Nontarget Screening of Novel Urinary Biomarkers for Occupational Exposure to Toxic Chemicals from Coking Industry Using HPLC-QTOF-MS.

High-resolution mass spectrometry is an advanced technique for comprehensive screening of toxic chemicals. In this study, urine samples were collected from both an occupationally exposed population at a coking site and normal inhabitants to identify novel urinary biomarkers for occupational exposure to coking contaminants. A coking-site-appropriate analytical method was developed for unknown chemical screening. Through nontarget screening, 515 differential features were identified, and finally, 32 differential compounds were confirmed as candidates for the current study, including 13 polycyclic aromatic hydrocarbon (PAH) metabolites. Besides monohydroxy-PAHs (such as 1-&2-naphthol, 2-&9-hydroxyfluorene, 2-&4-phenanthrol, and 1-&2-hydroxypyrene), many other PAH metabolites including dihydroxy metabolites, PAH oxide, and sulfate conjugate were detected, suggesting that the quantification based solely on monohydroxy-PAHs significantly underestimated the human exposure to PAHs. Furthermore, several novel compounds were recognized that could be considered as biomarkers for the exposure to coking contaminants, including quinolin-2-ol (1.10 ± 0.44 ng/mL), naphthylmethanols (11.4 ± 5.47 ng/mL), N-hydroxy-1-aminonaphthalene (0.78 ± 0.43 ng/mL), hydroxydibenzofurans (17.4 ± 7.85 ng/mL), hydroxyanthraquinone (0.13 ± 0.053 ng/mL), and hydroxybiphenyl (2.70 ± 1.03 ng/mL). Despite their lower levels compared with hydroxy-PAHs (95.1 ± 30.8 ng/mL), their severe toxicities should not be overlooked. The study provides a nontarget screening approach to identify chemicals in human urine, which is crucial for accurately assessing the health risks of toxic chemicals in the coking industry.

Spatial distribution, source identification, and human health risk assessment of PAHs and their derivatives in soils nearby the coke plants.

The coking industry can generate large amounts of polycyclic aromatic hydrocarbons (PAHs) and their derivatives, which may negatively impact the environment and human health. In this study, soils nearby a typical coking plant were sampled to assess the impact of coke production on the surrounding residential areas and human health. The mean concentration of PAHs and their derivatives in residential area soils nearby the coke plant was 4270 ng/g dw, which was 1 order of magnitude higher than that observed in areas far from the coke plant and approximately 4 times lower than that revealed the coke plant. In addition, the results showed that coking processing area was the most contaminant area of the coke plant (mean: 74.4 µg/g dw), where was also the main source of pollutants in residential areas. In terms of vertical soils in coking plant, the maximum levels of chemicals (mean: 205 µg/g dw) were presented at the leakage of underground pipelines, where were much higher than those in surface soils, and decreased with the increase of depth. The analysis of variance (ANOVA) results showed obvious differences in the concentrations of 6-nitrochrysene between the plant, residential areas and control areas. Meanwhile, 6-nitrochrysene had potential cancer risk (CR) for human in the coking site. Thus, 6-nitrochrysene was the most noteworthy PAH derivatives. Furthermore, the CR (mean: 5.94 × 10-5) and toxic equivalent quantities (TEQs) (mean: 14.8 µg·TEQ/g) of PAHs and their derivatives was assessed in this study. This finding suggested that PAHs and their derivatives especially those extremely toxic chemicals (Nitro-PAHs (NPAHs) and Br/Cl-PAHs (XPAHs)) might pose a potential health risk to residents nearby the coke plant. The current study provides further insights into the pollution characteristics of PAHs and their derivatives in coke plants and potential risks to the workers and surrounding residents.

Triticonazole enantiomers induced enantioselective metabolic phenotypes in Fusarium graminearum and HepG2 cells.

