固相萃取-气相色谱-三重四极杆串联质谱法测定茶叶中45种有机磷农药残留

采用气相色谱-三重四极杆串联质谱(GC-MS/MS)技术建立了茶叶中45种有机磷农药残留的检测方法。茶叶样品加水浸泡后采用乙腈提取,经TPT固相萃取柱净化,用V (乙腈) : V (甲苯) ＝ 3 : 1混合溶液洗脱,采用GC-MS/MS检测,基质匹配标准溶液外标法定量;分析了加水对茶叶中有机磷类农药提取效率的影响。结果表明:45种有机磷类农药在各自质量浓度范围内线性关系良好,相关系数均大于0.993。方法检出限为0.001~0.01 mg/kg,定量限为0.002~0.02 mg/kg。在0.05、0.5和5 mg/kg 3个添加水平下,45种农药平均回收率在71%~102%之间,相对标准偏差均小于9.4%。     

QuEChERS-气相色谱法检测苎麻及其土壤中8种有机磷农药残留

建立了采用QuEChERS-气相色谱快速检测苎麻及其土壤中8种有机磷农药残留的方法。样品用乙腈或乙腈-超纯水提取,苎麻叶样品用N-丙基乙二胺(PSA)配合石墨化炭黑(GCB)净化,土壤和苎麻杆样品经PSA净化,最后用丙酮置换乙腈,气相色谱-FPD检测,外标法定量。结果表明,在0.05～0.5 mg/kg添加水平范围内,8种农药在土壤、苎麻杆和苎麻叶样品中的平均添加回收率在71.1%～114.2%之间,相对标准偏差(RSD)为2.2%～14.6%,检出限(LOD)在0.006～0.016 mg/kg之间,定量限(LOQ)在0.020～0.050 mg/kg之间。该方法准确、快速、方便,适用于有机磷类农药在苎麻叶、苎麻杆和土壤中的残留检测。     

Interactions between S,S,S-tributyl phosphorotrithioate (DEF) and growth regulator herbicides. II.—Further studies

Additional results are presented from experiments with poplar and privet discussed in a previous paper. At 9 months after application mixtures of picloram and S,S,S-tributyl phosphorotrithioate (DEF) caused more severe effects than either compound applied alone. This result was in agreement with earlier assessments of phytotoxicity. The earlier synergism of DEF with the triethylamine salt of 2,4,5-T on privet diminished or disappeared, however. Mixtures of DEF with 2,4,5-T iso-octyl ester were antagonistic both in their shortand longer-term phytotoxic effects. Further experiments with guava plants grown in a glasshouse have demonstrated synergism in mixtures of DEF with salt or amine formulations ofpicloram, 2,4,5-T or mecoprop and antagonism between DEF and ester formulations of these herbicides. 2-Butyne-1,4-diol had no effect on the phytotoxicity of picloram ester. Xylene and a non-ionic emulsifying agent (Triton B 1956) did not appreciably increase the effects of picloram if a wetting agent was also present. Synergism was reduced when DEF was applied to guava leaves 2–4 h before or after picloram, and the compounds did not interact when they were introduced directly into the vascular system of this plant.     

Effects of insecticides on thermotropic lipid phase transitions

The organophosphorus insecticides, parathion and azinphos, significantly lower the midpoint temperature of thermotropic phase transitions of lipid bilayers, but malathion has no effect. The effects of parathion and azinphos are more pronounced on bilayers of short-chain lipids. These compounds induce phase separations in binary lipid mixtures containing dimyristoyl and distearoyl phosphatidylcholines, although similar effects could not be noticed in mixtures of dimyristoyl phosphatidylcholine and dimyristoyl phosphatidylethanolamine. Furthermore, the compounds partially restore phase transitions abolished by cholesterol. It is concluded that parathion and azinphos induce molecular disorders in lipid bilayers, either by having fluidizing effects or by promoting phase separations as a consequence of preferential interaction with more fluid lipids. The interaction of the compounds with cholesterol-containing bilayers produces phospholipid-rich lateral domains.     

