Integrated spectroscopic and computational analyses unravel the molecular interaction of pesticide azinphos-methyl with bovine beta-lactoglobulin.

Organophosphorus are typically hazardous chemicals used in the pharmaceutical, agricultural, and other industries. They pose a serious risk to human life and can be fatal upon direct exposure. Hence, studying the interaction between such compounds with proteins is crucial for environmental, health, and food safety. In this study, we investigated the interaction mechanism between azinphos-methyl (AZM) and ß-lactoglobulin (BLG) at pH 7.4 using a combination of biophysical techniques. Intrinsic fluorescence investigations revealed that BLG fluorescence was quenched in the presence of increasing AZM concentrations. The quenching mechanism was identified as static, as evidenced by a decrease in the fluorescence quenching constant (1.25 × 104, 1.18 × 104, and 0.86 × 104 M-1) with an increase in temperatures. Thermodynamic calculations (ΔH > 0; ΔS > 0) affirmed the formation of a complex between AZM and BLG through hydrophobic interactions. The BLG's secondary structure was found to be increased due to AZM interaction. Ultraviolet -visible spectroscopy data showed alterations in BLG conformation in the presence of AZM. Molecular docking highlighted the significant role of hydrophobic interactions involving residues such as Val43, Ile56, Ile71, Val92, Phe105, and Met107 in the binding between BLG and AZM. A docking energy of -6.9 kcal mol-1, and binding affinity of 1.15 × 105 M-1 suggest spontaneous interaction between AZM and BLG with moderate to high affinity. These findings underscore the potential health risks associated with the entry of AZM into the food chain, emphasizing the need for further consideration of its impact on human health.

Experimental and Established Oximes as Pretreatment before Acute Exposure to Azinphos-Methyl.

Poisoning with organophosphorus compounds (OPCs) represents an ongoing threat to civilians and rescue personal. We have previously shown that oximes, when administered prophylactically before exposure to the OPC paraoxon, are able to protect from its toxic effects. In the present study, we have assessed to what degree experimental (K-27; K-48; K-53; K-74; K-75) or established oximes (pralidoxime, obidoxime), when given as pretreatment at an equitoxic dosage of 25% of LD01, are able to reduce mortality induced by the OPC azinphos-methyl. Their efficacy was compared with that of pyridostigmine, the only FDA-approved substance for such prophylaxis. Efficacy was quantified in rats by Cox analysis, calculating the relative risk of death (RR), with RR=1 for the reference group given only azinphos-methyl, but no prophylaxis. All tested compounds significantly (p ≤ 0.05) reduced azinphos-methyl-induced mortality. In addition, the efficacy of all tested experimental and established oximes except K-53 was significantly superior to the FDA-approved compound pyridostigmine. Best protection was observed for the oximes K-48 (RR = 0.20), K-27 (RR = 0.23), and obidoxime (RR = 0.21), which were significantly more efficacious than pralidoxime and pyridostigmine. The second-best group of prophylactic compounds consisted of K-74 (RR = 0.26), K-75 (RR = 0.35) and pralidoxime (RR = 0.37), which were significantly more efficacious than pyridostigmine. Pretreatment with K-53 (RR = 0.37) and pyridostigmine (RR = 0.52) was the least efficacious. Our present data, together with previous results on other OPCs, indicate that the experimental oximes K-27 and K-48 are very promising pretreatment compounds. When penetration into the brain is undesirable, obidoxime is the most efficacious prophylactic agent already approved for clinical use.

Complete removal of the insecticide azinphos-methyl from water by the electro-Fenton method--a kinetic and mechanistic study.

The removal of organophosphorous insecticide azinphos-methyl (AZPM) from water has been investigated by the electro-Fenton method which produces hydroxyl radicals electro-catalytically. The reaction between these radicals and AZPM has led to the oxidation of AZPM up to carbon dioxide and inorganic ions indicating its complete removal from water. The oxidation kinetics was fitted to pseudo-first order reaction and absolute rate constant of the second order reaction kinetic was determined as (6.82 ± 0.18) × 10(9) M(-1) s(-1) by using competitive kinetics method. The oxidation of AZPM by hydroxyl radicals leads to the formation of different intermediates species; such as aromatic derivatives, short-chain carboxylic acids and inorganic ions as end products. The identification and quantification of these intermediates were deeply investigated by HPLC, GC-MS and ion chromatography analyses. Based on the identified intermediates, a general oxidation mechanism was proposed. Mineralization ability of the process was also tested by using aqueous AZPM solutions and its commercial formulation, Gusathion M WP 25 (GMWP25). Mineralization efficiency was followed by the chemical oxygen demand (COD) analysis of treated solutions. The overall mineralization of synthetic AZPM solution and that of commercial formulation at the end of treatment highlights the outstanding mineralization power of the electro-Fenton process.

