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.

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