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The widespread use of cyproconazole (CPZ) enhances food security but may pose potential risks to non-target organisms. Therefore, we applied Multi-omics techniques to reveal the response of the intestinal barrier to CPZ exposure and explore whether the Bifidobacterium intervention experiment can repair the damage. First, we found that exposure to CPZ at environmentally relevant concentrations led to intestinal injury phenotype, significantly down-regulated intestinal protein gene expression, and up-regulated pro-inflammatory gene expression, further causing intestinal dysbacteriosis and metabolic disorders. In particular, by combining analysis of gut microbiota and metabolites, we noticed acetate, a key metabolite, which decreased sharply after exposure to high concentration of CPZ. Expectedly, after supplementing with Bifidobacterium (a core bacterium that produces acetate), we noticed that the acetate content was quickly restored. Further, we also verified that the increase in acetate content after Bifidobacterium supplementation at least partially promoted IL-22 secretion, which in turn stimulated the secretion of β-defensins (zfbd-1, zfbd-2, zfbd-3), thereby repairing the intestinal damage. In conclusion, our work confirms the potential of Bifidobacterium to improve intestinal damage and metabolic dysbiosis caused by CPZ exposure. It provides directional recommendations for the application of probiotics to repair the toxicological risk of pesticide exposure., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that they could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

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.

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., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Amphibians are the most endangered class of vertebrates. In this study, Xenopus laevis frogs were exposed to 0, 1 and 10 mg/L of triadimefon or triadimenol. After 14 or 28 days of exposure, high levels of triadimefon or triadimenol obstructed the growth of frogs. However, low levels of triadimefon induced the growth of females after the longer period of exposure. We also found that the antioxidant enzyme activity and LDH levels in males were higher than those in females after 28-days exposure. In histopathology tests, triadimenol exerted more effect on the ovary while triadimefon exerted more effect on the testes. Additionally, the levels of Estradiol in all 14-day treatments, except 1 mg/L triadimenol, were significantly decreased, however, there was no difference in testosterone levels. Furthermore, triadimefon and triadimenol disrupted the expression of genes controlling hormone homeostasis and reproduction, and this effect depended on the exposure time and the gender of the organism. Our experiments explored the effect of triadimefon and its metabolite on the gonads of frogs and highlighted the role that pesticides are likely to play in the global decline of amphibians., Competing Interests: Declaration of competing interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In addition to their fungicidal activity, many triazole fungicides function as plant regulators, which might impose adverse effects on the growth and development of crops. For chiral triazole fungicides, these effects can be alleviated by applying stereoisomers with high fungicidal and low regulator activities. This study investigated the stereoselectivity of four stereoisomers and the racemate of metconazole (2.5 g/100 kg seeds) on emergence and growth of seedlings (BBCH 01-14) in wheat. Wheat seedlings, coated with cis -1 S ,5 R -metconazole, had a significantly lower seedling emergence ratio and shoot length than other metconazole treatments; however, the opposite effects were observed in the trans -1 S ,5 S -metconazole treatment. With regard to the hormonal level, enzyme activity, and gene transcription of gibberellin (GA) and jasmonic acid (JA), cis -1 S ,5 R -metconazole treatment inhibited GA biosynthesis while trans -1 S ,5 S -metconazole treatment promoted GA biosynthesis. Moreover, cis -1 S ,5 R -metconazole, trans -1 S ,5 S -metconazole, trans -1 R ,5 R -metconazole, and racemate treatments increased JA biosynthesis. The oxidative stress responses in trans -1 R ,5 R -metconazole and racemate treatments were more intensive. Therefore, compared with the control, treatment with cis -1 R ,5 S -metcoanzole exhibited minimal influence on wheat seedling growth. The results showed that the application of pure cis -1 R ,5 S -metcoanzole (instead of the racemate) in agricultural management could decrease the risks associated with crop growth and developmental damage.

Tebuconazole, a widely used fungicide, can severely disrupt the reproductive process of various organisms. In this study, we investigated the subacute effects of tebuconazole on the earthworm to fully understand its toxic implications. Herein, untargeted metabolomics, mRNA assay and biochemical approaches were adopted to evaluate the subacute effects of Eisenia fetida earthworms, when exposed to tebuconazole at three different concentrations (0.5, 5 and 50 mg/kg) for seven days. SOD enzyme activity test displayed that tebuconazole exposure interfered with the earthworms' ROS. ANN mRNA expression was down-regulated after tebuconazole exposure. 1 H nuclear magnetic resonance ( 1 H-NMR)-based untargeted metabolomics study showed that 5 mg/kg tebuconazole exposure interfered with earthworms' metabolism. Twelve significantly changed metabolites were identified. The pathway analyses indicate that tebuconazole can disrupt the earthworm's metabolism, particularly in the AMP pathway, which impact the reproduction. This may explain the tebuconazole's mechanism of action behind the down-regulation of the expression of ANN mRNA, which is related to the earthworm's reproductive process. We comprehensively evaluated the mRNA expression, enzyme activity, and metabolomics, and acquired sufficient information for evaluating the toxicity of tebuconazole., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Reptiles are an important part of vertebrates and are the primitive terrestrial vertebrates. However, lots of reptile species are endangered or susceptible to extinction. It is no doubt that contaminants are one of the important reasons for the decline of the lizard population. In this study, the selective metabolism of triadimenol (TN) in the male Eremias argus lizards and the toxic effects of TN on lizards were studied. TN chiral isomers were separated and detected by HPLC-MS/MS system with Lux Cellulose-1 column. Tissue distribution experiments showed the existence of stereoselectivity biotransformation of TN enantiomers among organs in lizards, and RR-TN preferentially emerged over the other enantiomers. The antioxidant enzymes (SOD, CAT, GST) activities and MDA content assays demonstrated that TN induced oxidative stress in most organs, especially in the liver, and the histopathology analysis showed the severe liver and testis damage caused by 14-day continuous TN gavage. The reproductive effects of TN-induced reflected in the increased sex hormone testosterone. This research confirms that TN could induce hepatic and reproductive toxicity of E. argus lizard.

