UPLC-MS/MS法测定阿加曲班中4个基因毒性杂质

目的：建立超高效液相色谱-串联质谱法（UPLC-MS/MS）同时测定阿加曲班中4个基因毒性杂质：3-甲基-8-喹啉磺酸、3-甲基-8-喹啉磺酸甲酯、3-甲基-8-喹啉磺酸乙酯、3-甲基-8-喹啉磺酰氯。方法：采用Waters ACQUITY UPLC BEH C₁₈（100 mm×2.1 mm，1.7 μm）色谱柱；以0.1%甲酸水溶液为流动相A，乙腈为流动相B进行线性梯度洗脱；流速0.2 mL·min^-1；采用ESI离子源正离子模式，多反应监测模式下，以外标法对4个基因毒性杂质同时进行定量测定。结果：除3-甲基-8-喹啉磺酰氯外，其他3个基因毒性杂质质量浓度在0.125~500 ng·mL^-1范围内线性关系良好；低、中、高3个浓度的加样回收率（n=3）范围为91.5%~104.2%，RSD范围为0.15%~4.6%；检测下限范围为0.042~1.667 ng·mL^-1，定量下限范围为0.125~5.000 ng·mL^-1。样品中均未检出杂质。结论：本方法操作简便，结果可靠，经方法学验证，可用于阿加曲班中4个基因毒性杂质的同时检测。

Objective: To establish a UPLC-MS/MS analytical method for the determination of genotoxic impurities (3-methyl-8-quinoline sulfonic acid, 3-methyl-8-methyl quinolinesulfonate, 3-methyl-8-ethyl quinolinesulfonate, and 3-methyl-8-quinolinesulphonyl chloride) in argatroban.Methods: The separation was performed on a Waters ACQUITY UPLC BEH C₁₈ (100 mm×2.1 mm, 1.7 μm) with the mobile phase consisting of 0.1% formic acid aqueous solution (mobile phase A) and acetonitrile (mobile phase B) by gradient elution at a flow rate of 0.2 mL·min^-1.Multiple reaction monitoring (MRM) was performed on a triple quadrupole mass spectrometer equipped with a ESI source in the positive mode.Results: Except for 3-methyl-8-quinolinesulphonyl chloride, the calibration curve was linear for other compounds in the range of 0.125-500 ng·mL^-1.The recoveries (n=3) of low, middle and high adding concentrations were 91.5%-104.2%, RSD were 0.15%-4.6%.The limit of detection was 0.042-1.667 ng·mL^-1, and the limit of quantification was 0.125-5.000 ng·mL^-1.No impurities were detected in the samples.Conclusion: This established method is simple, rapid and reliable, which is applicable for quantifications of four genotoxic impurities in argatroban.

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