Enantiomerization and Enantioselective Bioaccumulation of Metalaxyl in Tenebrio molitor Larvae

The enantiomerization and enantioselective bioaccumulation of metalaxyl by a single dose of exposure to Tenebrio molitor larvae under laboratory condition were studied by high‐performance liquid chromatography tandem mass spectroscopy (HPLC‐MS/MS) based on a ChiralcelOD‐3R [cellulosetris‐tris‐(3, 5‐dichlorophenyl‐carbamate)] column. Exposure of enantiopure R‐metalaxyl and S‐metalaxyl in Tenebrio molitor larvae exhibited significant enantiomerization, with formation of the R enantiomers from the S enantiomers, and vice versa, which might be attributed to the chiral pesticide catalyzed by a certain enzyme in Tenebrio molitor larvae. Enantiomerization was not observed in wheat bran during the period of 21 d. In addition, bioaccumulation of rac‐metalaxyl in Tenebrio molitor larvae was enantioselective with a preferential accumulation of S‐metalaxyl. These results showed that enantioselectivity was caused not only by actual degradation and metabolism but also by enantiomerization, which was an important process in the environmental fate and behavior of metalaxyl enantiomers. Chirality 26:88–94, 2014. © 2013 Wiley Periodicals, Inc.     

Enantioselective effects of metalaxyl on soil enzyme activity

The enantioselective effects of the chiral pesticide metalaxyl on soil enzyme activity were investigated. Incubation experiments were conducted to investigate the effects of metalaxyl enantiomers at different concentrations on the activities of urease, invertase, and catalase as well as the type of activity change (activation vs. inhibition) at different times during incubation. The results indicated that the effects of metalaxyl on the activity of soil enzymes were not only related to the concentration of the enantiomers and soil incubation time, but also to the chiral configuration, suggesting the effects were enantioselective. A pattern of inhibition‐recovery‐slight stimulation was observed in urease activity of the soil samples treated with metalaxyl enantiomers, but the effects of (–) ‐R‐metalaxyl were stronger than those of (+)‐S‐metalaxyl at the same concentration. Invertase activity in soil samples treated with metalaxyl enantiomers initially sharply decreased before finally returning to the normal level, and the effects of (+)‐S‐metalaxyl were stronger than those of (–) ‐R‐metalaxyl at the same concentration. Metalaxyl enantiomers influenced catalase activity in a pattern of slight stimulation‐inhibition‐recovery, and the effects of (–) ‐R‐metalaxyl were stronger than those of (+)‐S‐metalaxyl at the same concentration.     

Enantioselective Degradation of the Chiral Fungicides Metalaxyl and Furalaxyl by Brevibacillus brevis

For almost four decades, the chiral fungicides metalaxyl and furalaxyl have been in use in plant protection on a global scale. Both substances are distributed as racemic mixtures, yet the desirable interference in nucleic acid synthesis of harmful fungi only occurs by the (‐)‐R‐enantiomer. As enantioselective degradation in Scheyern (Germany) and Yaoundé (Cameroon) soils has been documented, the influence of 50 isolated microorganisms on the R/S ratio was investigated. A high‐pressure liquid chromatography method with a chiral column to separate enantiomers of metalaxyl and furalaxyl, and subsequent detection by tandem mass spectrometry, was employed. Only one of these microorganisms, a strain of Brevibacillus brevis, showed an enantioselective degradation pattern in liquid culture; the respective (‐)‐R‐enantiomers were preferably degraded. Moreover, (‐)‐R‐furalaxyl was degraded faster in cultures supplemented simultaneously with both fungicides of the same concentration. Chirality 25:336–340, 2013. © 2013 Wiley‐Liss Inc. Chirality 00:000‐000:, 2013. © 2013 Wiley Periodicals, Inc.     