Evaluation of the combined toxicity of 15 pesticides by uniform design

BACKGROUND: Environmental pesticides, including insecticides and herbicides, are frequently encountered as mixtures and threaten non‐target organisms in water. Evaluation of the combined toxicity of diverse pesticides with various concentration combinations is important, especially using limited experimental effort. Uniform design (UD) is one optimal experimental technique that can rationally arrange the concentrations of mixture components so that, with a minimum number of experimental runs, the combined toxicity of multiple pesticide mixtures can be evaluated. RESULTS: The concentration compositions of 18 pesticide mixture points designed by UD covered almost all possible concentration ranges of the mixture components on account of the two merits of ‘space filling’ and ‘multiple levels’. The combined toxicities of 18 mixture rays extended by using the fixed‐ratio ray design (FRRD) from 18 UD mixture points were evaluated by concentration addition (CA) and independent action (IA) models. It was found that the concentration–response curves (CRCs) predicted by CA were, on the whole, located between the 95% confidence intervals of the experimental CRCs, which implied that the combined toxicity of the pesticide mixture rays could be evaluated by CA. The CRCs predicted by IA were very similar to those from CA. CONCLUSION: The model developed from the UD mixture rays can effectively simulate mixtures with arbitrary concentration compositions of 15 pesticides. The CA model can accurately evaluate and predict the combined toxicity of the pesticides, which provides a useful tool for risk assessment of a mixture of multiple pesticides in the aquatic environment. Copyright © 2010 Society of Chemical Industry     

The joint action of mixtures of insecticides,or of insect growth regulators and insecticides,on susceptible and diflubenzuron-resistant strains of Spodoptera littoralis boisd

The joint action of insecticides, or of mixtures of insect growth regulators and insecticides, on the susceptible (S) strain and diflubenzuron-resistant (R_d) strain of the cotton leafworm Spodoptera littoralis Boisd. was investigated. The joint action of the insecticides and/or insect growth regulator mixtures was determined by mixing them in proportion to their activity equivalents at the LD₂₅ or ED₂₅ levels. A total of 15 mixtures of two synthetic pyrethroids, two organophosphorus, one carbamate and one organochlorine insecticides, were applied to the fourth-instar larvae of the S and R_d strains. The insecticide mixtures cypermethrin/methomyl and cypermethrin/endrin exhibited high and moderate levels of synergism on the S strain, respectively. However the mixtures chlorpyrifos/methomyl, phosfolan/methomyl, and phosfolan/endrin produced antagonism, while the other mixtures showed varying levels of additive effects. The response of the fourth-instar larvae of the S strain, to the joint action of diflubenzuron/juvenoid, diflubenzuron/insecticide, or insecticide/juvenoid mixtures, revealed that diflubenzuron produced high levels of synergism when combined with methoprene and progressively less with fenvalerate, methomyl and cypermethrin. On the other hand, the mixture diflubenzuron/triprene was antagonistic. Fenvalerate with the two juvenoids produced synergism while methomyl showed an additive effect with methoprene. However, the mixtures cypermethrin/methoprene, cypermethrin/triprene and methomyl/triprene produced antagonism. The mixtures that produced potentiation on the fourth-instar larvae of the S strain lost their high potency when tested against the R_d strain. The results also indicated that insecticide/juvenoid mixtures, when applied on 2-day-old pupae of the S strain, were synergistic, except in the case of cypermethrin/methoprene and methomyl/triprene mixtures, for which additive effects were observed. When the mixtures that had synergistic effects on the S strain were tested on the R_d strain, the results revealed that their synergistic effects were apparently reduced. This was attributed to the fact that the generalised levels of tolerance in the R_d strain towards various compounds may have influenced the several defence mechanisms to act against the synergistic action of the chemical mixtures.     

