LC-MS/MS法测定盐酸法舒地尔中2种基因毒性杂质的含量

Simultaneous determination of the contents of two genotoxic impurities in fashudil hydrochloride by LC-MS/MS

目的:建立液相色谱-质谱联用法(LC-MS/MS)测定盐酸法舒地尔中2种基因毒性杂质5-异喹啉磺酸甲酯(杂质A)和5-异喹啉磺酸乙酯(杂质C)的含量测定方法,并进行方法学验证。方法:采用LCMS/MS方法,使用Waters Sunfire C_(18)(100 mm×3 mm,3.5μm)色谱柱,以0.1%甲酸水-乙腈溶液为流动相,梯度洗脱,流速0.5 mL·min^(-1),柱温30℃,进样量10μL。采用电子喷雾源ESI,正离子模式采集分析。结果:杂质A和杂质C在线性范围内线性关系良好。杂质A和杂志C的检测限分别为0.2和0.02 ng·mL^(-1),平均回收率均在93.0%~98.0%之间,RSD<4.67%(n=9)。准确度、稳定性和耐用性的RSD均<10.0%。3批盐酸法舒地尔中均未检测到杂质A和杂质C。结论:所建立的方法快捷、准确且重复性好,可用于盐酸法舒地尔中杂质A和杂质C基因毒性杂质的控制。

Objective:To establish a liquid chromatography-tandem mass spectrometry method(LC-MS/MS method)for determination the contents of 5-methyl isoquinoline sulfonic acid(impurity A)and 5-ethyl isoquinoline sulfonic acid(impurity C)which are genotoxic impurities in fashudil hydrochloride.Methods:The LC-MS/MS method was achieved by a Waters Symmetry C_(18)(100 mm×3 mm,3.5μm)column utilizing a mobile phase of 0.1%formic acid aqueous solution-acetonitrile with gradient elution at a flow rate of 0.5 mL·min^(-1).The column temperature was set at 30℃,the injection volume was 10μL.The electrospray was operated with the source ESI under positive ionization scanning mode,and data were collected and analyzed.Results:Standard curves of impurity A and impurity C have a good linear relationship in the linear range.The limits of detection were 0.2 and 0.02 ng·mL^(-1),respectively,and the average recovery range was 93.0%~98.0%,RSD<4.67%(n=9).The RSDs of precision,accuracy,stability and durability tests were all less than 10.0%.Impurities A and C in the 3 batches of fashudil hydrochloride were not detected.Conclusion:The established method was fast,accurate and reproducible,which could be used for the control of genotoxic impurities A and C in fashudil hydrochloride.