The management of Fusarium head blight relies heavily on triazole fungicides. Most of triazole fungicides are chiral, and their enantioselective effects on metabolic phenotypes are poorly understood. Herein, we analyzed the bioactivity of triticonazole against Fusarium graminearum, and 1H-nuclear magnetic resonance-based metabolomics was used to assess the metabolic disturbances of triticonazole enantiomers in Fusarium graminearum and human hepatocarcinoma cells. Results indicated that the bioactivity of R-triticonazole was 4.28-fold higher than its antipode since it bound stronger with fungal CYP51B and induced more abnormal metabolic processes of Fusarium graminearum, including lipid metabolism, glycolysis, and amino acid metabolism. In human hepatocarcinoma cells, pathways of "alanine, aspartic acid and glutamate metabolism" and "pyruvate metabolism" were disturbed significantly by R-triticonazole; "phenylalanine metabolism" and "taurine-hypotaurine metabolism" were abnormal in the exposure of S-triticonazole. These results suggested that R- and S-triticonazole could affect different metabolic pathways of human hepatocarcinoma cells, and the massively use of inefficient S-triticonazole should be avoided. Our data will help to better understand the enantioselectivity of chiral pesticides and provide a reference for the development of green pesticides.

Enantioselective bioaccumulation, oxidative stress, and thyroid disruption assessment of cis-metconazole enantiomers in zebrafish (Danio rerio).

Chiral triazole pesticides may cause enantioselectively adverse effects to non-target organisms. In this work, we employed zebrafish as an aquatic organism model to explore stereoselective acute toxicity, bioaccumulation, oxidative stress, and thyroid disruption of cis-metconazole enantiomers. The median lethal concentration values of (1S, 5R)-metconazole, (1R, 5S)-metconazole, and the mixture of them against zebrafish were 4.01, 2.61 and 3.17 mg⋅L-1, respectively. (1R, 5S)-Metconazole was preferentially bioaccumulated in zebrafish than (1S, 5R)-metconazole, and the bioconcentration factor of (1R, 5S)-metconazole was 1.28-fold larger than that of (1S, 5R)-metconazole. Then, the activity order of catalase, superoxide dismutase, and glutathione-S transferase enzymes in zebrafish was expressed as (1S, 5R)-metconazole > the mixture > (1R, 5S)-metconazole, while the order of malondialdehyde content in zebrafish was (1R, 5S)-metconazole > the mixture > (1S, 5R)-metconazole. Moreover, cis-metconazole exhibited enantioselective regulation effects on the levels of triiodothyronine and thyroxine in zebrafish, and (1R, 5S)-metconazole possessed stronger thyroid disruption ability to zebrafish than the others. By virtue of molecular docking methodology, the binding affair and docking energy results supported that interactions between (1R, 5S)-metconazole and thyroid hormone receptors were much stronger than those between (1S, 5R)-metconazole and same receptors. This study of enantioselective evaluation of cis-metconazole in zebrafish can provide favorable information for risk assessments of chiral pesticides toward environment and health of aquatic organisms.

Risk Assessment of the Chiral Fungicide Triticonazole: Enantioselective Effects, Toxicity, and Fate.

The enantioselective toxicity of triticonazole (TRZ) to non-target organisms, the effect on wheat growth and quality, and the environmental fate of TRZ were investigated systematically in this study. The acute toxicity of S-TRZ to non-target aquatic and terrestrial organisms was greater than that of rac-TRZ and R-TRZ. The S-enantiomer significantly inhibited the growth and lodging resistance of wheat. S- and R-TRZ not only reduced the grain yield but also inhibited the activities of ADP-glucose pyrophosphorylase (AGPase) and starch synthase. The results of homology modeling and molecular docking further showed that the inhibition of AGPase activity by the two enantiomers hindered the accumulation of starch. By contrast, the racemate promoted the growth and development of wheat and improved grain quality. And the half-lives of the racemate in stems, grains, leaves, and soils were shorter than those of the enantiomers. The results of risk quotient (RQ) values showed that the application of TRZ enantiomers during wheat planting would bring a higher potential dietary risk to Chinese consumers. In comprehensive consideration of these results, the application of the racemate may be safer and more reasonable at the flowering stage of wheat.