Bioaccumulation and Excretion of Enantiomers of Myclobutanil in Tenebrio molitor Larvae Through Dietary Exposure

The bioaccumulation and excretion of enantiomers of myclobutanil in Tenebrio molitor larvae through dietary exposure under laboratory conditions were investigated using high‐performance liquid chromatography tandem mass spectrometry (HPLC‐MS/MS) based on a ChiralcelOD‐3R [cellulosetris‐tris‐(3, 5‐dichlorophenyl‐carbamate)] column. The wheat bran fed to Tenebrio molitor larvae was spiked with racemic myclobutanil at two dose levels of 20 mg/kg and 2 mg/kg (dry weight). The results showed that there was a significant trend of enantioselective bioaccumulation in the larvae with a preferential accumulation of (?)‐myclobutanil in 20 mg/kg dose exposure, but it was not obviously observed in the 2 mg/kg dose group. A kinetic model considering enantiomerization between the two enantiomers based on first‐order reactions was built and the rate constants were estimated to discuss the kinetic reason for the different concentrations of individual enantiomers in the larvae. The approximations implied an inversion between the two enantiomers with a relatively higher rate of the inversion from (?)‐myclobutanil to (+)‐myclobutanil. Meanwhile, analysis of data of excretion samples suggested the active excretion is probably an important pathway for the insect to eliminate myclobutanil rapidly with nonenantioselectivity as a passive transport process, which was consistent with the low accumulation efficiency of myclobutanil measured by BAF (bioaccumulation factor). Chirality 25:890–896, 2013. © 2013 Wiley Periodicals, Inc.     

Enantioselective Degradation of Metalaxyl in Grape,Tomato, and Rice Plants

Enantioselective biodegradation of chiral pesticide metalaxyl in grape, tomato, and rice plants under field conditions were studied. Metalaxyl enantiomers were completely separated with a resolution (Rs) of 5.01 by high‐performance liquid chromatography (HPLC) based on a cellulose tris (3‐chloro‐4‐methyl phenyl carbamate) chiral column (Lux Cellulose‐2). Metalaxyl enantiomers from matrixes were extracted by acetonitrile and purged using Cleanert Alumina‐A solid phase extraction (SPE). The linearity, recovery, precision, sensitivity, and matrix effect of the method were assessed. The result showed that significant stereoselectivity occurred in grape, tomato, and rice plants. In grape, (+)‐S‐metalaxyl with a half‐life of 5.5 d degraded faster than (–)‐R‐metalaxyl with that of 6.9 d, and the enantiomer fraction (EF) value reached 0.37 at 21 d. The same enantioselectivity was observed in tomato, and the half‐life was 2.2 d for the S‐enantiomer and 3.0 d for the R‐enantiomer. The EF values decreased from 0.49 of 0 d to 0.26 of 14 d. On the other hand, a preferential degradation of the R‐form was found in rice plants, with an EF value of 0.70 at 14 d, and the corresponding half‐life was 2.3 d for the R‐form and 2.8 d for the S‐form. Chirality 27:109–114, 2015. © 2014 Wiley Periodicals, Inc.     

Studies of Enantiomeric Degradation of the Triazole Fungicide Hexaconazole in Tomato,Cucumber, and Field Soil by Chiral Liquid Chromatography–Tandem Mass Spectrometry

Chiral pesticide enantiomers often show different bioactivity and toxicity; however, this property is usually ignored when evaluating their environmental and public health risks. Hexaconazole is a chiral fungicide used on a variety of crops for the control of many fungal diseases. This use provides opportunities for the pollution of food and soil. In this study, a sensitive and convenient chiral liquid chromatography coupled with tandem mass spectrometry (LC‐MS/MS) method was developed and validated for measuring hexaconazole enantiomers in tomato, cucumber, and soil. Separation was by a reversed‐phase Chiralcel OD‐RH column, under isocratic conditions using a mixture of acetonitrile‐2 mM ammonium acetate in water (60/40, v/v) as the mobile phase at a flow rate of 0.4 mL/min. Parameters including the matrix effect, linearity, precision, accuracy and stability were undertaken. Then the proposed method was successfully applied to investigate the possible enantioselective degradation of rac‐hexaconazole in plants (tomato and cucumber) and soil under field conditions. The degradation of the two enantiomers of hexaconazole proved to be enantioselective and dependent on the media: The (+)‐enantiomer showed a faster degradation in plants, while the (?)‐enantiomer dissipated faster than the (+)‐form in field soil, resulting in relative enrichment of the opposite enantiomer. The results of this work demonstrate that both the environmental media and environmental conditions influenced the direction and rate of enantioselective degradation of hexaconazole. Chirality 25:160–169, 2013. © 2013 Wiley Periodicals, Inc.     

