Chiral Fungicide Research Articles

Unraveling complexation and separation of novel asymmetric uranyl-5-methoxy-2-(4-methoxy-6’-(quinazolin-2-yl)-[2,2’-bipyridin]-6-yl) quinazoline to chiral fungicides R/S-metalaxyls and R/S-benalaxyls

ABSTRACT Chiral fungicides R/S-metalaxyls(R/S-MTLs) and R/S-benalaxyls(R/S-BNLs) have different biotoxicity. It is very important to unravel molecular recognition and enantiomeric separation of R/S-MTLs or R/S-BNLs. In this paper, we designed a novel asymmetric ligand: 5-methoxy-2-(4-methoxy-6”-(quinazolin-2-yl)-[2,2”−bipyridin]-6-yl)quinazoline(MMQBQ), further constructed a new asymmetric receptor Uranyl-MMQBQ by coordination of MMQBQ with uranyl. The complexation and separation of receptor Uranyl-MMQBQ with guests R/S-MTLs or R/S-BNLs were systematically studied using density functional theory (DFT) method. The respects of ESP, MBO, RFC, LOL, EDDM, EST-NOCV, QTAIM, FMOs, IGMH, and Gibbs free energy changes were calculated and analyzed. The results indicated that the MMQBQ could form a stable receptor Uranyl-MMQBQ with uranyl, which could selectively recognize and separate chiral R/S-MTLs or R/S-BNLs by receptor’s U coordinating to carbonyl oxygens of guests. For R/S-MTLs, separation factors(SF S/R) of Uranyl-MMQBQ were more than 16 in water, chlorobenzene, toluene, carbon tetrachloride and cyclohexane, enantioselectivity coefficients(ESCs) ranged of 99.92%-94.24% in the above four solvents. While for R/S-BNLs, ESCs of Uranyl-MMQBQ were over 99% and SF S/R values were a range of 533.97–2839.36 in the above organic solvents. The findings provide valuable information for design of novel uranyl ligand and its complexes, meanwhile supply useful guidance for experiments on separation of other chiral fungicides in environmental protection.

Enantioselectivity effects of energy metabolism in honeybees (Apis mellifera) by triticonazole

The heavy use of agrochemicals is considered a major factor contributing to the decline in wild honeybee populations. Development of low-toxicity enantiomers of chiral fungicides is the key to reducing the potential threats to honeybees. In this study, we evaluated the enantioselective toxic effects of triticonazole (TRZ) on honeybees and its molecular mechanisms. The results showed that after long-term exposure to TRZ, the content of thoracic ATP decreased significantly, by 41 % in R-TRZ treatments and by 46 % in S-TRZ treatments. Furthermore, the transcriptomic results indicated that S-TRZ and R-TRZ significantly altered the expression of 584 genes and 332 genes, respectively. Pathway analysis indicated that R- and S-TRZ could affect different genes expressed in GO terms and metabolic pathways, especially the transport GO terms (GO: 0006810) and pathways of alanine, aspartate and glutamate metabolism, drug metabolism - cytochrome P450, and pentose phosphate. Additionally, S-TRZ had a more pronounced effect on honeybee energy metabolism, disrupting a greater number of genes involved in the TCA cycle and glycolysis/glycogenesis, exerting a stronger effect on energy metabolic pathways, including nitrogen metabolism, sulfur metabolism, and oxidative phosphorylation. In summary, we recommend reducing the proportion of S-TRZ in racemate to minimize the threat to the survival of honeybees and protect the diversity of economic insects.

