活血通脉胶囊抗凝血酶活性生物效价测定法的建立

Establishment of a method to determine biological titer of antithrombin activity in Huoxuetongmai capsule

目的建立活血通脉胶囊抗凝血酶活性生物效价测定方法,用于评价和控制其质量。方法分别对Markwardt法、纤维蛋白原平板法(沉淀法)测定水蛭和活血通脉胶囊的生物效价进行比较研究;选择琼脂糖-纤维蛋白原平板法(沉淀法)作为活血通脉胶囊生物效价测定方法,考察试验系,并进行了方法学验证。结果琼脂糖-纤维蛋白原平板法(沉淀法)测定水蛭和活血通脉胶囊抗凝血酶生物效价的中间精密度RSD分别为3.5%、5.2%,优于Markwardt法(RSD分别为15.6%、12.9%)。本法的线性范围为5 BP^20 BP(R2=0.994),回收率为98.26%,RSD 2.3%(n=6),中间精密度RSD 3.2%,重复性RSD 4.7%。结论琼脂糖-纤维蛋白原平板法(沉淀法)可用于活血通脉胶囊中抗凝血酶活性生物效价的测定,以评价和控制其生物质量。

Aim To establish a method for determining the biological potency of antithrombin activity of Huoxuetongmai capsules for the evaluation and control of its quality.Methods The determination results of biological titers of leeches and Huoxuetongmai capsules by the Markwardt method and the fibrinogen plate method(precipitation method)were compared.Agarose-fibrinogen plate method(precipitation method)was selected as the method for determining the biological potency of Huoxuetongmai capsules,the experimental condition system was investigated,and methodological verification was carried out.Result The intermediate precision RSD of the agarose-fibrinogen plate method(precipitation method)for measuring the antithrombin biopotency of leech and Huoxuetongmai capsules was 3.5%and 5.2%,respectively,which was better than the Markwardt method(RSD was 15.6%,12.9%respectively).The linear range of this method was 5 BP^20 BP(R2=0.994),the recovery rate was 98.26%,RSD was 2.3%(n=6),precision RSD was 3.2%,and repeatability RSD was 4.7%.Conclusions The agarose-fibrinogen plate method(precipitation method)can be used for the determination of the biological potency of antithrombin activity in HuoxueTongmai capsules,and to evaluate and control its biological quality.