摘要
目的本研究对注射用兰索拉唑主要杂质进行了色谱条件优化与体外细胞毒性评价,建立了高灵敏、高特异性分析方法,以揭示其潜在生物安全风险。通过方法学改进与毒性验证相结合,为完善《中国药典》中兰索拉唑制剂质量控制标准提供科学依据。方法通过考察色谱条件,建立兰索拉唑主要杂质分析的新方法,并通过专属性、检测限/定量限、线性范围等指标进行方法学验证。采用MTT比色法构建L929细胞毒性评价模型,系统检测主要杂质对细胞增殖的抑制作用。最后应用建立的分析方法对多批次制剂进行有关物质含量测定和差异性分析。结果建立可同时测定兰索拉唑6种杂质的HPLC法,经方法学验证,系统适用性、专属性、线性、检测限以及定量限均符合要求,同时根据细胞毒性实验中各处理组的吸光度计算相对增殖率判断细胞分级及毒性反应程度,判定6种杂质的细胞毒性大小为Ⅱ>Ⅰ>Ⅴ>Ⅳ>Ⅵ>Ⅲ。结论与现行《中国药典》收录的兰索拉唑有关物质测定方法及限度要求相比,本方法优化了色谱条件。同时,在实际样品检测中,新方法显著提高了杂质检出率,揭示了市场上不同企业生产的注射用兰索拉唑样品在杂质含量上的差异。其中毒性最高的杂质Ⅱ含量普遍偏高,这一发现进一步凸显了对其进行严格监管的重要性。
Objective To optimize the chromatographic conditions for major impurities and conduct in vitro cytotoxicity evaluation of impurities in lansoprazole for injection,establish a highly sensitive and specific analytical method for related substances through methodological validation,and reveal its potential biosafety risks.By combinig methodological improvement and toxicity validation,this study provides scientific evidence for enhancing the quality control standards of lansoprazole preparations in the Chinese Pharmacopoeia.Methods By investigating chromatographic conditions,a novel analytical method for major impurities in lansoprazole was developed and validated through methodological evaluation of specificity,detection limit/quantitation limit,and linearity range.An L929 cytotoxicity evaluation model was established using the MTT colorimetric assay to systematically assess the inhibitory effects of major impurities on cell proliferation.The established analytical method was subsequently applied to determine related substance contents and analyze batch-to-batch variability in multiple production batches of the formulation.Results An high-performance liquid chromatography(HPLC)method for simultaneous determination of six lansoprazole impurities was established and validated for system suitability,specificity,linearity,limits of detection,and quantification.Cytotoxicity was ranked as Ⅱ>Ⅰ>Ⅴ>Ⅳ>Ⅵ>Ⅲ based on relative proliferation rates derived from absorbance measurements.Conclusion Compared with the current method and limit requirements for the determination of related substances of lansoprazole specified in the Chinese Pharmacopoeia,this method optimizes the chromatographic conditions.Furthermore,the established method significantly improved the impurity detection rate in practical sample testing and revealed differences in impurity contents among lansoprazole for injection products manufactured by different enterprises.Notably,impurity II,which exhibited the highest cytotoxicity,was found to have generally high contents in these samples,which further underscores the importance of stringent regulatory oversight.
作者
张浩
郑航
乔露
陈吉
ZHANG Hao;ZHENG Hang;QIAO Lu;CHEN Ji(Hubei Institute for Drug Control,Wuhan 430075,China)
出处
《药学前沿》
2026年第1期78-86,共9页
China Pharmacist
基金
湖北省药品监管工作政策研究课题项目(鄂药监函〔2025〕25号)。
关键词
兰索拉唑
杂质检测
高效液相色谱
细胞毒性
质量控制
方法学验证
中国药典
相对增殖率
Lansoprazole
Impurities detection
High-performance liquid chromatography
Cytotoxicity
Quality control
Method validation
Chinese Pharmacopoeia
Relative proliferation rates