The decrease in the population of amphibians all over the world has raised concerns. Adult X. laevis frogs were exposed to 0, 1 and 10 mg/L triadimefon and triadimenol. After 14 or 28 days exposure, we collected male and female specimens to study swimming activity, lactic dehydrogenase (LDH) and antioxidant enzyme activity in blood samples, histopathology of liver and thyroid tissue, thyroid hormone levels and thyroid hormone-related gene expression levels in brains. Our results showed that triadimefon and triadimenol could affect the swimming activity of frogs and that this was distinct at different levels of triadimenol. Moreover, triadimefon and triadimenol exposure produced a greater effect on superoxide dismutase (SOD) in females than in males, which was reverse to the finding for glutathione S-transferase (GST) and catalase (CAT). After 28 days exposure, triadimefon produced more toxic effects on the liver than observed for triadimenol. Besides this, triadimefon and triadimenol exposure exerted a greater effect on liver histology and thyroid hormone levels in male frogs than in the females. Our results also found that the expression of genes related to thyroid hormone in brains depended on the exposure level and time, as well as the sex of the treated individual. This study shed light on the relationships between the toxicity of metabolite products and their parent compounds and provided further understanding of the risk of pesticide use on amphibians., Competing Interests: Declaration of competing interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

(±) - PEN is a chiral fungicide widely used to control powdery mildew in agriculture. Currently, only a few studies have investigated the toxic effects of (±) - penconazole ((±) - PEN) on non-target organisms, and whether (±) - PEN from the enantiomeric level have toxic effects remains unclear. In this study, we systematically evaluated the effects of exposure to (±) - PEN, (+) - PEN and (-) - PEN on liver function in mice. Biochemical and histopathological analyses showed that exposure to (±) - PEN and (-) - PEN led to significant liver damage and inflammation. However, exposure to (+) - PEN treatment did not cause no adverse effects on liver function and inflammation. 1 H-NMR-based metabolomics revealed that exposure to (±) - PEN, (+) - PEN and (-) - PEN led to the animals developing liver metabolic disorder that was caused by changes in glycolipid metabolism. Quantitative analysis of genes regulating glycolipid metabolism revealed that expression of gluconeogenesis and glycolytic pathway genes were altered in individuals exposed to (±) - PEN, (+) - PEN and (-) - PEN. We also found that (±) - PEN, (+) - PEN and (-) - PEN have different effects on lipid metabolism of the liver. Exposure to (±) - PEN and (-) - PEN resulted in significant accumulation of lipids by regulating fatty acid synthesis, triglyceride synthesis, and fatty acid β oxidation pathways. In summary, we found different toxicological effects in individuals exposed to (±) - PEN, (+) - PEN and (-) - PEN. The results of this study are important for assessing the potential health risks of (±) - PEN., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The rational use and the environmental safety of chiral pesticides have attracted significant research interest. Here, enantioselective toxic effects and the selective toxic mechanism of triticonazole (TRZ) against the aquatic microalgae Chlorella pyrenoidosa were studied. The 96h-EC 50 values of rac-, (R)-(-)-, and (S)-(+)-TRZ were 1.939, 0.853, and 22.002 mg/L, respectively. At a concentration of 1 mg/L, the contents of photosynthetic pigments of C. pyrenoidosa exposed to (R)-(-)-TRZ were lower than if exposed to S-(+)-form and racemate. Transmission electron microscopic images showed that the R-(-)-form compromised the integrity of cells and disrupted the chloroplast structure. R-(-)-TRZ stimulated vast reactive oxygen species (ROS) and significantly increased superoxide dismutase (SOD) and catalase (CAT) activities, as well as malondialdehyde (MDA) content. For lipid accumulation experiments, nicotinamide adenine dinucleotide (NADH) and triacylglycerol (TAG) accumulations in algal cells treated with R-(-)-TRZ were 171.50% and 280.76%, respectively, compared with the control group. This far exceeded levels of algal cells treated with S-(+)- and rac-TRZ. Based on these data, R-(-)-TRZ was concluded to selectively affect the photosynthetic system, antioxidant system, and lipid synthesis of algal cells, thus causing enantioselective toxic effects of TRZ against C. pyrenoidosa, which indicating that the use of racemate may cause unpredictable environmental harm. Therefore, to reduce the hidden dangers of chiral pesticides for the ecological environment, the environmental risk of TRZ should be evaluated at the stereoselective level., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Triazole fungicides are widely used in agriculture production and have adverse impacts on aquatic organisms. As one of the triazole fungicides, prothioconazole has been reported to cause many toxicological effects, but its risks to aquatic organisms are unknown. In this study, we systematically explored the toxicity effects of prothioconazole exposure on zebrafish embryos (Danio rerio) involving in developmental toxicity, oxidative damage and metabolism disorders. The results showed that prothioconazole exposure to zebrafish embryos produced a series of toxic symptoms, including hatching inhibition, shortening of body length, pericardial cyst and yolk cyst. In addition, prothioconazole exposure caused significant lipid peroxidation and oxidative damage. Particularly, we also found that metabolites and genes involved in lipid metabolism also showed significant changes. This study may provide theoretical basis for systematically assessing the potential risks of zebrafish embryos with prothioconazole exposure., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Exposure to chiral pesticides poses many potential health risks. In this study, we examined the impacts of exposure to penconazole and its enantiomers on gut microbiota and metabolic profiles in mice. The relative abundance of microbiota in cecal content significantly changed following exposure to penconazole and its enantiomers. At the genus level, the relative abundances of seven gut microflora were altered following exposure to (-)-penconazole. Both (±)-penconazole and (+)-penconazole caused significant changes in the relative abundances of five gut microflora. In addition, targeted serum metabolomics analysis showed disturbed metabolic profiles following exposure. Respectively, (±)-penconazole, (+)-penconazole, and (-)-penconazole exposure significantly altered the relative levels of 29, 23, and 36 metabolites. In general, exposure to penconazole and its enantiomers caused disorders in gut microbiota and metabolic profiles of mice. The potential health risks of penconazole and its enantiomers now require further evaluation.

Hexaconazole and epoxiconazole are the worldwidely used fungicides. However, limited information is known about the toxicological effects of their enantiomers on aquatic organisms. In this study, zebrafish were separately exposed to 100 and 1000 μgL -1 hexaconazole and epoxiconazole enantiomers for 21 d 1 H NMR-based metabolomics analysis showed that the exposure of low and high dose of hexaconazole enantiomers altered energy metabolism, lipid metabolism and amino acid metabolism of zebrafish, with the different metabolic profiles resulted from the same dose of (+)-hexaconazole and (-)-hexaconazole. Similar to hexaconazole enantiomers, the metabolic profiles, including the changes related to energy metabolism, lipid metabolism and amino acid metabolism, were demonstrated in low and high dose epoxiconazole enantiomers treatment groups. There are differences in the metabolic profiles of zebrafish between exposed to (+)-epoxiconazole and (-)-epoxiconazole of the same dose. The results of histological examination revealed that the exposure of both enantiomers for hexaconazole and epoxiconazole resulted in the similar histopathological changes. The exposure of hexaconazole and epoxiconazole enantiomers at low and high dose resulted the vacuolization and swell in the liver of the female and male zebrafish. Compared to female zebrafish, more liver damage was found in male zebrafish in the hexaconazole enantiomers exposure groups. The reduction of spermatids was observed in hexaconazole and epoxiconazole enantiomers treatment groups of both doses. Hexaconazole enantiomers exposure of low and high dose resulted the increase in the number of mature eggs, while such effect was not observed in epoxiconazole enantiomers exposure groups. Hexaconazole and epoxiconazole enantiomers exposure resulted in no changes in brains of female and male zebrafish. As a result, both triazole-based chiral bactericides, hexaconazole and epoxiconazole, have similar toxicological effects but their mechanisms of action are not exactly the same. The above results will play an important part in making the differences in toxic effects of hexaconazole and epoxiconazole enantiomers clear. What's more, it is an indispensable part for an integrated environmental risk assessment., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Chiral fungicide prothioconazole has a wide range of antifungal spectrum; however, little research has been conducted to evaluate prothioconazole on an enantiomeric level. Five target pathogens and three common aquatic organisms were tested for the enantioselective bioactivity and toxicity of prothioconazole in this work. The antifungal activity of the enantiomers against wheat phytoalexin, rice blast fungus, exserohilum turcicum, Alternaria triticina, and Fusarium avenaceum was determined, and it was found that (-)-prothioconazole were 85 to 2768 times more active than (+)-prothioconazole toward these target organisms. In order to reflect the risk to aquatic ecosystem, the acute toxicity of the enantiomers to Daphnia magna, Chlorella pyrenoidosa, and Lemna minor L. was assessed. It was observed that the toxicity of (-)-prothioconazole to D. magna was 2.2 times higher than (+)-prothioconazole, but it was lower to C. pyrenoidosa and L. minor L. The toxicities of (+)-enantiomer and (-)-enantiomer to D. magna and C. pyrenoidosa were synergy, indicating that the racemate had higher threat to the organisms. It could be concluded that the effects of prothioconazole on target organisms and the acute toxicity to nontarget species were enantioselective with (-)-enantiomer possessing higher efficiency and lower toxicity. Such enantiomeric differences should be taken into consideration when assessing the performance of prothioconazole., (© 2019 Wiley Periodicals, Inc.)

