UPLC-MS/MS法测定奥美沙坦酯中7个亚硝胺类基因毒性杂质

Determination of seven N-nitrosamines genotoxic impurities in olmesartan medoxomil by UPLC-MS/MS

目的建立超高效液相色谱-串联质谱法(UPLC-MS/MS)测定奥美沙坦酯中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 poroshell PFP(100 mm×2.1 mm,2.7μm)色谱柱;流动相为0.1%甲酸水溶液(A)-甲醇(B)梯度洗脱;体积流量0.4 mL/min,柱温40℃;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。结果各杂质质量浓度在1~100 ng/mL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)为83%~117%,RSD值为0.8%~4.1%,平均加样回收率为87%~106%;检测限范围为0.02~0.19 ng/mL,定量限为0.06~0.65 ng/mL。4批奥美沙坦酯样品中均未检出杂质。结论该方法灵敏度高,专属性强,可用于测定奥美沙坦酯原料药中7个亚硝胺类杂质,为奥美沙坦酯的质量控制提供参考。

Objective To establish a 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-nitroso-diethylamine,N-nitroso-ethylisoprosylamine,N-nitroso-diisopropylamine,N-nitroso-dipropylamine and N-nitroso-dibutylamine)in olmesartan medoxomil.Methods The separation was performed on an Agilent poroshell PFP(100 mm×2.1 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 0.4 mL/min and the column temperature was 40℃.Multiple reaction monitoring(MRM)was performed on a triple quadripole mass spectrometer equipped with a APCI source in positive mode to quantitative detection of seven genotoxic impurities.Results The calibration curve was linear for seven compounds in the range of 1—100 ng/mL,r>0.995.The recoveries(n=3)of low,middle,high adding concentrations were 83%—117%,RSD were 0.8%—4.1%,average spike recoveries range were 87%—106%.The limit of detection was 0.02—0.19 ng/mL,and the limit of quantification was 0.06—0.65 ng/mL.No impurities were detected in four batches samples.Conclusion The method is sensitive,accurate,which is applicable for quantifications of seven NNitrosamines genotoxic impurities in olmesartan medoxomil.The method can provided a reference for the control of nitrosaminies impurities in olmesartan medoxomil.