Simulated solar light phototransformation of organophosphorus azinphos methyl at the surface of clays and goethite.

The photochemical behavior of the pesticide azinphos methyl at the surface of clays (kaolinite, bentonite) and goethite was studied using Suntest setup (λ > 300 nm). The quantum yield on the clays was found to be roughly three times lower than that in aqueous solution. However, the photochemical efficiency was much higher at the surface of goethite owing to its photocatalytic activity through the hydroxyl radical production. The added humic substances on kaolonite show an inhibition of azinphos methyl degradation while the incorporation of iron(III) aquacomplexes leads to an important increase of the disappearance together with the formation of iron(II). Hydroxyl radical species were found to be formed either by excitation of goethite or clays. The goethite support acts as a more efficient catalyst for the formation of these reactive oxygen species. The photodecomposition reactions observed were (i) hydrolysis process leading to the formation of benzotriazone and the oxidation of the P = S bond giving rise to the formation of the oxon derivative, and (ii) homolytic cleavage of the N-C and C-S bonds of the organophosphorus bridge leading to the formation of dimers that appear to be specific to the irradiation at the surface of solid supports since they were not observed when the irradiation was performed in aqueous media: a statement that is related to the presence of aggregates at the surface of solid supports.

Comparing the inhibitory effects of five protoxicant organophosphates (azinphos-methyl, parathion-methyl, chlorpyriphos-methyl, methamidophos and diazinon) on the spontaneously beating auricle of Sparus aurata: an in vitro study.

Organophosphates (OPs) can provoke toxicity by inhibiting acetylcholinesterase (AChE) in non-target organisms, like fish. In a previous pilot study, the anticholinesterase effects of paraoxon on the heart of Sparus aurata were examined [Tryfonos, M., Antonopoulou, E., Papaefthimiou, C., Chaleplis, G., Theophilidis, G., 2009. An in vitro assay for the assessment of the effects of an organophosphate, paraoxon, and a triazine, atrazine, on the heart of the gilthead sea bream (Sparus aurata). Pest. Biochem. Physiol. 93, 40-46]. The objective of the present study was to investigate the effects of the five protoxicant OPs, azinphos-methyl (MeAZP), parathion-methyl (MePS), chlorpyriphos-methyl (MeCCP), methamidophos (MET) and diazinon (DZ), on the spontaneously beating auricle of S. aurata. The results showed that: (1) MeAZP and MET induced exclusively cholinergic effects on auricle contractility. These effects were expressed as a significant decrease in the force and frequency of contractions and were fully reversible (140%) after the application of the muscarinic cholinergic receptor antagonist, atropine (15 microM). MeAZP was found to be the most effective anticholinesterase compound, with an IC(50) of 2.19+/-1.05 microM (n=6), while MET was less effective, with an IC(50) of 72.3+/-1.2 microM (n=6). (2) DZ and MePS, although classified as OPs, induced non-cholinergic effects. These effects were observed as an irreversible decrease in force and frequency of the auricle in all the concentrations examined; the depression is retained even after application of 15 microM atropine. (3) MeCCP was halfway between a typical OP and an OP lacking anticholinesterase properties, since there was a partial recovery in the force, but no recovery in the frequency of the auricle contractions. (4) The toxicity order, based on the IC(50), was as follows: MeAZP, 2.19+/-1.05 microM>paraoxon, 3.2+/-1.5 microM"MET, 72.3+/-1.2 microM>MePS, 80.3+/-1.03 microM>MeCPP, 93.7+/-1.01 microM>DZ, 164+/-1.01 microM. (5) There was a good correlation (r=0.779, p=0.04, n=5) between IC(50) and the previously determined logP (octanol:water partition coefficient) values for MeAZP, paraoxon, MeCCP, MePS and DZ. The results indicated that the increase in lipophilicity of MePS, MeCCP and DZ is accompanied by a decrease in their acute cardiotoxic properties in vitro. The non-cholinergic effects of these relatively high lipophilic OPs, might be caused by their tendency to distribute preferentially in the lipid bilayer of cardiac cells, affecting the proper functioning of the ionic channels which regulate the force (Ca(2+) channels) and the frequency (K(+) channels) of the spontaneous auricle contractions.

