凝血酶原复合物中肝素含量测定的两种方法的比较

Comparison of two methods for determination of heparin content in prothrombin complex concentrates

目的对凝血酶原复合物(prothrombin complex concentrate,PCC)中肝素含量测定的发色底物法和经典的硫酸鱼精蛋白中和肝素凝固法进行比较。方法对本公司制备的8批PCC实验样品,分别采用发色底物法和凝固法测定肝素含量,比较两种方法检测结果的差异和测定重复性。结果凝固法、发色底物法重复性均良好,凝固法检测有20%的误差,发色底物法CV在3.5%~9.2%之间;两种方法样品检测值的差值与均值比在4.4%~29.2%之间,检测值的相关系数为0.407,而仅选取用组分Ⅲ沉淀制备样品时,检测值的相关系数为0.938。结论凝血酶原复合物中肝素含量测定的两种方法中,发色底物法与经典的硫酸鱼精蛋白中和肝素凝固法相比,定量更精确,适用性更强。

Objective To compare the chromogenic substrate assay and classical protamine sulfate neutralization and clotting assay for determination of heparin content in prothrombin complex concentrates（PCC）. Methods Eight batches of experimental samples prepared by Shanghai RAAS Blood Products Co., Ltd. were determined for heparin content by chromogenic substrate assay and classical protamine sulfate neutralization and clotting assay respectively, and the difference and reproducibility of results were compared. Results Both the clotting assay and chromogenic substrate assay were reproducible, while an error of 20% error was observed in clotting assay. The coefficient of variation（CV） of determination results by chromogenic substrate assay was between 3. 5% and 9. 2%, and the ratio of the difference and the mean value of samples in two groups was between 4. 4% and 29. 2%. The correlation coefficient of determination results by two methods was 0. 407. However, when the samples prepared only by precipitation of fraction III were selected,the correlation coefficient of two methods was 0. 938. Conclusion For the determination of heparin content in PCC, the chromogenic substrate assay was more accurate and applicable than classical protamine sulfate neutralization and clotting assay.