盐酸普萘洛尔原料药及片剂有关物质检测方法的建立及未知杂质β-同分异构体的研究分析

Establishment of detection method for related substances of propranolol hydrochloride and study of an unknown impurity β-isomer

目的:建立新的盐酸普萘洛尔有关物质高效液相色谱分析方法,对《中华人民共和国药典》2020年版(二部)盐酸普萘洛尔原料药及片剂质量标准中有关物质检查方法进行改良,并对药品质量进行评价。方法:采用新型碱性色谱系统替代混合离子对方法,选择CERI色谱柱L-column3 C_(18),(250 mm×4.6 mm,5μm),筛选优化了缓冲液pH和有机相比例,在更为灵敏的230 nm波长进行检测,对新建方法进行方法学验证,并开展杂质分析,以及未知杂质的结构确定。结果:该方法不仅可对3种已知杂质(杂质A、B、C)进行有效分离,还解决了药典方法中α-萘酚峰与盐酸普萘洛尔峰重合的问题。在部分样品中发现的一较大的未知杂质,经分析确证并综合结构和合成工艺,确定该物质为盐酸普萘洛尔的β-同分异构体,是盐酸普萘洛尔的合成底物α-萘酚中混有β-萘酚而产生的合成副产物,目前各国药典所采用的方法均不能检出此杂质。结论:所建方法灵敏度高,专属性强,操作简便,实验结果准确可靠,能有效地测定盐酸普萘洛尔及其制剂中有关物质组成和含量,为该系列质量标准的修订提供数据参考。

Objective:To establish a new high performance liquid chromatography method for the determination of related substances of propranolol hydrochloride,to improve the related substance detection method in the quality standard of propranolol hydrochloride in the 2020 edition of the “Chinese Pharmacopoeia”(Part Two),and to evaluate drug quality. Methods:A new alkaline chromatography column system was used instead of the mixed ion pair method. CERI L-column3 C_(18)(250 mm×4.6 mm,5 μm) was chosen. The buffer pH and organic phase were optimized and 230 nm wavelength which was more sensitive than that of previous method was used. The new method was validated and used to analyze the impurities and determine the structure of the unknown impurities. Results:This method could not only effectively separate the three known impurities(impurity A,B and C),but also solve the overlapping problem of the α-naphthol peak and propranolol peak in the pharmacopoeia methods. A large unknown impurity found in some samples was confirmed with comprehensive structure and synthesis process analysis. This substance was determined to be the isomer of propranolol hydrochloride(named β-isoform). It was synthetic by-products produced by β-naphthol within the synthetic substrate of propranolol hydrochloride-α-naphthol. This β-isoform could not be detected by the methods currently used in pharmacopoeias from various countries. Conclusion:The improved method is sensitive,specific,simple and accurate. It can effectively investigate the composition and content of related substances of propranolol hydrochloride. It provides data reference for the revision of its quality standards.