血凝酶效价测定新方法的方法学研究

Methodology of a new potency assay of hemocoagulase

目的:本研究提供了一种新的血凝酶效价测定的方法。方法:采用凝血质控血浆为底物,以凝血分析仪进行测定,利用血凝酶可以直接水解血浆中纤维蛋白原为纤维蛋白使血浆凝固,并且在一定血凝酶浓度范围内,血浆的凝固时间与血凝酶效价成反比的原理,测得血凝酶效价。结果:本方法对血凝酶的精密度的相对标准偏差为2. 3%,准确度的相对标准偏差在4. 1%～4. 8%之间,定量线性范围为0. 4～2. 5 U·mL^(-1),并具有良好的专属性和耐用性,通过市售3个规格9批次注射用矛头蝮蛇血凝酶效价测定,能够较好的控制产品的效价。结论:本研究避免了检验人对凝固终点判定标准的不同和人-枸缘酸血浆存在的差异,所导致产品之间效价存在的差异,实现血凝酶高效、准确的测定。

Objective： To provide a new method for hemocoagulase potency assay. Methods： The coagulation ana- lyzer was used for the determination with the quality control coagulation plasma as substrate. The potency of hemo- coagulase was determined based on the principle that hemocoagulase hydrolyzed fibrinogen into fibrin and lead to plasma coagulation and coagulation time was inversely proportional to potency in a certain range of concentration Results： The relative standard deviation （RSD） of precision was 2.3%, the RSD of accuracy fell into 4. 1% to 4. 8%, the quantitative linear range fell into 0. 4 U·mL^-1 to 2.5 U·mL^-1, and the method had good specificity and robustness. The determination of 9 batches of 3 strengths showed that the potency of hemocoagulase bothrops atrox for injection could be controlled well. Conclusion ： This study avoided the differences of potency assay, which were caused by coagulation end point judgment by different persons, and different human plasma citrate. This study realized the efficient and accurate determination of hemocoagulase potency.