Enantioselective environmental behavior of the chiral herbicide fenoxaprop-ethyl and its chiral metabolite fenoxaprop in soil

The enantioselective degradation behavior of fenoxaprop-ethyl (FE) and its chiral metabolite fenoxaprop (FA) in three soils under native conditions was investigated. Two pairs of enantiomers were analyzed by high-performance liquid chromatography (HPLC) with an amylose tri-(3,5-dimethylphenylcarbamate) (ADMPC) chiral column. The degradation of racemic FE in three soils showed the herbicidally inactive S-(-)-enantiomer degraded faster than the active R-(+)-enantiomer. FE was configurationally stable in soils because no interconversion to the respective antipodes was observed during incubation of the enantiopure S-(-)- or R-(+)-FE. The main metabolites of FE were confirmed as FA and 6-chloro-2,3-dihydrobenzoxazol-2-one (CDHB), and the formation of the chiral metabolite FA showed enantioselectivity in soils. The degradation of rac-FA was also enantioselective with the S-(-)-FA preferentially degraded: the half-life (t(1/2)) of the S-form in the three soils ranged from 2.03 to 5.17 days, and that of R-form ranged from 2.42 to 20.39 days. The inversion of the S-(-)-enantiomer into the R-(+)-enantiomer occurred in two of the three soils when the enantiopure S-(-)- and R-(+)-FA were incubated. The data from sterilized control experiments indicated that the enantioselectivity of FE and FA was attributed to microbially mediated processes.     

Application of capillary electrophoresis to study the enantioselective transformation of five chiral pesticides in aerobic soil slurries

The enantiomers of five chiral pesticides of environmental interest, metalaxyl, imazaquin, fonofos (dyfonate), ruelene (cruformate), and dichlorprop, were separated analytically using capillary electrophoresis (CE) with cyclodextrin chiral selectors. For metalaxyl, imazaquin, and fonofos, aqueous slurries of soil samples from two sites in Georgia and one in Ohio were spiked with the racemate of each pesticide at 50-60 mg/L of aqueous phase of the slurry, and CE analyses were performed at various time intervals to determine enantiomer fractions (EF). Metalaxyl underwent enantioselective transformation; in one soil, the half-life of the target active R-(+)-enantiomer was 17 days while that for the S-(-)-enantiomer was 69 days. Transformation occurred more slowly in the other two soils but was still selective for the R-(+)-enantiomer. Imazaquin and fonofos exhibited nonselective enantiomer loss over their 3 months of incubation time; this could have been due to abiotic or nonselective microbial reactions. Ruelene and dichlorprop were transformed selectively in a variety of soils in a previously reported study (7) that showed the influence of environmental changes on the transformation of chiral pollutants in soils; analytical methods used in that study are reported here to further illustrate the application of CE. CE is shown to be a simple, efficient, and inexpensive way to follow the transformation of chiral pesticides in laboratory microcosms where concentrations can be made high enough (25-50 mg/L initial racemate concentration) for detection of residual parent enantiomers during most of the process.     

Enantiomeric resolution and growth-retardant activity in rice seedlings of uniconazole

The increasing application of chiral pesticides has enhanced interest in their enantioselectivity. However, little relevant information is currently available for enantioselective activity of chiral plant growth regulators. In an attempt to screen active enantiomers of uniconazole, this work investigated enantiomeric separation and the enantioselective effect of uniconazole on the growth of rice seedlings and cyanobacteria. Baseline resolution of uniconazole enantiomers was achieved on a Chiralpak AD column by chiral high-performance liquid chromatography (HPLC). The relationship among circular dichroism (CD), optical rotation (OR), and absolute configuration was successfully established by coupling of CD and OR detection. The t test at the 95% level of confidence indicated significant differences between the enantiomers in their retardant activity toward growth of rice seedlings and stimulation effect on growth of cyanobacteria, the natural biofertilizers in rice paddy fields. The S-(+)-enantiomer was more active than the R-(-)-enantiomer in retarding growth of rice seedlings and stimulating growth of Microcystis aeruginosa . This special enantiomeric selectivity was further elucidated by probing the binding mode of enantiomers to gibberellin (GA) 20-oxidase by molecular docking. The S-(+)-enantiomer was found to bind tightly with GA 20-oxidase. The results suggested that the S-(+)-enantiomer instead of a racemate of uniconazole should be used to improve rice seedling quality.     

