HPLC法测定氯法齐明原料药中有关物质的含量  

Determination of relevant substances in Clofazimine active pharmaceutical ingredients by HPLC method

作　　者：王文涛 高彦红 张升盛 王二兵[3] 赵正保 曲婷丽[1] WANG Wentao;GAO Yanhong;ZHANG Shengsheng;WANG Erbing;ZHAO Zhengbao;QU Tingli(School of Pharmacy,Shanxi Medical University,Taiyuan 030001,China;Shanxi Liye Pharmaceutical Company Limited;College of Chemical Engineering and Technology,Taiyuan University of Science and Technology)

机构地区：[1]山西医科大学药学院,太原030001 [2]山西立业制药有限公司 [3]太原科技大学化学工程与技术学院

出　　处：《山西医科大学学报》2024年第9期1207-1213,共7页Journal of Shanxi Medical University

基　　金：山西省重点研发项目(202102130501023);太原市科技局“双百攻关行动”首批关键核心技术攻关“揭榜挂帅”项目(2024TYJB0147);山西省技术创新中心项目(202104010911006);山西省科技厅基础研究项目(20210302124586)。

摘　　要：目的 建立测定氯法齐明原料药中有关物质的含量测定方法。方法 采用高效液相色谱(high performance liquid chromatography,HPLC)法,以Caprisil C8-P(250 mm×4.6 mm,5μm)为色谱柱;取十二烷基硫酸钠4.5 g,四丁基硫酸氢铵1.7 g和十二水合磷酸氢二钠1.77 g,加水1 000 mL溶解,用稀磷酸调节pH值至3.0,取上述溶液与乙腈以35:65混合作为流动相;流速1.0 mL/min;检测波长280 nm;柱温30℃;进样量20μL。对建立的HPLC方法进行方法学考察,对3批氯法齐明原料药进行有关物质检测,并比较外标法与加校正因子的主成分自身对照法测得结果。结果 氯法齐明、杂质A和杂质B分别在0.019~0.959μg/mL、0.017~0.858μg/mL和0.060~2.995μg/mL范围内线性关系良好(均R2>0.999);检测限分别为0.22 ng、0.27 ng、0.23 ng,定量限分别为0.72 ng、0.91 ng、0.76 ng;精密度、重复性、稳定性、耐用性实验的RSD值均小于5%,平均回收率为93.37%~98.17%。3批氯法齐明原料中均检测出杂质A和杂质B,杂质A的含量范围为0.013%~0.014%;杂质B的含量范围为0.025%~0.026%。加校正因子的主成分自身对照法与外标法测得的有关物质的含量无显著差异。结论 建立的HPLC方法便捷、高效、准确度高,可用于氯法齐明原料药中有关物质的含量测定,且加校正因子的主成分自身对照法成本低,实用性强。Objective To establish a method for determining the contents of substances in Clofazimine active pharmaceutical ingredient(API).Methods High performance liquid chromatography(HPLC)was performed on a Caprisil C8-P(250 mm×4.6 mm,5μm)column.And 4.5 g of sodium dodecyl sulfate,1.7 g of tetrabutylammonium hydrogensulfate and 1.77 g of disodium hydrogen phosphate dodecahydrate were dissolved in 1000 mL of water,then adjusted to pH=3.0 with dilute phosphoric acid,and then the above solution was mixed with acetonitrile in the ratio of 35∶65 as the mobile phase.The flow rate was 1.0 mL/min,the detection wavelength was 280 nm,the column temperature was 30℃,and the injection volume was 20μL.Methodological validation of this HPLC method was carried out.The contents of relevant substances in three batches of Chlorpheniramine API were determined,and the results measured by the external standard method were compared with the principal component self-control method with correction factor.Results There was a good linear relationship for Clofazimine,impurity A and impurity B in the range of 0.019-0.959μg/mL,0.017-0.858μg/mL,and 0.060-2.995μg/mL,respectively(all R2>0.999),the limits of detection were 0.22 ng,0.27 ng,and 0.23 ng,and the limits of quantification were 0.72 ng,0.91 ng,and 0.76 ng,respectively.The RSDs of precision,repeatability,stability and durability experi-ments were all less than 5%,and the average recoveries were 93.37%-98.17%.Impurity A and impurity B were detected in all three batches of Clofazimine raw material,with the content range of 0.013%-0.014%for impurity A,and 0.025%-0.026%for impurity B.There was no significant difference in the contents of related substances determined by the principal components self-control method with correction factor and the external standard method.Conclusion The established HPLC method is convenient,efficient and accurate,and can be used for determining the contents of the relevant substances in Clofazimine API.The principal component self-control method with correcti

关 键 词：高效液相色谱法 氯法齐明原料药 有关物质 含量测定 方法学验证 加校正因子的主成分自身对照法 

分 类 号：R927[医药卫生—药学]