有机磷化合物体外全血胆碱酯酶检测方法学研究

目的建立一种在体外快速测定乙酰胆碱酯酶（AChE）活性的方法,为有机磷毒剂中毒的诊断及救治提供依据。方法用改良的5,5-连硫代-双-2-硝基苯甲酸（DTNB）法测定AChE活性,以兔全血作为酶源进行方法学研究,并用不同种属动物全血进行了适用性考察,用SPECTRA MAX 190读板机测定412 nm处光密度的变化即可测定AChE活性。结果不同动物全血40倍稀释后作为酶源较为合适,批内差异为0.44%～0.67%,批间差异为1.03%～2.27%,该方法适用于兔、比格犬、豚鼠和猴全血AChE测定。梭曼和沙林对兔全血AChE活性抑制具有明显的量效关系。结论该方法简便、易行,适用于不同种属动物全血AChE活性的测定。

A blood cholinesterase assay method for organophosphorus compounds in vitro

Objective To establish a rapid method for determination of acetylcholinesterase（AChE） activity in vitro in order to facilitate the diagnosis and remedy of organophosphorus agent intoxication.Methods The activity of AChE was determined by modified DTNB method,rabbit blood was used as an enzyme when methodology was studied,and the applicability of the method was investigated with the blood of different animals.SPECTRA MAX 190 was used to determine the absorbance changes at 412 nm in order to determine the AChE activity.Results It was appropriate to choose whole blood diluted forty times as enzyme sources.The intra-and inter-assay coefficients of variation were 0.44%-0.67% and 1.03%-2.27%,respectively.This method was suitable for measuring AChE activity of blood in rabbits,beagle dogs,guinea pigs and monkeys.Soman and sarin could specifically suppress the AChE activity of rabbit blood and showed a significant dose-effect relationship.Conclusion This method is simple and suitable for determination of blood AChE activity of different animals.