UPLC-MS/MS测定酒石酸伐尼克兰中基因毒性杂质N-亚硝基伐尼克兰

Determination of Genotoxic Impurity N-nitrosovarenicline in Varenicline Tartrate by UPLC-MS/MS

关键词:  酒石酸伐尼克兰  基因毒性杂质  含量测定  N-亚硝基伐尼克兰  超高效液相色谱-串联三重四级杆质谱

Key words:  varenicline tartrate  genotoxic impurity  assay  N-nitrosovarenicline  UPLC-MS/MS

摘要:

目的 建立酒石酸伐尼克兰原料药和片剂中基因毒性杂质N-亚硝基伐尼克兰超高效液相色谱-串联三重四级杆质谱(UPLC-MS/MS)的检测方法。方法 采用ACQUITY UPLC^® CSH^TM Phenyl-Hexyl(150 mm×3.0 mm,1.7 μm)色谱柱;0.1%甲酸水溶液为流动相A,0.1%甲酸的甲醇溶液为流动相B,梯度洗脱;流速为0.45 mL·min^-1,柱温为50℃;采用ESI离子源正离子扫描,多反应监测(MRM)模式下,对基因毒性杂质进行定量检测。结果 杂质在0.10~10.04 ng·mL^-1具有良好的线性关系;原料药的低、中、高3个浓度的加样回收率(n=3)分别为103.58%(RSD=3.30%),98.65%(RSD=2.73%),92.00%(RSD=1.98%);片剂的低、中、高3个浓度的加样回收率(n=3)为91.53%(RSD=0.78%),96.76%(RSD=3.12%),93.01%(RSD=2.21%);检测限与定量限分别为0.014 ng·mL^-1和0.046 ng·mL^-1。结论 该方法灵敏度高,专属性强,可用于测定酒石酸伐尼克兰原料药和片剂中基因毒性杂质N-亚硝基伐尼克兰,为酒石酸伐尼克兰质量控制提供技术支持。

Abstract:

OBJECTIVE To eatablish a ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry(UPLC-MS/MS) method for the detection of genotoxic impurity N-nitrosovarenicline in varenicline tartrate active pharmaceutical ingredients(APIs) and tablets. METHODS The separation was performed on a ACQUITY UPLC^® CSH^TM Phenyl-Hexyl(150 mm×3.0 mm, 1.7 μm) column with the mobile phase consisting of 0.1% formic acid aqueous solution(mobile phase A) and 0.1% formic acid methanol solution(mobile phase B) by gradient elution at flow rate of 0.45 mL·min^-1 and the column temperature was 50℃. Multiple reaction monitoring(MRM) was performed to quantitatively detect genotoxicity impurities on triple quadrupole mass spectrometer equipped with ESI source in positive mode. RESULTS The impurity had a good linear relationshop in the range of 0.10-10.04 ng·mL^-1. The recoveries(n=3) in APIs of low, middle, high adding concentrations were 103.58%(RSD=3.30%), 98.65%(RSD=2.73%), 92.00%(RSD=1.98%), respectively. The recoveries(n=3) in tablets of low, middle, high adding concentrations were 91.53%(RSD=0.78%), 96.76%(RSD=3.12%), 93.01%(RSD=2.21%), respectively. The limit of detection and the limit of quantitation were 0.014 ng·mL^-1, 0.046 ng·mL^-1, respectively. CONCLUSION The method is sensitive, accurate, which is applicable for quantifications of genotoxic impurity N-nitrosovarenicline in varenicline tartrate APIs and tablets, providing technical support for the quality control of varenicline tartrate.