Chiral Separation of the Clinically Important Compounds Fucose and Pipecolic Acid Using CE: Determination of the Most Effective Chiral Selector

In this study, simple electrophoretic methods were developed for the chiral separation of the clinically important compounds fucose and pipecolic acid. In recent years, these analytes, and particularly their individual enantiomers, have attracted considerable attention due to their role in biological functions and disorders. The detectability and sensitivity of pipecolic acid and fucose were improved by reacting them with fluorenylmethyloxycarbonyl chloride (FMOC‐Cl) and 5‐amino‐2‐naphthalene‐sulfonic acid (ANSA), respectively. The enantioseparation conditions were optimized by initially investigating the type of the chiral selector. Different chiral selectors, such as polymeric surfactants and cyclodextrins, were used and the most effective ones were determined with regard to resolution and analysis time. A 10‐mM β‐cyclodextrin was able to separate the enantiomers of ANSA‐DL‐fucose and the polymeric surfactant poly(sodium N‐undecanoyl‐LL‐leucine‐valinate) was able to separate the enantiomers of FMOC‐DL‐pipecolic acid, with resolution values of 3.45 and 2.78, respectively. Additional parameters, such as the concentration and the pH of the background electrolyte (BGE), the concentration of the chiral selector, and the addition of modifiers were examined in order to optimize the separations. The addition of the chiral ionic liquid D‐alanine tert‐butyl ester lactate into the BGE was also investigated, for the first time, in order to improve resolution of the enantiomers. Chirality 25:556–560, 2013. © 2013 Wiley Periodicals, Inc.     

Enantioseparation and Determination of the Chiral Fungicide Furametpyr Enantiomers in Rice,Soil, and Water by High‐Performance Liquid Chromatography

The chiral fungicide furametpyr is widely used in the rice field to control rice sheath blight; however, furametpyr enantiomers are treated as just one compound in traditional achiral analysis, which gives only partial information. An effective chiral analytical method was developed for the resolution and determination of the fungicide furametpyr enantiomers in rice, soil, and water samples. Furametpyr enantiomers were excellently separated and determined on a Chiralpak AD‐H column with n‐hexane/ethanol (90:10, v/v) as mobile phase at a flow rate of 0.8 mL min^‐1 with UV detection at 220 nm. The resolution was up to 8.85. The first eluted enantiomer was (+)‐furametpyr and the second eluted one was (?)‐furametpyr. The effects of mobile‐phase composition and column temperature on the enantioseparation were evaluated. The method was validated for linearity, repeatability, accuracy, limit of detection (LOD), and limit of quantification LOQ. LOD was 2.0 µg kg^‐1 in water, 0.02 mg kg^‐1 in soil, and 0.07 mg kg^‐1 in rice with an LOQ of 6.7 µg kg^‐1 in water, 0.07 mg kg^‐1 in soil, and 0.23 mg kg^‐1 in rice. The average recoveries of the pesticide in all matrices ranged from 73.1 to 101.8% for all fortification levels. The precision values associated with the analytical method, expressed as relative standard deviation (RSD) values, were below 14.0% in all matrices. The methodology was successfully applied for the enantioselective analysis of furametpyr enantiomers in real samples. Chirality 25:904–909, 2013. © 2013 Wiley Periodicals, Inc.     

