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An integration strategy to measure enzyme activities for detecting irreversible inhibitors with dimethoate on butyrylcholinesterase as a model

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An integration strategy was investigated to measure initial rates of horse butyrylcholinesterase (BChE) at 50.0 µmol L-1 butyrylthiocholine (BTCh) for detecting irreversible inhibitors as pollutants in environment and foods with dimethoate as a model. In this integration strategy: (a) if BTCh consumption within 5.0 min was >60%, BChE initial rates were derived from maximal reaction rates, estimated by an improved integrated method, according to Michaelies-Menten kinetics at 47.0 µmol L-1 BTCh and Michaelis-Menten constant at 94.0 µmol L-1; (b) or else initial rates were determined by the classical initial rate method. Thus, the differences in BChE initial rates without and after dimethoate treatment indexed final dimethoate contents in reaction mixtures to treat BChE. Results supported that this integration strategy determined BChE activities with a linear range about two magnitudes and an upper limit about twice that by the classical initial rate method alone at 2.0 mmol L-1 BTCh. The coefficient of variation with this integration strategy was below 5%. The difference in BChE initial rates before and after dimethoate treatment was proportional to final dimethoate contents in reaction mixtures. By enzymatic analyses, the molar contents of dimethoate extracted from polluted cabbages were consistent with the summed molar contents of dimethoate and dimethoxon by gas-chromatography. Therefore, this integration strategy was effective to detect irreversible inhibitors as pollutants in environment and foods.
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Keywords: butyrylcholinesterase; dimethoate; enzymatic analysis; integration strategy; irreversible inhibitors

Document Type: Research Article

Affiliations: 1: Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, College of Pharmaceutical Sciences, Chongqing Medical University, Chongqing 400016, China 2: College of Public Health, Chongqing Medical University, Chongqing 400016, China

Publication date: April 1, 2011

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