Enantioselective acute toxicity, oxidative stress effects, neurotoxicity, and thyroid disruption of uniconazole in zebrafish (Danio rerio).

Uniconazole is a widely used plant growth retardant in the agricultural field. However, toxicological effects of uniconazole in aquatic ecosystem at chiral level are still unclear. Herein, acute toxicity, oxidative stress effects, neurotoxicity, and thyroid disruption of uniconazole enantiomers were investigated through using zebrafish as a model. (R)-Uniconazole possessed 1.16-fold greater acute toxicity to zebrafish than (S)-enantiomer. Then, integrated biomarker response values of oxidative stress parameters in zebrafish exposed to (R)-uniconazole were about 1.27~1.53 times greater than those treated by (S)-uniconazole, revealing that (R)-uniconazole could result in more significant adverse effects than (S)-uniconazole. Subsequently, the results of acetylcholinesterase activity of experimental fish demonstrated a state of inhibition-activation-inhibition after 14-day exposure to uniconazole, and a significant enantioselective neurotoxicity of uniconazole was observed in zebrafish after exposure for 4 and 7 days (p < 0.05). Moreover, thyroxine and triiodothyronine contents in (R)-uniconazole-exposed zebrafish were 0.89-fold (p=0.007) and 0.80-fold (p=0.007) than those in (S)-enantiomer-treated group, respectively. Furthermore, molecular docking results between uniconazole enantiomers and thyroid hormone receptors revealed that (R)-uniconazole was more tightly bound than (S)-uniconazole to the receptors. Briefly, our findings provide favorable information for ecological risk assessments of chiral agrochemicals in the environment and health of aquatic organisms.

Interactions of diclazuril enantiomers with serum albumins: Multi-spectroscopic and molecular docking approaches.

In this work, multi-spectroscopic and molecular docking methods have been conducted in the investigation of enantioselective interactions between diclazuril enantiomers and human/bovine serum albumins (HSA/BSA). The binding constants between serum albumins (SAs) and diclazuril enantiomers revealed that SAs exhibited stronger binding affinity for (R)-diclazuril than (S)-enantiomer. In addition, the fluorescence quenching of SAs induced by diclazuril enantiomers was ascribed to static quenching mechanism, in which hydrogen bonds and Van der Waals forces were the main interactions. According to the thermodynamic study, binding of diclazuril enantiomers and SAs was an exothermic process driven by enthalpy change. Then, circular dichroism spectroscopy of SAs with diclazuril enantiomers revealed that the SAs conformation had changed in the presence of diclazuril. Moreover, molecular docking technology was applied in exploration of interactions between SAs and diclazuril enantiomers. The docking energy between SAs and (R)-diclazuril was larger than (S)-diclazuril, which indicated that the affinity of SAs with (R)-diclazuril was stronger than (S)-enantiomer. This work may provide valuable information for explaining differences in pharmacokinetics and residue elimination of diclazuril enantiomers in living organisms.

Enantioselective analyses of chloroquine and hydroxychloroquine in rat liver microsomes through chiral liquid chromatography-tandem mass spectrometry.

