目的: 以丹红注射液的抗血小板聚集活性为切入点,旨在建立丹红注射液的生物活性测定法,以期量化丹红注射液的药理作用,并优化现有的中药注射液质量评价体系。方法: 以诱导剂为花生四烯酸(arachidonic acid),丹红注射液剂间比1∶0.8,血小板数500×109个·L-1,在体外抑制血小板聚集的生物作用为试验系,实施量效关系考察与效价定义,而后通过量反应平行线法计算效价,建立丹红注射液的生物活性测定法,同时进行方法学考察验证该方法的可行性与适用性。结果: 丹红注射液在64%~100%的浓度范围内呈现较显著的量效关系,且由效价定义得到的丹红注射液标准品效价为1 350 U·mL-1;其余7个不同批次的供试品效价分别为1 320.6、1 397.4、1 407.5、1 445.2、1 278.4及1 234.6 U·mL-1。结论: 基于抗血小板聚集活性建立的丹红注射液生物活性测定法操作简便,精确度高,同时以效价测定的方式量化了丹红注射液的关键活性,可为现有中药质量控制方法的改进与扩充提供借鉴。
Objective: Taking the anti-platelet aggregation activity of Danhong injection as the breakthrough point, to establish a bioassay method for Danhong injection, in order to quantify the pharmacological effect of Danhong injection and optimize the existing quality evaluation system of traditional Chinese medicine injection. Methods: Arachidonic acid was used as the inducer, the dose ratio of Danhong injection was 1∶0.8, and the platelet count was 500×109 L-1. The inhibition of platelet aggregation in vitro by Danhong injection was used as the experimental system, to examine the dose-effect relationship and define the valence, and then the valences were calculated by the dose-response parallel line method, and the bioassay method of Danhong injection was developed. At the same time, the feasibility and applicability of the method were verified by methodological investigation. Results: The results showed that Danhong injection displayed a significant dose-effect relationship within the concentration range of 64%-100%, and the valence of Danhong injection standard obtained from the valence definition was 1 350 U·mL-1. The valences of the remaining seven different batches of the test articles were 1 320.6, 1 397.4, 1 407.5, 1 445.2, 1 278.4 and 1 234.6 U·mL-1, respectively. Conclusion: The established bioassay method of Danhong injection based on anti-platelet aggregation activity is simple to operate and highly accurate, while quantifying the key activity of Danhong injection by valence determination, which provides a reference for the improvement and expansion of current quality control methods for traditional Chinese medicine.
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