Enantioselective Bioaccumulation of Diniconazole in Tenebrio Molitor Larvae

The enantioselective bioaccumulation of diniconazole in Tenebrio molitor Linne larva was investigated with liquid chromatography tandem mass spectrometry based on the ChiralcelOD‐3R[cellulose tri‐(3,5‐dimethylphenyl carbamate)] column. In this study we documented the effects of dietary supplementation with wheat bran contaminated by racemic diniconazole at two dose levels of 20 mg kg^‐1 and 2 mg kg^‐1 (dry weight) in Tenebrio molitor. The results showed that both doses of diniconazole were taken up by Tenebrio molitor rapidly in the first few days, the concentrations of R‐enantiomer and S‐enantiomer at high doses reached the highest level of 0.55 mg kg^‐1 and 0.48 mg kg^‐1, respectively, on the 1^st d, and the concentrations of them obtained a maxima of 0.129 mg kg^‐1 and 0.128 mg kg^‐1 at low dose, respectively, on the 3^rd d, which means that the concentration of diniconazole was proportional to the time of achieving the highest accumulated level. It afterwards attained equilibrium after a sharp decline at both 20 mg kg^‐1 and 2 mg kg^‐1 of diniconazole. The determination results from the feces of Tenebrio molitor demonstrated that the extraction recovery (ER) values of the high dose group were higher than that of the low dose group and the values were all above 1; therefore, it could be inferred that enantiomerization existed in Tenebrio molitor. Additionally, the biota accumulation factor was used to evaluate the bioaccumulation of diniconazole enantiomers, showing that the bioaccumulation of diniconazole in Tenebrio molitor was enantioselective with preferential accumulation of S‐enantiomer. Chirality 25:917–922, 2013. © 2013 Wiley Periodicals, Inc.     

Enantioselective Degradation and Chiral Stability of Metalaxyl‐M in Tomato Fruits

Metalaxyl is an important chiral acetanilide fungicide, and the activity almost entirely originates from the R‐enantiomer. Racemic metalaxyl has been gradually replaced by the enantiopure R‐enantiomer (metalaxyl‐M). In this study a chiral residue analysis method for metalaxyl and the metabolite metalaxyl acid was set up based on high‐performance liquid chromatography tandem mass spectroscopy (HPLC‐MS/MS). The enantioselective degradation and chiral stability of metalaxyl‐M in tomato fruits in two geographically distinct regions of China (Heilongjiang and Hunan Province) were evaluated and the enantioselectivity of metalaxyl acid was also investigated. Tomato plants grew under field conditions with a one‐time spray application of metalaxyl‐M wettable powder. It was found that R‐metalaxyl was not chirally stable and the inactive S‐metalaxyl was detected in tomato fruits. At day 40, S‐metalaxyl derived from R‐metalaxyl accounted for 32% and 26% of the total amount of metalaxyl, respectively. The metabolites R‐metalaxyl acid and S‐metalaxyl acid were both observed in tomato, and the ratio of S‐metalaxyl acid to the sum of S‐ and R‐metalaxyl acid was 36% and 28% at day 40, respectively. For both metalaxyl and metalaxyl acid, the half‐life of the S‐enantiomer was longer than the R‐enantiomer. The results indicated that the enantiomeric conversion should be considered in the bioactivity evaluation and environmental pollution assessment. Chirality 28:382–386, 2016. © 2016 Wiley Periodicals, Inc.     

Chiral Separation and Enantioselective Degradation of Vinclozolin in Soils

Vinclozolin is a chiral fungicide with potential environmental problems. The chiral separation of the enantiomers and enantioselective degradation in soil were investigated in this work. The enantiomers were separated by high‐performance liquid chromatography (HPLC) on Chiralpak IA, IB, and AZ‐H chiral columns under normal phase and the influence of the mobile phase composition on the separation was also studied. Complete resolutions were obtained on all three chiral columns under optimized conditions with the same elution order of (+)/(?). The residual analysis of the enantiomers in soil was conducted using accelerate solvent extraction followed by HPLC determination. The recoveries of the enantiomers ranged from 85.7–105.7% with relative standard deviation (SD) of 0.12–3.83%, and the limit of detection (LOD) of the method was 0.013 µg/g. The results showed that the degradations of vinclozolin enantiomers in the soils followed first‐order kinetics. Preferential degradation of the (?)‐enantiomer was observed only in one soil with the largest |ES| value of 0.047, and no obvious enantioselective degradation was observed in other soils. It was found that the persistence of vinclozolin in soil was related to pH values based on the half‐lives. The two enantiomers disappeared about 8 times faster in basic soils than that in neutral or acidic soils. Chirality 26:155–159, 2014. © 2014 Wiley Periodicals, Inc.     

