•  
  •  
 

Abstract

The esterase activity of Chrysemyswas determined by Ellman method, and the indole acetate was used as substrate to determine the esterase activity of Tupaiabelangeri. The optimum reaction conditions were determined by orthogonal array design.and the inhibition effects of several specific enzyme inhibitors were compared; The enzyme inhibition method was used to investigated the sensitivity of esterase form Chrysemyse and Tupaiabelangeri to organophosphorus and carbamate pesticides. The optimum conditions of esterase activity determination of Chrysemys were: incubation temperature 35 ℃, incubation time 15 min, buffer pH 7.5; The optimum conditions of esterase activity determination of Tupaiabelangeri were: incubation temperature 25 ℃, incubation time 15 min, buffer pH 8.0.The most sensitive to Chrysemysesterase was donepezil hydrochloride, the most sensitive to Tupaiabelangeri esterasewas bis (4-nitrophenyl) phosphate. The detection limits of esterase form Chrysemys and Tupaiabelangeri on trichlorfon, phoxim, dichlorvos, mevinphos, phosphamidon, metolcarb, carbofuran, methomyl, aldicarb and carbaryl were all lower than the maximum residue limits stipulated by national standards. The extracts of esterase from Chrysemys and Tupaiabelangeriwere showed high sensitivity to pesticide, which could be used as a new enzyme source in the enzyme inhibition method for rapid detection organophosphorus and carbamate pesticide.

Publication Date

11-28-2019

First Page

58

Last Page

63

DOI

10.13652/j.issn.1003-5788.2019.11.012

References

[1] 范宝光. 农药残留对食品安全的影响及其控制措施[J]. 中国农业信息, 2015, 3(6): 28-30.
[2] 张爱琳, 樊秀花, 郑敏. 鸡血清胆碱酯酶的提取纯化工艺研究[J]. 食品研究与开发, 2011, 32(5): 30-33.
[3] 余以刚, 卢志洪, 朱珍, 等. 广州市售蔬菜有机磷农药残留情况调查分析[J]. 现代食品科技, 2010, 26(7): 742-745.
[4] JAFARI M T, SARAJI M, SHERAFATM H. Polypyrrole/montmorillonite nanocomposite as a new solid phase microextraction fiber combined with gas chromatography-corona discharge ion mobility spectrometry for the simultaneous determination of diazinon and fenthion organophosphorus pesticides[J]. Analytica Chimica Acta, 2014, 814(7): 69-78.
[5] MENEZES FILHO A, DOS SANTOS P N, PEREIRA P A D P. Development, validation and application of a method based on DI-SPME and GC-MS for determination of pesticides of different chemical groups in surface and groundwater samples[J]. Microchemical Journal, 2010, 96(1): 139-45.
[6] FANG Jing, ZHAO Hong-zhi, ZHANG Yan-hao, et al. Performance of atmospheric pressure gas chromatography-tandem mass spectrometry for the analysis of organochlorine pesticides in human serum[J]. Analytical and Bioanalytical Chemistry, 2019, 411(18): 4 185-4 191.
[7] SANCHEZ M E, MENDEZ R, GOMEZ X, et al. Determination of diazinon and fenitrothion in environmental water and soil samples by HPLC[J]. Journal of Liquid Chromatography & Related Technologies, 2003, 26(8): 483-497.
[8] SONG N E, KIM D B, LIM T G, et al. Determining pesticide residues in wheat flour by ultrahigh-performance liquid chromatography/quadrupole time-of-flight mass spectrometry with QuEChERS extraction[J]. Food Additives & Contaminants 2019, 16(11): 1-11.
[9] SHAYEGHI M, KHOOBDEL M, VATANDOOST H. Determination of organophosphorus insecticides (malathion and diazinon) residue in the drinking water[J]. Pakistan Journal of Biological Sciences, 2007, 10(17): 2 900-2 904.
[10] SALM P, TAYLOR P J, ROBERTS D, et al. Liquid chromatography-tandem mass spectrometry method for the simultaneous quantitative determination of the organophosphorus pesticides dimethoate, fenthion, diazinon and chlorpyrifos in human blood[J]. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 2009, 877: 568-574.
[11] 麦昌青, 陈盛, 陈燕. 豇豆中有机磷和氨基甲酸酯类农药残留酶抑制率快速检测方法[J]. 现代农药, 2016, 15(1): 41-43.
[12] SUN Xia, WANG Xiang-you. Acetylcholinesterase biosensor based on prussian blue-modified electrode for detecting organophosphorous pesticides[J]. Biosens Bioelectron, 2010, 25(12): 2 611-2 614.
[13] VIOQUE-FERNANDEZ A, DE ALMEIDA E A, LOPEZ-BAREA J. Esterases as pesticide biomarkers in crayfish (Procambarus clarkii, Crustacea): Tissue distribution, sensitivity to model compounds and recovery from inactivation[J]. Comp Biochem Physiol C-Toxicol Pharmacol, 2007, 145(3): 404-412.
[14] LAGUERRE C, SANCHEZ-HERNANDEZ J C, KOHLER H R, et al. B-type esterases in the snail Xeropictaderbentina: An enzymological analysis to evaluate their use as biomarkers of pesticide exposure[J]. Environ Pollut, 2009, 157: 199-207.
[15] YANG Ling-ling, YANG Xiao, LI Guo-biao, et al. An integrated molecular docking and rescoring method for predicting the sensitivity spectrum of various serine hydrolases to organophosphorus pesticides[J]. Journal of the Science of Food and Agriculture, 2016, 96: 2 184-2 192.
[16] 高希武. Gorun等改进的Ellman胆碱酯酶活性测定方法介绍[J]. 昆虫知识, 1987(4): 245-246.
[17] 何绍志. 美洲蟑螂乙酰胆碱酯酶的分离纯化及性质研究[D]. 成都: 西华大学, 2014: 8-13.
[18] 刘永杰, 张金振, 曹明章, 等. 酶抑制法快速检测农产品农药残留的研究与应用[J]. 现代农药, 2004(2): 25-27.
[19] 苏鹤, 罗俊霞, 赵建波. 一种商品化酶抑制—比色法快速检测农药残留试剂盒对10种有机磷农药的敏感性测定[J]. 食品安全质量检测学报, 2019, 10(16): 5 512-5 516.

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.