Formic acid: A neurologically active, hydrolyzed metabolite of insecticidal formate esters

Formate esters have potential uses as insecticides in both grain fumigation and public health pest management. This research investigated the hydrolytic metabolism and neurological effects of formate ester compounds. Studies were conducted using Drosophila and houseflies as models for other dipteran pests of medical importance. Results indicated significant formic acid liberation from a broad range of formate esters, both in vivo and in vitro. Based on these initial observations, we subsequently investigated neurological effects of formic acid on the larval housefly nervous system. We found that formic acid caused significant neuroexcitation at concentrations lower than previously shown for inhibition of insect cytochrome c oxidase; however, this conclusion does not preclude that mitochondrial effects may also be occurring in non-nervous tissues. Finally, we investigated formate ester hydrolysis by A- and B-type esterases commonly considered in pesticide detoxification research; however, no significant interactions could be identified, suggesting that as yet unidentified carboxylesterases play roles in formate ester hydrolysis. These findings provide information on formate ester metabolism and modes of action, as well as rationale for further studies on formate ester neurotoxicology and mechanisms of selective toxicity.     

Mitochondrial impacts of insecticidal formate esters in insecticide‐resistant and insecticide‐susceptible Drosophila melanogaster

BACKGROUND: Previous research on insecticidal formate esters in flies and mosquitoes has documented toxicity profiles, metabolism characteristics and neurological impacts. The research presented here investigated mitochondrial impacts of insecticidal formate esters and their hydrolyzed metabolite formic acid in the model dipteran insect Drosophila melanogaster Meig. These studies compared two Drosophila strains: an insecticide‐susceptible strain (Canton‐S) and a strain resistant by cytochrome P450 overexpression (Hikone‐R). RESULTS: In initial studies investigating inhibition of mitochondrial cytochrome c oxidase, two proven insecticidal materials (hydramethylnon and sodium cyanide) caused significant inhibition. However, for insecticidal formate esters and formic acid, no significant inhibition was identified in either fly strain. Mitochondrial impacts of formate esters were then investigated further by tracking toxicant‐induced cytochrome c release from mitochondria into the cytoplasm, a biomarker of apoptosis and neurological dysfunction. Formic acid and three positive control treatments (rotenone, antimycin A and sodium cyanide) induced cytochrome c release, verifying that formic acid is capable of causing mitochondrial disruption. However, when comparing formate ester hydrolysis and cytochrome c release between Drosophila strains, formic acid liberation was only weakly correlated with cytochrome c release in the susceptible Canton‐S strain (r² = 0.70). The resistant Hikone‐R strain showed no correlation (r² < 0.0001) between formate ester hydrolysis and cytochrome c release. CONCLUSION: The findings of this study provide confirmation of mitochondrial impacts by insecticidal formate esters and suggest links between mitochondrial disruption, respiratory inhibition, apoptosis and formate‐ester‐induced neurotoxicity. Copyright © 2009 Society of Chemical Industry     

甲胺磷的薄层色谱及GC-MS定性检验

探讨了甲胺磷的定性分析方法。根据甲胺磷的特点,用薄层色谱法对甲胺磷的展开剂进行了实验选定,并对其GC-MS分析的色谱条件进行了探讨。 结果显示,甲胺磷的薄层色谱展开 剂中需含极性溶剂如乙醇、甲醇、乙酸等方能将其很好地展开,适合甲胺磷的展开剂为:氯仿-乙醇、氯仿-甲醇-环己烷等;用GC-MS更能准确定性甲胺磷,保留时间3.48 min,其质谱图与标准谱库相一致。     

Ethyl formate plus methyl isothiocyanate--a potential liquid fumigant for stored grains

BACKGROUND: Methyl bromide is being phased out for use on stored commodities, as it is listed as an ozone‐depleting substance, and phosphine is the fumigant widely used on grains. However, phosphine resistance occurs worldwide, and phosphine fumigation requires a long exposure period and temperatures of > 15 °C. There is an urgent requirement for the development of a fumigant that kills insects quickly and for phosphine resistance management. This paper reports on a new fumigant formulation of 95% ethyl formate plus 5% methyl isothiocyanate as an alternative fumigant for stored grains. RESULTS: The formulation is stable for at least 4 months of storage at 45 °C. A laboratory bioassay with the formulation showed that it controlled all stages of Sitophilus oryzae (L.), Sitophilus granarius (L.), Tribolium castaneum (Herbst), Rhyzopertha dominica (F.), Trogoderma variabile Ballion and Callosobruchus maculatus (Fabricius) in infested wheat, barley, oats and peas at 80 mg L^?1 for 5 days, and in canola at both 40 mg L^?1 for 5 days and 80 mg L^?1 for 2 days at 25 ± 2 °C. After an 8–14 day holding period, residues of ethyl formate and methyl isothiocyanate in wheat, barley, peas and canola were below the experimental permit levels of 1.0 and 0.1 mg kg^?1. However, fumigated oats needed an 18 day holding period. CONCLUSIONS: The findings suggest that the ethyl formate plus methyl isothiocyanate formulation has potential as a fumigant for the control of stored‐grain insect pests in various commodities. Copyright © 2011 Society of Chemical Industry     