Comparison of the partitioning of pesticides relative to the survival and behaviour of exposed amphipods.

Pesticides sprayed on farmlands can end up in rivers and be transported into estuaries, where they could affect aquatic organisms in freshwater and marine habitats. A series of experiments were conducted using the amphipod Corophium volutator Pallas (Amphipoda, Corophiidae) and single pesticides, namely atrazine (AT), azinphos-methyl (AZ), carbofuran (CA) and endosulfan (EN) that were added to sediments and covered with seawater. Our goal was to compare the concentrations affecting the survival of the animals relative to potential attractant or repellent properties of sediment-spiked pesticides. The avoidance/preference of contaminated/reference sediments by amphipods was examined after 48 and 96 h of exposure using sediments with different organic carbon content. The octanol-water partition coefficients (log K(ow)) ranked the pesticides binding to sediments as EN > AZ > AT > CA. LC(50) and LC(20) covered a wide range of nominal concentrations and ranked toxicity as CA-AZ > EN > AT. Under the experimental set up, only EN initiated an avoidance response and the organic carbon normalised concentration provided consistent results. Using the present data with wide confidence limits, >20% of a population of C. volutator could perish due to the presence of EN before relocation or detecting CA or AZ in sediments by chemical analysis.

Residual toxicity of two insecticides on three field populations of Lygus lineolaris (Hemiptera: Miridae) collected along the St Lawrence valley in eastern Canada.

Insecticides are still the single main pest control method employed today by most growers to mitigate damage done by the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae). In eastern Canada, the complex agricultural ecosystem, which may be described as a mosaic of farmlands dispersed among natural habitats (forest, prairies), allows tarnished plant bug adults to fly and move from sprayed to non-sprayed areas. In 2004 (late August to early September), three populations of L. lineolaris were collected from three mixed vegetation strips adjacent to orchards and vineyards along the St Lawrence valley: the Niagara Peninsula (Ontario), Dunham (Quebec) and La Pocatière (Quebec). Assays were done in the laboratory by confining adults in glass vials coated with dried residues. The estimated LC(50) values for the three populations varied from 11.2 to 16.8 x 10(-5) g L(-1) for azinphos-methyl and from 0.8 to 1.4 x 10(-5) g L(-1) for cypermethrin. In contrast to the Mississippi delta, no tolerance to insecticides was found in the populations collected. Possible explanations for this non-tolerance to insecticides includes a very low selection pressure as a result of the reduced number of insecticide treatments done in the context of the diversified agricultural landscapes encountered in eastern Canada which allow movements of adults from treated to non-treated areas.

Photodegradation of organophosphorus insecticides - investigations of products and their toxicity using gas chromatography-mass spectrometry and AChE-thermal lens spectrometric bioassay.

Four organophosphorus compounds: azinphos-methyl, chlorpyrifos, malathion and malaoxon in aqueous solution were degraded by using a 125 W xenon parabolic lamp. Gas chromatography-mass spectrometry (GC-MS) was used to monitor the disappearance of starting compounds and formation of degradation products as a function of time. AChE-thermal lens spectrometric bioassay was employed to assess the toxicity of photoproducts. The photodegradation kinetics can be described by a first-order degradation curve C=C0e(-kt), resulting in the following half lives: 2.5min for azinphos-methyl, 11.6 min for malathion, 13.3 min for chlorpyrifos and 45.5 min for malaoxon, under given experimental conditions. During the photoprocess several intermediates were identified by GC-MS suggesting the pathway of OP degradation. The oxidation of chlorpyrifos results in the formation of chlorpyrifos-oxon as the main identified photoproduct. In case of malathion and azinphos-methyl the corresponding oxon analogues were not detected. The formation of diethyl (dimethoxy-phosphoryl) succinate in traces was observed during photodegradation of malaoxon and malathion. Several other photoproducts including trimethyl phosphate esters, which are known to be AChE inhibitors and 1,2,3-benzotriazin-4(3H)-one as a member of triazine compounds were identified in photodegraded samples of malathion, malaoxon, and azinphos-methyl. Based on this, two main degradation pathways can be proposed, both result of the (P-S-C) bond cleavage taking place at the side of leaving group. The enhanced inhibition of AChE observed with the TLS bioassay during the initial 30 min of photodegradation in case of all four OPs, confirmed the formation of toxic intermediates. With the continuation of irradiation, the AChE inhibition decreased, indicating that the formed toxic compounds were further degraded to AChE non-inhibiting products. The presented results demonstrate the importance of toxicity monitoring during the degradation of OPs in processes of waste water remediation, before releasing it into the environment.

