[Determination of disulfoton and its metabolites in agricultural products by dispersive soild phase extraction-ultra high performance liquid chromatography-tandem mass spectrometry].

Disulfoton, an organophosphorus pesticide, is used to control cotton, beet, potato, and other seedling period aphids, leaf moths, underground pests, etc., with internal absorption, killing, gastric poisoning, and fumigation. Disulfoton is a highly toxic organophosphate pesticide, which can inhibit cholinesterase activity, resulting in neurophysiological disorders by inhalation, feeding, and transdermal absorption. Disulfoton is difficult to degrade in the environment, which leads to enrichment in organisms and interference with endocrine. This compound is harmful to the ecological environment and human health. To ensure the quality and safety of food, it is important to develop a detection method for disulfoton and its metabolites in agricultural products. A reliable method based on dispersive solid phase extraction (d-SPE) with ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed for the determination of disulfoton and its metabolites (disulfoton sulfone, disulfoton sulfoxide, demeton-S, demeton-S sulfone, and demeton-S sulfoxide) in agricultural products (pea, asparagus, wheat, coffee bean, and peanut). The optimal extraction method was as follows: 5.0 g the samples were extracted with acetonitrile (wheat, coffee bean, and peanut presoaked in 5 mL water) in a 50 mL centrifuge tube, followed by 10 min vortex. Before 30 s vortex, 4 g NaCl was added. After 5 min centrifugation, 1.5 mL of the supernatant was cleaned up with 50 mg octadecylsilane bonded silica (C18), 50 mg primary secondary amine (PSA), and 50 mg aminopropyl (NH2) adsorbents. The analytes were separated on a Thermo Syncronis C18 column (150 mm×2.1 mm, 5 µm) with gradient elution using water and acetonitrile at a column temperature of 40 ℃. The injection volume was 2 µL. Disulfoton and its metabolites were analyzed in multiple reaction monitoring (MRM) mode with positive electrospray ionization (ESI+) for the selective quantification. Qualitative and quantitative analyses were accorded to the retention times and characteristic ion pairs with one parent ion and two fragment ions. Quantitative analysis was performed by an external standard method using matrix-matched calibration curves. All the parameters that affected the extraction efficiencies were optimized. C18, PSA, and NH2 gave good recoveries of 87.9%-109.0%. Other adsorbents, multiwalled carbon nanotubes (MWCNTs), hydroxylated multiwalled carbon nanotubes (OH-MWCNTs), carboxylated multiwalled carbon nanotubes (COOH-MWCNTs), octylsilane bonded silica (C8), strong cation exchange (SCX) and neutral alumina (N-Al2O3), led to recoveries below 56.2%. The combination of adsorbents was also considered. Seven different combinations of 50 mg C18, 50 mg PSA, and 50 mg NH2 were chosen for the optimization experiments. There were no obvious differences in these combinations, and the target analytes recoveries ranged from 81.0% to 109.3% with relative standard deviations (RSDs) between 0.6% and 12.5%. The matrix effect could affect the extraction efficiency. The adsorbents of 50 mg C18, 50 mg PSA, and 50 mg NH2 showed weaker matrix effects as compared with other combinations of adsorbents in the instrument. The results for the matrix effect showed that peanuts and asparagus exceeded 20%, requiring matrix-matched calibration curves. Under the optimized conditions, disulfoton and its metabolites showed good linearities (R2≥0.9981) in the range of 2.0-200.0 µg/L. The average spiked recoveries of disulfoton and its metabolites in peas, asparagus, wheat, peanuts, and coffee beans ranged from 75.0% to 110.0%, with RSDs of 0.7% to 14.9%. The limits of detection (LODs) were between 0.02 and 2.0 µg/kg, and the limits of quantification (LOQs) were 5.0 µg/kg. The method was applied for the detection of 80 commercial productions, and neither disulfoton nor its metabolites were found. The proposed method is rapid, accurate, highly selective, and sensitive, and it is suitable for the simultaneous determination of disulfoton and its metabolites in grain, oil crops, vegetables, and other matrices.

Degradation of Praguicide Disulfoton Using Nanocompost and Evaluation of Toxicological Effects.