Modelling the kinetics of ethyl formate sorption by wheat using batch experiments

BACKGROUND: Ethyl formate formulations are being considered to replace methyl bromide for fast grain disinfestation. Grain adsorbs ethyl formate rapidly, which can result in inadequate fumigation concentrations and unacceptable grain residues. A model of ethyl formate sorption kinetics will enable fumigation approaches to be determined that meet disinfestation and food safety requirements. RESULTS: This paper identifies all mass transport processes involved in ethyl formate sorption by wheat from published and experimental evidence. The model accounts for reaction losses of ethyl formate in air and grain using first‐order kinetics, transport in the gas and solid phases with linear mass transfer coefficients and uses a linear partition relationship representation of sorption equilibrium. Batch experimental data were measured to determine model coefficients. Novel gaseous breakdown data for ethyl formate in air were measured, and first‐order kinetics was demonstrated, although the specific reactions involved were not identified. CONCLUSION: The model predicts air and grain fumigant concentrations relevant for grain disinfestation and food residue contamination successfully. The form of the model should be applicable to all fumigant–grain systems, as it accounts for the diffusion and reaction influences known to occur with all modern fumigants under concentration and exposure conditions relevant to industry. Copyright © 2009 CSIRO, Australia. Published by John Wiley & Sons, Ltd.     

Field evaluation of vaporised ethyl formate and carbon dioxide for fumigation of stored wheat

BACKGROUND: Vapormate? is a cylinderised non‐flammable mixture of ethyl formate (16.7% by weight) and carbon dioxide (CO₂) that has been developed as a rapid fumigant of stored grain. Four field trials were undertaken on wheat in 50 t farm silos to demonstrate the feasibility of dynamic application. To assess treatment efficacy, each trial tested mixed stages of Rhyzopertha dominica F. (>11 000), Tribolium castaneum (Herbst.) (>1500) and Sitophilus oryzae (L.) (>13 000) in mesh cages positioned through the centre of the grain bulk and on the grain surface. Ethyl formate and CO₂ concentrations were measured in the silo during fumigation and in ambient air outside the 6 m fumigation zone. Application rates of 420, 660 and 940 g m^?3 of ethyl formate/CO₂ formulation and exposure times of 24, 3 and 72 h, respectively, were examined using wheat of 10.4–11.7% moisture content and grain temperatures between 2 and 32 °C. RESULTS: All life stages of R. dominica and T. castaneum were fully controlled under all conditions tested, and mortality of all life stages of S. oryzae was greater than 98%. CONCLUSION: Dynamic application of vaporised ethyl formate and CO₂ to 50 t silos proved safe to operators and rapidly effective against stored‐grain insects on cold to warm grain. Copyright © 2009 CSIRO, Australia. Published by John Wiley & Sons, Ltd     

Extended summaries pesticides group symposium physicochemical properties as factors affecting the penetration of xenobiotics to the target site

Allyl alcohol esters (propionate, formate, and acetate) and structurally related compounds (3-methylallyl alcohol and vinyl acetate) were evaluated for their fumigation properties vis-a-vis Theba pisana (Mueller) snails. The esters were very effective in exterminating the snails, although less so than the parent alcohol. These results suggest that the esters might first have to undergo hydrolysis, and only thereafter demonstrate their potency. 3-Methylallyl alcohol (four-carbon member) and vinyl acetate (two-carbon member, after hydrolysis) were significantly less potent than the allyl esters. All three esters showed less phytotoxicity towards roses than did allyl alcohol itself. However, only allyl acetate caused no damage whatsoever to the entire plant tested (flower, leaves, sepals, and stem).     