Kinetics and thermodynamics of adsorption of azinphosmethyl from aqueous solution onto pyrolyzed (at 600 degrees C) ocean peat moss (Sphagnum sp.).

The removal of azinphosmethyl from aqueous solution onto pyrolyzed ocean peat moss (Sphagnum sp.), as a residue, from the Rhode Island coast (USA), has been investigated at different temperatures and initial concentrations. The ocean peat moss had been pyrolyzed at 600 degrees C in nitrogen atmosphere before the adsorption process. The kinetic data obtained from batch studies have been analyzed using pseudo-first order kinetic model. The rate constants were evaluated at different temperatures. The thermodynamic parameters (DeltaG degrees , DeltaH degrees , DeltaS degrees ) for the adsorption process were calculated and the results suggest that the nature of adsorption is endothermic and the process is spontaneous and favorable. The activation energy for adsorption process was estimated, about 18.3 kJ mol(-1). According to this value the adsorption of azinphosmethyl onto pyrolyzed ocean peat moss is in the range of physical adsorption. The experimental data have been modeled using Langmuir, Freundlich and Temkin isotherms. It was found that Langmuir and Freundlich isotherms give the best correlation with the experimental data.

Adsorption kinetics of azinphosmethyl from aqueous solution onto pyrolyzed Horseshoe sea crab shell from the Atlantic Ocean.

The adsorption behavior of azinphosmethyl on pyrolyzed Horseshoe Crab (Limulus polyphemus) outer shell, as a residue, from the Atlantic Ocean, collected along the Maine coast, USA, has been studied with regards to its kinetic and equilibrium conditions, taking into account adsorbate concentrations of 2 x 10(-3), 4 x 10(-3), 6 x 10(-3), and 8 x 10(-3), as well as temperatures of 30 degrees C, 40 degrees C, 50 degrees C, and 60 degrees C. The yield of adsorption of azinphosmethyl from aqueous solution ranged from 56.1% to 61% with temperature increasing. Kinetic studies showed that adsorption rate decreased as the initial azinphosmethyl concentration increased. It was found, that the adsorption reaction obeyed first-order kinetics. The overall rate constants were estimated for different temperatures. The activation energy for adsorption was about 1.52 kJmol(-1), which implies that azinphosmethyl mainly adsorbed physically onto Horseshoe Crab outer shell. Langmuir and Freundlich isotherms were applied to the experimental data and isotherm constants were calculated. The thermodynamic parameters DeltaG0, DeltaH0 and DeltaS0 for the adsorption reaction were evaluated based on equilibrium data and in connection with this result the thermodynamic aspects of adsorption reaction were discussed. The adsorption was found to be endothermic in nature. The adsorbent used in this study proved highly efficient for the removal of azinphosmethyl.

Removal of azinphos methyl by alfalfa plants (Medicago sativa L.) in a soil-free system.

The objective of this study was to investigate the removal of azinphos methyl assisted by alfalfa plants, with special emphasis on the effects of this compound on some plant's physiological parameters. Hydroponic cultures of alfalfa (Medicago sativa L., var Romagnola) were employed as a model system. These cultures were exposed to a nutrient medium containing 10 mg/l of azinphos methyl. A first-order kinetic approach was used to describe the removal of azinphos methyl from the solution. After 20 days of culture, the initial amount of azinphos methyl was reduced to non-detectable levels in the presence of plants. In the absence of plants, 20% of azinphos methyl remained in the solution after 30 days of treatment. The half-life of the pesticide was reduced from 10.8 to 3.4 days in the presence of plants. The growth index of alfalfa plants exposed to azinphos methyl was negatively affected. Chlorophyll contents were reduced after 24 h of treatment and thereafter the levels were comparable to that of control plants. The peroxidase activity of alfalfa roots was not affected by the presence of azinphos methyl. In conclusion, alfalfa plants were able to survive when exposed to an effective concentration of 10 mg/l of azinphos methyl in the root zone, with some alterations on their physiological parameters.

A study on the environmental degradation of pesticides azinphos methyl and parathion methyl.