Enantiomeric separation of type I and type II pyrethroid insecticides with different chiral stationary phases by reversed‐phase high‐performance liquid chromatography

The enantiomeric separation of type I (bifenthrin, BF) and type II (lambda‐cyhalothrin, LCT) pyrethroid insecticides on Lux Cellulose‐1, Lux Cellulose‐3, and Chiralpak IC chiral columns was investigated by reversed‐phase high‐performance liquid chromatography. Methanol/water or acetonitrile/water was used as mobile phase at a flow rate of 0.8 mL/min. The effects of chiral stationary phase, mobile phase composition, column temperature, and thermodynamic parameters on enantiomer separation were carefully studied. Bifenthrin got a partial separation on Lux Cellulose‐1 column and baseline separation on Lux Cellulose‐3 column, while LCT enantiomers could be completely separated on both Lux Cellulose‐1 and Lux Cellulose‐3 columns. Chiralpak IC provided no separation ability for both BF and LCT. Retention factor (k) and selectivity factor (α) decreased with the column temperature increasing from 10°C to 40°C for both BF and LCT enantiomers. Thermodynamic parameters including ?H and ?S were also calculated, and the maximum R_s were not always obtained at lowest temperature. Furthermore, the quantitative analysis methods for BF and LCT enantiomers in soil and water were also established. Such results provide a new approach for pyrethroid separation under reversed‐phase condition and contribute to environmental risk assessment of pyrethroids at enantiomer level.     

Indirect synchronous fluorescence spectroscopy and direct high‐performance thin‐layer chromatographic methods for enantioseperation of zopiclone and determination of chiral‐switching eszopiclone: Evaluation of thermodynamic quantities of chromatographic separation

Economic and enantioselective synchronous fluorescence spectroscopy and high‐performance thin‐layer chromatography methods have been developed and validated as per ICH guidelines for the separation of zopiclone enantiomers using L‐(+)‐tartaric acid as a chiral selector, followed by determination of the chiral‐switching eszopiclone. Synchronous fluorescence spectroscopy was successfully applied for chiral recognition of R & S enantiomers of zopiclone at ?λ = 110 nm based on creating of diastereomeric complexes with 0.06M tartaric acid in an aqueous medium containing 0.2M disodium hydrogen orthophosphate. Synchronous fluorescence intensities of eszopiclone were recorded at 296 nm in concentration range 0.2‐ to 4‐μg/mL eszopiclone. High‐performance thin‐layer chromatography method depends on resolution of zopiclone enantiomers on achiral HPTLC silica‐gel plates using acetonitrile:methanol:water (8:2:0.25, v/v/v) containing L‐(+)‐tartaric acid as a chiral mobile‐phase additive followed by densitometric measurements at 304 nm in concentration range of 1 to 10 μg/band of eszopiclone. The effect of chiral‐selector concentration, pH, and temperature on the resolution have been studied and optimized for the proposed methods. The cited procedures were successfully applied to determine eszopiclone in commercial tablets of pure and racemic forms. Enantiomeric excess was evaluated using optical purity test and integrated peak area to describe the enantiomeric ratio. Thermodynamics of chromatographic separation, enthalpy, and entropy were evaluated using the Van't Hoff equation. The proposed methods were found to be selective for identification and determination of the eutomer in drug substances and products.     

Effects of metalaxyl enantiomers stress on root activity and leaf antioxidant enzyme activities in tobacco seedlings

The objective of this experiment was to study the effects of metalaxyl enantiomers on the activity of roots and antioxidative enzymes in tobacco seedlings. Water culture experiment was conducted to analyze the effects of different concentrations of metalaxyl enantiomers (30 and 10 mg L^?1) on root activity and leaf superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities and malondialdehyde (MDA) content of tobacco seedlings. The results showed that metalaxyl significantly inhibited root activity and significantly improved leaf SOD, POD, and CAT activities and MDA content. A better physiological response in tobacco seedlings was observed at 30 mg L^?1 than at 10 mg L^?1 metalaxyl. The stereoselectivity for different enantiomers had no obvious effect on root activity and the leaf POD activity, but it affected significantly the SOD and CAT activities and MDA content. The SOD activity was promoted more by R‐enantiomer than by S‐enantiomer at 30 mg L^?1 metalaxyl, and the same effect was observed on CAT activity from the beginning to the end of the stress period. The MDA content under the stress by R‐enantiomer was higher than that under the stress by S‐enantiomer at 10 mg L^?1 metalaxyl.     

