Validation and kinetic of enzymatic method for the detection of organophosphate insecticides based on cholinesterase inhibition

IF 2.8 4区医学 Q2 TOXICOLOGY

Toxicology Mechanisms and Methods Pub Date : 2020-02-01 DOI:10.1080/15376516.2019.1669248

S. Badawy

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引用次数: 2

Abstract

Abstract Kinetic and validation of the enzymatic method for the determination of fenitrothion organophosphorus based on cholinesterase inhibition were studied. A Linear relationship was obtained with a determination coefficient R2 of 0.9989 suggesting that the noncompetitive inhibition kinetic equation is suitable to represent the enzymatic assay of fenitrothion. The value of the inhibition constant KI was 0.374 mg/ml/min. The analytical logarithmic curve for the determination of fenitrothion concentration using the percentage of cholinesterase inhibition presented good linear relations at concentrations of 0.05–2 mg/ml (R2 = 0.9889). The maximum inhibition 83% was observed at 2.0 mg/ml final assay concentration. The lower inhibition 3.3% was observed at 0.05 mg/ml detection limit. The experimental measurement condition was optimized. The enzymatic method exhibited detection limits (LOD) in the range of 0.05–2.0 mg/ml. The limit of quantification (LOQ) was 0.06 mg/ml with inhibition 13%. The concentration of fenitrothion that inhibited the hydrolysis of substrate by 50% (IC50 value) was 0.4 mg/ml. Standard deviations and coefficients of variation indicate a good precision of the enzymatic method for the detection of organophosphate insecticides at an incubation time of 20 min.

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基于胆碱酯酶抑制的酶法检测有机磷杀虫剂的验证和动力学

摘要研究了胆碱酯酶抑制法测定有机磷非硝硫磷的动力学和验证。结果表明，非竞争抑制动力学方程与非竞争抑制动力学方程具有良好的线性关系，决定系数R2 = 0.9989。抑制常数KI值为0.374 mg/ml/min。在0.05 ~ 2 mg/ml浓度范围内，用胆碱酯酶抑制率测定非硝硫磷浓度的分析对数曲线呈现良好的线性关系(R2 = 0.9889)。最终测定浓度为2.0 mg/ml时，抑制作用最大，达到83%。在0.05 mg/ml检出限下，抑制率为3.3%。优化了实验测量条件。该方法的检出限为0.05 ~ 2.0 mg/ml。定量限为0.06 mg/ml，抑制率为13%。抑制底物水解50% (IC50值)的非硝硫磷浓度为0.4 mg/ml。标准偏差和变异系数表明，在孵育时间为20 min时，酶法检测有机磷杀虫剂具有良好的精度。

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来源期刊

Toxicology Mechanisms and Methods TOXICOLOGY-

自引率

3.10%

发文量

期刊介绍： Toxicology Mechanisms and Methods is a peer-reviewed journal whose aim is twofold. Firstly, the journal contains original research on subjects dealing with the mechanisms by which foreign chemicals cause toxic tissue injury. Chemical substances of interest include industrial compounds, environmental pollutants, hazardous wastes, drugs, pesticides, and chemical warfare agents. The scope of the journal spans from molecular and cellular mechanisms of action to the consideration of mechanistic evidence in establishing regulatory policy. Secondly, the journal addresses aspects of the development, validation, and application of new and existing laboratory methods, techniques, and equipment. A variety of research methods are discussed, including: In vivo studies with standard and alternative species In vitro studies and alternative methodologies Molecular, biochemical, and cellular techniques Pharmacokinetics and pharmacodynamics Mathematical modeling and computer programs Forensic analyses Risk assessment Data collection and analysis.