基于HPLC指纹图谱和L-02细胞/CCK-8测定法的复方牛胎肝提取物片质量评价研究  

Quality evaluation of compound embryonic bovine liver extract tablets by HPLC fingerprint and L-02 cells/CCK-8 method

作　　者：武晗燕[1] 段艳 周长明[1] WU Han-yan;DUAN Yan;ZHOU Chang-ming(Beijing Institute for Drug Control,NMPA Key Laboratory for Research and Evaluation of Generic Drugs,Beijing Key Laboratory of Analysis and Evaluation on Chinese Medicine,Beijing 102206,China)

机构地区：[1]北京市药品检验所,国家药品监督管理局仿制药研究与评价重点实验室,中药成分分析与生物评价北京市重点实验室,北京102206

出　　处：《药物分析杂志》2021年第5期839-845,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：国家药品标准提高课题资助项目(2017103)。

摘　　要：目的:建立复方牛胎肝提取物片高效液相色谱(HPLC)指纹图谱,结合L-02细胞/CCK-8测定法,为复方牛胎肝提取物片的质量评价提供一种新方法。方法:选择3家生产企业生产的9批复方牛胎肝提取物片样品。采用HPLC法,以0.1%甲酸为流动相A,0.1%甲酸甲醇溶液为流动相B,进行梯度洗脱;流速1.0 mL·min^(-1);检测波长260 nm;柱温30℃;进样量20μL。用中药色谱指纹图谱相似度评价系统(2012版)建立复方牛胎肝提取物片的指纹图谱,进行相似度分析,并用L-02细胞/CCK-8测定法对产品进行活性测定。结果:建立了复方牛胎肝提取物片的HPLC指纹图谱,相似度计算结果在0.963~1.000范围内,3家企业生产工艺稳定,同企业产品批间一致性良好。L-02细胞/CCK-8测定法检测3家企业9批复方牛胎肝提取物片的活性,产品对细胞的修复率在49%~147%范围内,不同企业产品活性存在较大差异。结论:所建立的指纹图谱方法专属性强,高效快速,可用于考察复方牛胎肝提取物片的批间一致性。采用L-02细胞/CCK-8测定法测定产品对细胞的修复率,明显提升了指标的科学性,改善现标准的指标钝化的现状,为复方牛胎肝提取物片的质量评价提供参考。Objective:To establish an HPLC fingerprint of compound embryonic bovine liver extract tablets,and to provide a new method for the quality control of compound embryonic bovine liver extract tablets using L-02 cells/CCK-8 method.Methods:A total of 9 batches of compound embryonic bovine liver extract tablets supplied by 3 pharmaceutical manufacturers were applied in this study.HPLC analysis was performed with 0.1%formic acid solution(A)and 0.1%formic acid methanol solution(B)as the mobile phase at a flow rate of 1.0 mL·min^(-1).The detection wavelength was 260 nm.The column temperature was 30℃.The injection volume was 20μL.The fingerprint common pattern and similarity calculation were established using traditional Chinese medicine chromatographic fingerprint similarity evaluation system(version 2012).The L-02 cells/CCK-8 method was established to perform activity determination analysis.Results:An HPLC fingerprint of compound embryonic bovine liver extract tablets was established.The similarities ranged from 0.963 to 1.000.The production processes of these 3 pharmaceutical manufacturers were stable.The results indicated components in each batch of compound embryonic bovine liver extract tablets were consistent for each manufacturer.However,there were differences among the 9 different batches in the activities as determined by L-02 cells/CCK-8 method.As showed in the results,the cell repair rate were from 49%to 147%.There were significant differences in productions in the 3 pharmaceutical manufacturers.Conclusion:The established fingerprint method is specific,efficient and rapid,which could be used as a reference for the inter batch consistency of compound embryonic bovine liver extract tablets.The L-02 cells/CCK-8 method used to measure the cell repair rate significantly improved the scientificity and index passivation,which provided a reference for the quality evaluation of compound embryonic bovine liver extract tablets.

关 键 词：HPLC指纹图谱 L-02细胞/CCK-8测定法 相似度分析 细胞修复率 多组分生化药 复方牛胎肝提取物片 质量评价 

分 类 号：R917[医药卫生—药物分析学]