羟胺三氯化铁法测定人血清胆碱酯酶方法的优化

目的： 鉴于羟胺三氯化铁法（WS/T66-1996）测定人血清胆碱酯酶（ChEs）时效率较低以及容易产生沉淀等缺陷，本研究旨在对该方法进行优化，以使测定更加高效。方法： 首先验证羟胺三氯化铁法测定AChE及BChE的适用性。在此基础上，进一步研究微孔板羟胺三氯化铁法测定BChE的适用性。最后，以改良Ellman法为参考，对优化前后的准确性及可靠性进行验证。结果： 羟胺三氯化铁法测定人血清中AChE酶活性的灵敏度较差，但是对人血清中BChE酶活性的测定结果较为理想。羟胺三氯化铁法与改良Ellman法测定BChE酶活性测定结果之间呈正相关关系（r=0.652，P < 0.01）；微孔板羟胺三氯化铁法与羟胺三氯化铁法测定BChE酶活性测定结果之间也呈正相关关系（r_s=0.653，P < 0.01）。与羟胺三氯化铁法方法相比，微孔板羟胺三氯化铁法的准确性及可靠性略佳，主要体现在最低检测浓度及变异系数上。结论： 微孔板羟胺三氯化铁法是一种通量较高、经济性较好的测定人血清BChE酶活性的方法，有一定的推广应用前景。

Optimization for determination of cholinesterase in human serum using the hydroxylamine ferric chloride method

OBJECTIVE:In view of the low efficiency and easy to precipitate in the determination of human serum cholinesterase(ChEs) by the hydroxylamine ferric chloride method(WS/T66-1996), the method was optimized to make the determination more efficient. METHODS:The determination of AChE and BChE by the hydroxylamine ferric chloride method was verified. On this basis, the determination of BChE by the microporous plate hydroxylamine ferric chloride method was further studied. Finally, taking the improved Ellman method as a reference, the accuracy and reliability before and after optimization were verified.RESULTS: The hydroxylamine ferric chloride method for determination of AChE enzyme activities in human serum was poor, while it was ideal for determination of BChE enzyme activities. There was a positive correlation between the results of BChE enzymatic activities as determined by the hydroxylamine ferric chloride method and the improved Ellman method(r=0.652,P<0.01). In addition,there was also a positive correlation between the results of BChE enzymatic activities as determined by the microporous plate hydroxylamine ferric chloride method and the hydroxylamine ferric chloride method(r_s=0.653, P<0.01). Compared with the hydroxylamine ferric chloride method,the accuracy and reliability of the microporous plate hydroxylamine ferric chloride method was slightly better,mainly reflected in the lowest detection concentration and the coefficient of variation. CONCLUSION: The microporous plate hydroxylamine ferric chloride method is a high flux and economical method for the determination of BChE enzymatic activity in human serum.