Different enantioselective degradation of pyraclofos in soils

This study investigated the enantioselective degradation behavior of pyraclofos in three soils (NC, HZ, and ZZ) under native and sterilized conditions. The absolute configuration of pyraclofos enantiomers has been determined by the combination of experimental and calculated electronic circular dichroism spectra. S-(+)- and R-(-)-Pyraclofos were separated and determined on a cellulose tri-(4-chloro-3-methylphenylcarbamate) (Lux Cellulose-4) chiral column by reversed-phase high-performance liquid chromatography-tandem mass spectrometry. Pyraclofos enantiomers were configurationally stable in three soils and no interconversion was observed during the incubation of enantiopure S-(+)- or R-(-)-pyraclofos under native conditions. The enantioselective degradation behavior of chiral pyraclofos was dramatically different in three soils under native conditions, with half-lives (t(1/2)) of pyraclofos in NC, HZ, and ZZ soils of 2.6, 13.4, and 7.8 days for S-(+)-pyraclofos and 9.2, 9.3, and 8.2 days for R-(-)-pyraclofos. Compared to the half-lives (t(1/2)) of rac-pyraclofos of 21.5, 55.9, and 14.4 days in sterilized NC, HZ and ZZ soils, the degradation velocity was greatly improved in native soils, indicating that degradation was greatly attributed to microbially mediated processes in agricultural cultivating soils.     

Single and joint acute toxicity of isocarbophos enantiomers to Daphnia magna

Although enantioselectivity in the toxicity of chiral pesticides has received considerable attention over recent years, how coexisting enantiomers interact with each other during their toxic action remains unknown. In this study, we attempted to resolve the enantiomers of a chiral organophosphate insecticide, isocarbophos, and investigated the acute toxicity of individual enantiomers and various enantiomer mixtures. Baseline enantiomeric separation of isocarbophos was achieved on a Chiralcel OD column with the mobile phase of n-hexane/isopropanol (90/10, v/v) at a flow rate of 0.8 mL/min. The resolved enantiomers were differentiated by their responses on a circular dichroism detector. The median lethal concentrations (LC 50) of racemate, (+)-enantiomer, and (-)-enantiomer of isocarbophos toward Daphnia magna were 13.9, 7.08, and 353 microg/L, respectively, after 48 h of static exposure, displaying a 50-fold difference between the enantiomers. Toxic unit (TU) analysis was employed to evaluate the joint toxicity of isocarbophos enantiomer mixtures. The calculated TU mix for the acute toxicity (48 h test) of various binary mixtures ranged from 0.83 to 1.04, suggesting a mode of additive effect. Further evaluation of available literature data for chiral organophosphorus insecticides showed that the joint toxicity of enantiomers may be additive, synergistic, and antagonistic. Therefore, when significant enantioselectivity exists for a chiral pesticide, it is important to also evaluate the interaction of enantiomers in the joint toxicity effect when enantiomers are present in a mixture.     

Determination of enantiomers of synthetic pyrethroids in water by solid phase microextraction - enantioselective gas chromatography

Solid phase microextraction (SPME) is an ideal sample preparation technique because of its speed and solvent-free features. Sampling by SPME is selective and only the dissolved concentration is measured, which allows measurement of the bioavailable fraction of a contaminant in aqueous media. One potential application of SPME is for analysis of enantiomers of chiral contaminants in environmental samples. In this study, a method was developed for determining enantiomers of (Z)-cis-bifenthrin and cis-permethrin in water using coupled SPME and enantioselective gas chromatography (GC). Following SPME sampling, enantiomers of (Z)-cis-bifenthrin and cis-permethrin were separated at the baseline on a beta-cyclodextrin-based enantioselective column, and analyte enrichment onto the SPME fiber was not enantioselective. The GC response increased as sampling time was increased from 0 to 240 min, and as sampling temperature was increased from 20 to 40 degrees C. Organic solvents such as methanol, acetone, and acetonitrile enhanced, while soil extracts slightly decreased, the GC response. The integrated SPME-enantioselective GC method was used to analyze surface runoff samples. The analysis showed preferential degradation of the 1S-3S enantiomer over the 1R-3R enantiomer for both (Z)-cis-bifenthrin and cis-permethrin. The concentrations detected by SPME-GC were substantially smaller than those determined following solvent extraction, suggesting that SPME-enantioselective GC analysis selectively measured the dissolved fraction.     

