盐酸小檗碱原料药及其片剂杂质鉴定及分析方法建立

The structure identification and analytical procedure establishment of impurities in berberine hydrochloride and berberine hydrochloride tablets

目的:探讨植物来源盐酸小檗碱及其片剂主要杂质并鉴定未知杂质化学结构,建立有关物质分析方法,为《中国药典》2020年版盐酸小檗碱及盐酸小檗碱片有关物质分析方法的修订/增加提供参考。方法:采用LC-MS对盐酸小檗碱中主要未知杂质进行鉴定,优化《中国药典》2020年版盐酸小檗碱有关物质项下分析方法,并进行验证,同时测定多批次原料药及片剂有关物质的含量。结果:除《中国药典》2020年版盐酸小檗碱有关物质项下规定的已知杂质盐酸药根碱、盐酸巴马汀外,首次鉴定出其余3个含量超出鉴定限(0.1%)的未知杂质去亚甲基小檗碱、小檗红碱、芬氏唐松草定碱;建立可同时测定去亚甲基小檗碱、小檗红碱、芬氏唐松草定碱、盐酸药根碱、盐酸巴马汀的高效液相色谱法,经方法学验证,系统适用性、专属性、线性、检测限、定量限、重复性、中间精密度、准确度、耐用性均符合要求,并完成多批次样品检测。结论:在现行《中国药典》2020年版盐酸小檗碱的有关物质测定方法及限度要求基础上,优化色谱条件,同时增加对已知杂质的控制限度,更好地对盐酸小檗碱及盐酸小檗碱片进行质量控制。

Objective:To explore the impurities of berberine hydrochloride extracted from plant and its tablets,identify the structure of unknown impurity,establish the analytical procedure of related substances and provide reference for the revision and addition of the related substances of berberine hydrochloride and berberine hydrochloride tablets in the Chinese Pharmacopoeia 2020.Methods:The major unknown impurities in berberine hydrochloride were identified by LC-MS,and the analytical procedure of related substances of berberine hydrochloride in the Chinese Pharmacopoeia 2020 was optimized and verified.The related substances of multiple batches of active pharmaceutical ingredients and its tablets were determined.Results:In addition to the known impurities of jatrorrhizine hydrochloride and palmatine hydrochloride listed in the related substances of berberine hydrochloride in the Chinese Pharmacopoeia 2020,three unknown impurities of demethyleneberberine,berberrubine and thalifendine beyond the identification limit(0.1%)were identified for the first time.The HPLC method was established for simultaneous determination of those impurities,and the methodology validation was made.The system suitability,specificity,linearity,limit of detection,limit of quantitation,repeatability,intermediate precision,accuracy and robustness met the requirements.The multiple batches of samples were determined.Conclusion:On the basis of the current analytical method and limit requirements of related substances of berberine hydrochloride in the Chinese Pharmacopoeia 2020,the chromatographic condition is optimized and the control limit of known impurities is set to better control the quality of berberine hydrochloride and berberine hydrochloride tablets.