Enantiomeric Fraction Values Research Articles

Enantioselective Degradation of Indoxacarb in Cabbage and Soil under Field Conditions

The enantioselective degradation of indoxacarb in cabbage and soil has been investigated in Beijing and Anhui under open conditions. Indoxacarb enantiomers in samples were extracted with acetonitrile, cleaned up by florisil SPE column, separated on high performance liquid chromatography with a cellulose-tris-(3, 5-dimethylphenylcarbamate)-based chiral stationary phase (CDMPC-CSP), and determined by a photodiode array detector. The validation of the developed method by fortification rac-indoxcarb in cabbage and soil showed good accuracy and precision. The results of field trials indicated that the dissipation of indoxacarb enantiomers followed pseudo-first-order kinetics or first-order kinetics in cabbage and soil at two locations. The half-lives of two enantiomers in cabbage ranged from 2.8 to 4.6 d which were shorter than those in soil ranging from 23 to 35 d. The changes of enantiomeric fraction values proved that enantioselective degradation of indoxacarb happened in cabbage and soil. The (-)-indoxacarb showed faster degradation in the Beijing cabbage, whereas in the Anhui cabbage, (+)-indoxacarb preferentially degraded. In soil, preferential degradation of (+)-indoxacarb was observed at two locations.

Trace analysis of fluoxetine and its metabolite norfluoxetine. Part II: Enantioselective quantification and studies of matrix effects in raw and treated wastewater by solid phase extraction and liquid chromatography–tandem mass spectrometry

The isotope-labeled compounds fluoxetine-d5 and norfluoxetine-d5 were used to study matrix effects caused by co-eluting compounds originating from raw and treated wastewater samples, collected in Uppsala, Sweden. The matrix effects were investigated by the determination of matrix factors (MF) and by a post-column infusion method. The matrix factors were determined to be 38–47% and 71–86% for the enantiomers of norfluoxetine-d5 and fluoxetine-d5, respectively. The influence of matrix effects when quantifying the enantiomers of the active pharmaceutical ingredient and the metabolite in wastewater samples with LC–MS/MS is discussed and methods to overcome the problem are presented. The enantiomeric concentrations of fluoxetine and its human metabolite norfluoxetine, quantified by a one-point calibration method, were 12–52pM (3.5–16ngL−1) in raw wastewater and 4–48pM (1.2–15ngL−1) in treated wastewater. Furthermore, the calculated enantiomeric fractions (EF) of the substances were found to be between 0.68 and 0.71 in both matrices. Neither the EF values for fluoxetine nor those for norfluoxetine were significantly different in the raw wastewater compared to the treated wastewater. Interestingly, the concentration of (S)-fluoxetine was found to be higher than the concentration of (R)-fluoxetine in both raw and treated wastewater. These results are different from other results presented in the literature, which shows that the relative concentrations of the enantiomers of a chiral active pharmaceutical ingredient might be significantly different in wastewater samples from different treatment systems. We report, for the first time, the concentrations of the enantiomers of norfluoxetine in wastewater samples. The concentrations of (S)-norfluoxetine were found to be higher than the concentration of (R)-norfluoxetine in the raw as well as in the treated wastewater samples.

Hexabromocyclododecanes in Surface Soils from E-Waste Recycling Areas and Industrial Areas in South China: Concentrations, Diastereoisomer- and Enantiomer-Specific Profiles, and Inventory

Hexabromocyclododecanes (HBCDs) are raising concern because of their potential persistence, bioaccumulation, and toxicity. In this study, we investigated the concentrations, diastereoisomer- and enantiomer-specific profiles, and mass inventories of HBCDs in 90 surface soils from two e-waste recycling areas (Qingyuan, Guiyu) and from industrial areas in South China. The mean concentrations of total HBCDs in the surface soils ranged from 0.22 to 0.79 and from 0.31 to 9.99 ng/g dw for two surrounding e-waste recycling sites and industrial areas, respectively. The highest total HBCD concentration (284 ng/g dw) was found at the e-waste recycling site in Qingyuan, while total HBCD levels fell dramatically with increasing distance from the recycling site, suggesting that the e-waste recycling activities were an important source of HBCDs. The diastereoisomer profiles in 75 of the 90 soil samples differed from those of the commercial products. The mean enantiomeric fraction values for α-, β-, and γ-HBCDs in soils ranged from 0.503(0.010) to 0.507(0.003), 0.494(0.003) to 0.506(0.009), and 0.502(0.003) to 0.511(0.006), respectively, suggesting that there was no stereoselective transformation of the three diastereoisomers. The mass inventories of HBCDs gave preliminarily estimates of 3.42 kg and 1.84 tonnes for the e-waste recycling areas and industrial areas, respectively. It is notable that the diasteroisomer and enantiomer profiles of this study failed to distinguish definitely that the isomeric transformation occurred during the product processing or in the environmental matrix. Further laboratory studies on abiotic and biotic transformation are needed to clarify this issue.