The effect of environmental parameters (temperature and relative humidity) on the degradation rate of azinphos methyl and parathion methyl was studied. Proprietary emulsifiable concentrates were diluted and added to each of 90 glass Petri dishes for each pesticide and were left overnight to dry. Petri dishes were placed in 18 air-tight containers (9 for each pesticide) in which were created environments with relative humidity (RH) of 60, 82, and 96%. The containers were stored at 0, 20, and 40 degrees C. From the experimental results best fit curves, kinetic equations, rate constants, and half-lives were calculated. Half-lives of azinphos methyl for the RH studied were, from 124 to 267 days at 0 degrees C, from 89 to 231 days at 20 degrees C, and from 25 to 71 days at 40 degrees C. Corresponding half-lives for parathion methyl were from 48 to 57 days at 0 degrees C, from 9.2 to 10.5 days at 20 degrees C and from 1.3 to 1.5 days at 40 degrees C. The results were correlated with relevant results from the decomposition of the same or similar pesticides on apples both, on the trees and during refrigerated storage. These correlations are suggesting that biological factors strongly affected the decomposition rate of azinphos methyl. On the contrary the decomposition of parathion methyl was mainly affected by environmental rather than biological factors.

Fate and effects of azinphos-methyl in a flow through wetland in South Africa.

Our knowledge about the effectiveness of constructed wetlands in retaining agricultural nonpoint-source pesticide pollution is limited. A 0.44-ha vegetated wetland built along a tributary of the Lourens River, Western Cape, South Africa, was studied to ascertain the retention, fate, and effects of spray drift-borne azinphos-methyl (AZP). Composite water samples taken at the inlet and outlet during five spray drift trials in summer 2000 and 2001 revealed an overall reduction of AZP levels by 90 +/- 1% and a retention of AZP mass by 61 +/- 5%. Samples were collected at the inlet outlet, and four platforms within the wetland to determine the fate and effect of AZP in the wetland after direct spray drift deposition in the tributary 200 m upstream of the inlet. Peak concentrations of AZP decreased, and the duration of exposure increased from inlet (0.73 microg/L; 9 h) via platforms 1 and 4 to outlet (0.08 microg/L; 16 h). AZP sorbed to plants or plant surfaces, leading to a peak concentration of 6.8 microg/kg dw. The living plant biomass accounted for 10.5% of the AZP mass initially retained in the wetland, indicating processes such as volatilization, photolysis, hydrolysis, or metabolic degradation as being very important AZP was not detected in sediments. Water samples taken along two 10-m transects situated perpendicular to the shore indicated a homogeneous horizontal distribution of the pesticide: 0.23 +/- 0.02 and 0.14 +/- 0.04 microg/L (n = 5), respectively. Both Copepoda (p = 0.019) and Cladocere (p = 0.027) decreased significantly 6 h postdeposition and remained at reduced densities for at least 7 d. In parallel, the chlorophyll a concentration showed an increase, although not significant, within 6 h of spray deposition. The study highlights the potential of constructed wetlands as a risk-mitigation strategy for spray drift-related pesticide pollution.

Degradation of the insecticide azinphos methyl in orange and peach juices during storage at different temperatures.

The degradation of azinphos methyl in orange and peach juice was studied. The insecticide was aseptically added to packed orange and peach juices and stored at 40, 20 and 0 degrees C. Samples were removed at regular intervals and were analysed for residues of azinphos methyl. Residues were determined with a simple gas-chromatographic method using a 30 m glass capillary column and an NP detector. The recoveries of azinphos methyl were 87-110% for orange juice and 92-108% for peach juice, and the limit of determination was 0.004 mg/kg for both juices. From the experimental data, rate constants, half-lives, and times needed to reach legal limits for the decomposition of azinphos methyl in orange and peach juices were evaluated. Half lives for orange and peach juices were 6.5 and 7.8 days at 40 degrees C, 86.6, and 92.4 days at 20 degrees C, and 407.7 and 495.1 days at 0 degrees C, respectively.

Reduction of azinphos-methyl, chlorpyrifos, esfenvalerate, and methomyl residues in processed apples.

McIntosh, Red Delicious, and Golden Delicious from two years of experimental spray programs using azinphos-methyl, chlorpyrifos, esfenvalerate, and methomyl were processed into frozen apple slices, applesauce, single-strength juice, and juice concentrate. Residue levels were expressed as micrograms per 150 g of apple or the equivalent amount of apple product to calculate the percentage change in these pesticides brought about by processing. Producing single-strength apple juice reduced azinphos-methyl, chlorpyrifos, esfenvalerate, and methomyl residues by 97.6, 100, 97.8, and 78.1%, respectively. Production of applesauce reduced all four compounds by >/=95%. Azinphos-methyl, chlorpyrifos, esfenvalerate, and methomyl residues were reduced in apple slices by 94.1, 85.7, 98.6, and 94.7%, respectively. Processing is shown to be very effective in reducing the levels of these pesticides.

