小鼠耳廓蓝染类过敏检测方法用于注射益气复脉(冻干)的质控研究

Quality control study on Yiqi Fumai Lyophilized Injection for Mouse ear blue staining pseudo-allergic reactions

目的对小鼠耳廓蓝染类过敏检测法进行方法学的验证和优化,建立注射用益气复脉(冻干,YQFM)质控方法。方法小鼠1次性尾iv含0.4%依文思蓝(EB)的受试物,以耳廓蓝染发生率和EB渗出量作为类过敏评价指标,以Compound48/80为阳性对照,考察了方法的重复性、鼠耳萃取时间等影响因素,并对20140105和20140804两个批次YQFM(1 800 mg/kg)进行了耳廓蓝染类过敏试验。结果 Compound 48/80具有明显的致类过敏反应,模型稳定,重复性好;鼠耳在加入甲酰胺后室温下浸泡2~5 d进行萃取,EB含量未发生显著变化;YQFM 20140105和20140804两个批次均未发生类过敏反应。结论小鼠耳廓蓝染类过敏检测方法较稳定,可用于YQFM质量控制。

Objective To develop and validate the detection method of mouse ear blue staining pseudo-allergic reactions, and establish the method of quality control for Yiqi Fumai lyophilized injection（YQFM）. Method Mouse anaphylactoid reaction model was developed by tail iv injection of test substance solutions containing Evans blue（EB）. Scores of ear blue staining and quantitation of ear EB exudation were the parameters for the pseudo-allergy reaction. The repeatability of the method and the extraction time of the mouse ear were investigated with Compound 48/80 as the positive control. The auricle blue anaphylactoid tests were applied to investigate two batches of YQFM（1800 mg/kg）, which are 20140105 and 20140804. Results Compound 48/80 caused severe ear bluing and EB exudation by inducing obvious vascular hyperperme ability, which indicated that they can induce mouse pseudo-allergy. The model of mouse pseudo-allergy induced by YQFM was stable and repeatable. Ears can be soaked for 2 ~ 5 days in formamide, and the content of ear EB of mouse had no significant change. Moreover, no anaphylaxis was found in YQFM20140105 and 20140804 batches. Conclusion The detection method of mouse ear blue staining pseudo-allergic reactions is stable and can be used for the quality control of YQFM.