Insights on the isolation, identification, and degradation characteristics of three bacterial strains against mandipropamid and their application potential for polluted soil remediation

Bacteria-induced biodegradation techniques have become an effective approach for removing pesticide residues from polluted soils. However, their effect on chiral fungicides must be systematically evaluated and the efficiency and risk of each chiral enantiomer must be better understood. In this study, we isolated and enriched seven bacterial strains that are able to degrade mandipropamid from contaminated soil samples. Three bacterial strains with high degradation efficiency (63.6%–73.4%) were screened and identified as Pseudomonas sp. (M01), Mycolicibacterium parafortuitum (MW05), and Stenotrophomonas maltophilia (MW09) by morphological and 16S rRNA gene sequencing analyses. The degradation characteristics of three strains (M01, MW05, and MW09) was investigated and it was revealed that pH, temperature, and initial concentration of mandipropamid significantly impacted their degradation efficiency. The optimal conditions for degradation were a nutrient source of mandipropamid and an inoculation amount of 5%. We used a Box-Behnken model experiment and an analysis of variance to determine the most suitable conditions for degrading mandipropamid at various pH, temperature, and initial concentration levels. A response surface methodology analysis showed that the three strains had the highest mandipropamid degradation efficiency (> 96%) under various conditions (pH: 7.15–7.71, temperature: 28.61–30.76 °C, initial concentration: 5.524–5.934 mg/L). Mycelial, intracellular, and extracellular enzymes also had an impact on the degradation of mandipropamid enantiomers by the three strains. In soil remediation trials, the three bacterial strains could effectively enantioselectively degrade rac-mandipropamid residues in polluted sterilized and natural soil samples (R-enantiomer was degraded faster) and influence the activity of urease and β-glucosidase in the soil. The results revealed several candidate bacterial strains for mandipropamid biodegradation and provide information on mandipropamid biological detoxification in soil environments.

Reduction in the Residues of Penthiopyrad in Processed Edible Vegetables by Various Soaking Treatments and Health Hazard Evaluation in China

Tomato and cucumber are two vital edible vegetables that usually appear in people's daily diet. Penthiopyrad is a new type of amide chiral fungicide, which is often used for disease control of vegetables (including tomato and cucumber) due to its wide bactericidal spectrum, low toxicity, good penetration, and strong internal absorption. Extensive application of penthiopyrad may have caused potential pollution in the ecosystem. Different processing methods can remove pesticide residues from vegetables and protect human health. In this study, the penthiopyrad removal efficiency of soaking and peeling from tomatoes and cucumbers was evaluated under different conditions. Among different soaking methods, heated water soaking and water soaking with additives (NaCl, acetic acid, and surfactant) presented a more effective reduction ability than other treatments. Due to the specific physicochemical properties of tomatoes and cucumbers, the ultrasound enhances the removal rate of soaking for tomato samples and inhibits it for cucumber samples. Peeling can remove approximately 90% of penthiopyrad from contaminated tomato and cucumber samples. Enantioselectivity was found only during tomato sauce storage, which may be related to the complex microbial community. Health risk assessment data suggests that tomatoes and cucumbers are safer for consumers after soaking and peeling. The results may provide consumers with some useful information to choose better household processing methods to remove penthiopyrad residues from tomatoes, cucumbers, and other edible vegetables.

Systemic enantioselectivity study of penthiopyrad: enantioselective bioactivity, acute toxicity, degradation and influence on tomato.

In order to promote the green development of agriculture, it is important to study the enantioselective effect of chiral pesticides. The bioactivity of the chiral fungicide penthiopyrad (PEN) racemate and enantiomers against phytopathogens, toxicity to non-target organisms, effect on tomato fruit growth and maturation, and environmental fate in tomato cultivation were evaluated at an enantioselective level in this study. The results indicated that at the same efficacy, the optically pure S-(+)-PEN could lower the dosage of racemate by 20-96%. The S-enantiomer had low toxicity to earthworms. Besides, the S-(+)-PEN did not cause significant abiotic stress to the tomato and increased fruit fresh weight and size via modulating the contents of plant hormones. However, the content of hydrogen peroxide (H2 O2 ), superoxide (O2 - ) and malondialdehyde in the R-enantiomer treatment group was significantly higher than the control group. The effect of the racemate on tomato fruit was between the enantiomers. Furthermore, compared to R-(-)-PEN and racemate, the S-enantiomer degraded more quickly in tomato fruit, leaves, and soil, reducing the danger of human exposure. The S-enantiomer is highly effective and less toxic. The development of enantiomer pure S-(+)-PEN products might be an efficient and low-risk strategy. The results lay the foundation for comprehensive evaluation and proper application of PEN. © 2023 Society of Chemical Industry.