Investigation of source apportioning for α-HCH using enantioselective analysis

Enantiomeric analysis can be used as a complementary tool for source apportionment of chiral compounds, particularly for α-HCH. In this study we used archived samples from studies related to the distribution of POPs in air–water and air–soil–grass systems. Such approach is based on the behaviour of chiral compounds released into the atmosphere from a primary source, when they are expected to show racemic or close to racemic composition. Contrarily, when chiral compounds have been reemitted from secondary sources (e.g. water or soil), their enantiomeric signatures are frequently non-racemic and are similar to the signature of the secondary source. To show such evidence, extracts from passive air samples deployed throughout Europe were analyzed for the enantiomers of α-HCH. The proximity to a large water body showed a high impact on the enantiomeric signatures: Baltic air had enantiomeric fractions (EFs) <0.500, while Mediterranean air had predominantly EFs >0.500. Similarly, Atlantic air showed a latitude influence: above 50°N most EFs <0.500, whereas at latitudes below 50°N, EFs were >0.500. A similar trend was also observed for EFs of α-HCH measured in air samples from a latitudinal transect during an Atlantic cruise. This transect shows that samples from higher latitudes (above 40°N) have EF <0.500, whereas in the more southern samples (African coast and Southern Atlantic), there is no clear trend for EFs. Inland air samples showed a large range in EF values, with racemic signatures for samples with the highest α-HCH concentrations and an increasing spread in the EFs for lower α-HCH concentrations. As expected, the EF values of α-HCH in air, soils and grass were also impacted by latitude. Correlations between EFs and geographic characteristics of the sampling locations, as well as α-HCH concentrations, α-/γ-isomer ratios, or temperature suggest that enantioselective analysis can give additional information on the distribution and sources of α-HCH in the environment.

The effect of humic acid and ash on enantiomeric fraction change of chiral pollutants

The aim of the study was to determine changes of enantiomeric fraction (EF) of chiral compounds interacting with components of soil and to describe the effect of amount of soil components on EF value. Soil components (humic acid and ash) were isolated from soil using method recommended by the International Humic Substance Society (IHSS). Dichloromethane was used as an extraction solvent for extraction of polychlorinated biphenyls (PCBs), mecoprop and its methyl ester enantiomers from humic acid (HA) and ash. The extracts were analyzed by a high performance liquid chromatography (HPLC). HPLC columns filled with silica gel modified by permethylated cyclodextrine acting as a chiral selector were used for analytical separation of enantiomers. The results confirm the stereoselective effect of soil components on extractable amount of individual enantiomers. Significant changes of EF of chiral pollutants were observed for investigated PCBs enantiomers (HA 0.49–0.45, ash 0.50–0.58) and for mecoprop-methyl ester (HA 0.55–0.59, ash 0.49–0.34), smaller changes of EF were observed for mecoprop (HA 0.52–0.50, ash 0.49–0.50). The changes of EF were dependent on an amount of humic acid and ash. Inverse trends of humic acid EF changes were obtained comparing to trends of EF changes of ash.

Can Physicochemical and Microbial Soil Properties Explain Enantiomeric Shifts of Chiral Organochlorines?

Enantiomeric fractions (EF) of PCB 95, 132, 149, and 174, alpha-HCH, o,p'-DDD, and o,p'-DDT were analyzed in 112 soil samples using two-dimensional gas chromatography and triple-quadrupole mass spectrometry. To assess the soil conditions that facilitate enantioselective fractionation of chiral compounds, EF values of selected PCBs were further correlated with a wide range of physicochemical and microbial soil parameters in an attempt to identify the influential factors and their mutual relations. It was evident that soils where nonracemic ratios of investigated compounds were found were more carbon rich but they also contained significantly more humic and fulvic acids and total nitrogen. These specific physicochemical properties were accompanied by significantly increased values of all key biotic variables, the amount of microbial biomass, and its respiration activity (both basal and substrate-induced). Therefore, the shifts from racemic ratios appeared to be associated with more sustainable and active soil microflora. Among other abiotic characteristics, most significant differences were detected in the soil texture. Soil samples with significant shifts contained increased amount of clay component and correspondingly decreased proportion of sand fraction. These differences can also be associated with more intensive microbial activity, because clay content and texture with an increased amount of microaggregates are known to be favorable for soil microflora and its viability.