Enantioselective residue dissipation of hexaconazole in cucumber (Cucumis sativus L.), head cabbage (Brassica oleracea L. var. caulorapa DC.), and soils

In this study, the enantioselective dissipation behavior of hexaconazole was investigated in cucumber fruit, head cabbage, and two different types of agricultural soils. The dissipation kinetics was determined by reverse-phase liquid chromatography-tandem mass spectrometry on a cellulose tris (3-chloro-4-methylphenylcarbamate) chiral column. Dissipation rates of hexaconazole enantiomers followed first-order kinetics; the residues of (+)-enantiomer decreased more rapidly than (-)-enantiomer in cucumber and head cabbage, resulting in relative enrichment of the (-)-form, while the two enantiomers showed similar degradation rates in the tested soils. These results indicate substantial enantioselectivity in the residue dissipation of hexaconazole enantiomers in cucumber and head cabbage; however, nonenantioselective dissipation was observed in the tested soils.     

Determination of gossypol enantiomer ratio in cotton plants by chiral higher-performance liquid chromatography

A celulose tris(3,5-dimethylphenylcarbamate) coated onto APS silica (Nucleosil, particle size, 7 microm; pore size, 500 A) was used under a reversed-phase condition to measure the enantiomeric ratios of gossypol enantiomers in cottonseeds, flowers, and roots in a number of cultivars samples of different Gossipium species. While unidimensional chromatography was used for measuring the enantiomeric ratio of all the samples of G. hirsutum, G. mustelinum, and in the seeds of G. barbadense, multidimensional chromatography was necessary for the analysis of samples of roots and flowers of G. barbadense. In the latter case, an ODS Hypersil column was used in the first dimension for sample clean up, and the enantiomers were resolved on the second dimension by the chiral column. As expected, all the seed samples of G. hirsutum and G. mustelinum showed the (P)-(+)-enantiomer in excess, whereas the seeds of G. barbadense showed the (M)-(-)-enantiomer. However, (P)-(+)-gossypol was found in enantiomeric excess in three samples examined of roots and in one of flower of G. barbadense. These results are discussed in this paper.     

Enantioselective separation and phytotoxicity on rice seedlings of paclobutrazol

The environmental significance of enantioselectivity in chiral insecticides and herbicides has been widely studied. However, little information is currently available on the enantioselective behavior of chiral plant growth regulators. In this study, paclobutrazol enantiomers were resolved and prepared by chiral high-performance liquid chromatography with a Sino-chiral OJ column. The relationship among absolute configuration, optical activity and circular dichroism of paclobutrazol enantiomers was established. The enantioselective behavior of paclobutrazol, including enantioselective effect of paclobutrazol on the growth of rice seedlings and cyanobacteria and enantioselective loss of paclobutrazol in rice seedling growth media, in rice culture system was studied. The (2S,3S)-(-)-enantiomer was almost 3.1 times more active than the (2R,3R)-(+)-enantiomer toward shoot growth as measured by 7 day EC50 values. Enantioselectivity could not be determined with respect to root growth of rice seedlings because a typical dosage response was not observed in the range of the concentrations studied. The dissipation of paclobutrazol in rice growth medium is not enantioselective. Enantiomers and diastereoisomer of paclobutrazol all facilitated the growth of cyanobacteria, which increase the effectiveness of rice biofertilizers. The (2S,3S)-(-)-enantiomer showed stronger stimulatory activity on Microcystis aeruginosa cyanobacteria than the (2R,3R)-(+)-enantiomer, whereas the latter was a more potent stimulator of Anabaena sp. growth. These observations indicate that application of the (2S,3S)-(-)-enantiomer of paclobutrazol and Microcystis aeruginosa in rice cultivation is a good strategy for improving rice seedling performance.     