Fertilization can modify the enantioselective persistence of penthiopyrad in relation to the co-influence on soil ecological health

Penthiopyrad is a widely used chiral fungicide for controlling rust and Rhizoctonia diseases. Development of optically pure monomers is an important strategy to realize amount reduction and increment effects of penthiopyrad, wherein, fertilizers as the co-exiting nutrient supplement may alter the enantioselective residues of penthiopyrad in soil. In our study, influences of urea, phosphate, potash, NPK compound, organic granular, vermicompost and soya bean cake fertilizers on enantioselective persistence of penthiopyrad were fully evaluated. This study demonstrated that R-(−)-penthiopyrad dissipated faster than S-(+)-penthiopyrad during 120 days. High pH, available nitrogen, invertase activities and reduced available phosphorus, dehydrogenase, urease, catalase activities were situated to benefit removing the concentrations of penthiopyrad and weakening enantioselectivity in soil. With respect to the impact of different fertilizers on soil ecological indicators, vermicompost contributed to enhanced pH. Urea and compound fertilizer played an absolute advantage in promoting available nitrogen. All fertilizers didn't go against available phosphorus. Dehydrogenase responded negatively to phosphate, potash and organic fertilizers. Urea increased invertase, besides, it and compound fertilizer both diminished urease activity. The catalase activity was not activated by organic fertilizer. Based on all the findings, soil application of urea and phosphate fertilizers was recommended and considered as a better option to exhibit high efficiency for the dissipation of penthiopyrad. The combined environmental safety estimation can effectively guide the treatment of fertilization soils in line with the nutrition requirements and pollution regulation from penthiopyrad.

Enantioselective Degradation and Bioactivity Mechanism of a New Chiral Fungicide Fluindapyr in Paddy Ecosystems.

Fluindapyr is a novel chiral succinate dehydrogenase inhibitor used to control fungal diseases. The enantioselective effects of fluindapyr in paddy ecosystems are unknown. We developed a new chiral determination method of fluindapyr using ultrahigh performance liquid chromatography tandem mass spectrometry. The absolute configuration of the fluindapyr enantiomers was identified by an electron circular dichroism model. A new husk-based biochar material was used to optimize and establish a QuEchERs method for paddy soil determination. Under anaerobic conditions, the half-lives of R-fluindapyr and S-fluindapyr in paddy soil were 69.6 and 101.8 days, respectively. R-fluindapyr degraded more rapidly than S-fluindapyr. S-fluindapyr was 87.8 times more active against Rhizoctonia solani than R-fluindapyr. The enantioselective bioactivity mechanism was illustrated by molecular docking between the fluindapyr enantiomers and SDH of R. solani. The binding powers of R-fluindapyr and S-fluindapyr to proteins were -32.12 and - 42.91 kcal/mol, respectively. This study reports the stereoselectivity of fluindapyr about determination, degradation, bioactivity, and its mechanism. It provides a foundation for an in-depth study of fluindapyr at the enantiomer level.

Subzero-temperature homogeneous liquid-liquid extraction for the stereoselective determination of chiral triadimefon and its metabolite in water, fruit juice, vinegar, and fermented liquor by HPLC.