Enantioselective Bioaccumulation of Hexabromocyclododecane and Congener-Specific Accumulation of Brominated Diphenyl Ethers in an Eastern Canadian Arctic Marine Food Web

The extent of trophic transfer of the three diastereoisomers of hexabromocyclododecane (HBCD) and seven brominated diphenyl ether (BDE) congeners was examined in components of an Arctic marine food web from eastern Canada. Alpha and gamma-HBCD diastereoisomers were detected in all species and total (sigma) HBCD concentrations ranged from 0.6 +/- 0.2 pg/g (geometric mean +/- 1 x standard error (SE), lipid weight (lw)) in arctic cod to 3.9 +/- 0.9 ng/g (lw) in narwhal. beta-HBCD was below method detection limits in all the samples. Mean sigmaBDE (sum of seven congeners) concentrations ranged from 0.4 +/- 0.2 ng/g (lw) in walrus to 73 +/- 10 ng/g (lw) in zooplankton. The relative trophic status of biota was determined by nitrogen stable isotopes (delta15N), and results indicated clear differences in HBCD isomer and BDE congener profiles with trophic level (TL). Trophic magnification was observed for the alpha-diastereoisomer and BDE-47 as concentrations increased with increasing TL in the food web, whereas there was trophic dilution of gamma-HBCD and BDE-209 through the food web. Only the (-)alpha-enantiomer showed a strong positive relationship between concentration and TL (p < 0.01) with a trophic magnification factor (TMF) value of 2.2. A small but significant increase in the enantiomeric fraction value of the alpha-enantiomers with TL was also observed (r2 = 0.22, p < 0.005), implying that there is an overall preferential enrichment of the (-)alpha-enantiomer relative to the (+)alpha-enantiomer likely due to the greater bioaccumulation potential of the (-)alpha-enantiomer and/or to the greater susceptibility of the (+)alpha-enantiomer to metabolism.

Enantiomeric Enrichment of 2,2′,3,3′,6,6′-Hexachlorobiphenyl (PCB 136) in Mice After Induction of CYP Enzymes

Several PCB congeners, present in commercial PCB formulations, are chiral. These PCBs can undergo enantiomeric enrichment in many animal species and in humans due to currently uncharacterized enantioselective biotransformation processes. To investigate if certain cytochrome P-450 enzymes (CYPs), such as CYP2B's, are responsible for this enantiomeric enrichment, we investigated the enantioselective disposition of (+/-)-PCB 136 in female mice after induction of different CYP enzymes by pretreatment with corn oil alone, beta-naphthoflavone (CYP1A's), phenobarbital (CYP2B's), or dexamethasone (2B's and 3A's), followed by oral PCB administration. PCB 136 levels were significantly lower in phenobarbital- and, to a lesser extent, in dexamethasone-pretreated animals, presumably due to the induction of PCB 136 metabolizing enzymes. Although (+)-PCB 136 was enriched in all tissues, none of the pretreatments altered the enantioselective disposition of PCB 136 in a manner that suggests a particular CYP subfamily as the cause of the enrichment of (+)-PCB 136. Fecal PCB levels and enantiomeric fraction values changed over time in a manner consistent with slower digestive motility in the mice pretreated with phenobarbital and dexamethasone. Overall, this study does not support the hypothesis that metabolism by CYP2B enzymes is responsible for the enrichment of (+)-PCB 136 in mice.

Enantioselective Microbial Transformation of the Phenylpyrazole Insecticide Fipronil in Anoxic Sediments

Fipronil, a chiral insecticide, was biotransformed initially to fipronil sulfide in anoxic sediment slurries following a short lag period. Sulfidogenic or methanogenic sediments transformed fipronil with half-lives of approximately 35 and 40 days, respectively. In all microbially active sediment slurries tested, the transformation of fipronil to fipronil sulfide was enantioselective. In the sulfidogenic sediment slurry, the enantiomeric fraction (EF) of fipronil decreased from an initial racemic EF value of 0.46 to a value of 0.22 during the incubation period of active fipronil transformation, indicating preferential transformation of the S-(+)-enantiomer. A previously unidentified product, 5-amino1-[2,6-dichloro-4-(trifluoromethyl)-phenyl]-4-(trifluoromethylthio)-1-H-pyrazole-3-carboxyamide, or fipronil sulfide-amide, was detected in the sulfidogenic slurries and coincided with the loss of fipronil sulfide. Biota from methanogenic freshwater sediment slurries also transformed fipronil enantioselectively but with a preference for the R-(-)-enantiomer. In all microbially inhibited (autoclaved) sediment slurries tested, no changes in the enantiomeric fractions of fipronil were observed and only low levels (< 5% of the added fipronil) of the fipronil sulfide metabolite were detected. In defined (model) chemical experiments, solutions of pyrite (FeS2) and iron sulfide (FeS) non-enantioselectively transformed fipronil primarily to either 2,6-dichloro-4-(trifluoromethyl)-aniline or to fipronil sulfide and fipronil amide, respectively. This report provides the first experimental evidence of enantioselective microbial transformation of fipronil in a natural environment (soil, water, and sediment) as well as identification of a novel fipronil biotransformation product.