An efficient, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) chiral analysis method was established for determination of chloroquine and hydroxychloroquine enantiomers in rat liver microsomes. Effects of polysaccharide chiral stationary phases and basic additives on chiral separations of two analytes were discussed in detail. Amylose tris(3, 5-dimethylphenylcarbamate)-coated chiral stationary phase showed the best separation performance for them with acetonitrile-diethylamine-ethanol-diethylamine mixture (90:0.1:10:0.1, v/v/v/v) among four chiral stationary phases. Then, multiple reaction monitoring mode was selected as the data acquisition for determination of two pairs of enantiomers. The proposed LC-MS/MS chiral analysis method was validated in terms of linearity, accuracy, precision, and specificity. Good linearity with correlation coefficient over 0.998 was obtained in the concentration range of 0.05-5 µM. Limits of quantification for chloroquine and hydroxychloroquine enantiomers were 5.0 and 1.0 nM, respectively. The recoveries ranged from 81.14% to 111.09%. The intra-day and inter-day relative standard deviation were less than 6.5%. Moreover, concentrations of chloroquine and hydroxychloroquine enantiomers in rat liver microsomes were determined through the proposed LC-MS/MS analysis method. After incubated with rat liver microsomes for 10 min, the enantiomeric factor of hydroxychloroquine decreased from 0.50 to 0.45 (p < 0.001). In brief, our developed determination method for chloroquine and hydroxychloroquine enantiomers through LC-MS/MS spectrometry showed the characteristics of high-efficiency, fast speed, and very low detection limit, and would be greatly beneficial for screening and quantitation of them in biological matrices.

Stereochemistry of chiral pesticide uniconazole and enantioselective metabolism in rat liver microsomes.

In this work, stereochemistry of uniconazole enantiomers and their metabolism behaviors in rat liver microsomes have been researched. Significance analysis has been applied in data processing. Absolute configurations of uniconazole enantiomers were identified through vibrational circular dichroism spectroscopy. According to their elution order from the chiral column using the CO2-methanol (80:20, v/v) mixture, two eluted fractions were determined to be (R)-uniconazole and (S)-uniconazole, respectively. A high-efficient and sensitive LC-MS/MS chiral analysis method was established for investigating the metabolism of uniconazole enantiomers in rat liver microsomes. The metabolic half-life of (R)-uniconazole (38.7 min) in rat liver microsomes was half that of (S)-enantiomer (74.5 min), and maximum velocity of metabolism, Michaelis constant of metabolism as well as the intrinsic metabolic clearance of (R)-uniconazole were significantly higher than (S)-enantiomer (p < 0.05), which indicated that (R)-uniconazole was preferentially metabolized in rat liver microsomes. By the virtue of molecular docking, (R)-uniconazole exhibited a higher binding affinity to cytochrome CYP2D2 than (S)-enantiomer, which corroborated well with the metabolism results. This work will shed light on the risk assessment of uniconazole toward human health and the ecological environment.

Systematic investigation of stereochemistry, stereoselective bioactivity, and antifungal mechanism of chiral triazole fungicide metconazole.

In this study, the stereochemistry, stereoselective fungicidal bioactivity, and antifungal mechanism of chiral triazole fungicide metconazole were investigated. The configurations of metconazole stereoisomers were determined to be (1R, 5R)-metconazole, (1R, 5S)-metconazole, (1S, 5S)-metconazole, and (1S, 5R)-metconazole through using electronic circular dichroism spectroscopy. The bioactivities of four stereoisomers and their stereoisomer mixture toward Fusarium graminearum Schw and Alternaria triticina were found to be in the following order: (1S, 5R)-metconazole > the stereoisomer mixture > (1S, 5S)-metconazole > (1R, 5R)-metconazole > (1R, 5S)-metconazole. In addition, the fungicidal activities of (1S, 5R)-metconazole against two tested pathogens was 13.9-23.4 times higher than those of (1R, 5S)-metconazole. Molecular docking methodology was applied to characterize the docking energy and distances between Cytochrome P450 CYP51B and the metconazole stereoisomers, and (1S, 5R)-metconazole showed the strongest binding energy and the shortest distance binding to CYP51B than the other three stereoisomers. Moreover, enantioselective metabolisms of (1S, 5R)-metconazole and (1R, 5S)-metconazole by Fusarium graminearum Schw were investigated through NMR-based metabolomics. The amounts of alanine, arginine, acetate, ethanol, and dimethylamine produced in the presence of (1R, 5S)-metconazole were significantly higher than corresponding amounts in the presence of (1S, 5R)-metconazole, whereas the amounts of glucose, glycerol, glutamate, methionine, and trimethylamine formed in the presence of (1R, 5S)-metconazole were much less than those in the presence of (1S, 5R)-metconazole. This systematic investigation of metconazole stereoisomers would provide a new perception of metconazole in stereoisomeric level, including bioactivities, metabolic behaviors and antifungal mechanism.