A novel method based on homogeneous liquid-liquid extraction with deep eutectic solvents (DES) under subzero-temperature conditions in combination with high performance liquid chromatography (HPLC) for the determination of chiral fungicide triadimefon (TF) and its metabolite triadimenol (TN) in water, fruit juice, vinegar, and fermented liquor was developed in this study. The method involved using deep eutectic solvents (DES) under subzero-temperature conditions in combination with high performance liquid chromatography (HPLC). This novel technique, known as subzero-temperature homogeneous liquid-liquid extraction (STHLLE), offers several advantages, including high efficiency, time-saving, low-cost, and eco-friendliness. The enantiomers of chiral TF and TN were simultaneously separated and quantified using HPLC coupled with a Daicel Chiralpak OD-RH column. Various experimental parameters such as DES composition and volume, freezing condition, salt concentration, and pH were optimized to enhance the recoveries of the target analytes. Under the optimized conditions, spiked recoveries of six enantiomers (i.e., S-TF, R-TF, SR-TN, RS-TN, SS-TN, and RR-TN) in the water, fruit juice, vinegar, and fermented liquor samples were 82.2-100.1% with relative standard deviations of 0.4-10.1%. The current method demonstrated a detection range of 0.03-0.06 mg L-1 in the target analytes. This established technique exhibits potential for efficient and precise extraction and quantification of the enantiomers of TF and TN in water phase samples.

Stereoselective behavior and residues of the imazalil during strawberry growth and strawberry wine production

Imazalil is a chiral fungicide widely used to protect strawberries against gray mold, which may pose threats to food safety. This study aims to investigate the stereoselective behavior of imazalil during strawberry growth and strawberry wine production. A method was proposed and validated for the extraction and quantitative analysis of imazalil residues in strawberry, strawberry pomace, and strawberry wine by using ultra-high performance liquid chromatography-tandem mass spectrometry. The method exhibited mean recoveries ranging from 86.2% to 119.7% with relative standard deviations of 0.1–11.3%. The dissipation curve of imazalil during strawberry growth followed the first-order kinetic model with a half-life ranging from 6.5 to 7.1 days. Significant enantioselectivity of imazalil was observed in strawberry grown under field conditions and strawberry wine production process, with enantiomeric fraction values ranging from 0.51 (2 h) to 0.42 (27d) and from 0.48 (0d) to 0.52 (10d), respectively. (+)-imazalil was preferentially degraded in strawberry under field conditions, while (−)-imazalil was preferentially degraded during the fermentation process. The processing factor was lower than 1 for each procedure, indicating that the wine-making process can reduce imazalil residue in strawberry. These findings may facilitate a more accurate risk assessment of imazalil and provide important guidance for the safe and efficacious use of imazalil in agriculture.

Effects of Household Processing on Residues of the Chiral Fungicide Mandipropamid in Four Common Vegetables.

The study aimed to detect the content of mandipropamid enantiomers in unprocessed and processed tomato, cucumber, Chinese cabbage, and cowpea samples and assess the health risks to Chinese consumers. Data showed that washing and soaking with an acidic solution reduced the mandipropamid residue from vegetable samples by 54.1-82.2%. The pickling process resulted in a 6.2-65.2% loss of mandipropamid from cucumber, Chinese cabbage, and cowpea samples. Peeling and juicing were the best removing techniques for mandipropamid residues in tomato and cucumber (removal rate (RR) value > 91%), and cooking for 5 min could effectively reduce the levels of mandipropamid in Chinese cabbage and cowpea (RR values of 81.4-99.7%). The values of processing factor for the processed vegetable samples are all less than one. No significant enantioselectivity of mandipropamid was found in the vegetables during processing. Health risk data showed that samples of four types of mandipropamid-contaminated vegetables were safe for consumption after processing.

Stereoselective accumulation and biotransformation of chiral fungicide epoxiconazole and oxidative stress, detoxification, and endogenous metabolic disturbance in earthworm (Eisenia foetida)

>80 % of applied pesticides in agriculture will enter the soil and be exposed to soil animals. Little is known about the stereoselective metabolic effects of epoxiconazole (EPO) on soil animals. In this study, EPO-mediated stereoselective enrichment, biotransformation, oxidative stress, detoxification, and global metabolic profiles in earthworms were investigated by exposure to EPO and its enantiomers at 1 mg/kg and 10 mg/kg doses. Preferential enrichment of (−)-EPO was observed, and the five transformation products (TPs) exhibited the chemically specific stereoselective accumulation with inconsistent configurations. Biochemical markers related to reactive oxygen species (ROS) and detoxification (·OH− content, SOD, CAT, GST, and CYP450 enzymes) showed a significant stereoselective activation overall at the low-level exposure (p-value <0.05). Based on untargeted metabolomic analysis, the steroid biosynthesis and ROS-related biotransformation, glutathione metabolism, TCA cycle, amino acid metabolism, purine and pyrimidine metabolism of earthworms were significantly interfered with by EPO and its enantiomer exposure. More pronounced stereoselectivity was observed at the level of the global metabolic profile, while comparable levels of metabolic perturbations were identified at the individual metabolite level. This study provides novel insights into the stereoselective effects of the chiral fungicide EPO, and valuable evidence for soil environmental risk assessments.