Influence of dietary fat on the enantioselective disposition of 2,2′,3,3′,6,6′-hexachlorobiphenyl (PCB 136) in female mice

Although ingestion is the major route of exposure to polychlorinated biphenyls (PCBs), dietary factors altering their absorption and excretion are only poorly understood. In the present study, (±)-PCB 136 was administered orally to female C57BL/6 mice fed an unrefined (URD, 10% fat) or high fat (HFD, 40% fat) diet to investigate the effect of the dietary fat content on the disposition of PCBs. Three days after administration, PCB levels in the adipose tissue were significantly lower in HFD animals than URD animals, partly due to a higher excretion rate of PCB 136 in the HFD group. (+)-PCB 136 was enriched in all organs and in feces. In both groups, enantiomeric fractions in feces increased each day after administration. We hypothesize that low EF (enantiomeric fraction) values in feces excreted within 24 h of exposure are due to the presence of undigested, racemic PCB. Higher EF values in feces excreted after two and three days are due to excretion of previously absorbed PCBs. Overall, our study suggests that the EF value may be a good tool to investigate the absorption and excretion of PCBs in vivo.

Factors influencing enantiomeric fractions of hexabromocyclododecane measured using liquid chromatography/tandem mass spectrometry

Hexabromocyclododecane (HBCD), the most heavily produced of the cycloaliphatic brominated flame retardants (BFRs), is a mixture of three predominant diastereomers (alpha-, beta-, and gamma-HBCD), each with a corresponding pair of enantiomers. We have investigated the liquid chromatography/mass spectrometry (LC/MS) behaviour of the HBCD enantiomers, and demonstrated that enantiomeric fractions (EFs) calculated using data uncorrected for instrument and/or matrix effects can result in potentially inaccurate EF values. However, use of labelled surrogates effectively corrects for these effects. Experiments with racemic HBCD standards indicate that chromatographic factors, including mobile phase composition and column bleed from chiral stationary phases, may be contributors to variations in the mass spectrometric response of the HBCD enantiomers.

Chiral organochlorine pesticides in the atmosphere

We measured the enantiomeric fractions (EF) of various organochlorine pesticides in air samples from Indiana, Arkansas, and Louisiana collected in 2002–2003. Enantiomeric fractions for o,p′-DDD are reported here for the first time, and they suggest preferential depletion of one enantiomer over the other, with a mean value of 0.44. For the chlordanes, the EFs followed the same trends as reported in previous studies. EF values for trans-chlordane (TC) were slightly below 0.5, and values for cis-chlordane (CC) were slightly above 0.5. For cis-heptachlor epoxide (HEPX), the EF values averaged 0.65, indicating a clear preferential depletion of the (−)-enantiomer. For TC and CC, no clear distinction could be made between the values measured among the three sites, suggesting that it is not possible to use these EF values as “source signatures”. For HEPX, the Indiana site had significantly higher EF values than did the Arkansas site; this difference may reflect differences in the historic use of heptachlor in these two regions.

Persistent organochlorine contaminants and enantiomeric signatures of chiral pollutants in ringed seals (Phoca hispida) collected on the east and west side of the Northwater Polynya, Canadian Arctic.

To examine the influence of diet and age on organochlorine contaminant (OC) concentrations in two closely related ringed seal (Phoca hispida) populations enantiomeric fractions (EFs) of chiral contaminants and stable isotopes of nitrogen (delta15N) and carbon (delta13C) were measured along with OCs in ringed seals collected from the east and west side of the Northwater Polynya. Seals from these two locations were feeding at the same trophic level based on delta15N values in muscle but had slightly different sources of carbon based on delta13C measurements in muscle. After removing the influence of age, sex, and blubber thickness, OC concentrations did not vary between ringed seals from the east and west side of the polynya. SigmaPCB, SigmaDDT, and Sigmachlordane were found to increase with age for both male and female seals. The inclusion of older (>20 years) female seals, which may have a reduced reproductive effort, may influence the relationships in females. Stable isotopes failed to describe OC concentrations in ringed seals suggesting that diet was not a major factor in variation of OC concentrations within this ringed seal population. Cis- and trans-chlordane, oxychlordane, and heptachlor epoxide were all nonracemic in the ringed seal blubber but did not vary with age, sex, or collection site. Alpha-HCH appeared racemic (enantiomeric fraction = 0.50 +/- 0.01) in the seals, although this EF is different than those previously observed in their prey species, and was found to vary significantly with age. EF values in the ringed seals varied considerably from other Arctic marine mammals and seabirds, providing addition evidence that the type(s) and characteristic(s) of the enzymes involved in biotransformation of chiral OCs vary between these organisms.