Toxicity of formic acid to red imported fire ants, Solenopsis invicta Buren

BACKGROUND: Ants often compete with other ants for resources. Although formic acid is a common defensive chemical of formicine ants, it does not occur in any other subfamilies in Formicidae. No information on toxicity of formic acid to red imported fire ants, Solenopsis invicta, is available. This study examined its contact and fumigation toxicity to S. invicta in the laboratory. RESULTS: In a contact toxicity bioassay, 24 h LD₅₀ values of formic acid for workers ranged from 124.54 to 197.71 µg ant⁻¹. Female alates and queens were much less sensitive to formic acid than workers. At a concentration of 271.72 µg ant⁻¹, which killed 81.09 ± 16.04% of workers, the 24 h mortality was up to 39.64% for female alates and 38.89% for queens. In fumigation bioassays, 24 h LC₅₀ values ranged from 0.26 to 0.50 µg mL⁻¹ for workers, 0.32 µg mL⁻¹ for male alates and 0.70 µg mL⁻¹ for female alates. Complete mortality (100%) in queens occurred 24 h after they had been exposed to 1.57 µg mL⁻¹ of formic acid. At a concentration of 2.09 µg mL⁻¹, KT₅₀ values ranged from 23.03 to 43.85 min for workers, from 37.84 to 58.37 min for male alates, from 86.06 to 121.05 min for female alates and from 68.00 to 85.92 min for queens. CONCLUSION: When applied topically, formic acid was significantly less toxic than bifenthrin to red imported fire ants. Although its fumigation toxicity was lower than that of dichlorvos, formic acid had about an order of magnitude higher toxicity to S. invicta than to other insects studied so far. It may be worth investigating the use of formic acid for managing imported fire ants. Published 2012 by John Wiley & Sons, Ltd.     

Effects of aliphatic acids on the metabolism and potency of fenitrothion in flour beetles

Formic, acetic, or n-propionic acid applied with fenitrothion to Tribolium castaneum, inhibited the production of both fenitroxon and O-desmethyl fenitrothion, but did not appear to affect metabolic attack on the P-O-phenyl link. These effects are comparable to those of some pyrethrum synergists. The toxicity of fenitrothion was synergised by formic or acetic acid, and antagonised by propionic acid, but these effects on potency were slight.     

The pyrethrins and related compounds; Part XXIII:Kinetic control in the formation of pyrethroidal estersb

The unequal amounts of the two diastereomers of (RS)-1-(3-phenoxyphenyl)ethyl (RS)-2-(4-chlorophenyl)-3-methylbutyrate, obtained from equimolar proportions of the alcohol and acyl chloride under normal conditions, imply stereoselectivity in the reaction. The extent to which this depends on the nature of the α-substituent for arange of 3-phenoxybenzyl esters, on the nature of the acid, and on the method of esterification is investigated. Selectivity arises from kinetic, rather than thermodynamic control.     

赤霉酸·6-BA 3.6%液剂高效液相色谱分析方法研究

本文采用高效液相色谱法,以甲醇＋甲酸水溶液为流动相,使用ODSC18、51μm为填料的不锈钢柱和二极管阵列检测器。在210nm波长下对赤霉酸·6-BA进行分离和定量分析。结果表明,该分析方法的线性相关系数赤霉酸为O．9999;6-BA为0．998;标准偏差赤霉酸为0．01;6-BA为0．02;变异系数赤霉酸为0．46％;6-BA为0．30％;赤霉酸平均回收率为97．7％;6-BA平均回收率为98．7％。     

Diphenamid removal from water and metabolism by aquatic plants

Several aquatic plants and algae were effective for removal of N, N-dimethyl-2,2-diphenylacetamide ([¹⁴C]diphenamid) from water and degradation to various metabolites. Parrotfeather (Myriophyllum brasilience L.) and water hyacinth (Eichornis crossiper L.) removed large quantities of diphenamid from water while algae (Ulothrix sp., Oedogonium sp., Gloeocystis sp., and Scenedesmus sp. mixed culture) and waterthread pondweed (Potamogeton diversifolius Raf.) were less effective in reducing diphenamid concentration in the water. A somewhat stable metabolite in algae, water hyacinth, and parrotfeather was N-methyl-2,2-diphenylacetamide. Algae and water hyacinth degraded diphenamid further to unknown products. One of these compounds moved more slowly than N-methyl-2,2-diphenylacetamide on thin-layer chromatography using benzene, n-heptane saturated with water and methanol solvent. While most solvents did not resolve 2,2-diphenylacetic acid from the base, petroleum ether, ethyl ether, and formic acid (50:50:2, v/v/v) was found to move 2,2-diphenylacetic acid further than diphenamid or N-methyl-2,2-diphenylacetamide on thin-layer chromatography.     