Triazole resistance in Aspergillus fumigatus exposed to new chiral fungicide mefentrifluconazole.

Triazole resistance in the human fungal pathogen Aspergillus fumigatus has been a growing challenge in clinic treatment with triazole drugs such as itraconazole. The fast evolvement of triazole resistance in A.fumigatus in the ecosystem has drawn great attention, and there has been a possible link between the application of triazole fungicides in agriculture and triazole resistance in A.fumigatus. The change in susceptibility of A.fumigatus exposed to the new chiral triazole fungicide mefentrifluconazole was investigated in this study. The results indicated that triazole resistance in A.fumigatus was acquired with exposure to mefentrifluconazole at a level of greater than or equal to 2mg L-1 in liquid medium and soil (not at 0.4 nor 1mg L-1 ). Interestingly, stereoselectivity was found in the acquisition of triazole resistance in A.fumigatus when exposed to mefentrifluconazole. R-mefentrifluconazole, which is very active on plant pathogens, exhibited stronger possibility in the development of the resistance in A.fumigatus than its antipode. Overexpression of cyp51A, AtrF, AfuMDR1 and AfuMDR4 were associated with the acquired resistance in A.fumigatus with hereditary stability. The results suggest that triazole resistance in A.fumigatus could be resulted from the selection of mefentrifluconazole at concentrations larger than 2mg L-1 . Mefentrifluconazole should be applied within the dosage recommended by good agricultural practice to avoid the resistance in A.fumigatus in soil. This also may be applicable to other triazole fungicides. © 2022 Society of Chemical Industry.

Toxicokinetics of Two Oxathiapiprolin Enantiomers in Rats and Their Stereoselective Interaction with Oxysterol Binding Protein.

Oxathiapiprolin is a chiral fungicide, and it can affect the metabolism of the cholesterol compounds by inhibiting oxysterol binding protein (OSBP) to exert its fungicidal effect. The application of oxathiapiprolin in agriculture is widespread, and its residue in the environment is a threat to both human and animal health. The two oxathiapiprolin enantiomers differ in their fungicidal activity, biotoxicity, and degradation by environmental forces. However, their biotoxicity has not been reported in animals. The toxicokinetics of a pesticide should be a crucial component for the evaluation of its toxicity in vivo. In this study, we investigated the absorption, bioavailability, tissue distribution, and excretion of the two oxathiapiprolin enantiomers in rats to verify their toxicokinetic process in animals. An ultrahigh-performance liquid chromatography triple quadrupole tandem mass spectrometry (UHPLC-QQQ/MS) method was established to quantify the two oxathiapiprolin enantiomers in vivo. The two oxathiapiprolin enantiomers were found to have approximately the same absorption rate and bioavailability, and both were excreted mainly in the feces. The half-life of R-(-)-oxathiapiprolin was nearly twice that of S-(+)-oxathiapiprolin. R-(-)-oxathiapiprolin also had greater distribution than S-(+)-oxathiapiprolin in the liver, lungs, heart, spleen, kidneys, stomach, large intestine, small intestine, brain, and pancreas, supporting the notion that R-(-)-oxathiapiprolin could better bind with OSBP. The stereoselectivity of S-(+)-oxathiapiprolin in these tissues may be responsible for it being readily metabolized in vivo. The molecular docking technique was subsequently used to verify the more superior binding between R-(-)-oxathiapiprolin and OSBP compared with the binding between S-(+)-oxathiapiprolin and OSBP. The findings of this study could provide more reliable data for determining the toxicokinetics of a single enantiomer of oxathiapiprolin in animals, thereby providing some theoretical basis for its subsequent toxicological study.