Metconazole (MEZ) is a broad-spectrum fungicide with four optical stereoisomers. Compared to traditional fungicides, it achieves better control effect at lower dosages. However, its toxicity to non-target organisms has rarely been investigated. This study investigated the stereoselective toxicity of metconazole to Chlorella pyrenoidosa (C. pyrenoidosa). The results indicate that the presence of the racemate and four stereoisomers of MEZ caused a sudden increase of reactive oxygen species (ROS). This in turn stimulated antioxidant defense, impaired photosynthesis and responses of subcellular structure, and eventually inhibited cell growth. The 96 h-EC 50 of the racemate, cis-1R,5S-MEZ, cis-1S,5R-MEZ, trans-1S,5S-MEZ, and trans-1R,5R-MEZ were 0.058, 0.182, 0.129, 0.032, and 0.038 mg/L, respectively. Furtheromre, the generation of ROS, antioxidant response, and the loss of photosynthetic function in C. pyrenoidosa were all preferentially trans-1S,5S-MEZ induced. These results aid the understanding of the stereoselective effects of chiral pesticides on C. pyrenoidosa. Such stereoselective differences must be considered when assessing the risk of metconazole to environment., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Pesticides are a major cause of reduction in the global population of amphibians. This study investigates the effect of varying concentrations of cyproconazole (1 and 10 mg/L) on Rana nigromaculata during a chronic 90 days exposure period. High levels of cyproconazole (10 mg/L) induced declined body weight, short snout-vent length, slow metamorphic development and abnormal behavioral endpoints in R. nigromaculata tadpoles. Tadpoles exposed to 10 mg/L did not survive beyond 42 days. Abnormal behaviors were observed more frequently with exposure to the highest concentration of cyproconazole. Compared with controls, the concentrations of dismutase (SOD), catalase (CAT) and glutathione (GSH) were significantly increased in tadpoles exposed to 1 mg/L cyproconazole. However, when the concentration of cyproconazole increased to 10 mg/L, concentrations of SOD, GSH and CAT activity began to decline. In addition, thyroid and gonad development were also affected at the gene and hormone level, with varied effects observed with different exposure levels and days. Exposure to cyproconazole at the lower level of 1 mg/L induced damage to histological structures of the thyroid gland. Stereoselective tissue distribution and bioaccumulation of cyproconazole was observed in tadpoles. The ranked order of bioaccumulation was: enantiomer -4 > 3> 2 > 1, with the level of cyproconazole highest in the gut. These findings reflect the toxicity of cyproconazole to R. nigromaculata and further our understanding of the effects of pesticide exposure on global amphibian population declines., (Copyright © 2018. Published by Elsevier B.V.)

Numerous studies suggested that reptiles are sensitive to environmental pollution and the abundance of many species are in decline. Our research is aim to assess the toxic effects of pesticide in reptiles. And we also want to supply some data about nondestructive samples for environmental risk assessment in reptiles. Lizards were orally administered a single-dose of beta-cypermethrin (BCP) or myclobutanil (MC) at the concentration of 20 mg/kg body weight (bw). The results showed that pesticides could induce changes in enzymatic activities (SOD, CAT, LDH, AChE) and MDA levels in organs or tissues of lizards. BCP could cause more severe oxidative damage than that of MC. Salivary enzymes activities showed sensitivity changes to the toxicity of pesticides. We could use saliva to reflect whether the reptiles are toxic by pesticides. We also agree that buccal swabs could be used as a tool for saliva sampling., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The environmental contaminant, especially pesticides, threatened the amphibian population. In this assay, the enantioselective behavior of cyproconazole on Rana nigromaculata was studied. We found LC50 (lethal concentration causing 50% mortality) of 4-enantiomers was nearly twice as 3-enantiomers in 96 h acute toxicity test. Besides, the significant considerable variation of oxidative stress and LDH (lactic dehydrogenase) induced by the four enantiomers indicated that cyproconazole could enantioselectively affect enzymes in tadpoles. Bioaccumulation experiments showed the order of cyproconazole in the tadpoles was 4-enantiomers>3- enantiomers>2- enantiomers>1- enantiomers during the exposure for 28d. In tissue distribution test, cyproconazole was formed and accumulated in order of 4-enantiomers>2-enantiomers>3- enantiomers>1- enantiomers, except that in the gut. During the elimination experiment, cyproconazole was rapidly eliminated by 95% within the only 24 h. These results suggested that the influence of enantioselective behavior should consider when assessing ecological risk of chiral pesticides to amphibians., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Pesticides are one of major causes for amphibian population declines and the behavior of pesticide metabolite products to amphibians has become a rising concern. In this study, the acute toxicity and the chronic effects of triadimefon and triadimenol (the metabolite of triadimefon) on Rana. nigromaculata were investigated. In the acute assay, significant differences were observed in antioxidant enzyme activities and malondialdehyde levels between the triadimefon and triadimenol. The 96 h-acute toxicity of triadimefon (25.97 mg/L) and triadimenol (34.55 mg/L) to tadpoles was low. In 28d-chronic exposure, we studied the relative expression of tadpoles genes related to thyroid hormone-dependent metamorphic development, histological examination of liver and some biological index, including wet weight, snout-to-vent length (SVL) and development stages. The results revealed that the effects of triadimefon and triadimenol on tadpole development are driven by a disruption of the hormonal pathways involved in metamorphosis. Interestingly, triadimefon was more harmful on R. nigromaculata than triadimenol at high dose, whereas the reverse result was observed at low doses. According to the relative expression of thyroid hormone-dependent genes, we also found that the two compounds may have different mechanisms of toxic action on R. nigromaculata. Our study developed a pragmatic approach for use in the risk assessment of pesticide and its metabolite，and increased the information and understanding of the impacts of fungicides and other potential endocrine disrupting environmental contaminants on amphibians., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In recent years, serious environmental pollution has caused a decrease in the abundance of many species worldwide. Reptiles are the most diverse group of terrestrial vertebrates. There are large amounts of toxicological data available regarding myclobutanil, but the adverse effects of myclobutanil on lizards has not been widely reported. In this study, treatment groups were orally administered a single-dose of myclobutanil (20mg/kg body weight (bw)). Subsequently, it was found that there were differences in myclobutanil levels between the different tissues and concentrations also changed with degradation time. The tissue concentrations of myclobutanil decreased in the order of: stomach > liver > lung > blood > testis > kidney > heart > brain. Based on our results, the liver and testis were considered to be the main target organs in lizards, indicating that the myclobutanil could induce potential hepatic and reproductive toxicity on lizards. Meanwhile, it was also demonstrated that the toxic effects of myclobutanil was different in different species, and the distribution of different pesticides in lizards were different., (Copyright © 2017. Published by Elsevier Inc.)