The pyrethrins and related compounds. XIX Geometrical and optical isomers of 2,2-dimethyl-3-(2,2-dichlorovinyl)-cyclopropanecarboxylic acid and insecticidal esters with 5-benzyi-3-furylmethyi and 3-phenoxybenzyl alcohols

Esters of 2,2-dimethyl-3-(2,2-dichlorovinyl)-cyclopropanecarboxylic acid with appropriate alcohols are more active insecticides than the corresponding 3-isobutenyl compounds (chrysanthemates). (±)-Cis and (±)-trans forms of the dichloro acid are obtained by fractional crystallisation of the mixed acids or by hydrolysis of the ethyl esters separated by fractional distillation. The (±)-cis and (±)-trans acids are resolved into their (+)- and (-)-forms with α-methyl-benzylamine and threo-l-p-nitrophenyl-2-N,N-dimethyiamino-propane-1,3-diol respectively. As for the corresponding chrysanthemates, the (+)-cis and (+)-trans acids give esters more active as insecticides than their enantiomers.     

Plant volatile aldehydes as natural insecticides against stored-product beetles

BACKGROUND: Infestation by stored-product pests causes serious losses in food and feed commodities. Among possible strategies against these pests, which aim to reduce the use of synthetic insecticides, including fumigants, natural insecticides produced by plants represent one of the most promising approaches for their ecochemical control. Three six-carbon and nine-carbon aldehydes, natural plant volatiles produced by the plant lipoxygenase pathway, were tested for their insecticidal activity against five species of stored-product beetles in feeding, fumigation and combined bioassays. RESULTS: The compounds (2E,6Z)-nonadienal, (2E)-nonenal and (2E)-hexenal were incorporated into feeding discs in feeding bioassays or evaporated from filter paper in closed glass chambers in fumigation tests. Beetle sensitivity to aldehydes differed according to the different treatments. The highest activity was obtained by (2E)-hexenal in fumigation tests, with the LC(50) ranging from 4 to 26 mg L(-1), while (2E, 6Z)-nonadienal was the most effective in feeding tests, giving LD(50)s ranging from 0.44 to 2.76 mg g(-1) when applied to feeding discs. Fumigation tests in the presence of wheat grains confirmed that (2E)-hexenal was the most effective compound, with a calculated LC(99) ranging from 33 to 166 mg L(-1). CONCLUSION: The results of both feeding and fumigation tests indicated that natural plant aldehydes are potential candidates to control stored-product beetles.     

New conjugated metabolites of 3-phenoxybenzoic acid in plants

The metabolism of 3-phenoxybenzoic acid, a common plant metabolite of deltamethrin, cypermethrin and fenvalerate, has been studied in abscised leaves of cabbage, cotton, cucumber, kidney bean and tomato plants. The [¹⁴C]-acid was readily converted into more polar conjugates by esterification with glucose, 6-O-malonylglucose, gentiobiose, cellobiose, glucosylxylose and two types of triglucose with different isomerism. Other metabolites identified were the glucosyl ether of 3-(4-hydroxyphenoxy)benzoic acid, and a 3-(2-hydroxyphenoxy)benzoic acid derivative with a total of two molar equivalents of glucose linked to the carboxyl and phenolic -OH groups. The conjugation pathways were somewhat plant-specific. The glucosylxylose ester was found only in cotton, and the cellobiose and triglucose esters were found only in tomato. All of the conjugates except the glucose and glucosylxylose esters were plant metabolites that had not been identified previously. Furthermore, this is the first report to show the presence of cellobiose and triglucose conjugates in plants. However, neither of the acetyl derivatives of the [¹⁴C]-triglucoside was identical with the synthetic deca-acetyl derivative of [1→6]-triglucoside.     

Improved efficacy of ethyl formate against stored grain insects by combination with carbon dioxide in a 'dynamic' application

Ethyl formate is being evaluated as a fumigant for stored grain as it is a potential alternative to the ozone-depleting fumigant methyl bromide and to phosphine, which is under pressure owing to the development of strong resistance in stored grain insects. However, use of ethyl formate faces significant challenges, such as poor penetration through grain, significant losses to grain sorption, high concentrations of fumigant required to control insects, and flammability risks, which have limited its further development. In this study it was found that the combination of carbon dioxide (5-20%) with ethyl formate significantly enhanced efficacy of the fumigant against external living stages of the rice weevil, Sitophilus oryzae (L.), the lesser grain borer, Rhyzopertha dominica F., and the flour beetle, Tribolium castaneum (Herbst). Dynamic application of ethyl formate and carbon dioxide mixture (100 mg litre-1 ethyl formate, 20% CO2) pumped through a model silo containing wheat (50 kg) for one gas exchange was also investigated. A flow rate of 6 litres min-1 gave a relatively even distribution of fumigant throughout the grain column and similar mortality levels among cultures of S. oryzae and T. castaneum placed at three positions, the top, middle and bottom of the column. Mortality of 99.8% of mixed stage cultures of T. castaneum and 95.1% of S. oryzae was achieved in 3 h exposures to 111 and 185 mg ethyl formate h litre-1 respectively applied by the dynamic method. It is concluded that the combination of carbon dioxide with ethyl formate and dynamic application enhances distribution and efficacy of the fumigant against stored grain insects.     

