HPLC-MS/MS法检测利伐沙班中痕量基因毒性杂质  

Detection of trace genotoxic impurities in rivaroxaban by HPLC-MS/MS

作　　者：李萍[1,2] 张凤妹 周萍 曹粤锋[2] 王建 LI Pingg;ZHANG Feng-mei;ZHOU Ping;CAO Yue-feng;WANG Jian(Zhejiang University of Technology,Hangzhou 310014,China;Zhejiang Institute for Food and Drug Control,Key Laboratory for Core Technology of Generic Drug Evaluation National Medical Product Administration&Key Laboratory of Drug Contacting Materials Quality Control of Zhejiang Province,Hangzhou 310051,China)

机构地区：[1]浙江工业大学,杭州310014 [2]浙江省食品药品检验研究院国家药品监督管理局仿制药评价关键技术重点实验室浙江省药品接触材料质量控制研究重点实验室,杭州310051

出　　处：《药物分析杂志》2024年第1期101-108,共8页Chinese Journal of Pharmaceutical Analysis

基　　金：浙江省药品监管系统科技计划项目(202201)。

摘　　要：目的:建立HPLC-MS/MS法检测利伐沙班中基因毒性杂质(S)-5-氯-N-(3-氯-2-羟丙基)噻吩-2-酰胺和4-(3-氧代-4-吗啉基)硝基苯的方法,系统考察影响杂质分离和质谱响应的因素。方法:采用ZORBAX SB-C_(18)(150 mm×2.1 mm, 3.5μm)色谱柱,以10 mmol·L^(-1)甲酸铵溶液-甲醇为流动相,梯度洗脱,流速0.4 mL·min^(-1);电喷雾正离子化MRM检测。结果:(S)-5-氯-N-(3-氯-2-羟丙基)噻吩-2-酰胺和4-(3-氧代-4-吗啉基)硝基苯质量浓度分别在0.008~3.4 ng·mL^(-1)和0.2~3.4 ng·mL^(-1)范围内线性关系良好,平均回收率(n=9)分别为97.9%和98.9%,RSD分别为0.94%和0.68%;定量限分别为0.008 ng·mL^(-1)和0.2 ng·mL^(-1);15批利伐沙班样品中基因毒性杂质残留量检测结果均远低于限度值。结论:该方法灵敏度高,专属性强,可有效检测和控制利伐沙班中痕量基因毒性杂质的含量。Objective:To establish an HPLC-MS/MS method for the determination of the genotoxic impurities(S)-5-chloro-N-(3-chloro-2-hydroxypropyl) thiophene-2-amide and 4-(3-oxy-4-morpholine) nitrobenzene in rivaroxaban. The factors affecting impurities separation and the response of mass spectrometry were systematically investigated. Methods: A ZORBAX SB-C_(18) column(150 mm×2.1 mm, 3.5 μm) was used with 10 mmol·L^(-1 )ammonium formate solution-methanol as mobile phase at flow rate of 0.4 mL·min^(-1). And MRM detection by electrospray positive ionization was used. Results:(S)-5-chloro-N-(3-chloro-2-hydroxypropyl) thiophene-2-amide and 4-(3-oxy-4-morpholine) nitrobenzene had good linear relationships in the range of 0.008-3.4 ng·mL^(-1) and 0.2-3.4 ng·mL^(-1), respectively. The average recoveries(n=9) were 97.9% and 98.9%, RSD were 0.94% and 0.68% respectively. The limits of quantitation were 0.008 ng·mL^(-1) and 0.2 ng·mL^(-1) respectively. The results of genotoxic impurity residue determination in 15 batches of rivaroxaban samples were far below the limit value. Conclusion: The method is sensitive and specific, and can effectively detect trace genotoxic impurities in rivaroxaban.

关 键 词：利伐沙班 基因毒性杂质 液相色谱-质谱联用法 (S)-5-氯-N-(3-氯-2-羟丙基)噻吩-2-酰胺 4-(3-氧代-4-吗啉基)硝基苯 

分 类 号：R917[医药卫生—药物分析学]