Although epoxiconazole is the worldwidely used fungicide, limited information is known about its toxic effects and bioaccumulation behavior in freshwater ecosystems. In this study, zebrafish were exposed to epoxiconazole at concentrations of 100 and 1000 μg L -1 for 21 d. 1 H NMR-based metabolomics analysis showed that low- and high-dose epoxiconazole exposure resulted in two similar but not identical patterns for the change of endogenous metabolites related to energy, lipid and amino acid metabolism. The expression of genes associated with mitochondrial respiratory chain, ATP synthesis and fatty acid β-oxidation were further measured to explore the reason for the disturbed energy metabolism, finding epoxiconazole had an inhibition effect on the genes expression of the above ways. Significant enantioselectivity was observed with (+)-epoxiconazole enrichment in the bioaccumulation process. These results will be of great importance in understanding the toxic effects induced by epoxiconazole and provide important basis for its comprehensive environmental assessment., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Research on the enantioselective behavior of chiral pesticides on amphibians has received growing attention, because amphibians are experiencing a population decline and amphibian metamorphosis shares many similarities with human fetal development. In this study, the enantioselective behavior of myclobutanil on Pelophylax nigromaculatus tadpole was studied. The antioxidant enzyme (SOD, GST) activities and malondialdehyde (MDA) content were investigated to assess the different toxic effects when tadpoles were exposed to myclobutanil enantiomers for 96 h. In the chronic exposure experiment, the bioaccumulation concentration of (-)-myclobutanil in tadpoles is significantly higher than that of (+)-myclobutanil, and the concentration of myclobutanil in tadpole intestine and liver was higher compared with other tissues. During the elimination experiment, about 95% of myclobutanil in tadpoles was eliminated within only 24 h. On the basis of these data, the enantiomeric differences should be taken into consideration in the risk assessment of myclobutanil.

Food safety problems such as damage to immune, nervous, and endocrine systems leading to cancer and malformations have received increasing attention. To achieve the maximum residue limits, the most discussed method of high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) is widely used with a advantage of high precision and resolution. Prothioconazole is a broad-spectrum thiocarbamate fungicide. It can rapidly metabolize to prothioconazole-desthio in different matrixes. Rapid and effective methods for the determination of prothioconazole-desthio in five kinds of different animal food were developed. Samples were extracted with acetonitrile or acetonitrile/water and determined by HPLC-MS/MS. The limit of detection and limit of quantification values of prothioconazole-desthio were 0.015 and 0.05 mg/kg for porcine liver and kidney, 0.0015 and 0.005 mg/kg for pork, and 0.003 and 0.01 mg/kg for eggs, together with 0.0012 and 0.004 mg/kg for milk of the detected method, respectively. A good linear regression trend can be observed in a certain concentration range for all of the animal food. At fortified levels, recoveries were between 83.6 and 105%, with relative standard deviations of 1.5-10.3%. A sample survey of 150 samples with 30 samples for each kind of animal food across the country was conducted and found that there was no prothioconazole-desthio detected in all samples.

As a systemic triazole fungicide, limited information is known about diniconazole. In this study, toxicological effects and bioaccumulation behavior of diniconazole in zebrafish were both evaluated to perform an overall assessment of its environmental risk towards aquatic organisms. The 1 H NMR-based metabolomics analysis revealed that 70 μg L -1 diniconazole exposure increased valine, leucine, isoleucine, alanine, lactate and choline, accompanied by decreased glucose, creatine and taurine, in comparison to the control. In the exposure group of 300 μg L -1 diniconazole, only down-regulated glucose and creatine were observed. The above results indicated that diniconazole disturbed energy metabolism, amino acid metabolism and lipid metabolism. Histological examination showed that the swell and vacuolization in the liver, as well as the reduction of spermatids in the diniconazole exposure groups. No enantioselectivity was observed in the bioaccumulation process of both 70 and 300 μg L -1 diniconazole exposure groups. The above results play a vital role for a comprehensive environmental assessment of diniconazole., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Enantioselectivity in ecotoxicity, digestion and uptake of chiral pesticide cyproconazole to Chlorella pyrenoidosa was studied. The 96h-EC50 values of rac- and the four enantiomers were 9.005, 6.616, 8.311, 4.290 and 9.410 mg/L, respectively. At the concentrations of 8 mg/L and 14 mg/L, the contents of pigments exposed in rac-, enantiomer-2 and 4 were higher than that exposed in enantiomer-1 and 3. The superoxide dismutase (SOD) and catalase (CAT) activity of algae exposed to enantiomer-1 and 3 was higher than that exposed to the rac-, enantiomer-2 and 4 at three levels. In addition, the malondialdehyde (MDA) concentrations in algae disposed with enantiomer-1 and 3 were increased remarkably at three levels. For the digestion experiment, the half-lives of four enantiomers in algae suspension were 28.06, 19.10, 21.13, 15.17 days, respectively. During the uptake experiment, the order of the concentrations of cyproconazole in algae cells was enantiomer-4, 2, 3 and 1. Based on these data, we concluded that ecotoxicity, digestion and uptake of chiral pesticide cyproconazole to C. pyrenoidosa were enantioselective, and such enantiomeric differences must be taken into consideration when assessing the risk of cyproconazole to environment., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Little is known about the bioaccumulation and toxicity of hexaconazole (HEX) in spite of the fact that they are indispensable parts for a comprehensive assessment of its environmental behavior and toxic effects in organisms of freshwater ecosystems. In this study, adult zebrafish were used to study the enantioselective bioaccumulation of HEX and its effect endpoints in liver, including oxidative stress and the regulation of apoptosis-related gene expression. Significant enantioselective bioaccumulation was demonstrated when exposed to HEX of 100 and 200 μg L(-)(1), finding that the (-)-enantiomer tended to accumulate in zebrafish more easily than (+)-enantiomer. Activities of antioxidant enzymes (SOD, CAT and GPx) and GSH content were all significantly decreased when zebrafish were exposed to 50 and 200 μg L(-1) HEX for 21 d. A series of genes of the apoptosis pathway were examined in groups treated with 50 and 200 μg L(-)(1) HEX for 21 d using real-time PCR. Significant up-regulation of p53, Puma, Apaf-1, caspase-3 and caspase-9 expression and down-regulation of Bcl-2/Bax expression ratio were proved. The overall results indicated that waterborne HEX was able to produce oxidative stress and induce apoptosis through the involvement of caspases in adult zebrafish. The above information will play a vital role in the integrated environmental risk assessment of HEX and make its toxic mechanism in fish clear., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In the present study, the enantioselective metabolism, cytotoxicity of hexaconazole and its influence on tryptophan metabolism in rat hepatocytes in vitro were investigated. Following the exposure of primary rat hepatocytes to rac-hexaconazole, the concentrations of its enantiomers in the media were determined by chiral high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The half-lives (t1/2) of (+)-hexaconazole and (-)-hexaconazole were 5.17 h and 19.80 h, respectively, indicating that the metabolic process was enantioselective with (-)-hexaconazole enrichment. Using the MTT method, the EC50 values of rac-hexaconazole, (+)-hexaconazole and (-)-hexaconazole after 12h of exposure were determined to be 71.62, 62.71 and 67.94 μM, respectively. Tryptophan metabolism was monitored using metabolomics profiling techniques. Hexaconazole and its enantiomers caused the down-regulation of tryptophan levels and the up-regulation of kynurenine (KYN) levels, suggesting a role for hexaconazole in the activation of the KYN pathway and providing information for the mechanism of its toxicity., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Myclobutanil, (RS)-2-(4-chlorophenyl)-2-(1H-1, 2, 4-triazol-1-ylmethyl) hexanenitrile is a widely used triazole fungicide. In this study, enantioselective metabolism and cytotoxicity were investigated in rat hepatocytes by chiral HPLC-MS/MS and the methyl tetrazolium (MTT) assay, respectively. Furthermore, tryptophan metabolism disturbance in rat hepatocytes after myclobutanil exposure was also evaluated by target metabolomics method. The half-life (t1/2) of (+)-myclobutanil was 10.66 h, whereas that for (-)-myclobutanil was 15.07 h. Such results indicated that the metabolic process of myclobutanil in rat hepatocytes was enantioselective with an enrichment of (-)-myclobutanil. For the cytotoxicity research, the calculated EC50 (12 h) values for rac-myclobutanil, (+)- and (-)-myclobutanil were 123.65, 150.65 and 152.60 µM, respectively. The results of tryptophan metabolites profiling showed that the levels of kynurenine (KYN) and XA were both up-regulated compared to the control, suggesting the activation effect of the KYN pathway by myclobutanil and its enantiomers which may provide an important insight into its toxicity mechanism. The data presented here could be useful for the environmental hazard assessment of myclobutanil., (© 2015 Wiley Periodicals, Inc.)

