基于抗凝血活性的丹红注射液生物效价测定方法的建立  

Quality evaluation of Danhong Injection based on anticoagulant biological potency

作　　者：张宇航[1] 杨静[1] 吴宿慧[1] 李寒冰[1] 唐进法[2] ZHANG Yu-hang;YANG Jing;WU Su-hui;LI Han-bing;TANG Jin-fa(Henan Health-Aging Engineering Research Center,Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Province Engineering Research Center for Clinical Application,Evaluation and Transformation of Traditional Chinese Medicine,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学河南省健康衰老产业工程研究中心,河南郑州450046 [2]河南中医药大学第一附属医院河南省中药临床应用、评价与转化工程研究中心,河南郑州450000

出　　处：《中草药》2022年第8期2348-2355,共8页Chinese Traditional and Herbal Drugs

基　　金：河南省中医药科学研究专项(20-21ZY1001)。

摘　　要：目的 以凝血酶时间(thrombin time,TT)为指标,建立丹红注射液(Danhong Injection,DHI)抗凝血生物效价测定方法,以评价其质量。方法 收集10批次合格DHI,同时采用光照、高温、暴露进行稳定性加速处理,共得样品40批。工作参照物的制备采用丹酚酸B、羟基红花黄色素A与辅料1∶1∶8混合,并调制成40 mg/mL。按照“量反应平行线(3.3)法”设计抗凝血活性测定实验,所用的血浆为4只家兔等比混合的血浆,药物剂间距为1∶0.8,半自动凝血分析仪测定TT值,对线性关系、可靠性、精密度、重复性、稳定性进行考察,优化并确定检测条件,即反应体系为100μL血浆+工作参照物或样品溶液50μL+凝血酶试剂100μL,对样品开展生物效价检测。结果 工作参照物和DHI在20~40 mg/mL时,与TT值线性关系良好,r值分别为0.982 7、0.969 6。生物效价结果表明工作参照物和合格的DHI均具有显著的抗凝血活性,且结果能通过可靠性检验,10批次DHI的效价值范围为3700~4300 U/mL;与加速处理的样品生物效价测定结果比较,暴露、光照处理DHI样品可被全部检出,高温处理DHI检出率80%。结论 所建立的生物效价方法用于DHI的质量评价,具有精密度高、重复性好、结果可靠的特点。Objective To establish a method for determining the anticoagulant biological potency of Danhong Injection(DHI) with thrombin time(TT) as an index, so as to evaluate its quality.Methods Ten batches of qualified DHI were collected and subjected to stability acceleration under illumination, high temperature and exposure, yielding 40 batches of samples.The working reference substances were prepared by mixing salvianolic acid B, hydroxysafflor yellow A and excipients(1:1:8), and preparing to a concentration of 40 mg/mL.According to the “quantitative reaction parallel line(3.3) method” to design anticoagulant activity determination experiment, the plasma used was the plasma of four rabbits mixed in an equal ratio, the interval between drugs was 1:0.8, and the TT value was measured by a semi-automatic coagulation analyzer.The linear relationship, reliability, precision, repeatability and stability were investigated, and the detection conditions were optimized and determined.That is, the reaction system was 100 μL plasma + working reference substances or sample solution 50 μL + 100 μL thrombin reagent, and the biological potency of the samples was detected.Results The linear relationship with TT was good when the working reference substance and DHI were in the range of 20—40 mg/mL, with the r values being 0.982 7 and 0.969 6.The biological activity test results showed that the working reference and qualified DHI had significant anticoagulant activity, and the results could pass the reliability test.The biological potency values of 10 batches of DHI were in the range of 3700—4300 U/mL.Compared with the results of biological potency of samples after accelerated treatment, the samples treated with exposure and light could be detected completely, and the detection rate of high temperature treatment of DHI was 80%.Conclusion The established biological potency method can be used for the determination of biological potency and quality evaluation of DHI.The method has high precision, good repeatability and reliable re

关 键 词：生物效价 丹红注射液 量反应平行线 凝血酶时间 质量评价 

分 类 号：R283.6[医药卫生—中药学]