Enantiomeric resolution and biotoxicity of methamidophos

Despite the fact that the biological processes of chiral pesticides are enantioselective, the biotoxicity of methamidophos with respect to enantioselectivity has so far received limited research. In this study, the enantiomeric separation and biotoxicity of the two enantiomers of methamidophos were investigated. Successful enantioseparation of methamidophos using high-performance liquid chromatography on a Chiralcel OD column was achieved. As indicated by the optical rotation and circular dichroism detection, (+)-methamidophos was eluted prior to (-)-methamidophos. The t test at the 95% level of confidence indicated significant differences between the enantiomers in their in vitro inhibition toward acetylcholinesterases of bovine erythrocytes and Electrophorus electricus and in vivo acute aquatic toxicity to Daphnia magna. The in vitro assays showed that (-)-methamidophos was about 8.0-12.4 times more potent to the enzymes than its (+)-form. In contrast, the (+)-enantiomer was 7.0 times more toxic to D. magna in 48 h tests. The toxicity of racemic methamidophos was intermediate in both in vitro and in vivo bioassays. These results suggest that the biotoxicity of chiral OPs to nontargeted organisms is enantioselective and therefore should be reevaluated with their pure enantiomers.     

Toxicity of (+)- and (-)-gossypol to the plant pathogen,Rhizoctonia solani

The dimeric sesquiterpene gossypol occurs naturally in cottonseed and other parts of the cotton plant. Gossypol exists as enantiomers because of the restricted rotation around the central binaphthyl bond. The (-)-enantiomer is toxic to nonruminant animals while the (+)-enantiomer exhibits little, if any, toxicity to these animals. Developing cotton plants with low levels of the (-)-gossypol could expand the use of cottonseed as a feed source. Gossypol also may play a role in protecting the plant from pathogens. The relative toxicity of (+)- and (-)-gossypol to plant pathogens has not been reported. We measured the concentration of (+)- and (-)-gossypol in roots from cotton seedlings that were treated with the biocontrol agent Trichoderma virens that induces biosynthesis of gossypol and related terpenoids in cotton roots. (-)-Gossypol was the minor enantiomer in control and treated roots, but levels were slightly higher in roots from T. virens-treated seed. We also determined the toxicity of the gossypol enantiomers and the racemate to the seedling disease pathogen Rhizoctonia solani. The (+)- and (-)-enantiomers of gossypol and the racemate are equally effective in inhibiting growth of this pathogen. The lethal doses of the gossypols required to kill the pathogen appeared to be similar, but their toxicities are significantly less than those of related cotton and kenaf sesquiterpenes. The results indicate that altering the enantiomeric ratio in cotton roots will not adversely affect the resistance of seedlings to the seedling pathogen R. solani.     

Enantioselective degradation and chiral stability of pyrethroids in soil and sediment

Synthetic pyrethroids contain two or three chiral centers, making them a family of chiral pesticides with a large number of stereoisomers. Recent studies showed significant differences in aquatic toxicity between enantiomers from the same diastereomers of pyrethroids. To better understand the ecotoxicological effect and fate of pyrethroid insecticides, chirality in biodegradation must also be considered. In this study, we examined enantiomer compositions of selected pyrethroids in field sediment samples taken from various locations in southern California. Enantioselective degradation was frequently observed for cis-bifenthrin, permethrin, and cyfluthrin under field conditions. We further conducted long incubation experiments under laboratory-controlled conditions using single enantiomers of cis-bifenthrin, cis-permethrin, and cypermethrin. The half-lives for individual enantiomers were calculated to be 277-770 days for cis-bifenthrin enantiomers, 99-141 days for cis-permethrin enantiomers, and 52-135 days for cypermethrin enantiomers, respectively. The direction and degree of enantioselectivity in degradation were found to closely depend on the specific compound as well as experimental conditions. Because no significant difference in degradation was observed after samples were sterilized, the observed enantioselectivity may be attributed to preferential biological transformations.     

Enantioselective syntheses and sensory properties of the 3-mercapto-2-methylpentanols

3-Mercapto-2-methylpentanol, a new powerful flavor compound, exhibits two stereocenters giving rise to two pairs of diastereomers. To determine differences in the sensory properties, all four diastereomers and enantiomers were stereo- and enantioselectively synthesized. A highly diastereoselective aldol reaction using a chiral auxiliary was one of the key steps in the synthesis. Further derivatization yielded the enantiopure compounds. Odor thresholds in air and water were determined.     