Stereoselective bioaccumulation and dissipation of pyrisoxazole in earthworm-soil microcosm

Pyrisoxazole is a chiral fungicide with high sterilizing activity to the plant pathogenic bacteria. It has two chiral C atoms, which bring four stereoisomers. The present work was the first time to explore the stereoselective bioaccumulation behavior of pyrisoxazole in earthworms by chiral liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS). The absolute configurations of pyrisoxazole stereoisomers were confirmed by circular dichroism (CD) coupled with calculated electronic circular dichroism (ECD) method, and the elution order in Lux Cellulose-3 column was as follows: (−)-3S, 5R-pyrisoxazole, (+)-3R, 5S-pyrisoxazole, (+)-3S, 5S-pyrisoxazole and (−)-3R, 5R-pyrisoxazole. The recoveries of pyrisoxazole stereoisomers in earthworm and soil samples ranged from 80.8 % to 101 % with the RSD lower than 6.3 %. In bioaccumulation progress, (+)-3R, 5S-pyrisoxazole was accumulated preferentially in earthworms, and the bioaccumulation concentrations of high-activity (−)-3S, 5R-pyrisoxazole were the lowest. There were no stereoselective bioaccumulation between (+)-3S, 5S-pyrisoxazole and (−)-3R, 5R-pyrisoxazole, while there was diastereoselectivity between Z-pyrisoxazole and E-pyrisoxazole with higher E-pyrisoxazole concentrations. In the whole bioaccumulation process, the BAF values of (+)-3R, 5S-pyrisoxazole were significantly higher than (−)-3S, 5R-pyrisoxazole, and the BAF values of (−)-3S, 5R-pyrisoxazole were the lowest. The dissipation of pyrisoxazole stereoisomers in the artificial soil was very slow and had no stereoselectivity, and the existence of earthworms had little effects on the dissipation of pyrisoxazole stereoisomers, which indicated that the stereoselective behaviors of pyrisoxazole in earthworms were caused by the stereoselective enrichment and dissipation of earthworms themselves. Taken together, (−)-3S, 5R-pyrisoxazole was recommend as a commercial product. This study played a positive role in guiding the development of environmentally friendly pesticides and provided database for the environmental and biological risk assessment of pyrisoxazole.

Environmental behavior of the chiral fungicide epoxiconazole in earthworm-soil system: Enantioselective enrichment, degradation kinetics, chiral metabolite identification, and biotransformation mechanism.

The environmental impact of the chiral fungicide epoxiconazole and its chiral transformation products (TPs) on non-target organisms and the environment has become a significant concern due to its widespread use in agricultural practice. Enantioselectivity studies of parent contaminants cannot adequately assess the complexity of its chiral TPs in the environment. This study aimed to investigate the environmental behavior of epoxiconazole in an earthworm-soil system. 2S,3R-(-)-epoxiconazole was preferentially enriched in earthworms during the accumulation phase (p<0.05), but no enantioselectivity was observed during the elimination phase. One methoxylated and four hydroxylated chiral TPs were identified in soil, earthworm, and excrement. The epoxy ring hydroxylated TP and methoxylated TP of epoxiconazole were discovered for the first time in the environment. The chemically specific enantioselectivity with enantiomer fraction (EF)>0.8 was observed for the TPs in different matrices. The CYP450 monooxygenase of earthworm was significant activated. In vitro enzyme metabolism experiments (earthworm microsomes and recombinant CYP450 enzymes CYP2A6, CYP 2C9, and CYP 3A4) were carried out to further explain the biotransformation mechanism of epoxiconazole in earthworm. This study provides new evidence of enantiomeric biotransformation of chiral fungicide epoxiconazole in the earthworm-soil system and could provide valuable insights into their environmental risk assessment.