1. The enantiomeric enrichment or degradation of epoxiconazole has been reported in grape, soil, tubifex and mealworm beetle. But, little is known about its enantioselective behaviors in mammals. 2. To further understand differences in the distribution, degradation and excretion of epoxiconazole enantiomers in vivo, male CD-1 mice were selected as the test model to investigate the enantioselective behaviors after a single oral gavage. Mice were sacrificed after 1 h, 3 h, 7 h, 12 h, 24 h, 48 h, 72 h treatment, blood, tissues and excretions were collected for epoxiconazole analysis by LC-MS/MS. 3. On the Lux-Cellulose-1 chiral column, an enrichment of the second eluting (+)-epoxiconazole was generally observed, and feces and urine showed similar EF with major tissues. 4. To elucidate the potential role of intestinal bacterial flora in stereospecific degradation of epoxiconazole, mice fecal flora were cultured in vitro and incubated with epoxiconazole for 48 h. Results showed that (-)-epoxiconazole was preferentially degraded by intestinal bacterial. 5. These results may provide useful information for risk assessment of epoxiconazole on non-target animals.

The enantioselective bioactivity against pathogens (Cercospora arachidicola, Fulvia fulva, and Phytophthora infestans) and acute toxicity to Daphnia magna of the fungicide myclobutanil enantiomers were studied. The (+)-enantiomer in an antimicrobial activity test was about 1.79-1.96 times more active than the (-)-enantiomer. In the toxicity assay, the calculated 24-h LC50 values of the (-)-form, rac-form and (+)-form were 16.88, 13.17, and 11.91 mg/L, and the 48-h LC50 values were 10.15, 9.24, and 5.48 mg/L, respectively, showing that (+)-myclobutanil was more toxic. Meanwhile, the enantioselective metabolism of myclobutanil enantiomers following a single intravenous (i.v.) administration was investigated in rabbits. Total plasma clearance value (CL) of the (+)-enantiomer was 1.68-fold higher than its antipode. Significant differences in pharmacokinetics parameters between the two enantiomers indicated that the high bioactive (+)-enantiomer was preferentially metabolized and eliminated in plasma. Consistent consequences were found in the tissues (liver, brain, heart, kidney, fat, and muscle), resulting in a relative enrichment of the low-activity (-)-myclobutanil. These systemic assessments of the stereoisomers of myclobutanil cannot be used only to investigate environmental and biological behavior, but also have human health implications because of the long persistence of triazole fungicide and enantiomeric enrichment in mammals and humans., (© 2014 Wiley Periodicals, Inc.)

Triadimefon, a chiral fungicide, could be metabolized to triadimenol which has two chiral centers. In this work, Tubifex tubifex was exposed to triadimefon through the aqueous and soil phase to explore the relative importance of the routes of uptake. Bioaccumulation of triadimefon in tubifex was detected in both treatments, and the kinetics of the accumulation processes were significantly different in these two experiments. In spiked water treatment, (S)-triadimefon was preferentially accumulated over the (R)-triadimefon, whereas the enantioselective bioaccumulation was not detected in the spiked soil microenvironment. Simultaneously, four stereoisomers of triadimenol were also found in the tubifex tissue. Although the amount of these stereoisomers were different from each other with relatively more accumulation of the most fungi-toxic stereoisomer (1S,2R), the abundance ratios in the two exposure treatments were similar at the same sampling, following the order (1S,2S) > (1R,2S) > (1R,2R) > (1S,2R). The bioaccumulation factor was calculated for parent compound triadimefon and metabolite enrichment factor for metabolite. The results showed that both uptake routes, epidermal contact in the aqueous phase and ingestion of solid particles in soil, were important to the bioaccumulation of the triadimefon and triadimenol in tubifex.

