肝素钠效价测定方法的筛选

Screening the determination method of heparin sodium

为研究简单、快速、方便和准确的肝素钠效价测定方法,利用天青A和肝素钠反应生成络合物使天青A吸收波长蓝移的原理,首先筛选最佳缓冲液,并研究比较单波长法、双波长法和一阶导数法测定肝素钠效价的适宜性和可行性,确定最佳的测定方法。结果表明:Tris-HCl缓冲液完全可以替代巴比妥缓冲液;高效价测定时,三种方法测定肝素钠效价准确性由低到高的顺序分别为单波长法、双波长和一阶导数法;低效价测定时,只能用一阶导数法。其中一阶导数法的最佳测定条件为Tris-HCl缓冲液浓度0.05mol/L,pH=8.6,天青A浓度0.2mg/mL。这种方法的相关系数平方为0.9980,变异系数为0.43%。

In order to explore easy,frequent,convenient and accurate method for measuring heparin sodium potency,based on the reaction of heparin sodium with Azure A,the best buffer for the reaction was firstly screened,single-wavelength method,dual-wavelength method and the first order derivative method were further compared. According to the suitability and feasibility,the best method was finally determined. The results showed that Tris-HCI buffer could replace barbiturates buffer totally,when measured low potency,three methods for measurement accuracy heparin sodium potency were feasible and the accuracy order from low to high was that..single-wavelength method ,dual-wavelength and the first order derivative method. When measured low potency,only the first order derivative worked. The first order derivative method of optimum conditions were Tris-HCI buffer concentration 0.05mol/L, pH=8.6,Azure A concentration 0.2mg/mL. The squared correlation coefficient was 0.9980,the variation coefficients of this method was 0.43%.