Organophosphates (OPPs) are an important element of modern agriculture; however, because they are being used excessively, their residues are leaching and accumulating in the soil and groundwater, contaminating aquatic and terrestrial food chains. An important OPP called disulfoton is frequently used to eradicate pests from a wide range of crops, including Brazil's coffee crops. Additionally, it does not easily degrade in the environment, and as such, this compound can slowly build up in living organisms such as humans. Moreover, this compound has been classified as "extremely hazardous" by the World Health Organization. This study evaluated the degradation efficiency of disulfoton using a Fenton-like reaction catalyzed by magnetite nanoparticles and determined the toxicity of the by-products of the degradation process using the bioindicator Allium cepa. Further, the removal efficiency of disulfoton was determined to be 94% under optimal conditions. On the other hand, the Allium cepa bioassay showed different toxic, cytotoxic, genotoxic, and mutagenic outcomes even after the remediation process. In conclusion, the Fenton process catalyzed by magnetite nanoparticles presents great efficiency for the oxidation of disulfoton. However, it is important to highlight that the high degradation efficiency of the Fenton-based process was not sufficient to achieve detoxification of the samples.

Determination of 60 pesticides in hen eggs using the QuEChERS procedure followed by LC-MS/MS and GC-MS/MS.

An analytical method involving QuEChERS (quick, easy, cheap, effective, rugged, and safe) sample preparation, followed by LC-MS/MS and GC-MS/MS was developed and validated for the determination of 60 pesticides in eggs. Recoveries of 70-120% were achieved for selected pesticides and relative standard deviations <20% were obtained for most analytes at three concentrations. The limit of quantification was <10 µg kg-1 for 83% of the total pesticides. This method was used to analyze 58 egg samples and the residues of seven pesticides (disulfoton, fipronil sulfone, cyromazine, o,p-DDT, p,p-DDD, p,p-DDT, and permethrin) were quantified in 16 egg samples at levels of 5-10 µg kg-1, which was below the corresponding the maximum residue levels, as established by Korean Ministry of Food and Drug Safety. We demonstrated that LC-MS/MS and GC-MS/MS in combination with QuEChERS can be used to routinely monitor multiple pesticide residues in egg samples.

Comparative analysis of the metal-dependent structural and functional properties of mouse and human SMP30.

Senescence Marker Protein (SMP30) is a metalloenzyme that shows lactonase activity in the ascorbic acid (AA) biosynthesis pathway in non-primate mammals such as a mouse. However, AA biosynthesis does not occur in the primates including humans. Several studies have shown the role of SMP30 in maintaining calcium homeostasis in mammals. In addition, it is also reported to have promiscuous enzyme activity with an organophosphate (OP) substrate. Hence, this study aims to recombinantly express and purify the SMP30 proteins from both mouse and human, and to study their structural alterations and functional deviations in the presence of different divalent metals. For this, mouse SMP30 (MoSMP30) as well as human SMP30 (HuSMP30) were cloned in the bacterial expression vector. Proteins were overexpressed and purified from soluble fractions as well as from inclusion bodies as these proteins were expressed largely in insoluble fractions. The purified proteins were used to study the folding conformations in the presence of different divalent cations (Ca2+, Co2+, Mg2+, and Zn2+) with the help of circular dichroism (CD) spectroscopy. It was observed that both MoSMP30 and HuSMP30 acquired native folding conformations. To study the metal-binding affinity, dissociation constant (Kd values) were calculated from UV-VIS titration curve, which showed the highest affinity of MoSMP30 with Zn2+. However, HuSMP30 showed the highest affinity with Ca2+, suggesting the importance of HuSMP30 in maintaining calcium homeostasis. Enzyme kinetics were performed with γ-Thiobutyrolactone and Demeton-S in the presence of different divalent cations. Interestingly, both the proteins showed lactonase activity in the presence of Ca2+. In addition, MoSMP30 and HuSMP30 also showed lactonase activity in the presence of Co2+ and Zn2+ respectively. Moreover, both the proteins showed OP hydrolase activities in the presence of Ca2+ as well as Zn2+, suggesting the metal-dependent promiscuous nature of SMP30.