Stereoselective in vitro metabolism of cyproconazole in rat liver microsomes and identification of major metabolites.

The vast usage of agrochemicals enhances food security globally but may pose challenge to understand the risk assessment to non-target organisms and human beings, and liver microsomes are responsible for metabolism of these agrochemicals in vivo. In this study, stereoselective metabolism of chiral triazole fungicide cyproconazole in rat liver microsomes has been investigated through chiral LC-MS/MS technique. The half-lives of four cyproconazole stereoisomers were different ranging from 95 to 187 min, and (2S, 3R)-cyproconazole preferentially metabolized in rat liver microsomes. In addition, the results from metabolism kinetic study indicated that rat liver microsomes showed the stronger potency to deplete (2S, 3R)-cyproconazole than the others. Then, homology modeling and molecular docking results revealed that the docking energy between (2S, 3R)-cyproconazole and the cytochrome P450 CYP3A1 (-7.46 kcal⋅mol-1) was higher than the others, meaning that (2S, 3R)-cyproconazole exhibited the strongest binding ability to this enzyme. Moreover, two main metabolites of cyproconazole coming from hydroxylation and dehydration were observed, and possible metabolic reactions of cyproconazole in rat liver microsomes were identified through using an LCQ ion trap mass spectrometer. This kind of systematic metabolic investigation of cyproconazole at chiral level would provide valuable information for ecological and human health risk assessment of chiral pesticides.

Stereoselective Physiological Effects of Metconazole on Seed Germination and Seedling Growth of Wheat.

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-1S,5R-metconazole, had a significantly lower seedling emergence ratio and shoot length than other metconazole treatments; however, the opposite effects were observed in the trans-1S,5S-metconazole treatment. With regard to the hormonal level, enzyme activity, and gene transcription of gibberellin (GA) and jasmonic acid (JA), cis-1S,5R-metconazole treatment inhibited GA biosynthesis while trans-1S,5S-metconazole treatment promoted GA biosynthesis. Moreover, cis-1S,5R-metconazole, trans-1S,5S-metconazole, trans-1R,5R-metconazole, and racemate treatments increased JA biosynthesis. The oxidative stress responses in trans-1R,5R-metconazole and racemate treatments were more intensive. Therefore, compared with the control, treatment with cis-1R,5S-metcoanzole exhibited minimal influence on wheat seedling growth. The results showed that the application of pure cis-1R,5S-metcoanzole (instead of the racemate) in agricultural management could decrease the risks associated with crop growth and developmental damage.

Pharmacokinetics, Activity, and Residue Elimination of R- and S-Diclazuril in Broiler Chickens.

Diclazuril (DIC) is widely used as a racemic mixture to prevent and treat coccidiosis in farm animals, while the pharmacokinetics, bioactivity, and toxicity of DIC enantiomers are not known at all. This study first established a simple, sensitive, and reliable liquid chromatography tandem mass spectrometry method for separation of R-DIC and S-DIC and their analyses. Then, it was applied to investigate the stereoselective pharmacokinetics and residual elimination of individual enantiomers, and their anticoccidial activity was also evaluated in broiler chickens. The results indicated that the area under the concentration-time curve (AUC) and elimination half-life (t1/2ß) were significantly different (p < 0.05) for two enantiomers in chicken plasma. The AUC and t1/2ß of S-DIC were approximately 2 and 1.4 times those of R-DIC, respectively. The residual elimination of DIC enantiomers in chicken tissues was also stereoselective. The concentrations of S-DIC in chicken muscle and liver were greater than those of R-DIC, and it is the opposite in the kidney. There was no significant difference (p > 0.05) in the anticoccidial activity of racemate and enantiomers when a single enantiomer in feed was added above 0.5 mg kg-1. However, the anticoccidial activity of R-DIC (0.25 mg kg-1) was significantly higher (p < 0.05) than that of S-DIC (0.25 mg kg-1) in the diet. It should be mentioned that in chicken small intestine and cecum, the enantiomerization rate of each enantiomer in the infection group was faster than that in the uninfected group.

