HPLC法测定阿哌沙班及其12个有机杂质  

Determination of apixaban and its 12 organic impurities by HPLC

作　　者：金云 王俊 张显华 张桐露 陈应芝 赵龙山[1] JIN Yun;WANG Jun;ZHANG Xian-hua;ZHANG Tong-lu;CHEN Ying-zhi;ZHAO Long-shan(College of Pharmaceutical Scinces,Shenyang Pharmaceutical University,Shenyang 110016,China;Zhejiang Huahai Pharmaceuticals Co.,Ltd.,Linhai 317024,China)

机构地区：[1]沈阳药科大学药学院,沈阳110016 [2]浙江华海药业股份有限公司,临海317024

出　　处：《药物分析杂志》2024年第12期2088-2094,共7页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立HPLC法检测阿哌沙班原料药中的有关物质。方法:采用ACE Excel3 C_(18)-PFP(150 mm×4.6 mm,3μm)色谱柱,以缓冲液(30 mmol·L^(-1)乙酸铵溶液)-乙腈(90∶10)为流动相A,缓冲液(30 mmol·L^(-1)乙酸铵溶液)-乙腈(5∶95)为流动相B,梯度洗脱,流速1.2 mL·min^(-1),检测波长280 nm,柱温40℃,进样量10μL,样品温度5℃。结果:阿哌沙班与各杂质及降解产物能够完全分离(分离度>2.0),供试品溶液在48 h内稳定性良好;阿哌沙班和甲酯化物、乙酯化物、氯代杂质、脱氢杂质、双水解杂质、开环甲酯化物、环合物、杂质D、水解杂质、开环酸杂质、开环酰胺杂质、5-氯己酰氯衍生杂质的定量限均为0.05%;阿哌沙班和甲酯化物、乙酯化物、水解杂质、开环酸杂质、开环酰胺杂质、5-氯己酰氯衍生杂质线性相关系数均>0.99,范围为杂质含量的LOQ到指标浓度的150%;甲酯化物、乙酯化物、水解杂质、开环酸杂质、开环酰胺杂质、5-氯己酰氯衍生杂质平均回收率(RSD)(n=9)分别为102.0%(2.7%)、106.4%(2.2%)、111.2%(4.0%)、104.4%(2.9%)、102.9%(2.7%)、101.8%(2.9%);重复性和中间精密度符合规定。经检测,3批阿哌沙班原料药6个月加速稳定性各个杂质结果均符合质量标准。结论:该方法简便快速,灵敏度高,专属性强,可用于阿哌沙班原料药有关物质的测定。Objective:To establish an HPLC method for determination of related substances in apixaban API.Methods:The analytical column was an ACE Excel3 C_(18)-PFP(150 mm×4.6 mm,3μm).The mobile phase A was buffer(30 mmol·L^(-1)ammonium acetate in water)-acetonitrile(90∶10)and the mobile phase B was buffer(30 mmol·L^(-1)ammonium acetate in water)-acetonitrile(5∶95).The whole run carried out by gradient elution at a flow rate of 1.2 mL·min^(-1).The detection wavelength was set at 280 nm,the column temperature was 40℃and the injection volume was 10μL.Results:Apixaban was separated completely from the impurities and degradation products(the resolution>2.0).The test solution was stable for at least 48 h.The LOQs of apixaban,methyl ester product,ethyl ester product,chlorine impurity,dehydrogenation impurity,bihydrolytic impurity,ringopen methyl ester product,cyclate,impurity D,hydrolytic impurity,ringopen acid impurity,ringopen amide impurity and 5-chlorhexyl chloride derived impurity,were all 0.05%.The linear correlation coefficients of apixaban,methyl ester product,ethyl ester product,hydrolytic impurity,ringopen acid impurity,ringopen amide impurity and 5-chlorhexyl chloride derived impurity were all more than 0.99.The range were from the LOQ for impurity content to 150%of the target concentration.The average recoveries(RSD)(n=9)of methyl ester product,ethyl ester product,hydrolytic impurity,ringopen acid impurity,ringopen amide impurity and 5-chlorhexyl chloride derived impurity were 102.0%(2.7%),106.4%(2.2%),111.2%(4.0%),104.4%(2.9%),102.9%(2.7%),101.8%(2.9%).The repeatability and intermediate precision completely met the requirements.The impurities contents in three batches of apixaban API 6 months accelerate stability test completely met the requirements,respectively.Conclusion:This method is simple,rapid,sensitive and specific to be used for the determination of related substances in apixaban API.

关 键 词：阿哌沙班 阿哌沙班原料药 有关物质 高效液相色谱 质量控制 分析方法 分离 

分 类 号：R917[医药卫生—药物分析学]