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兔血清乙酰胆碱酯酶用于农药残留速测的最佳条件研究

作  者:
王文祥;胡美英;刘萱清;蒲小明;钟国华
单  位:
华南农业大学昆虫毒理研究室农药与化学生物学教育部重点实验室
关键词:
农药残留;快速检测;乙酰胆碱酯酶;丙硫磷
摘  要:
研究了磷酸缓冲液pH值、反应时间、反应温度、底物、酶量对兔血清乙酰胆碱酯酶(AChE)活力测定的影响,结果表明,供试AChE活力最佳条件是:磷酸缓冲液(pH8.0)3.0ml,显色剂0.6%二硫双对硝基苯甲酸(DTNB)50μl,0.75%酶液(pH8.0)100μl,反应温度35℃,混匀稳定后加入底物1%碘化硫代乙酰胆碱(BTCI)50μl,反应180s后在412nm处测定光密度。在上述条件下,测定表明有机磷杀虫剂丙硫磷(Prothiofos)浓度在0.0125~0.4μg/ml,保育时间5~30min之间与AChE抑制率之间存在良好的相关性,可根据检测需要调整保育抑制时间。本试验结果将为应用兔血清AChE研制、改进有机磷和氨基甲酸酯类农药残留速测卡,实现农产品、食品农药残留快速检测提供重要参考。
译  名:
Study on Optimal Conditions of Assaying Pesticides Residue through Inhibition of Rabbit Serum Acetylcholinesterase
作  者:
WANG Wen-xiang,HU Mei-ying,LIU Xuan-qing,PU Xiao-ming,ZHONG Guo-hua*(Lab of Insect Toxicology, South China Agricultural University, Key Lab of Pesticide Chemical Biology,Ministry of Education, Guangzhou 510640, China)
关键词:
pesticide residue;rapid detection;acetylcholinesterase;prothiofos
摘  要:
The conditions were studied for assaying of acetylcholinesterase (AChE) activity from rabbit serum, as the targetenzyme of organophosphate and carbamate pesticide. The results showed that the optimal reaction conditions were: 3.0ml of pH8.0buffer solution of phosphoric acid, 50μl of 0.6% DTNB and 100μl of 0.75% AChE were mixed with 50μl of 1% BTCI at 35℃,and then value detected of OD412 after 180s. The AChE activity showed good inhibition effect on the prothiofos, anorganophosphate pesticide, with 0.0125~0.4μg/ml as the concentration range and / or 5 ̄30 min as incubation time under the abovecondition. It indicated that the assay time could be adjusted. These results would be important to the R&D of rapid detectioncard for assaying pesticide residue by rabbit serum AChE and improveing test accuracy.

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