Abstract

Rabeprazole sodium is a widely used drug for gastrointestinal disorders. Several analytical methods for identifying rabeprazole sodium and its impurities have been reported. However, the genotoxicity of rabeprazole sodium and its impurities is still unclear. Thus, it is necessary to develop analytical methods that can identify the structures of its impurities and evaluate their genotoxicity. Here, we used high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry for identifying the impurities in rabeprazole sodium enteric-coated tablets. Impurities in the samples were matched with synthesized impurities based on the exact mass and secondary mass spectrometry characteristics and then subjected to in silico analysis using the Derek and Sarah software, as well as in vitro genotoxicity evaluations. Our method successfully identified the impurities as 2-[[4-(3-methoxy propane)-3-methyl-N-oxido-2-pyridyl] methyl sulfonyl]-1H-benzimidazole (impurity I), 2-[[4-(3-methoxy propane)-3-methyl-2-pyridyl]methyl sulfonyl]-benzimidazole (impurity II), 2-[[4-(3-methoxy propane)-3-methyl-2-pyridyl] methionyl]-1H-benzimidazole (impurity III), and 2-mercapto benzimidazole (impurity IV). In silico analysis predicted that impurity III demonstrated a structural alert; thus, this impurity was evaluated for in vitro genotoxicity using the Ames test and chromosomal aberration assay. Impurity III at concentrations of 7.5–30 μg/mL had an aberration rate of over 5% with or without S-9 mix. Furthermore, impurity III at concentrations of 40–1000 μg/plate significantly increased the number of mutagenic colonies with or without S-9 mix. These results indicated that impurity III should be regulated to the limit of 0.01%.

摘要

雷贝拉唑钠是一种广泛用于胃肠道疾病的药物。已经报道了几种鉴定雷贝拉唑钠及其杂质的分析方法。然而，雷贝拉唑钠及其杂质的遗传毒性仍不清楚。因此，有必要开发能够识别其杂质结构并评估其遗传毒性的分析方法。在这里，我们使用高效液相色谱-四极杆飞行时间质谱法鉴定雷贝拉唑钠肠溶片中的杂质。根据精确的质量和二级质谱特性将样品中的杂质与合成的杂质进行匹配，然后使用 Derek 和 Sarah 软件进行计算机分析以及体外分析遗传毒性评估。我们的方法成功地将杂质鉴定为2-[[4-(3-甲氧基丙烷)-3-甲基-N-氧化-2-吡啶基]甲基磺酰基]-1H-苯并咪唑(杂质I)、2-[[4- (3-甲氧基丙烷)-3-甲基-2-吡啶基]甲基磺酰基]-苯并咪唑(杂质II)，2-[[4-(3-甲氧基丙烷)-3-甲基-2-吡啶基]甲硫酰基]-1H -苯并咪唑（杂质 III）和 2-巯基苯并咪唑（杂质 IV）。计算机分析预测杂质 III 表现出结构警报；因此，该杂质在体外进行了评估使用 Ames 试验和染色体畸变试验测定遗传毒性。浓度为 7.5–30 μg/mL 的杂质 III 在有或没有 S-9 混合物的情况下具有超过 5% 的畸变率。此外，浓度为 40–1000 μg/板的杂质 III 显着增加了有或没有 S-9 混合物的诱变菌落数量。这些结果表明杂质III应控制在0.01%的限度内。