Isolation of Sphingomonas sp. AJ-1 and its enantioselective S-methylation of the triazole fungicide prothioconazole

Prothioconazole is a widely used chiral triazole fungicide, and its residue pollution has attracted wide attention in recent years. However, little is known about microbial metabolic processes of prothioconazole enantiomers. In this study, a prothioconazole-degrading strain, Sphingomonas sp. AJ-1, was isolated from activated sludge. The optimal temperature and pH for prothioconazole degradation by strain AJ-1 were 30 °C and 6.0, respectively, and the degradation rate of prothioconazole by strain AJ-1 was negatively correlated with the initial concentration. When supplemented with additional carbon source, the degradation rates of 10 mg/L (Rac)-/(S)-/(R)-prothioconazole by strain AJ-1 were 76.0 %, 100.0 % and 64.8 % within 6 d, respectively. The CS bond of prothioconazole was methylated to produce (S)-/(R)-prothioconazole-S-methyl by strain AJ-1, but the degradation rate of prothioconazole by strain AJ-1 with (S)-enantiomer was 2.54-fold of that with (R)-enantiomer. Moreover, the toxicity of (Rac)-prothioconazole-S-methyl was 5.57 times lower than that of (Rac)-prothioconazole to Pseudokirchneriella subcapitata. The results showed that strain AJ-1 had obvious enantioselective metabolism for prothioconazole, and this metabolism was a detoxification process. This study provides new insights into the enantioselective metabolism of the chiral fungicide prothioconazole in microorganisms.

Systematic evaluation of chiral fungicide penflufen for the bioactivity improvement and input reduction using alphafold2 models and transcriptome sequencing

Traditional risk assessment of pesticide concludes at the racemic level, which is often incomprehensive. In this study, systematic studies on environmental stability, bioactivity, and ecotoxicological effects of fungicide penflufen were carried out at the enantiomeric level. The single-enantiomer of penflufen was successfully separated and prepared, and their stability was verified in different environmental matrices. Meanwhile, bioactivity test indicated that S-(+)-penflufen had increased bioactivity with its bioactivities against Rhizoctonia solani, Fusarium oxysporum, and Fusarium moniliforme being factors of 7.8, 1.8, and 4.7, respectively greater than those of R-(-)-penflufen. Molecular docking results showed the strong hydrogen bond interactions with Leu300, enantiomer-specific hydrophobic interactions with Cys299, Arg91, and His93, and the greater binding energy between S-(+)-penflufen and succinate dehydrogenase of Rhizoctonia solani caused the selective bioactivity. Additionally, two enantiomers showed low acute toxicity whereas selective sub-chronic toxicity to earthworms. In sub-chronic toxicity test, the accumulated enantiomers caused abnormalities in intestinal tract structure, enzyme activities, and gene expression of earthworms, especially in the S-(+)-penflufen treatment. The selective interactions between penflufen enantiomers and key proteins were elucidated using molecular docking, which may be the main reason of stereoselective subchronic toxicity. S-(+)-penflufen has high bioactivity and low acute risk, it has great potential for development.

Enantioselective aquatic toxicity and degradation in soil of the chiral fungicide oxathiapiprolin.

Oxathiapiprolin is an efficient and chiral fungicide for peronosporomycetes. The enantioselective environmental behavior and ecotoxicity of oxathiapiprolin are still unclear. The enantioselectivity of oxathiapiprolin enantiomers was explored, including their acute toxicity toward aquatic plants (Auxenochlorella pyrenoidosa and Soirodela polyrhiza) along with their influence on photosynthetic pigment production, the acute toxicity and morphological differences for the embryos, larvae and adult stages of zebrafish (Danio rerio), and the degradation in four typical soils (aerobic, anaerobic and sterilized conditions). The enantioselective toxicity of oxathiapiprolin showed that the toxicity of R-oxathiapiprolin was 1.8-2.1 times higher than that of S-oxathiapiprolin toward the two aquatic plants. In particular, the content of photosynthetic pigments decreased significantly stronger after exposure to R-oxathiapiprolin compared with S-oxathiapiprolin. The LC50 values of R-oxathiapiprolin in zebrafish in the different life stages were 1.6-2.1 times higher than those of S-oxathiapiprolin. The zebrafish embryos were most sensitive to the oxathiapiprolin enantiomers. After exposure to R-oxathiapiprolin, zebrafish embryos showed noticeable hatching delays, inhibition or deformation. R-oxathiapiprolin degraded preferentially in all four soils, with an enantiomeric fraction (EF) ranging from 0.28 to 0.42 under aerobic conditions. Enantioselective degradation was not found under anaerobic and sterilized conditions. The enantioselectivity of new chiral pesticides should be fully considered in risk assessments to provide a basis for the development and preparation of pure optical enantiomers.