Occurrence of (+)- and (-)-gossypol in wild species of cotton and in Gossypium hirsutum Var. marie-galante (Watt) Hutchinson

Gossypol occurs as a mixture of enantiomers in cottonseed. These enantiomers exhibit different biological activities. The (-)-enantiomer is toxic to animals, but it has potential medicinal uses. Therefore, cottonseed with >95% (-)-gossypol could have biopharmaceutical applications. The (+)-enantiomer shows little, if any, toxicity to nonruminant animals. Thus, cottonseed with >95% (+)-gossypol could be more readily utilized as a feed for nonruminants. The (+)- to (-)-gossypol ratio in commercial Upland (Gossypium hirsutum) cottonseed is usually about 3:2, whereas that in commercial Pima cottonseed (Gossypium barbadense) is approximately 2:3. Herein are reported the (+)- to (-)-gossypol ratios in the seed from 28 wild species of cotton (194 accessions), 94 accessions of G. hirsutum var. marie-galante, and 3 domesticated species (11 accessions). It was found that some or all of the accessions of Gossypium darwinii, Gossypium sturtianum, Gossypium areysianum, Gossypium longicalyx, Gossypium harknessii, and Gossypium costulatum produce an excess of (-)-gossypol but none >65%. At least one accession of Gossypium anomalum, Gossypium mustelinum, Gossypium gossypioides, and Gossypium capitis-viridis contained >94% (+)-gossypol. One of the 94 accessions of G. hirsutum var. marie-galante (i.e., no. 2469) contained 97% (+)-gossypol.     

手性异丙甲草胺在土壤中的选择性降解

Separation of chiral enantiomers and the dissipation of rac-metolachlor and S-metolachlor in soil were evaluated using achiral high-performance liquid chromatography （HPLC） and chiral gas chromatography （GC） methods. Under the experimental conditions the possible metabolite was considered to be N-（2-ethyl-6-methyl-phenyl）-2-hydroxy-acetamide. Because of the presence of two chiral elements （asymmetrically substituted carbon and chiral axis）, the baseline separation of metolachlor enantiomers was not achieved. S-metolachlor degraded faster in soil than rac-metolachlor. After a 42-day incubation, 73.4% of rac-metolachlor and 90.0% of S-metolachlor were degraded. However, due to the absence of biological processes the degradation process in sterilized soil showed no enantioselectivity. The results indicated that enantioselective degradations could greatly affect the environmental fate of metolachlor and should be considered when the environmental behavior of these compounds was assessed.     

Enantioselective analysis of triazole fungicide myclobutanil in cucumber and soil under different application modes by chiral liquid chromatography/tandem mass spectrometry

A sensitive and enantioselective method was developed and validated for the determination of myclobutanil enantiomers by chiral liquid chromatography coupled with tandem mass spectrometry. The separation and determination were performed using reversed-phase chromatography on a Chiralcel OD-RH column, with ACN-water (70/30, v/v) as the mobile phase under isocratic conditions at 0.5 mL/min flow rate. The matrix effect, linearity, precision, accuracy, and stability were evaluated. The proposed method then was successfully applied to the study of enantioselective degradation of rac-myclobutanil in cucumber and soil under different application modes. The results showed that the preferential degradation of (+)-myclobutanil resulted in an enrichment of the (-)-myclobutanil residue in plant and soil. Moreover, in cucumber, the stereoselective intensity of myclobutanil under root douche treatment was stronger than that under foliar spraying treatment, whereas in soil, the intensity was exactly opposite. The probable reasons underlying these enantioselective effects were also discussed. This study highlighted the importance of examining the fate of both enantiomers in the greenhouse system for the correct use of chiral pesticides.     

In vivo studies on the metabolism of the monoterpenes S-(+)- and R-(-)-carvone in humans using the metabolism of ingestion-correlated amounts (MICA) approach

The major in vivo metabolites of S-(+)- and R-(-)-carvone in a metabolism of ingestion correlated amounts (MICA) experiment were newly identified as alpha,4-dimethyl-5-oxo-3-cyclohexene-1-acetic acid (dihydrocarvonic acid), alpha-methylene-4-methyl-5-oxo-3-cyclohexene-1-acetic acid (carvonic acid), and 5-(1,2-dihydroxy-1-methylethyl)-2-methyl-2-cyclohexen-1-one (uroterpenolone) on the basis of mass spectral analysis in combination with syntheses and NMR experiments. Minor metabolites were identified as reduction products of carvone, namely, the alcohols carveol and dihydrocarveol. The previously identified major in vivo metabolite in rabbits, 10-hydroxycarvone, could not be detected, indicating either concentration effects or interspecies differences. Metabolic pathways for carvone in humans including oxidation of the double bond in the side chain and, to a minor extent 1,2- and 1,4 + 1,2-reduction of carvone, are discussed. No differences in metabolism between S-(+)- and R-(-)-carvone were detected.     