Air Activated Self-Decontaminating Polydicyclopentadiene PolyHIPE Foams for Rapid Decontamination of Chemical Warfare Agents.

The threat of chemical warfare agents (CWA) compels research into novel self-decontaminating materials (SDM) for the continued safety of first-responders, civilians, and active service personnel. The capacity to actively detoxify, as opposed to merely sequester, offending agents under typical environmental conditions defines the added value of SDMs in comparison to traditional adsorptive materials. Porous polymers, synthesized via the high internal phase emulsion (HIPE) templating, provide a facile fabrication method for materials with permeable open cellular structures that may serve in air filtration applications. PolyHIPEs comprising polydicyclopentadiene (polyDCPD) networks form stable hydroperoxide species following activation in air under ambient conditions. The hydroperoxide-containing polyDCPD materials react quickly with CWA simulants, Demeton-S and 2-chloroethyl ethyl sulfide, forming oxidation products as confirmed via gas chromatography mass spectrometry. The simplicity of the detoxification chemistry paired with the porous foam form factor presents an exciting opportunity for the development of self-decontaminating filter media.

Determination of Organothiophosphate Insecticides in Environmental Water Samples by a Very Simple and Sensitive Spectrofluorimetric Method.

A simple, rapid and sensitive spectrofluorimetric method was developed for the determination of di-syston, ethion and phorate in environmental water samples. The procedure is based on the oxidation of these pesticides with cerium (IV) to produce cerium (III), and its fluorescence was monitored at 368 ± 3 nm after excitation at 257 ± 3 nm. The variables effecting oxidation of each pesticide were studied and optimized. Under the experimental conditions used, the calibration graphs were linear over the range 0.2-15, 0.1-13, 0.1-13 ng mL(-1) for di-syston, ethion and phorate, respectively. The limit of detection and quantification were in the range 0.034-0.096 and 0.112-0.316 ng mL(-1), respectively. Intra- and inter-day assay precisions, expressed as the relative standard deviation (RSD), were lower than 5.2 % and 6.7 %, respectively. Good recoveries in the range 86 %-108 % were obtained for spiked water samples. The proposed method was applied to the determination of studied pesticides in environmental water samples.

Study of the correlation between blood cholinesterases activity, urinary dialkyl phosphates, and the frequency of micronucleated polychromatic erythrocytes in rats exposed to disulfoton

Organophosphates (OPs) are widely used as pesticides, and its urinary metabolites as well as the blood cholinesterases (ChEs) activity have been reported as possible biomarkers for the assessment of this pesticide exposure. Moreover, the OPs can induce mutagenesis, and the bone marrow micronucleus test is an efficient way to assess this chromosomal damage. This paper reports a study carried out to verify the correlation among the disulfoton exposure, blood ChEs activity, urinary diethyl thiophosphate (DETP), and diethyl dithiophosphate (DEDTP), as well as micronucleated polychromatic erythrocytes (MNPCEs) frequency. Four groups of rats (n=12) were exposed to disulfoton at 0, 2.8, 4.7, and 6.6 mg kg-1 body weight. The blood ChEs activity, urinary DETP and DEDTP concentrations, and MNPCEs frequency were determined. It was observed that the plasmatic and erythrocytary ChEs activity decreased from 2.9% to 0.5% and from 35.9 to 3.3%, respectively, when the disulfoton dose was increased from 0 to 6.6 mg kg-1 (correlation of 0.99). Urinary DETP and DEDTP concentrations, as well as the MNPCEs frequency, increased from 0 to 6.58 µg mL-1, from 0 to 0.04 µg mL-1, and from 0 to 1.4%, respectively, when the disulfoton dose was increased from 0 to 6.58 mg kg-1 body weight.

