HPLC法检测醋氯芬酸有关物质及主要杂质鉴别  

HPLC determination of related substances and identification of main impurity in aceclofenac

作　　者：侯晓蓉[1] 张焕鑫 叶永臻 任立琴 夏春年[1] 单伟光[1] 李成平[1,3] HOU Xiao-rong;ZHANG Huan-xin;YE Yong-zhen;REN Li-qin;XIA Chun-nian;SHAN Wei-guang;LI Cheng-ping(College of Pharmaceutical Science,Zhejiang University of Technology,Hangzhou 310014,China;Taizhou GreenChem Pharma-Tech Co.,Ltd.,Taizhou 225300,China;College of Biology and Environmental Engineering,Zhejiang Shuren University,Hangzhou 310015,China)

机构地区：[1]浙江工业大学药学院,杭州310014 [2]泰州葛林美克医药科技有限公司,泰州225300 [3]浙江树人大学生物与环境工程学院,杭州310015

出　　处：《药物分析杂志》2018年第7期1217-1225,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：科技部中国-白俄罗斯政府间科技合作项目(CB11-3)

摘　　要：目的:建立HPLC法检测醋氯芬酸有关物质,并采用NMR、MS法对主要杂质进行结构鉴定。方法:采用Shim-pack VP-ODS(250 mm×4.6 mm,5μm)色谱柱,以1.12 mg·m L^(-1)磷酸溶液(用1.0 mol·L^(-1)氢氧化钠溶液调节p H至7.0)为流动相A,乙腈为流动相B,梯度洗脱,流速1.0 m L·min^(-1),检测波长276 nm,柱温25℃。用半制备液相色谱分离纯化主要杂质,采用Sun Fire C_(18)色谱柱,0.50 mg·m L^(-1)磷酸溶液-甲醇为流动相梯度洗脱,通过NMR、ESI-MS确定结构。结果:主要杂质为双氯芬酸乙酯、醋氯芬酸乙酯、醋氯芬酸叔丁酯。建立的HPLC有关物质分析方法专属性强,各杂质在1.6~17.6μg·m L^(-1)范围内呈良好的线性关系,回收率在92%~105%之间,准确度高。对5批醋氯芬酸样品进行检测,结果符合规定。结论:建立的方法可作为醋氯芬酸原料药的质控方法。Objective：To establish an HPLC method for determination of related substances in aceclofenac and to identify the structure of the main impurity by NMR and MS.Methods：A Shim-pack VP-ODS（250 mm×4.6mm,5μm）was adopted for the study.The mobile phase A was 1.12 mg·m L^（-1） phosphoric acid solution（adjusted to p H 7.0 using 1.0 mol·L^（-1） sodium hydroxide solution）,and the mobile phase B was acetonitrile with gradient elution at the flow rate of 1.0 m L·min^（-1）,the detection wavelength was 276 nm,and the column temperature was25℃.The main impurity was separated and purified by semi-preparative HPLC.The determination was performed on a Sun Fire C_（18） column and using 0.50 mg·m L^（-1） phosphoric acid solution-methanol as the mobile phase with gradient elution.The structure was confirmed by NMR and ESI-MS.Results：The main impurity was ethyl ester of diclofenac,ethyl ester of aceclofenac and benzeneacetic acid,respectively.The established method for the determination of related substances in aceclofenac was specific,and the impurities showed a good linearity in the range of 1.6-17.6μg·m L^（-1）.The recovery rates were 92%-105%.The results of 5 batches of aceclofenac sample met with the requirement.Conclusion：The established HPLC method can be applied in the quality control of the aceclofenac bulk drug.

关 键 词：醋氯芬酸 抗炎镇痛药物 有关物质 杂质结构鉴定 高效液相色谱 核磁共振波谱 质谱 

分 类 号：R917[医药卫生—药物分析学]