation of Prothioconazole and Its Metabolite Prothioconazole-Desthio in Rice Fields and Risk Assessment of Its Dietary Intake.

Rice is an important food crop, with a long history of cultivation in China, and is one of the important staple foods for Chinese today. However, the occurrence of rice diseases has had a substantial impact on its yield. At present, chemical control is the main means of prevention and control of rice diseases. As a high-efficacy and broad-spectrum fungicide, prothioconazole shows an ideal effect on the prevention and control of common rice diseases, but the residuals that remain after its use may have an impact on human health. In this paper, ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to develop a residue analysis method and explore the dissipation of prothioconazole and its metabolite prothioconazole-desthio in rice grain, rice husk, and rice plants, and the risks of chronic and acute dietary intake of prothioconazole in different Chinese populations were evaluated. The results showed that at a concentration range of 0.005-5.0 µg/mL, prothioconazole and its metabolites showed good linear correlations, and the correlation coefficients were all above 0.9992. The average recoveries of prothioconazole in three matrixes ranged from 80.0% to 99.7%, the RSD was between 1.54% and 11.0%, and the limit of quantitation was less than 0.01 µg/g. The sensitivity, accuracy, and precision of the established method all met the measurement requirements. The dissipation experimental results showed that the parent compound prothioconazole was rapidly metabolized to prothioconazole-desthio in the rice field environment, and the dissipation half-life in rice plants after application was 2.5-10.1 d. According to the results of dietary risk assessment, under the standardized residue test conditions, the residual level of rice at the sampling interval of 7 days was within the acceptable range for the chronic and acute dietary risks of different populations in China. Our research is important for directing the scientific application of pesticides in China.

Residue and intake risk assessment of prothioconazole and its metabolite prothioconazole-desthio in wheat field.

In the environment, plants and animals in vivo, pesticides can be degraded or metabolized to form transformation products (TPs) or metabolites, which are even more toxic than parent pesticides. Hence, it was necessary to evaluate residue and risk of pesticides and their TPs (or metabolites). Here, a rapid, simple, and reliable method using QuEChERS and LC-MS/MS had been developed for simultaneous analysis of prothioconazole and its toxic metabolite, prothioconazole-desthio, in soil, wheat plant, straw, and grain. The average recoveries of prothioconazole and prothioconazole-desthio in four matrices ranged from 86 to 108% with relative standard deviations (RSDs) of 0.53-11.87% at three spiking levels. The method was successfully applied to investigate the dissipation and terminal residues of the two compounds in wheat field. It was shown that prothioconazole was rapidly degraded to prothioconazole-desthio, with half-lives below 5.82 days. Prothioconazole-desthio was slowly dissipated in soil and plant. The terminal residues of prothioconazole in wheat grain with a pre-harvest interval (PHI) of 21 or 28 days were below the maximum residue limits (MRLs) (0.1 mg/kg, Codex Alimentarius Commission (CAC)). We also evaluated the intake risk of prothioconazole-desthio residues in wheat grain in China. For long-term intake assessment, the hazard quotients (HQ) ranged from 1.30 to 5.95%. For short-term intake assessment, the acute hazard indexes (aHI) ranged from 1.94 to 18.2%. It indicated that the intake risk of prothioconazole-desthio in wheat consumption was acceptable. Thus, the prothioconazole application on wheat with the scientific practices would not pose public health risk.