A green and effective method for the determination of metalaxyl enantiomers in tobacco and soil by supercritical fluid chromatography–tandem mass spectrometry

We reported a new methodology for the stereoselective determination of metalaxyl enantiomers in tobacco and soil. The QuEChERS (quick, easy, cheap, effective, rugged, and safe) method was used for the extraction and clean-up of the tobacco and soil samples. Separation of the metalaxyl enantiomers was performed on an ACQUITY UPC2 Trefoil CEL1 chiral column coupled with supercritical fluid chromatography with tandem mass spectrometry (SFC-MS/MS), and the run time was only 5 minutes. Under the optimized conditions, the recoveries for the enantiomers were between 78.2% and 93.3% with intraday relative standard deviations (RSDs) ranging from 1.1% to 5.4%. The limit of detection (LOD) for the enantiomers in tobacco and soil varied from 0.005 to 0.007 mg/kg, and the limit of quantitation (LOQ) ranged from 0.017 to 0.020 mg/kg. In this method, only a small amount of methanol was consumed to obtain a rapid stereoselective separation. This proposed method showed good accuracy and precision and might be suitable for fast enantioselective determination of metalaxyl in food and environmental samples. The developed method was further validated by application to the analysis of authentic samples.     

Enantioselective Acute Toxicity Effects and Bioaccumulation of Furalaxyl in the Earthworm (Eisenia foetida)

The enantioselectivities of individual enantiomers of furalaxyl in acute toxicity and bioaccumulation in the earthworm (Eisenia foetida) were studied. The acute toxicity was tested by filter paper contact test. After 48 h of exposure, the calculated LC₅₀ values of the R‐form, rac‐form, and S‐form were 2.27, 2.08, and 1.22 µg cm^‐2, respectively. After 72 h of exposure, the calculated LC₅₀ values were 1.90, 1.54, and 1.00 µg cm^‐2, respectively. Therefore, the acute toxicity of furalaxyl enantiomers was enantioselective. During the bioaccumulation experiment, the enantiomer fraction of furalaxyl in earthworm tissue was observed to deviate from 0.50 and maintained a range of 0.55–0.60; in other words, the bioaccumulation of furalaxyl was enantioselective in earthworm tissue with a preferential accumulation of S‐furalaxyl. The uptake kinetic of furalaxyl enantiomers fitted the first‐order kinetics well and the calculated kinetic parameters were consistent with the low accumulation efficiency. Chirality 26:307–312, 2014. © 2014 Wiley Periodicals, Inc.     

A New Chiral Residue Analysis Method for Triazole Fungicides in Water Using Dispersive Liquid‐Liquid Microextraction (DLLME)

A rapid, simple, reliable, and environment‐friendly method for the residue analysis of the enantiomers of four chiral fungicides including hexaconazole, triadimefon, tebuconazole, and penconazole in water samples was developed by dispersive liquid–liquid microextraction (DLLME) pretreatment followed by chiral high‐performance liquid chromatography (HPLC)‐DAD detection. The enantiomers were separated on a Chiralpak IC column by HPLC applying n‐hexane or petroleum ether as mobile phase and ethanol or isopropanol as modifier. The influences of mobile phase composition and temperature on the resolution were investigated and most of the enantiomers could be completely separated in 20 min under optimized conditions. The thermodynamic parameters indicated that the separation was enthalpy‐driven. The elution orders were detected by both circular dichroism detector (CD) and optical rotatory dispersion detector (ORD). Parameters affecting the DLLME performance for pretreatment of the chiral fungicides residue in water samples, such as the extraction and dispersive solvents and their volume, were studied and optimized. Under the optimum microextraction condition the enrichment factors were over 121 and the linearities were 30–1500 µg L^?1 with the correlation coefficients (R²) over 0.9988 and the recoveries were between 88.7% and 103.7% at the spiking levels of 0.5, 0.25, and 0.05 mg L^?1(for each enantiomer) with relative standard deviations varying from 1.38% to 6.70% (n = 6) The limits of detection (LODs) ranged from 8.5 to 29.0 µg L^?1(S/N = 3). Chirality 25:567‐574, 2013. © 2013 Wiley Periodicals, Inc.     