One- and two-dimensional direct chiral liquid chromatographic determination of mixtures of diclofop-Acid and diclofop-methyl herbicides

Simple one- and two-dimensional high-performance liquid chromatography (HPLC) methods for the simultaneous enantiomeric determination of alkyloxyphenoxypropionic acid herbicides is presented. Compounds studied were ( R, S)-2-[4-(2,4-dichlorophenoxy)phenoxy]propionic acid (diclofop-acid) and ( R, S)-2-[4-(2,4-dichlorophenoxy)]methyl propionate (diclofop-methyl). Mobile phases necessary to separate their enantiomers on an alpha1-acid glycoprotein chiral stationary phase are different; therefore, the simultaneous separation by an isocratic mode is not possible. The chiral separation method proposed involves a one-step gradient allowing for the simultaneous determination of both racemic enantiomers. Detection limits of the method were 0.03 mg/L for both diclofop-acid enantiomers and 0.14 and 0.15 mg/L for diclofop-methyl enantiomers, respectively. The two-dimensional method involves the use of two chromatographs in one achiral-chiral coupling. The LC-LC method is more suitable for complex samples because it involves an online cleanup effect. Detection limits were 1.25 and 1.87 mg/L for diclofop-acid and 2.70 and 3.02 mg/L for diclofop-methyl enantiomers, respectively. Accuracy, repeatability, and reproducibility have been studied in standard samples and a technical product.     

Enantiomeric resolution of chiral pesticides by high-performance liquid chromatography

Successful enantiomeric separation of 10 chiral pesticides by high-performance liquid chromatography (HPLC) using cellulose-tris(3,5-dimethylphenylcarbamate) (CDMPC) chiral stationary phase (CSP) was performed. The mobile phase was n-hexane modified by ethanol, propanol, 2-propanol (IPA), butanol, or isobutanol. The effects of mobile phase composition and column temperature on the separation were investigated. Baseline separation was obtained with ethofumesate, fluroxypyr-meptyl, malathion, benalaxyl, diclofop-methyl, methamidophos, vinclozolin, and lactofen, whereas near baseline separation was obtained with profenofos and acetochlor. Butanol was the best modifier for benalaxyl; isobutanol was the best modifier for lactofen, malathion, diclofop-methyl, and ethofumesate; and IPA was the best modifier for the other five. Better separations were not always at low temperature. The elution orders of the eluting enantiomers were determined by a circular dichroism (CD) detector. The quantitative analysis methods for the enantiomers of ethofumesate, benalaxyl, and diclofop-methyl were established. Validation parameters include linearity, precision, and limit of detection (LOD). The enantiomeric residual analysis procedures in soil and water samples were also developed using acetone extraction and C(18) solid phase extraction. The methods were reliable for residual analysis of the enantiomers.     

Enantiomeric separation and toxicity of an organophosporus insecticide, pyraclofos

Despite the fact that the biological processes of chiral pesticides are enantioselective, knowledge of the toxicities of pyraclofos due to enantiospecificity is scarce. In this study, the optical isomers of pyraclofos were separated and their toxicities to butyrylcholinesterase (BChE) and Daphnia magna were assessed. Baseline resolution of the enantiomers was obtained on both Chiralcel OD and Chiralpak AD columns. The effect of the mobile phase composition and column temperature were then discussed. The resolved enantiomers were characterized by their optical rotation and circular dichroism signs. The anti-BChE tests demonstrated that (-)-pyraclofos was about 15 times more potent than its (+)-form. However, acute aquatic assays suggested that (+)-pyraclofos was about 6 times more toxic than its antipode. Moreover, the joint toxicity of pyraclofos enantiomers to D. magna was found to be an additive effect. These results demonstrated that the overall toxicity of pyraclofos should be assessed using the individual enantiomers.