Os organofosforados (OPs) são amplamente usados como praguicidas e a atividade da colinesterase sanguínea bem como os metabólitos urinários desses praguicidas têm sido reportados como biomarcadores eficazes para avaliar casos de exposição. Além disso, os OPs podem induzir mutagênese e o teste de micronúcleo de medula óssea é uma boa alternativa para avaliar os danos cromossômicos. Esse artigo reporta um estudo sobre a correlação entre a exposição a dissulfoton, a atividade da colinesterase sanguínea, a excreção urinária de dietil tiofosfato e dietil ditiofosfato e a frequência de micronúcleos em eritrócitos policromáticos. Quatro grupos de ratos (n=12) foram expostos a dissulfotom nas doses de 0, 2,8, 4,7, e 6,6 mg kg-1 de peso corpóreo. A atividade da colinesterase sanguínea as concentrações urinárias de dietil tiofosfato e dietil ditiofosfato e a frequência de micronúcleos foram determinadas. Os resultados demonstraram que as atividades da colinesterase plasmática e eritrocitária diminuíram de 2,9 para 0,5% e de 35,9 para 3,3% , respectivamente, quando a dose de dissulfoton foi aumentada de 0 para 6,6 mg kg-1 (correlação de 0,99). As concentrações urinárias de dietil tiofosfato e dietil ditiofosfato bem como a frequência de micronúcleos aumentaram de 0 a 6,56 µg mL-1, 0 a 0.04 µg mL-1 e de 0 a 1.4%, respectivamente, quando a dose de dissulfotom foi aumentada de 0 a 6,58 mg kg-1.

Rapid determination of disulfoton and its oxidative metabolites in human whole blood and urine using QuEChERS extraction and liquid chromatography-tandem mass spectrometry.

A liquid chromatography-tandem mass spectrometry method was developed and validated for simultaneous determination of disulfoton and five of its oxidative metabolites (disulfoton-sulfoxide, disulfoton-sulfone, demeton-S, demeton-S-sulfoxide and demeton-S-sulfone) in human whole blood and urine. Extraction was undertaken using a QuEChERS method, which is commonly used in food analysis. D10-Disulfoton was used as the internal standard. Separation was carried out using a CAPCELL-PAK MG II column (35×2.0 mm i.d., 5 µm, Shiseido) with a mobile phase of 10 m mol/L ammonium formate and methanol. This method was applied in an autopsy case, and disulfoton and its oxidative metabolites were successfully detected in both blood and urine. The concentrations of disulfoton in the blood and urine were 360 and 23.8 ng/mL, respectively. There was a relatively low concentration of demeton-S in both the blood (4.0 ng/mL) and urine (45.7 ng/mL). To date, there have been no reported cases of detection of demeton-S in human samples.

Detections of eleven organophosphorus insecticides and one herbicide threatening Pacific salmonids, Oncorhynchus spp., in California, 1991-2010.

California's surface water monitoring results from 1991 through 2010 were analyzed to determine whether 12 organophosphorus insecticides and herbicides (i.e., azinphos methyl, bensulide, dimethoate, disulfoton, ethoprop, fenamiphos, methamidophos, methidathion, methyl parathion, naled, phorate, and phosmet) and their degradates have been detected above maximum concentration limits (MCLs) in Pacific salmonid habitats. Methidathion, methyl parathion, phorate, phosmet, and the oxygen analogue of naled (DDVP) detections exceeded MCLs. Methyl parathion detections may be accounted for by monthly use trends, while methidathion detections may be explained by yearly use trends. There were inadequate phorate, phosmet, or DDVP data to evaluate for correlations with use.

Development of a solid phase extraction method for agricultural pesticides in large-volume water samples.

An analytical method using solid phase extraction (SPE) and analysis by gas chromatography/mass spectrometry (GC-MS) was developed to determine trace levels of a variety of 41 agricultural pesticides and selected transformation products in high-elevation surface waters. Large-volume water sampling (up to 100L) was employed because it was anticipated that pesticide contamination, if present, would be at very low levels. The target compounds comprise pesticides (and selected oxygen transformation products) known to have been extensively used in agriculture in the San Joaquin Valley, CA, USA. Solid phase extraction using the polymeric resin Abselut Nexus was optimized to extract the pesticide analytes from water samples. A single determinative method using GC-MS with electron ionization was used for all the analytes. Recoveries from 100L of reagent water at 100pg/L and 1ng/L concentrations were generally greater than 75%, although dimethoate, disulfoton, and phorate were not recovered. Analysis of the extracts without cleanup yielded detection limits for the remaining 38 analytes between 0.1 and 30ng/L. A silica cleanup with separate analysis of 3 eluant fractions improved detection limits for 37 of the compounds to between 6 and 600pg/L in high-elevation surface waters.