气相色谱法快速检测粮谷中的有机磷类、有机氮类和氨基甲酸酯类农药残留

建立了稻米、小麦面粉中8种有机磷类、3种氨基甲酸酯类、3种有机氮类农药的毛细管气相色谱氮磷检测器的快速分析方法。试样用超声波丙酮提取,硅镁吸附剂净化,丙酮定容,GC—NPD测定。结果表明,该方法简单、快速,灵敏度高、分离度好,14种农药测定方法的最低检出限为0．24—1．98μg／kg．平均添加回收率为71．4％～117．2％,变异系数〈11．6％．此方法是一种快速有效的检测方法,已成功应用于稻米和小麦面粉试样中痕量农药残留分析。     

Fibre-reactive insect-resist agents for wool: The synthesis and effectiveness of some organophosphorus compounds

Fibre-reactive derivatives of 10 organophosphorus compounds were synthesised and evaluated as insect-resist agents for wool. Some were very effective against wool pests and, when the fibre-reactive group was a 2-bromoacrylic ester group. these compounds also exhibited good fastness to washing and dry-cleaning.     

Effect of the combined treatment of insecticides and an attractant for the control of Phloeotribus scarabaeoides,a pest of Olea europea

Different insecticides have been tested for the control of the olive bark beetle, Phloeotribus scarabaeoides Bern. This scolytid can be managed at two points in its biological cycle: in pruned logs, where it excavates reproduction galleries, or in living trees, after emergence from the logs, where it digs feeding galleries. In mortality laboratory bioassays, the efficiency of organophosphorus insecticides has been ranked as follows: chlorpyrifos + dimethoate < formothion < methidathion. Formothion and methidathion, the two most efficient, were sprayed on olive logs together with a pyrethroid insecticide, deltamethrin, and a formulation which combined an organophosphorus (fenitrothion) and a pyrethroid (cypermethrin) insecticide. Deltamethrin inhibited the excavation of new reproduction galleries and induced a repellent effect on the olive pest. In contrast, none of the organophosphorus insecticides or the combination, fenitrothion + cypermethrin, were able to control the olive bark beetle. In olive trees, deltamethrin controlled this olive pest without showing the repellent effect observed for logs. Ethylene, a plant hormone, has been reported as an attractant for the olive bark beetle. The use of dispensers which released ethylene increased the number of P. scarabaeoides approaching the treated olive trees, thus favouring its use in a lure-and-trap control system.     

Interactions between tri-S-butyl phosphorotrithioate (DEF) and growth regulator herbicides: I.—A preliminary report

Mixtures of tri-S-butyl phosphorotrithioate (DEF) emulsion and the potassium salt of picloram were strongly synergistic when applied as foliar sprays to pot-grown privet, poplar, guava and Eucalyptus plants. A similar but less marked effect was observed in privet, using mixtures of DEF and a water-soluble amine formulation of 2,4,5-T. By contrast, mixtures of an oil-soluble ester formulation of 2,4,5-T with DEF were antagonistic with privet.     

固相萃取-气相色谱-串联质谱法测定大蒜中19种有机磷农药残留量

建立了采用固相萃取(SPE)结合气相色谱-三重四级杆串联质谱(GC-MS/MS)测定大蒜中19种有机磷农药残留量的方法。采用V(乙酸)∶V(乙酸乙酯)=1∶99混合溶液提取,Carbon/NH2固相萃取小柱净化,在GC-MS/MS的多反应(MRM)模式下进行外标法定量。结果表明:在0.04~0.8 mg/L范围内,19种农药的色谱峰面积与其相应的质量浓度间均呈良好的线性关系;所有供试农药检测方法的定量限(LOQ)均低于0.01 mg/kg;在0.01~0.2 mg/kg添加水平下,19种农药的平均回收率在68.0%~130%之间,相对标准偏差(RSD)≤15.6%。该方法背景干扰少,灵敏度高,适合基质复杂的大蒜样品中有机磷农药残留量的检测。     

Kinetic efficiency of mutant carboxylesterases implicated in organophosphate insecticide resistance