Identification, Quantification, and Stereoselective Degradation of Triazole Fungicide Cyproconazole in Two Matrixes through Chiral Liquid Chromatography-Tandem Mass Spectrometry.

Systematic investigation of cyproconazole, including absolute stereochemistry, fungicidal activity, quantification in two matrixes, and stereoselective degradation in cucumber, are conducted in this study. By virtue of vibrational circular dichroism (VCD) spectroscopy, absolute configurations of four stereoisomers were identified to be (2R,3R)-(+)-, (2R,3S)-(+)-, (2S,3S)-(-)-, and (2S,3R)-(-)-cyproconazoles. Then four stereoisomers exhibited stereoselective fungicidal activities against Fusarium graminearum Schw and Magnaporthe oryzae, and the order of fungicidal activity was (2S,3S)-(-)-stereoisomer > the stereoisomer mixture > (2S,3R)-(-)-stereoisomer > (2R,3R)-(+)-stereoisomer > (2R,3S)-(+)-stereoisomer. Moreover, chiral liquid chromatography-tandem mass spectrometry was used to identify and quantify cyproconazole stereoisomers in soil and cucumber matrixes. Good linearity (R2 ≥ 0.99) and recoveries (86.79-92.47%, RSD ≤ 3.94%) for them were achieved, individually. Furthermore, stereoselective degradation of four cyproconazole stereoisomers was observed in cucumber and the order of degradation rate was (2R,3R)-(+)-cyproconazole > (2S,3S)-(-)-cyproconazole > (2R,3S)-(+)-cyproconazole > (2S,3R)-(-)-cyproconazole. We envision that such systematic assessments of chiral fungicides at an enantiomeric level would provide valuable information in future studies involving enantioselective physiological, metabolic, and toxicological activities.

Fast enantioselective determination of triadimefon in different matrices by supercritical fluid chromatography.

A fast analytical method through supercritical fluid chromatography was developed for enantioselective determination of triadimefon in soil and apple samples. Effects of chiral stationary phases, co-solvents, column temperature, and back pressure on chiral separation of triadimefon were discussed in detail. Chiral stationary phases containing electron-donating units located at phenylcarbamate moieties showed better enanitonrecognition abilities in triadimefon than those with electron-withdrawing units. Then, the modified pretreatment procedure was applied in enantioselective analysis of triadimefon in two matrices. When using cellulose tris(3,5-dimethylphenylcarbamate)-coated silica gel as the chiral stationary phase and the CO2-methanol (95/5, v/v) mixture as the mobile phase, the LOD and LOQ were 0.38 mg/kg and 2.00 mg/kg, respectively, much lower than the others chiral stationary phases. Good linearity (R2 = 0.9999) and recoveries (92.90-105.56%，RSD ≤ 1.52%) for them were achieved.

High-fast enantioselective determination of prothioconazole in different matrices by supercritical fluid chromatography and vibrational circular dichroism spectroscopic study.

Herein, we developed a rapid supercritical fluid chromatography (SFC) method for chiral separation and enantioselective determination of prothioconazole in soil and tomatoes. The potential effects of chiral stationary phases, co-solvents, column temperature, and back pressure on enantioseparation of prothioconazole have been studied in detail. Two prothioconazole enantiomers were best separated on cellulose tris(3, 5-dimethylphenylcarbamate)-coated chiral stationary phase with CO2-2-propanol (80:20, v/v) as the mobile phase, and the run time through SFC (about 4 min) was five-fold shorter than HPLC. Then, through comparing the experimental vibrational circular dichroism spectrum of the later-eluted component with the calculated pattern based on the (R)-configuration, it should be (R)-(-)-prothioconazole. Moreover, the modified QuEChERS (quick, easy, cheap, effective, rugged and safe) extraction and cleanup procedures were applied in enantiomeric analysis of prothioconazole in two matrices. Good linearity (R2 ≥ 0.9992) and recoveries (91.84-101.66%, RSD ≤ 3.98%) for two enantiomers were achieved. This proposed method showed good accuracy and precision, and might be suitable for fast enantioselective determination and residual quantitative analysis of prothioconazole in food and environmental samples.