Deleterious effect of soil‐applied metalaxyl and mancozeb on the mycoparasite Pythium oligandrum

Pythium oligandrum was recovered, identified and quantified from air‐dried soil plated on 1.5% water agar containing 0.1% glucose. Isolations of P. oligandrum over 2 years from soils treated with single applications of metalaxyl plus mancozeb were consistently lower than those from untreated soil from the same fields. In three fields in the first year P. oligandrum was reduced from a range of 43.3–115.0 to 17.0–43.2 isolates g^‐1 soil. In the second year, results from 11 fields showed reductions from 27.8–141.8 to 2.8–44.5 isolates g^‐1 soil. P. oligandrum was sensitive to both metalaxyl and mancozeb, with median effective dose (ED₅₀) values of 0.13 ± 0.02 μg m^‐1 and 3.33 ± 0.12 μg ml^‐1, respectively. In a pot test with three soils treated with metalaxyl, mancozeb or the combination of fungicides, levels of P. oligandrum declined over 3 months, with effects first recorded 2 weeks after treatment. Levels of P. oligandrum were reducd by differing degrees in the three soils. Isolate counts from untreated soils declined from a mean of 58.0 g^‐1 soil at the start of the experiment to 27.5 g^‐1 after 3 months, whereas fungicide treatments caused further reductions to 11.1 (metalaxyl), 9.7 (mancozeb) and 4.8 isolates g^‐1 (metalaxyl plus mancozeb).     

Effect of Chromatographic Conditions on Enantioseparation of Bovine Serum Albumin Chiral Stationary Phase in HPLC and Thermodynamic Studies

Twelve chiral compounds were enantiomerically resolved on bovine serum albumin chiral stationary phase (BSA‐CSP) by high‐performance liquid chromatography (HPLC) in reversed‐phase modes. Chromatographic conditions such as mobile phase pH, the percentage of organic modifier, and concentration of analyte were optimized for separation of enantiomers. For N‐(2, 4‐dinitrophenyl)‐serine (DNP‐ser), the retention factors (k) greatly increase from 0.81 to 6.23 as the pH decreasing from 7.21 to 5.14, and the resolution factor (R_s) exhibited a similar increasing trend (from 0 to 1.34). More interestingly, the retention factors for N‐(2, 4‐dinitrophenyl)‐proline (DNP‐pro) decrease along with increasing 1‐propanol in mobile phase (3%, 5%, 7% and 9% by volume), whereas the resolution factor shows an upward trend (from 0.96 to 2.04). Moreover, chiral recognition mechanisms for chiral analytes were further investigated through thermodynamic methods. Chirality 25:487–492, 2013. © 2013 Wiley Periodicals, Inc.     

Enantioselective kinetics of α-hexachlorocyclohexane in earthworm (Eisenia fedtia) and forest soil

The enantioselective bioaccumulation and elimination behaviors of α-hexachlorocyclohexane (α-HCH) enantiomers in earthworm and soil were investigated by chiral gas chromatography. Enantiomer fraction values were calculated as indicators of the enantioselectivity. The mature earthworms were exposed to 0.10 μg g(-1)(wwt) (0.14 μg g(-1)(dwt)) spiked soil continuously for the bioaccumulation, and the elimination was conducted after an enrichment period in the soil. The results showed that both the bioaccumulation and elimination processes followed monophasic kinetics, body residues of α-HCH in earthworm increased to high level at the fifth day, and enantioselectivity was found in the bioaccumulation process with the rate constant (k) of 0.80 d(-1) for (+)-α-HCH and 0.74 d(-1) for (-)-α-HCH. The half life (t(1/2)) of the enantiomers obtained in the elimination process was within one day. The bioaccumulation factors of steady state of α-HCH enantiomers were 2.82 for (+)-α-HCH and 2.75 for (-)-α-HCH. The enantiomer fractions of earthworm and soil obviously below 0.5 during uptake and elimination processes indicate significant enantioselectivity and preferential depuration of (+)-α-HCH in earthworm. However, earthworms do not have a great capacity for getting rid of α-HCH in polluted soil shown by a contradistinctive experiment.