Resistance to organophosphorus (OP) insecticides in Lucilia cuprina arises from two mutations in carboxylesterase E3 that enable it to hydrolyse the phosphate ester of various organophosphates, plus the carboxlyester in the leaving group in the case of malathion. These mutations are not found naturally in the orthologous EST23 enzyme in Drosophila melanogaster. We have introduced the two mutations (G137D and W251L) into cloned genes encoding E3 and EST23 from susceptible L. cuprina and D. melanogaster and expressed them in vitro with the baculovirus system. The ability of the resultant enzymes to hydrolyse the phosphate ester of diethyl and dimethyl organophosphates was studied by a novel fluorometric assay, which also provided a sensitive titration technique for the molar amount of esterase regardless of its ability to hydrolyse the fluorogenic substrate used. Malathion carboxylesterase activity was also measured. The G137D mutation markedly enhanced (>30-fold) hydrolysis of both classes of phosphate ester by E3 but only had a similar effect on the hydrolysis of dimethyl organophosphate in EST23. Introduction of the W251L mutation into either gene enhanced dimethyl (23-30-fold) more than diethyl (6-10-fold) organophosphate hydrolysis and slightly improved (2-4-fold) malathion carboxylesterase activity, but only at high substrate concentration.     

Herbicidal cyanoacrylates with antimicrotubule mechanism of action

The herbicidal mode of action of the new synthetic cyanoacrylates ethyl (2Z)-3-amino-2-cyano-4-ethylhex-2-enoate (CA1) and its isopropyl ester derivative CA2 was investigated. For initial characterization, a series of bioassays was used indicating a mode of action similar to that of mitotic disrupter herbicides such as the dinitroaniline pendimethalin. Cytochemical fluorescence studies including monoclonal antibodies against polymerized and depolymerized tubulin and a cellulose-binding domain of a bacterial cellulase conjugated to a fluorescent dye were applied to elucidate effects on cell division processes including mitosis and microtubule and cell wall formation in maize roots. When seedlings were root treated with 10 microM of CA1 or CA2, cell division activity in meristematic root tip cells decreased within 4 h. The chromosomes proceeded to a condensed state of prometaphase, but were unable to progress further in the mitotic cycle. The compounds caused a complete loss of microtubular structures, including preprophase, spindle, phragmoplast and cortical microtubules. Concomitantly, in the cytoplasm, an increase in labelling of free tubulin was observed. This suggests that the herbicides disrupt polymerization and microtubule stability, whereas tubulin synthesis or degradation appeared not to be affected. In addition, cellulose labelling in cell walls of root tip cells was not influenced. The effects of CA1 and CA2 were comparable with those caused by pendimethalin. In transgenic Arabidopsis plants expressing a green fluorescent protein-microtubule-associated protein4 fusion protein, labelled arrays of cortical microtubules in living epidermal cells of hypocotyls collapsed within 160 min after exposure to 10 microM CA1 or pendimethalin. Moreover, a dinitroaniline-resistant biotype of goosegrass (Eleusine indica (L) Gaertn) with a point mutation in alpha-tubulin showed cross-resistance against CA1 and CA2. The results strongly indicate that the cyanoacrylates are a new chemical class of herbicide which possess the same antimicrotubule mechanism of action as dinitroanilines, probably including interaction with the same binding site in alpha-tubulin.     

Synergism of organophosphorus insecticides by diethyl maleate and related compounds in house flies

The toxicity of several organophosphorus and one carbamate insecticide for house flies is enhanced by simultaneous administration of diethyl maleate. Synergism factors vary from 2 to 116 and are strongly dependent on the combination of strain and insecticide studied. In general, it seems that thiono compounds are less synergized than their oxon analogs. Comparison of some analogs of maleic acid esters showed that the diethyl ester was not the most active compound. trans-Phenylbutenone was found to be a good alternative for diethyl maleate. Both diethyl maleate and trans-phenylbutenone deplete glutathione in the flies, but this depletion per se is certainly not the most important mode of action. In vitro experiments made clear that sufficient glutathione remained to permit a substantial insecticide degradation rate. Moreover, treatment of the flies with diethyl maleate 2–4 hr before the insecticide application resulted in a partial or complete disappearance of synergism in a period in which the glutathione concentration in the flies was still very low. Apart from this effect on the glutathione levels in the insect tissues it appeared from further experimental work in vitro that both compounds had a direct inhibitory effect on glutathione-dependent transferase(s) as well as on oxidative enzymes involved in insecticide degradation. The contribution of these reactions to the eventual synergism factor is discussed.     