Myclobutanil, (RS)-2-(4-chlorophenyl)-2-(1H-1, 2, 4-triazol-1-ylmethyl)hexanenitrile is a broad-spectrum systemic triazole fungicide which consists of a pair of enantiomers. The stereoselective degradation of myclobutanil was investigated in rat liver microsomes. The concentrations of myclobutanil enantiomers were determined by high-performance liquid chromatography (HPLC) with a cellulose-tris-(3,5-dimethyl-phenylcarbamate)-based chiral stationary phase (CDMPC-CSP) under reversed phase condition. The t(1/2) of (+)-myclobutanil is 8.49 min, while the t(1/2) of (-)-myclobutanil is 96.27 min. Such consequences clearly indicated that the degradation of myclobutanil in rat liver microsomes was stereoselective and the degradation rate of (+)-myclobutanil was much faster than (-)-myclobutanil. In addition, significant differences between two enantiomers were also observed in enzyme kinetic parameters. The V(max) of (+)-myclobutanil was about 4-fold of (-)-myclobutanil and the CL(int) of (+)-myclobutanil was three times as much as (-)-myclobutanil after incubation in rat liver microsomes. Corresponding consequences may shed light on the environmental and ecological risk assessment for myclobutanil and may improve human health., (© 2013 Wiley Periodicals, Inc.)

In the present study we investigated the enantioselective disappearance of hexaconazole in rat liver microsomes system prepared from both genders. High-performance liquid chromatography (HPLC) was used for identification and quantification. The degradation of the (+)-hexaconazole was faster than that of the (-)-hexaconazole in racemic hexaconazole and single enantiomer incubation in both sexes. The degradation half-life of the (+)-hexaconazole or (-)-hexaconazole was also gender-related. The metabolism of (+)-hexaconazole and (-)-hexaconazole were faster in male rat hepatic microsomes than that in female, suggesting that at least one of the cytochrome P450s (CYP) in the male rat liver microsomes system responsible for hexaconazole metabolism was male-specific or considerably more active. Kinetic assays showed that the intrinsic clearance in male rat liver microsomes was higher than that in female. All these results strongly suggest that sexual dimorphic metabolism of hexaconazole exists in rats. The inhibition experiments with CYP inhibitors showed that the inhibitory effect of inhibitors was enantioselective and affected by sex. The results suggest that the enantioselective metabolism of hexaconazole was determined by the amount of hepatic cytochrome P450 and the expression of individual isoforms of CYPs., (© 2013 Wiley Periodicals, Inc.)

Research on the enantioselective environmental behavior of chiral pesticides has been a hot spot of environmental chemistry recently. In this study, the acute toxicity of myclobutanil enantiomers was investigated with the aquatic algae Scendesmus obliquus. After exposure for 96 h, the EC50 values for (-)-myclobutanil, rac-myclobutanil and (+)-myclobutanil were 3.951, 2.760, and 2.128 mg/L, respectively. The photosynthetic pigment (chlorophyll a, chlorophyll b, and carotenoids) and antioxidant enzyme activities catalase (CAT) were determined to evaluate the different toxic effects when S. obliquus were exposed to 1.5, 5 and 15 mg/L of rac-myclobutanil, (-)-myclobutanil, and (+)-myclobutanil for 96 h, respectively. In addition, the degradation of myclobutanil enantiomers in S. obliquus was also studied. Myclobutanil in the medium inoculated with algae degraded faster than in the uninoculated medium. The degradation of (-)-myclobutanil was faster than that of (+)-myclobutanil at a concentration of 3 mg/L. On the basis of these data, the acute toxicity and toxic effects of myclobutanil against S. obliquus were concluded to be enantioselective, and such enantiomeric differences should be taken into consideration in pesticide risk assessment., (© 2013 Wiley Periodicals, Inc.)

In this study, the bioactivity, acute toxicity and dissipation in vegetables of the individual enantiomers of the fungicide hexaconazole had been investigated. The optical pure single enantiomers were prepared and the bioactivity of (+)-, (-)- and rac-hexaconazole was tested using four target fungi including Colletotrichum gloeosporioides Penz, Alternaria solani, Alternaria mali Roberts and Monilinia fructicola. The results showed (-)-hexaconazole was always more active than (+)-hexaconazole with the fungicidal activity 11–13-fold higher to A. solani, A. mali Roberts and Monilinia fructicola, and 1.26-fold higher to C. gloeosporioides Penz. (-)-Hexaconazole also showed 1.3-fold higher acute toxicity to aquatic species Daphnia magna based on the 48 h EC50 values. There was obvious enantioselectivity in the dissipation in tomato with (-)-hexaconazole degraded faster resulting an enrichment of (+)-form, and the half-lives of (-)-hexaconazole and (+)-hexaconazole in tomato were 2.96 d and 3.38 d respectively, while it was not enantioselective in green pepper, in which the both enantiomers had the half-lives about 4.36 d. The findings are helpful for better environmental and ecological risk assessment of hexaconazole on an enantiomeric level.

Racemic triadimefon (TF) was applied to wheat and soil at three sites (Beijing, Huaibei, and Zhengzhou in China) under open field conditions. Its enantioselective degradation and stereoselective transformation to the major metabolite, triadimenol (TN), in wheat straw, grain and soil were investigated. At all sites, the degradation of TF enantiomers in straw and soil followed first-order kinetics. In soil from Beijing and Zhengzhou R-(-)-TF was preferentially degraded; however, preferential enantioselective degradations were not recorded in soil from Huaibei or in the straw from all sites. There were noticeable differences in the stereoselective formation of TN stereoisomers in all straw and soil samples. TN diastereomer A with high animal toxicity was preferentially produced via a reductive reaction in straw. In contrast, diastereomer B, was preferential in soil across the experimental period. Different TN concentrations were found in the order of SR-(-)-TN>RR-(+)-TN>RS-(+)-TN>SS-(-)-TN in straw, and RR-(+)-TN>SS-(-)-TN>SR-(-)-TN>RS-(+)-TN in soil. Neither TF nor TN was found in wheat grain at harvest. Because of differences in degradation, formation, and toxicity, the characterization of enantiomers and stereoisomers in this study contributes toward comprehensively assessing the fate and risk of chiral agrochemicals in the environment and food., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A rapid, simple, reliable, and environment-friendly method for the residue analysis of the enantiomers of four chiral fungicides including hexaconazole, triadimefon, tebuconazole, and penconazole in water samples was developed by dispersive liquid-liquid microextraction (DLLME) pretreatment followed by chiral high-performance liquid chromatography (HPLC)-DAD detection. The enantiomers were separated on a Chiralpak IC column by HPLC applying n-hexane or petroleum ether as mobile phase and ethanol or isopropanol as modifier. The influences of mobile phase composition and temperature on the resolution were investigated and most of the enantiomers could be completely separated in 20 min under optimized conditions. The thermodynamic parameters indicated that the separation was enthalpy-driven. The elution orders were detected by both circular dichroism detector (CD) and optical rotatory dispersion detector (ORD). Parameters affecting the DLLME performance for pretreatment of the chiral fungicides residue in water samples, such as the extraction and dispersive solvents and their volume, were studied and optimized. Under the optimum microextraction condition the enrichment factors were over 121 and the linearities were 30-1500 µg L(-1) with the correlation coefficients (R(2)) over 0.9988 and the recoveries were between 88.7% and 103.7% at the spiking levels of 0.5, 0.25, and 0.05 mg L(-1) (for each enantiomer) with relative standard deviations varying from 1.38% to 6.70% (n = 6) The limits of detection (LODs) ranged from 8.5 to 29.0 µg L(-1) (S/N = 3)., (© 2013 Wiley Periodicals, Inc.)

