UPLC-MS/MS法同时测定氯沙坦钾和缬沙坦中7个亚硝胺类基因毒性杂质

Determination of seven N-nitrosamines genotoxic impurities in losartan potassium and valsartan by UPLC-MS/MS

目的:建立超高效液相色谱-串联质谱法(UPLC-MS/MS)同时测定氯沙坦钾和缬沙坦中7个基因毒性杂质(N-亚硝基二甲胺、N-亚硝基-4-甲基-4-氨基丁酸、N-亚硝基二乙胺、N-亚硝基乙基异丙基胺、N-亚硝基二异丙胺、N-亚硝基二丙胺、N-亚硝基二丁胺)。方法:采用Agilent Infinity Lab Poroshell 120 SB-AQ(3.0 mm×150 mm,2.7μm)色谱柱,泵混合器与进样器之间接Agilent Poroshell 120 EC-C_(18)(4.6 mm×50 mm,2.7μm)色谱柱,以0.1%甲酸水溶液为流动相A,甲醇为流动相B进行梯度洗脱,流速0.5m L·min^(-1),柱温50℃;采用APCI离子源正离子扫描,多反应监测(MRM)模式下,对7个基因毒性杂质同时进行定量检测。结果:在氯沙坦的水溶系统中,各杂质质量浓度在1-100 ng·mL^(-1)范围内具有良好线性关系,r>0.995;低、中、高3个浓度的加样回收率(n=3)范围为79.0%-108.8%,RSD范围为0.57%-4.8%;检测下限范围为0.03-0.22 ng·mL^(-1),定量下限范围为0.09-0.76 ng·mL^(-1)。在缬沙坦的甲醇溶系统中,各杂质质量浓度在1-100 ng·mL^(-1)范围内具有良好线性关系,r>0.995;低、中、高3个浓度的加样回收率(n=3)范围为81.0%-105.7%,RSD范围为0.89%-4.1%;检测下限范围为0.05-0.29 ng·mL^(-1),定量下限范围为0.17-0.95 ng·mL^(-1)。样品中均未检出杂质。结论:该方法灵敏度高,专属性强,可同时用于测定氯沙坦钾原料药和缬沙坦原料药中7个亚硝胺类杂质,为化学药品中亚硝胺类杂质控制方法提供参考。

Objective:To establish an UPLC-MS/MS analytical method for the determination of seven N-nitrosamines genotoxic impurities(N-nitroso-dimethylamine,N-nitroso-N-methyl-4-aminobutyric acid,N-nitrosodiethylamine,N-nitroso-ethyl-isoprosylamine,N-nitroso-diisopropylamine,N-nitroso-dipropylamine and N-nitroso-dibutylamine)in losartan potassium and valsartan.Methods:The separation was performed on an Agilent InfinityLab Poroshell 120 SB-AQ(3.0 mm×150 mm,2.7μm)column with the mobile phase consisting of 0.1%formic acid aqueous solution(mobile phase A)and methanol(mobile phase B)by gradient elution at a flow rate of 0.5 mL·min^(-1) and the column temperature was 50℃.Another Agilent Poroshell 120 EC-C_(18)(4.6 mm×50 mm,2.7μm)column was installed between pump mixer and injector.Multiple reaction monitoring(MRM)was performed on a triple quadripole mass spectrometer equipped with a APCI source in positive mode.Results:In the test of losartan potassium,the calibration curve was linear for seven compounds in the range of 1-100 ng·mL^(-1),r>0.995.The recoveries(n=3)of low,middle,high adding concentrations were 79.1%-108.8%.RSDs were 0.57%-4.8%;the limit of detection was 0.03-0.22 ng·mL^(-1),and the limit of quantification was 0.09-0.76 ng·mL^(-1).In the test of valsartan,the calibration curve was linear for seven compounds in the range of 1-100 ng·mL^(-1),with r>0.995.The recoveries(n=3)of low,middle,high added amounts were 81.0%-105.7%.RSDs were 0.89%-4.1%.The limit of detection was 0.05-0.29 ng·mL^(-1),and the limit of quantification was 0.17-0.95 ng·mL^(-1).No impurities were detected in the samples.Conclusion:The method is sensitive,accurate,which is applicable for quantifications of seven N-nitrosamines genotoxic impurities in losartan potassium and valsarten.The method can provied a reference for the control of nitrosaminies impurities in chemical drugs.