Interaction between benzoylprop ethyl, flamprop methyl or flamprop isopropyl and herbicides used for broadleaved weed control

A marked antagonism of A. fatua control was found when 2,4-D, dicamba, 2,4-D-dicamba-mecoprop or bromoxynil were added to benzoylprop ethyl, flamprop methyl or flamprop isopropyl. Because of the much poorer control of A. fatua by these mixtures obtaining broad spectrum weed control with them in one spray operation would not be possible. The A. fatua herbicides did not affect the activity of the added herbicides on Fagopyrum lataricum. In eight out of nine mixtures tested there was no significant antagonism in the field conditions when MCPA or bromoxynil-MCPA was added to the A. fatua herbicides. There were significant antagonistic effects, however, in six out of nine such mixtures in glasshouse experiments. This suggests that under farm use conditions benzoylprop ethyl, flamprop methyl or fiamprop isopropyl can be mixed with MCPA amine or ester or with bromoxynil-MCPA to obtain broad spectrum weed control from one spray operation, though slightly poorer A. fatua control may be expected with these mixtures.     

Pesticide action: Different response of erythrocyte membrane acetylcholinesterase to inhibition by organophosphorus compounds under varied dietary conditions

The inhibitory effect of malathion in conjunction with maloxon or other derivatives on the activity of rat erythrocyte membrane-bound acetylcholinesterase has been investigated after feeding diets known to affect the lipid composition of the membrane. The inhibition of AchE by preincubation of membrane preparations with this organophosphorus compound, possibly with contaminants, differed with the diet: the curve was sigmoidal after supplementation with corn oil and hyperbolic after supplementation with lard. In the latter case the kinetics of the inhibition became sigmoidal by assaying in the presence of cortisol or isoproterenol or by previous solubilization of the enzyme. These results indicate that some organophosphorus compounds can differentiate conformational states of acetylcholinesterase present in the erythrocyte membrane.     

Synergism of pyrethroid — organophosphorus insecticide mixtures in insects and their toxicity againstspodoptera littoralis larvae

Synergism of mixtures of pyrethroids with organophosphorus (OP) compounds in insects is reviewed, and the toxicity of such combinations againstSpodoptera littoralis (Boisd.) larvae is reported. Mixtures of one of the pyrethroids cypermethrin, fenvalerate or deltamethrin with one of the OP compounds monocrotophos, profenofos, azinphos-methyl or acephate were assayed at different ratios as 24-h-old dipping residues on alfalfa, which was fed toS. littoralis larvae for 48 h. With most of the binary mixtures containing various OP concentrations in excess of those of the pyrethroids, synergism was demonstrated. In the pairs fenvalerate — azinphos-methyl, deltamethrin — azinphos-methyl and deltamethrin — profenofos, however, no synergism was found. In a detailed investigation with pyrethroid concentrations causing 20% mortality and OP concentrations giving a kill of no higher than ;10%, the above findings on synergism were amply confirmed. A cypermethrinmonocrotophos mixture showed synergism also on cotton leaves sprayed in the field. Synergism could not be demonstrated by topical application of pyrethroid — OP mixtures.     

Resynthesis of multiple resistance to organophosphorus insecticides from strains with factors of resistance isolated from the SKA strain of house flies