Responses to clomazone and 5-ketoclomazone by Echinochloa phyllopogon resistant to multiple herbicides in Californian rice fields.

BACKGROUND: Late watergrass [Echinochloa phyllopogon (Stapf.) Koss.] is a major weed of Californian rice that has evolved P450-mediated metabolic resistance to multiple herbicides. Resistant (R) populations are also poorly controlled by the recently introduced herbicide clomazone. The authors assessed whether this cross-resistance was also P450 mediated, and whether R plants also had reduced sensitivity to photooxidation. Understanding mechanism(s) of resistance facilitates the design of herbicide management strategies to delay resistance evolution. RESULTS: Ratios (R/S) of R to susceptible (S) GR(50) were near 2.0. [(14)C]Clomazone uptake was similar in R and S plants. Clomazone and its metabolite 5-ketoclomazone reduced chlorophyll and carotenoids in S more than in R plants. The P450 inhibitors disulfoton and 1-aminobenzo-triazole (ABT) safened clomazone in R and S plants. Disulfoton safened 5-ketoclomazone only in S plants, while ABT synergized 5-ketoclomazone mostly against S plants. Paraquat was more toxic in S than in R plants. CONCLUSION: Cross-resistance to clomazone explains failures to control R plants in rice fields, and safening by P450 inhibitors suggests that oxidative activation of clomazone is needed for toxicity to E. phyllopogon. Clomazone resistance requires mitigation of 5-ketoclomazone toxicity, but P450 detoxification may not significantly confer resistance, as P450 inhibitors poorly synergized 5-ketoclopmazone in R plants. Responses to paraquat suggest research on mechanisms to mitigate photooxidation in R and S plants is needed.

Cycloheximide and disulfoton are positive in the photoclastogencity assay but do not absorb UV irradiation: another example of pseudophotoclastogenicity?

There is considerable concern regarding the biological plausibility of the response of certain chemicals in the in vitro photoclastogenicity assay, suggesting that this assay is oversensitive and lacks specificity. To explore this further, four coded compounds (aminotriazole, propantheline bromide, cycloheximide and disulfoton) were evaluated for their potential response in a photoclastogenicity assay in cultured Chinese hamster ovary (CHO) cells. None of the four compounds were shown to absorb ultraviolet radiation (UVR) or visible light in the 290- to 700-nm region of the electromagnetic spectrum. A fifth coded compound, tetracycline, which absorbs UVR, was also tested as this has previously been shown to be phototoxic in vitro (3T3-NRU assay) and is cytotoxic, but not genotoxic, at high concentrations in standard 'dark' genotoxicity assays in mammalian cells. The results showed that cycloheximide, disulfoton and tetracycline were clastogenic in CHO cells following UVR exposure (solar-simulated light at 700 mJ/cm(2)) but not in the absence of UVR. Aminotriazole and propantheline were negative in the presence and absence of UVR exposure. Follow-up testing showed that neither cycloheximide nor disulfoton was positive in the 3T3-NRU assay, the standard in vitro regulatory test for phototoxicity, a result consistent with their inability to absorb UVR. These data suggest that both cycloheximide and disulfoton are pseudophotoclastogens, like zinc oxide. Together, these data question the specificity of the in vitro photoclastogencity assay in CHO cells and raises further concern regarding its use for the assessment of chemical photosafety for regulatory purposes. At the very least, a review of the current guidance documents for the photosafety evaluation of pharmaceuticals and cosmetics should be undertaken urgently.

Determination of organophosphorous pesticides in wastewater samples using binary-solvent liquid-phase microextraction and solid-phase microextraction: a comparative study.