Enantioselectivity in ecotoxicity of chiral pesticides in the aquatic environment has been a subject of growing interest. In this study, the toxicological impacts of hexaconazole enantiomers were investigated with freshwater algae Scenedesmus obliquus. After 96 h of exposure, the EC(50) values for rac-hexaconazole, (+)-hexaconazole, and (-)-hexaconazole were 0.178, 0.355, and 0.065 mg l(-1) , respectively. Therefore, the acute toxicities of hexaconazole enantiomers were enantioselective. In addition, the different toxic effects were evaluated when S. obliquus were exposed to 0.2, 0.5, and 1.0 mg l(-1) of rac-hexaconazole, (+)-hexaconazole, and (-)-hexaconazole during 96 h, respectively. The chlorophyll a and chlorophyll b contents of S. obliquus treated by (-)-hexaconazole were lower than those exposed to (+)-hexaconazole, whereas the malondialdehyde contents of S. obliquus treated by (-)-form were higher than those exposed to (+)-form at higher concentrations. In general, catalase activities were significantly upregulated by exposure to (-)-enantiomer than (+)-enantiomer at all three concentrations. However, superoxide dismutase activities exposed to (-)-hexaconazole were lower than that exposed to (+)-hexaconazole at 0.2 mg l(-1) and 0.5 mg l(-1) . On the basis of these data, the acute toxicity and toxic effects of hexaconazole against S. obliquus were enantioselective, and such enantiomeric differences must be taken into consideration in pesticide risk assessment., (Copyright © 2012 Wiley Periodicals, Inc.)

Stereoselective dissipation of epoxiconazole had been studied in grape and soil during plant growing under field conditions in this paper. A sensitive and rapid chiral method was developed and validated for the determination of epoxiconazole stereoisomers in grape and soil based on liquid chromatography coupled with triple quadrupole mass spectrometry (LC-MS/MS). Phenomenex Lux Cellulose-1 column was used for enantioseparation with a mixture of acetonitrile/water (90/10, v/v) as mobile phase at flow rate of 0.3 mL min(-1). Fortified recoveries in grape and soil samples ranged from 76.0% to 91.9% and relative standard deviations were less than 11.4% with fortified levels of 0.025-1.0 mg kg(-1). The limits of detection and quantification were 0.005 mg kg(-1) and 0.025 mg kg(-1), respectively, with linear calibration curves extending up to 5.0 mg kg(-1). The field experimental results showed that dissipations of epoxiconazole stereoisomers in grape followed first-order kinetics (R(2)>0.92) and stereoselectivity occurred in 2 h after spraying. The (-)-stereoisomer with half-life of 9.3 d degraded faster than (+)-stereoisomer with that of 13.2 d, and resulted in relative enrichment of (+)-stereoisomer. However, the stereoisomeric dissipations in soil were triphasic ("increase-decrease-steady") with lower dissipation rates, and also occurred with preferential degradation of (-)-stereoisomer under field condition. The results for stereoselective dissipations can be applied for food and environmental assessments of chiral pesticides., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Hexaconazole [(RS)-2-(2,4-dichlorophenyl)-1-(1H-1,2,4-triazol-1-yl) hexan-2-ol] is a potent triazole fungicide and consists of a pair of enantiomers. Enantioselective degradation of hexaconazole was investigated in rat hepatic microsomes in vitro. Concentrations of (-)- and (+)-hexaconazole and enantiomer fraction were determined by high performance liquid chromatography with a cellulose-tris-(3,5-dimethylphenylcarbamate)-based chiral stationary phase. The t(1/2) of (-)-hexaconazole and (+)-hexaconazole were 23.70 and 13.95 min for rac- hexaconazole and 44.18 and 23.54 for enantiomers examined separately. Furthermore, hexaconazole is configurationally stable in rat hepatic microsomes, demonstrating no chiral inversion from the (-)-hexaconazole to (+)-hexaconazole or vice versa. Intrinsic metabolic clearance of (+)-hexaconazole is 1.12 times than that of (-)-hexaconazole. Interaction study revealed that there was competitive inhibition between (-)-hexaconazole and (+)-hexaconazole. In addition, there was a significant difference between the inhibitory concentration (IC(50)) of (-)- to (+)-hexaconazole and (+)- to (-)-hexaconazole [IC(50)(-)/(+)/IC(50)(+)/(-) = 1.88]. These results may have potential implications for better environmental and ecological risk assessment for hexaconazole., (Copyright © 2012 Wiley Periodicals, Inc.)

A sensitive and rapid analytical method was developed for simultaneous determination of triadimefon (TF) and triadimenol (TN) stereoisomers in wheat, straw, and soil by liquid chromatography coupled with triple quadrupole mass spectrometry (LC-MS/MS). The direct enantioseparation of TF and TN was performed on a Lux cellulose-1 column packed with cellulose-tris-(3,5-dimethylphenylcarbamate). The effects of mobile-phase composition on the separation were investigated and stereoisomeric elution orders were confirmed with a polarimeter detector. The pesticides were extracted from samples with acetonitrile and cleaned up by solid-phase extraction or activated carbon. Based on the developed stereoselective LC-MS/MS method, for TF and TN stereoisomers, good linearities were obtained over the concentration range of 0.003-4 mg/L; recoveries were 84.2-102.7% in wheat, 84.0-104.0% in straw, and 85.2-106.8% in soil at spiked concentrations of 0.007-2.0 mg/kg; intra-day and inter-day assay precisions were below 12.2%. Limits of detection (LODs) and limits of quantification (LOQs) in wheat, straw, and soil were 0.001-0.005 mg/kg and 0.007-0.02 mg/kg, respectively. Finally, the method was successfully applied to detect TF and TN stereoisomers in wheat, straw, and soil samples from residual trials in farm., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The aim of this study was to assess the stereoselectivity of two tebuconazole [(RS)-1-p-chlorophenyl-4,4-dimethyl-3-(1H-1,2,4-triazol-1-ylmethyl)pentan-3-ol] enantiomers in in vitro system (rat liver microsomes). The analytes were extracted with acetic ether and concentrations were determined by high performance liquid chromatography (HPLC) with a cellulose tris(3,5-dimethylphenylcarbamate)-based chiral stationary phase. The degradation of rac-tebuconazole (15 μM) followed first-order kinetics, and the degradation of the S-tebuconazole (t(1/2) = 22.31 min) was faster than that of the R-tebuconazole (t(1/2) = 48.76 min), but no significant difference between the enantiomers was found in the respective incubation (7.5 μM for each). Kinetic assays showed that the K(m) was different between the two enantiomers (K(mR) = 14.83 ± 2.19, K(mS) = 12.23 ± 2.72). The interaction results revealed that there was competitive inhibition between S- and R-form, and there was a significant difference between the IC(50) of R- to S-tebuconazole and S- to R-tebuconazole (IC(50R/S)/IC(50S/R) = 4.98)., (Copyright © 2011 Wiley Periodicals, Inc.)