固相萃取-高效液相色谱-串联质谱法检测稻田中硝磺草酮和2甲4氯钠残留

建立了一种同步检测稻田中硝磺草酮和2甲4氯残留的固相萃取-高效液相色谱-串联质谱（SPE-LC-MS/MS）分析方法。样品经乙酸乙酯提取后,用PSA吸附、C₁₈固相萃取小柱净化,C₁₈色谱柱分离,0.1%甲酸水-0.1%甲酸甲醇溶液为流动相,梯度洗脱分离,在电喷雾ESI离子源正、负离子模式下,采用质谱多反应监测（MRM）模式定性分析。结果表明:硝磺草酮和2甲4氯的检出限（LOD）分别为0.005和0.025 ng,定量限（LOQ）分别为0.01和0.05 mg/L。在0.001~2 mg/L和0.005~5 mg/L范围内,硝磺草酮和2甲4氯的质量浓度与对应的峰面积间呈良好的线性关系,其相关系数均大于0.999 7。在0.01~2 mg/L添加水平下,硝磺草酮在田水、土壤、植株、稻壳和糙米中的平均回收率为75%~103%,相对标准偏差（RSD）为0.1%~8.2%;在0.05~10 mg/L添加水平下,2甲4氯在田水、土壤、植株、稻壳和糙米中的平均回收率为90%~113%,RSD为0.5%~7.4%。说明该方法高效、快捷、精确度高,能够满足硝磺草酮和2甲4氯钠在稻田中的残留量检测分析要求。     

不同时期嫁接感染甘薯病毒病害SPVD对甘薯产量的影响

以苹果腐烂病菌、苹果轮纹病菌、棉花枯萎病菌、柑橘炭疽病菌与白菜黑斑病菌为供试菌,采用生长速率法测定25种中药材甲醇粗提物的抑菌活性。结果表明,在浓度为5．0 mg／mL 时,大部分中药材的甲醇粗提物对至少一种植物病原真菌都表现出不同程度的抑制活性,其中知母、青蒿、细辛与白鲜皮对3种供试植物病原菌的抑制率大于75％,而茜草与姜黄对4种供试植物病原菌的抑制率均在80％以上,具有广泛的抑菌谱。进一步采用系统溶剂法得到的茜草不同溶剂萃取物中,水饱和正丁醇相抑菌活性明显优于石油醚及乙酸乙酯相,表明茜草抑菌活性成分主要为中等及大极性化合物。     

甲酸乙酯对面粉中赤拟谷盗的熏蒸效果

本文研究了甲酸乙酯对面粉中赤拟谷盗不同虫态的熏蒸作用。通过对熏蒸期间熏蒸剂浓度检测,计算浓度时间乘积(CT)计算不同时间下甲酸乙酯对赤拟谷盗各虫态达到99%控制的处理指标(Lct_(99))。结果表明甲酸乙酯熏蒸能有效防治面粉中的赤拟谷盗各虫态,赤拟谷盗不同虫态对甲酸乙酯的敏感性由强到弱依次为:卵>低龄幼虫>成虫>高龄幼虫>蛹。在23℃条件下处理6h和12h对面粉中赤拟谷盗的Lct_(99)分别为448.42和599.34mg·h/L     

茜草等25种中药材对5种植物病原真菌的抑制活性

以苹果腐烂病菌、苹果轮纹病菌、棉花枯萎病菌、柑橘炭疽病菌与白菜黑斑病菌为供试菌,采用生长速率法测定25种中药材甲醇粗提物的抑菌活性。结果表明,在浓度为5.0mg/mL时,大部分中药材的甲醇粗提物对至少一种植物病原真菌都表现出不同程度的抑制活性,其中知母、青蒿、细辛与白鲜皮对3种供试植物病原菌的抑制率大于75%,而茜草与姜黄对4种供试植物病原菌的抑制率均在80%以上,具有广泛的抑菌谱。进一步采用系统溶剂法得到的茜草不同溶剂萃取物中,水饱和正丁醇相抑菌活性明显优于石油醚及乙酸乙酯相,表明茜草抑菌活性成分主要为中等及大极性化合物。