A simple and efficient binary solvent-based two-phase hollow fiber membrane (HFM)-protected liquid-phase microextraction (BN-LPME) technique for moderately polar compounds was developed. Six organophosphorous pesticides (OPPs) (triethylphosphorothioate, thionazin, sulfotep, phorate, disulfoton, methyl parathion and ethyl parathion) were used as model compounds and extracted from 10-mL wastewater with a binary-solvent (toluene:hexane, 1:1) mixture. Some important extraction parameters, such as extraction time, effect of salt, sample pH and solvent ratio composition were optimized. BN-LPME combined with gas chromatography/mass spectrometric (GC/MS) analysis provided repeatability (R.S.D.s < or = 12%, n = 4), and linearity (r < or = 0.994) and solid-phase microextraction provides comparable of R.S.D.s < or = 13%, n = 4 and linearity (r < or = 0.966) for spiked water samples. The limits of detection (LODs) were in the range of 0.3-11.4 ng L(-1) for BN-LPME and 3.1-120.5 ng L(-1) for SPME at (S/N = 3) under GC/MS selective ion monitoring mode. In addition to high enrichment, BN-LPME also served as a sample cleanup procedure, with the HFM act as a filtering medium to prevent large particles and extraneous materials from being extracted. To investigate and compare their applicability, the BN-LPME and SPME procedures were applied to the detection of OPPs in domestic wastewater samples.

Potential management strategies for the linden borer (Coleoptera: Cerambycidae) in urban landscapes and nurseries.

The linden borer, Saperda vestita Say (Coleoptera: Cerambycidae), is a native insect species that is common throughout north central and northeastern North America. Over the past decade, increasing occurrence of damage associated with the linden borer has been reported on Tilia spp. in city street trees and nurseries throughout Wisconsin, probably because of increased use of these trees. Our objective was to gain a better understanding of the seasonal biology and potential management strategies for this important pest. We evaluated the effectiveness of three systemic insecticides, imidacloprid, thiamethoxam, and disulfoton, and a mechanical control method of chipping linden borer-infested wood as a means of reducing S. vestita larval survival, subsequent emergence, and oviposition. Autumn and spring soil injections of imidacloprid to linden borer-infested Tilia cordata'Greenspire' nursery stock (< 11.4 cm in diameter at breast height [dbh]) provided >90% control. Autumn soil injections of imidacloprid and thiamethoxam and a spring granular soil application treatment of disulfoton applied to larger (>22 cm dbh) Tilia spp. did not effectively control linden borer at the application rates tested. Chipping infested Tilia spp. effectively destroyed linden borer larvae, pupae, and adults. Arborists and landscape managers should consider chipping felled Tilia spp. trees infested with S. vestita to prevent adults from potentially attacking nearby susceptible trees.

Characterization of a phosphodiesterase capable of hydrolyzing EA 2192, the most toxic degradation product of the nerve agent VX.

Glycerophosphodiesterase (GpdQ) from Enterobacter aerogenes is a nonspecific diesterase that enables Escherichia coli to utilize alkyl phosphodiesters, such as diethyl phosphate, as the sole phosphorus source. The catalytic properties of GpdQ were determined, and the best substrate found was bis(p-nitrophenyl) phosphate with a kcat/Km value of 6.7 x 10(3) M-1 s-1. In addition, the E. aerogenes diesterase was tested as a catalyst for the hydrolysis of a series of phosphonate monoesters which are the hydrolysis products of the highly toxic organophosphonate nerve agents sarin, soman, GF, VX, and rVX. Among the phosphonate monoesters tested, the hydrolysis product of rVX, isobutyl methyl phosphonate, was the best substrate with a kcat/Km value of 33 M-1 s-1. The ability of GpdQ to hydrolyze the phosphonate monoesters provides an alternative selection strategy in the search of enhanced variants of the bacterial phosphotriesterase (PTE) for the hydrolysis of organophosphonate nerve agents. This investigation demonstrated that the previously reported activity of GpdQ toward the hydrolysis of methyl demeton-S is due to the presence of a diester contaminant in the commercial material. Furthermore, it was shown that GpdQ is capable of hydrolyzing a close analogue of EA 2192, the most toxic and persistent degradation product of the nerve agent VX.