Biophysical perturbations induced by ethylazinphos in lipid membranes.

Perturbations induced by ethylazinphos on the physical organization of dipalmitoylphosphatidylcholine (DPPC) and DPPC/cholesterol membranes were studied by differential scanning calorimetry (DSC) and fluorescence polarization of 2-, 6-, 12-(9-anthroyloxy) stearic acids and 16-(9-anthroyloxy) palmitic acid. Ethylazinphos (50 and 100 microM) increases the fluorescence polarization of the probes, either in the gel or in the fluid phase of DPPC bilayers, and this concentration dependent effect decreases from the surface to the bilayer core. Additionally, the insecticide displaces the phase transition to a lower temperature range and broadens the transition profile of DPPC. A shifting and broadening of the phase transition is also observed by DSC. Furthermore at insecticide/lipid molar ratios higher than 1/7, DSC thermograms, in addition to the normal transition centered at 41 degrees C, also display a new phase transition centered at 45.5 degrees C. The enthalpy of this new transition increases with insecticide concentration, with a corresponding decrease of the main transition enthalpy. Ethylazinphos in DPPC bilayers with low cholesterol (< or = 20 mol%) perturbs the membrane organization as described above for pure DPPC. However, cholesterol concentrations higher than 20 mol% prevent insecticide interaction, as revealed by fluorescence polarization and DSC data. Apparently, cholesterol significantly modulates insecticide interaction by competition for similar distribution domains in the membrane. The present results strongly support our previous hypothesis that ethylazinphos locates in the cooperativity region, i.e. the region of C1-C9 atoms of the acyl chains, and extends to the lipid-water interface, where it increases lipid packing order sensed across all the thickness of the bilayer. Additionally, and, on the basis of DSC data, a lateral regionalization of ethylazinphos is here tentatively suggested.

Characterization of P-S bond hydrolysis in organophosphorothioate pesticides by organophosphorus hydrolase.

The extensive use of organophosphorothioate insecticides in agriculture has resulted in the risk of environmental contamination with a variety of broadly based neurotoxins that inhibit the acetylcholinesterases of many different animal species. Organophosphorus hydrolase (OPH, EC 3.1.8.1) is a broad-spectrum phosphotriesterase that is capable of detoxifying a variety of organophosphorus neurotoxins by hydrolyzing various phosphorus-ester bonds (P-O, P-F, P-CN, and P-S) between the phosphorus center and an electrophilic leaving group. OPH is capable of hydrolyzing the P-X bond of various organophosphorus compounds at quite different catalytic rates: P-O bonds (kcat = 67-5000 s-1), P-F bonds (kcat = 0.01-500 s-1), and P-S bonds (kcat = 0.0067 to 167 s-1). P-S bond cleavage was readily demonstrated and characterized in these studies by quantifying the released free thiol groups using 5,5'-dithio-bis-2-nitrobenzoic acid or by monitoring an upfield shift of approximately 31 ppm by 31P NMR. A decrease in the toxicity of hydrolyzed products was demonstrated by directly quantifying the loss of inhibition of acetylcholinesterase activity. Phosphorothiolate esters, such as demeton-S, provided noncompetitive inhibition for paraoxon (a P-O triester) hydrolysis, suggesting that the binding of these two different classes of substrates was not identical.

[Gas chromatographic determination of azinphos-methyl in imported crops].

A gas chromatographic method for the quantitative analysis of azinphos-methyl in agricultural products was studied. Azinphos-methyl was extracted with acetone. The extract was cleaned up by charcoal column chromatography for the treatment of wheat and soybean and by using coagulating reagent for the treatment of lemon and orange, then determined by a gas chromatograph equipped with a flame photometric detector (FPD). The mean recovery of azinphos-methyl added to wheat, soybean and lemon at the level of 0.1 or 0.2 microgram/g was 96.6-101.0%. The detection limit of azinphos-methyl was 0.04 microgram/g in the case of wheat and soybean extracted from 10 g of each sample and 0.02 microgram/g in the case of lemon and orange extracted from 20 g of each sample. Azinphos-methyl was not detected in any of the imported, 6 wheat, 6 soybean, 5 lemon and 4 orange tested using the above mentioned method.