高效液相色谱-质谱联用法同时测定盐酸二甲双胍肠溶片中6种亚硝胺类基因毒性杂质

Simultaneous determination of 6 nitrosamine genotoxic impurities in metformin hydrochloride enteric-coated tablets by high performance liquid chromatography-mass spectrometry

建立高效液相色谱–质谱联用法同时测定盐酸二甲双胍肠溶片中6种N-亚硝胺类基因毒性杂质(N-亚硝基二甲胺、N-亚硝基-N-甲基-4-氨基丁酸、N-亚硝基二乙胺、N-亚硝基乙基异丙胺、N-亚硝基二异丙胺和N-亚硝基二丁胺)。采用ACE C_(18)柱(150 mm×4.6 mm,5μm),流动相为0.1%甲酸水–甲醇,流量为0.6 mL/min,梯度洗脱,柱温为40℃,采用正离子多反应监测(MRM)模式,以大气压化学电离源为离子源。6种N-亚硝胺类基因毒性杂质的质量浓度在各自的线性范围内与色谱峰面积线性关系良好,相关系数均不小于0.9995。检出限和定量限分别为0.0337~0.03389 ng/mL和0.10076~0.10268 ng/mL。样品平均加标回收率为97.83%~103.33%,测定结果的相对标准偏差为1.19%~4.45%(n=9)。该方法专属性强,简单、快速、灵敏,适用于测定盐酸二甲双胍肠溶片中的N-亚硝胺类基因毒性杂质。

A high performance liquid chromatography-mass spectrometry method for the determination of 6 N-nitrosamine genotoxic impurities(N-nitrosodimethylamine,N-nitrosoethylisopropylamine,N-nitrosodiethylamine,N-nitroso-N-methyl-4-aminobutyric acid,N-nitrosodibutylamine,N-nitroso diisopropylamine)in metformin hydrochloride enteric-coated tablets simultaneously was established.The method was carried out on ACE C_(18) column(150 mm×4.6 mm,5μm).And mobile phase was 0.1%formic acid water(A)-methanol(B)with gradient elution at a flow rate of 0.6 mL/min,and the column temperature was 40℃.The positive ion multiple reaction monitoring(MRM)mode was used,and atmospheric pressure chemical ionization source is used as ion source The mass concentration of 6 N-nitrosamine genotoxic impurities had good linear relationships within their respective linear ranges(r^(2)≥0.9995).The overall recoveries were between 97.83%and 103.33%,and the RSD was 1.19%-4.45%(n=9).The LODs and LOQs were 0.0337-0.03389 ng/mL,0.10076-0.10268 ng/mL,respectively.The established method is specific,simple,rapid,and sensitive,which is suitable for the determination of N-nitrosamine genotoxic impurities in metformin hydrochloride enteric-coated tablets.