HPLC-MS/MS分析阿哌沙班中基因毒性杂质及基质效应消除研究

Study on Genotoxic Impurities and Elimination of Matrix Effect in Apixaban by HPLC-MS/MS

关键词:  阿哌沙班  基因毒性杂质  基质效应  液相色谱-质谱联用法

Key words:  apixaban  genotoxic impurities  matrix effect  HPLC-MS/MS

摘要:

目的 建立HPLC-MS/MS测定阿哌沙班中基因毒性杂质胺化物、开链酰胺和肼基物的方法。方法 色谱柱为Zobax Eclipse XDB C₁₈(3.0 mm×150 mm,3.5μm);胺化物和开链酰胺测定采用梯度洗脱法,流动相A为10 mmol·L^-1乙酸铵溶液,流动相B为乙腈;肼基物测定采用等度法,流动相为水-甲醇(30∶70)。电喷雾离子化法在正离子模式下进行多重反应离子监测结构确认。结果 胺化物、开链酰胺和肼基物平均回收率分别为99.6%,92.2%和97.5%,RSD分别为3.8%,1.9%和7.2%(n=9);定量限分别为0.05,0.05,0.05 ng·mL^-1。结论 该方法有效地消除了分析中出现的基质效应,可用于阿哌沙班中基因毒性杂质的定量检测。

Abstract:

OBJECTIVE To establish an HPLC-MS/MS method for the determination of amines,open-chain amides and hydrazine based compound in apixaban.METHODS Separation was performed on a Zobax Eclipse XDB C₁₈(3.0 mm×150 mm,3.5μm).Amines and open-chain amides were separated with the mobile phase of 10 mmol·L^-1 ammonium acetate solution and acetonitrile in gradient elution.Hydrazine based compound was separated with the mobile phase of water-methanol(30:70) in isocratic elution.The electrospray ionization source with positive ion mode was used for structure confirmation in the multiple reaction monitoring(MRM) mode.RESULTS The average recoveries for the determination of amines,open-chain amides and hydrazine based compounds were 99.6%,92.2%and 97.5%,RSDs were 3.8%,1.9%and 7.2%(n=9),respectively.Limits of quantitation were 0.05,0.05 and 0.05 ng·mL^-1.CONCLUSION The method can effectively eliminate the matrix effect and can be used for the quantitative determination of amines,open-chain amides and hydrazine based compound in apixaban.