Individual factors of resistance to insecticides attributable to chromosomes II, III and ^V of the SKA strain of houseflies (Musca domestica L) were combined in pairs to determine how their presence affects resistance. The re-synthesised strains with resistance factors on chromosomes II and V, and on chromosomes III and V, were tested with several organophosphorus insecticides and DDT. The penetration delaying mechanism Pen on chromosome III, which alone gives little or no resistance, slightly increased the resistance of flies with the microsomal detoxifying factor Ses on chromosome V to diazinon and malaoxon-ethyl (c. × 1.5), but was more effective in increasing resistance to DDT (× 6). There was no effect on the response to other insecticides tested. The combined effect of the mechanisms of resistance on chromosome II (glutathione S-ethyl transferase and phosphatase) and on chromosome V (microsomal detoxication) approximated to the product of the resistance conferred by each of these mechanisms singly, suggesting that the mechanisms of resistance on the two chromosomes act independently. Therefore, most of the strong resistance to organophosphorus insecticides in the SKA strain results from the interaction between delayed penetration (chromosome III) and the factors of resistance on chromosome II, and the independent action of the resistance factors on chromosomes II and V.     

Two‐component cultivar mixtures reduce rice blast epidemics in an upland agrosystem

The effect of two‐component rice cultivar mixtures on the control of rice blast disease was studied in three different experiments under rainfed upland conditions in the Madagascar Highlands. The mixtures involved a susceptible cultivar (either susceptible or very susceptible) and a resistant cultivar in different mixture arrangements (random or row mixtures) and with different proportions of the susceptible cultivar (50, 20 and 16·7%), which were compared to the susceptible cultivar grown in a pure stand. The effect of these mixtures on the incidence and severity of leaf and panicle blast was measured weekly, and on yield and yield components at harvest time. The mixture effect was more efficient in reducing disease with a proportion of 16·7% susceptible component than with a proportion of 50%. Blast epidemic was significantly reduced in all three experiments. However, under high blast pressure, there was no reduction in the disease by the end of the epidemic and yields of the susceptible cultivar were almost zero whatever the mixture. In two other experiments performed under lower blast pressure, disease incidence and severity were significantly lower in mixtures, and yields of the susceptible cultivars grown in mixtures were higher than those of their respective pure stands. Cultivar mixtures are a promising strategy that could contribute to a more sustainable cultivation of rice under upland conditions in the context of subsistence agriculture in Madagascar, where all cropping operations are manual.     

Control of Aspergillus growth and aflatoxin production using natural maize phytochemicals under different conditions of water activity

The effects of the natural phytochemicals trans-cinnamic acid (CA) and ferulic acid (FA) alone at concentrations of 1-25 mM and in 16 combinations (M: mixtures) on growth and aflatoxin B(1) production by Aspergillus flavus Link and A. parasiticus Speare were evaluated. Studies on growth rate and aflatoxin B(1) production were carried out in vitro in relation to a water activity a(w) of 0.999, 0.971, 0.955 and 0.937. Overall, CA at concentrations of 10 and 20 mM and FA-CA mixtures M3 (20 + 5 mM respectively), M8 (25 + 5 mM), M9 (1 + 10 mM), M10 (10 + 10 mM), M11 (20 + 10 mM), M12 (25 + 10 mM), M13 (1 + 20 mM), M14 (10 + 20 mM), M15 (20 + 20 mM) and M16 (25 + 20 mM) were the treatments most effective at inhibiting growth of the four species assayed. All strains were much more sensitive to all natural phytochemicals tested on growth rate at a(w) = 0.937. CA and the FA-CA mixtures M1 (1 + 1 mM respectively), M4 (25 + 1 mM), M5 (1 + 5 mM), M6 (10 + 1 mM), M7 (20 + 1 mM), M8 (25 + 5 mM), M9 (1 + 10 mM), M10 (10 + 10 mM), M11 (20 + 10 mM), M12 (25 + 10 mM), M13 (1 + 20 mM), M14 (10 + 20 mM), M15 (20 + 20 mM) and M16 (25 + 20 mM) completely inhibited aflatoxin B(1) production by all strains at a(w) = 0.999, 0.971, 0.955 and 0.937. Decreased aflatoxin B(1) levels in comparison with the control were observed with FA at 1, 10, 20 and 25 mM with the strains RCM89, RCM108 and RCM38 at a(w) = 0.971, 0.955 and 0.999 respectively. The data show that CA and FA can be considered as effective fungitoxicants for A. flavus and A. parasiticus in in vitro assay. The information obtained is part of an ongoing study to determine their application at the storage level.