Mutant of Bungarus fasciatus acetylcholinesterase with low affinity and low hydrolase activity toward organophosphorus esters.

Enzymes hydrolysing highly toxic organophosphate esters (OPs) are promising alternatives to pharmacological countermeasures against OPs poisoning. Bungarus fasciatus acetylcholinesterase (BfAChE) was engineered to acquire organophosphate hydrolase (OPase) activity by reproducing the features of the human butyrylcholinesterase G117H mutant, the first mutant designed to hydrolyse OPs. The modification consisted of a triple mutation on the (122)GFYS(125) peptide segment, resulting in (122)HFQT(125). This substitution introduced a nucleophilic histidine above the oxyanion hole, and made space in that region. The mutant did not show inhibition by excess acetylthiocholine up to 80 mM. The k(cat)/K(m) ratio with acetylthiocholine was 4 orders of magnitude lower than that of wild-type AChE. Interestingly, due to low affinity, the G122H/Y124Q/S125T mutant was resistant to sub-millimolar concentrations of OPs. Moreover, it had hydrolysing activity with paraoxon, echothiophate, and diisopropyl phosphofluoridate (DFP). DFP was characterised as a slow-binding substrate. This mutant is the first mutant of AChE capable of hydrolysing organophosphates. However, the overall OPase efficiency was greatly decreased compared to G117H butyrylcholinesterase.

Reaction of thiometon and disulfoton with reduced sulfur species in simulated natural environment.

The reactions of thiometon and its ethyl analogue, disulfoton, with reduced sulfur species [e.g., bisulfide (HS-), polysulfide (S(n)2-), thiophenolate (PhS-), and thiosulfate (S2O3(2-))] were examined in well-defined aqueous solutions under anoxic conditions. The role of reduced sulfur species was investigated in the abiotic degradation of thiometon and disulfoton. Experiments at 25 degrees C demonstrated that HS-, S(n)2-, PhS-, and S2O3(2-) promoted the degradation of thiometon to a great extent while only S(n)2- and PhS- showed a small accelerating effect in the degradation of disulfoton. Reactions were monitored at varying concentrations of reduced sulfur species to obtain the second-order rate constants. The reactivity of the reduced sulfur species decreased in the following order: S(n)2- > PhS- > HS- approximately S2O3(2-). Transformation products were confirmed by standards or characterized by gas chromatography mass spectrometry. The results illustrate that multiple pathways occur in the reactions with reduced sulfur species, among which the nucleophilic attack at the alpha-carbon of the alkoxy group was the predominant pathway. Activation parameters of the reaction of thiometon and disulfoton with HS- were also determined from the measured second-order rate constants over a temperature range. DeltaH( not equal) values indicated that the reactivity of thiometon toward HS- was much greater than for disulfoton. Nucleophilic attack at the alkoxy group was more important for thiometon than disulfoton. When the measured second-order rate constants at 25 degrees C are multiplied by [HS-] and Sigma[S(n)2-] reported in saltmarsh porewaters, predicted half-lives show that reduced sulfur species present at environmentally relevant concentrations may present an important sink for thiometon in coastal marine environments.

[Analysis of disulfoton sulfoxide in chingentsuai by pulsed splitless mass spectrometry using programmed temperature vaporization (PTV) inlet].

A peak considered to be due to disulfoton sulfoxide as a metabolite of disulfoton was detected in the analysis of the chingentsuai extraction solution of vegetables by GC-FPD. In the analysis by GC/MS for identification, though the retention time and the mass spectrum were the same as those of the standard, the spectrum was different from MS library data. It appeared that decomposition of disulfoton sulfoxide occurred at the inlet. Therefore, we used a programmed temperature vaporization (PTV) inlet in the pulsed splitless mode to achieve a low inlet temperature and high injection pressure. As a result, the expected mass spectrum of disulfoton sulfoxide was obtained. Under this condition, the peak obtained from chingentsuai was identified as disulfoton sulfoxide. Disulfoton sulfoxide was detected from 2 of 25 chingentsuai samples, at concentrations of 0.66 microg/g and 0.14 microg/g.