Enantioselective determination of metconazole in multi matrices by high-performance liquid chromatography.

A reliable and effective HPLC analytical method has been developed to stereoselectively quantify metconazole in soil and flour matrices. Effects of polysaccharide chiral stationary phase, type and content of alcoholic modifier on separation of racemic metconazole have been discussed in detail. Resolution and quantitative determination of metconazole stereoisomers were performed by using an Enantiopak OD column, with the n-hexane-ethanol mixture (97:3, v/v) at the flow rate of 1.0mL/min. Then, extraction and cleanup procedures followed by the modified QuEChERS (quick, easy, cheap, effective, rugged and safe) method were used for metconazole racemate in soil and flour matrices. The residual analysis method was validated. Good linearity (R2 ≥ 0.9997) and recoveries (94.98-104.89%, RSD ≤ 2.0%) for four metconazole stereoisomers were obtained. In brief, this proposed method showed good accuracy and precision, which might be applied in enantioselective determination, residual quantitative analysis, and degradation of metconazole in food and environmental matrices.

Reliable HPLC separation, vibrational circular dichroism spectra, and absolute configurations of isoborneol enantiomers.

Resolution of chiral compounds has played an important role in the pharmaceutical field, involving detailed studies of pharmacokinetics, physiological, toxicological, and metabolic activities of enantiomers. Herein, a reliable method by high-performance liquid chromatography (HPLC) coupled with an optical rotation detector was developed to separate isoborneol enantiomers. A cellulose tris(3, 5-dimethylphenylcarbamate)-coated chiral stationary phase showed the best separation performance for isoborneol enantiomers in the normal phase among four polysaccharide chiral packings. The effects of alcoholic modifiers and column temperature were studied in detail. Resolution of the isoborneol racemate displayed a downward trend along with an increase in the content of ethanol and column temperature, indicating that less ethanol in the mobile phase and lower temperature were favorable to this process. Moreover, two isoborneol enantiomers were obtained via a semipreparative chiral HPLC technique under optimum conditions, and further characterized by analytical HPLC, and experimental and calculated vibrational circular dichroism (VCD) spectroscopy, respectively. The solution VCD spectrum of the first-eluted component was consistent with the Density Functional Theory (DFT) calculated pattern based on the SSS configuration, indicating that this enantiomer should be (1S, 2S, 4S)-(+)-isoborneol. Briefly, these results have provided reliable information to establish a method for analysis, preparative separation, and absolute configuration of chiral compounds without typical chromophoric groups.

Stereoselective quantification of triticonazole in vegetables by supercritical fluid chromatography.

A highly fast analytical method though supercritical fluid chromatography (SFC) has been developed to quantify triticonazole enantiomers in cucumbers and tomatoes. Effects of organic modifier type and concentration on chiral separation and quantification of standard solution as well as matrix-matched standard solutions have been studied in detail. Among three organic modifiers, better separation of triticonazole racemate was achieved with 20% ethanol (v/v). The run time in SFC (ca 3min) with CO2-ethanol (80:20, v/v) as the mobile phase was six-fold shorter than HPLC analysis (about 18min). Then, QuEChERS (quick, easy, cheap, effective, rugged and safe) extraction procedure was used for triticonazole in vegetables. The residue analysis method was validated. Good linearity (R2≥0.9988) and recoveries (81.62-106.21%, RSD≤7.30%) for the two enantiomers were achieved. This developed method described herein is convenient and reliable for enantioselective detection of triticonazole in vegetables, which might provide additional information for reliable risk assessment of chiral pesticides.