Evaluation of Chiral Fungicide Penflufen in Legume Vegetables: Enantioseparation and Its Mechanism, Enantioselective Behaviors, and Risk Assessment.

Illustrating the enantioselective behaviors of the novel chiral fungicide penflufen was extremely important for ecological safety and human health. For penflufen enantiomers, an excellent separation method including a short analysis time (4 min), a high sensitivity (2 ng/g), and lesser consumption of an organic solvent was first established through supercritical fluid chromatography-tandem mass spectrometry. The enantioseparation mechanism was explained by computational chemistry, and the stronger binding ability of S-(+)-penflufen with cellulose tris-(3-chloro-4-methylphenylcarbamate) (the chiral stationary phase OZ-3 column) contributed to the posterior elution. In legume vegetables, penflufen dissipation was the fastest in Pisum sativum Linn plants (half-life, 1 day) and the slowest in Glycine max plants (half-lives, 11.3-12.9 days). After 30, 50, and 40 days, the rac-penflufen residues were lower than the maximum residue level value in the Electronic Code of Federal Regulations (10 ng/g) in G. max, P. sativum Linn, and Vigna unguiculata, respectively. Abundant S-(+)-penflufen was found in these plants with stereoisomeric excess (se) changes being >10% in the initial stage, so the risk assessment might be driven by S-(+)-penflufen. However, the se changes were <10% in V. unguiculata plants, and the risk assessment might be calculated based on rac-penflufen. Moreover, penflufen enantiomers could be transferred from legume vegetables to soils, and the concentrations increased with time. The high persistence and medium mobility of penflufen in soils might lead to potential groundwater contamination, which was noteworthy. These results could contribute to a more accurate risk assessment of penflufen in legume vegetables.

Simultaneous enantioselective determination of two succinate- dehydrogenase-inhibitor fungicides in plant-origin foods by ultra-high performance liquid chromatography−tandem mass spectrometry

Fluindapyr and penthiopyrad are two new succinate-dehydrogenase-inhibitor fungicides both employed as racemic mixtures of enantiomers to control various fungal pathogens. In the present work, a robust and highly-sensitive method for simultaneous determination of fluindapyr and penthiopyrad enantiomers in plant-origin foods (cereals, fruits and vegetables) was developed using UPLC-MS/MS combined with a chiral stationary phase. Rapid baseline chiral separation of four stereoisomers of fluindapyr and penthiopyrad was obtained within 4.2 min on chiral MX(2)-RH column under reversed-phase conditions (with the eluent of acetonitrile/0.1% formic acid in water =70/30 (V:V) and column temperature maintained at 30 °C). The plant-origin samples were extracted quickly with acetonitrile and purified with multi-walled carbon nanotubes. Excellent linearity for the target analytes was observed in the concentration ranging from 1 to 250 µg/L with regression coefficient no less than 0.9967. The mean recoveries of fluindapyr and penthiopyrad enantiomers from six matrices were 77.1–107.2%, with all relative standard deviations values lower than 9.1%. The limit of quantification of four stereoisomers of two target chiral fungicides was 5 µg/kg. The analysis of real samples reveal that the developed method is suitable for the simultaneous chiral determination of fluindapyr and penthiopyrad residues in cereals, fruits and vegetables samples at enantiomeric level and can support their further investigation on enantioselective environmental behaviors and residue surveillance.