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山羊体内血液胆碱酯酶的电测法验证

Validation of an electrometric blood cholinesterase measurement in goats.

作者信息

Al-Jobory M M H, Mohammad F K

机构信息

Department of Physiology, Division of Pharmacology and Toxicology, College of Veterinary Medicine, University of Mosul, Mosul, P. O. Box 11136, Iraq.

出版信息

J Vet Sci. 2005 Dec;6(4):299-303.

Abstract

A modified electrometric method was described and validated for measurement of plasma and erythrocyte cholinesterase activities in 6-18 months old goats. The enzymatic reaction mixture contained 3 ml distilled water, 3 ml barbital-phosphate buffer (pH 8.1), 0.2 ml plasma or erythrocytes and 0.1 ml acetylthiocholine iodide (7.5%) as a substrate. The mixture was incubated at 37 degrees C for 40 minutes. The pH of the reaction mixture was determined by a pH meter before and after the incubation. The initial pH was measured before the substrate addition. The enzyme activity was expressed as DeltapH/40 min. The coefficients of variation of the described method in measuring plasma and erythrocyte cholinesterase activities were 4 and 2%, respectively. Preliminary reference values (n = 14) of the mean cholinesterase activity (DeltapH/40 min) and 95% confidence interval in the plasma were 0.194 and 0.184-0.204, respectively, and those of the erythrocytes were 0.416 and 0.396-0.436, respectively. The pseudocholinesterase activity of the plasma cholinesterase was 63.5% as determined by quinidine sulfate inhibition. The organophosphorus insecticides dichlorvos and diazinon at 0.5-4 microM and the carbamate insecticide carbaryl at 5-20 microM in the reaction mixture significantly inhibited plasma (13.7-85.5%) and erythrocyte (16.4-71.9%) cholinesterases in vitro in a concentration-dependent manner. The results suggest that the described electrometric method is simple, precise and efficient in measuring blood cholinesterase activity in goats.

摘要

描述并验证了一种改良的电测法,用于测定6至18月龄山羊的血浆和红细胞胆碱酯酶活性。酶促反应混合物包含3毫升蒸馏水、3毫升巴比妥 - 磷酸盐缓冲液(pH 8.1)、0.2毫升血浆或红细胞以及0.1毫升碘化硫代乙酰胆碱(7.5%)作为底物。混合物在37℃孵育40分钟。孵育前后用pH计测定反应混合物的pH值。初始pH在添加底物前测量。酶活性以ΔpH/40分钟表示。所述方法在测量血浆和红细胞胆碱酯酶活性时的变异系数分别为4%和2%。血浆中胆碱酯酶活性(ΔpH/40分钟)的初步参考值(n = 14)及95%置信区间分别为0.194和0.184 - 0.204,红细胞的分别为0.416和0.396 - 0.436。通过硫酸奎尼丁抑制法测定,血浆胆碱酯酶的假性胆碱酯酶活性为63.5%。反应混合物中浓度为0.5至4微摩尔的有机磷杀虫剂敌敌畏和二嗪农以及浓度为5至20微摩尔的氨基甲酸酯杀虫剂西维因在体外以浓度依赖方式显著抑制血浆(13.7 - 85.5%)和红细胞(16.4 - 71.9%)胆碱酯酶。结果表明,所述电测法在测量山羊血液胆碱酯酶活性方面简单、精确且高效。

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