The stereoselective pharmacokinetics of flutriafol were investigated in male and female adult Japanese white rabbits. Following intravenous administration of rac-flutriafol to rabbits at 5 mg/kg (bd wt), the concentrations of the enantiomers in plasma were determined by a HLPC-UV method using a CDMPC-CSP chiral column. R-Flutriafol exhibited a shorter distribution half-life but a longer elimination half-life than the S-isomer. In female rabbits, the distribution half-lives of R- and S-flutriafol were found to be 0.09 and 0.18 h, respectively, significantly shorter than those in male rabbits, but the volume of distribution and elimination half-life for flutriafol enantiomers in both sexes of rabbit showed no significant differences. Female rabbits had a higher clearance for both flutriafol enantiomers. The protein binding value was high for both isomers, with enantioselectivity, but no gender difference. It was an important factor in modulating the disposition of flutriafol. Flutriafol concentrations in kidney, liver, fat, and lung were higher than in other tissues at 10 h after administration, and the concentrations of R-flutriafol were higher in all tissues than those of its antipode. However, gender difference in flutriafol residues in tissues was not observed. It is concluded that the stereoselectivity of flutriafol on distribution and elimination in rabbits mainly depends upon gender.

A series of novel chiral triazole compounds were synthesized. They were separated into enantiomers by liquid chromatography on an amylose tris(3,5-dimethylphenylcarbamate) (ADMPC) chiral stationary phase (CSP). The absolute configuration of each enantiomer of the investigated compounds was established by combined use of chemical correlation, chiral HPLC and circular dichroism (CD) spectra analysis methods. The influence of the mobile-phase modifiers and the structure of chiral triazole compounds on the chiral separation and retention were investigated. Reversal of the elution order of some enantiomeric pairs upon using different mobile-phase modifier was observed. The temperature effect on the chiral separation and the thermodynamic properties including enthalpy and entropy change of binding to the ADMPC-CSP were also investigated., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Diniconazole [(E)-(RS)-1-(2,4,-dichlorophenyl)-4,4-dimethyl-2-(1H-1,2,4-triazole-1-yl)pent-1-en-3-ol)] is a potent triazole fungicide. The enantioselective pharmacokinetics of diniconazole enantiomers in rabbits was studied via intravenous (i.v.) injection. The pharmacokinetics and the enantiomer fraction (EF) were determined using normal high-performance liquid chromatography with diode array detection and a cellulose-tris-(3,5-dimethylphenylcarbamate)-based chiral stationary phase (CDMPC-CSP). The time-concentration curves in plasma were fitted by a two-compartment open mode. The results showed that the concentration of S-diniconazole in plasma decreased faster than that of R-diniconazole, and EFs increased with time after administration of racemic diniconazole (rac-diniconazole). The R-/S-enantiomer ratio of the area under the time-plasma concentration curve (AUC(0-infinity)) after administration was 1.52. The total plasma clearance value of S-enantiomer was 1.57-fold higher than that of the R-diniconazole. These results indicate substantial stereoselectivity in the kinetics of diniconazole enantiomers in rabbit., (2008 Wiley-Liss, Inc.)

The amylose-tris(3,5-dimethylphenylcarbamate) chiral stationary phase was synthesized and used to separate the enantiomers of triazole pesticides by high-performance liquid chromatography. The mobile phase was n-hexane-isopropanol applying a flow rate of 1.0 mL/min. Six triazole pesticides were enantioselectively separated. Myclobutanil, paclobutrazol, tebuconazole, and uniconazole obtained complete separation with the resolution factors of 5.73, 2.99, 1.72, and 2.07, respectively, and imazalil and diniconazole obtained partial separation with the resolution factors of 0.79 and 0.77 under the optimized conditions. The effect of the content of isopropanol as well as column temperature on the separation was investigated. A circular dichroism detector was used to identify the enantiomers and determine the elution orders. The results showed the low temperature was good for the chiral separation except for diniconazole. The thermodynamic parameters calculated based on linear Van't Hoff plots showed the chiral separations were controlled by enthalpy.

Methods for the enantiomeric quantitative determination of 3 chiral pesticides, paclobutrazol, myclobutanil, and uniconazole, and their residues in soil and water are reported. An effective chiral high-performance liquid chromatographic (HPLC)-UV method using an amylose-tris(3,5-dimethylphenylcarbamate; AD) column was developed for resolving the enantiomers and quantitative determination. The enantiomers were identified by a circular dichroism detector. Validation involved complete resolution of each of the 2 enantiomers, plus determination of linearity, precision, and limit of detection (LOD). The pesticide enantiomers were isolated by solvent extraction from soil and C18 solid-phase extraction from water. The 2 enantiomers of the 3 pesticides could be completely separated on the AD column using n-hexane isopropanol mobile phase. The linearity and precision results indicated that the method was reliable for the quantitative analysis of the enantiomers. LODs were 0.025, 0.05, and 0.05 mg/kg for each enantiomer of paclobutrazol, myclobutanil, and uniconazole, respectively. Recovery and precision data showed that the pretreatment procedures were satisfactory for enantiomer extraction and cleanup. This method can be used for optical purity determination of technical material and analysis of environmental residues.

The enantiomeric separations of three pesticides fipronil (asymmetric nitrogen), isocarbophos (asymmetric phosphorus) and carfentrazone-ethyl (asymmetric carbon) were studied on cellulose-tri(3,5-dimethylphenylcarbamate) chiral stationary phase using high-performance liquid chromatography under normal phase. The mobile phase was n-hexane with alcohols including ethanol, n-propanol, iso-propanol, n-butanol and iso-butanol as polar modifiers. The flow rate was 1.0 mL/min with UV detection at 280, 225 and 230 nm for fipronil, isocarbophos and carfentrazone-ethyl respectively. The influence of the modifiers and their volume content and temperature from 0 to 50 degrees C on the separations was investigated. The chiral stationary phase showed excellent stereoselectivity for the two enantiomers of fipronil and isocarbophos and certain chiral recognition for carfentrazone-ethyl. Iso-propanol was more suitable for the chiral separation of isocarbophos and carfentrazone-ethyl, and iso-butanol was better for fipronil. The resolutions increased with the decreasing modifier content and temperature for all the three chiral pesticides., (Copyright 2005 John Wiley & Sons, Ltd.)

Cellulose-tris (3,5-dimethylphenylcarbamate; CDMPC) was synthesized and coated on aminopropylsilica to prepare chiral stationary phase (CSP). Normal-phase high-performance liquid chromatography (HPLC) methods for the resolutions of five chiral triazole pesticides, diniconazole, tebuconazole, hexaconazole, triadimefon and flutriafol, on the CSP were developed. Several operating parameters such as mobile phase composition, modifier and column temperature were studied for the optimization of the resolutions. Better separations were achieved using 2% iso-butanol for diniconazole, 2% ethanol for tebuconazole, 2% iso-propanol for hexaconazole, 1% n-butanol for triadimefon and 2% n-propanol for flutriafol as modifiers in n-hexane at 0 degrees C with the resolution factors (Rs) of 1.62, 1.66, 2.46, 1.68 and 1.98, respectively. Low temperature was better for the resolutions. Validation of the methods included linearity and precision.