正相高效液相色谱法测定IMH中氯代烷烃类遗传毒性杂质氯代物  

Determination of chloroalkane genotoxic impurities chloride in new drug IMH by NP-HPLC

作　　者：王越 吴松 邵会会 张文轩 张坤 杨庆云 WANG Yue;WU Song;SHAO Hui-hui;ZHANG Wen-xuan;ZHANG Kun;YANG Qing-yun(State Key Laboratory of Bioactive Substance and Function of Natural Medicines,Institute of Materia Medica,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100050,China)

机构地区：[1]中国医学科学院&北京协和医学院药物研究所天然药物活性物质与功能国家重点实验室,北京100050

出　　处：《药物分析杂志》2022年第4期653-659,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：中国医学科学院医学与健康科技创新工程(2016-12M-3-010)。

摘　　要：目的:建立抗脂肪肝治疗药4,4’-二甲氧基-5,6,5’,6’-双(亚甲二氧基)-2’-吗啉亚甲基联苯-2-甲酸甲酯甲磺酸盐(简称IMH)中氯代烷烃类遗传毒性杂质4,4’-二甲氧基-5,6,5’,6’-双(亚甲二氧基)-2’-氯甲基-2-甲酸甲酯(简称氯代物)的分析方法。方法:采用正相高效液相色谱(NP-HPLC)法,选用Phenomenex Luna■Silica色谱柱(250 mm×4.6 mm,5μm),以正己烷-异丙醇(95∶5)为流动相,流速0.8 mL·min^(-1),检测波长226 nm,柱温35℃,进样量50μL;采用外标法对氯代物进行定量。结果:在水相中易分解的氯代物在该正相色谱方法条件下能够达到有效分离,专属性良好;氯代物质量浓度在0.03~3.0μg·mL^(-1);的范围内呈现良好的线性关系,检测限和定量限分别为0.15 ng和0.31 ng;平均回收率处于99%~103%,精密度及溶液稳定性良好。3批实测样品中均未检出氯代物,表明后续工艺能有效清除本品中的氯代物。结论:该色谱方法快速、准确、灵敏,能够对IMH中的氯代物进行准确的定性定量分析,有效控制药物安全。同时,可为含有卤代烷烃类遗传毒性杂质的其他药物的质量控制提供参考方法。Objective:To establish an analytical method for 4,4’-dimethoxy-5,6,5’,6’-bis(methylenedioxy)-2’-chloromethyl-2-methyl formate(chloride)that was genotoxic impurity of chloroalkanes in 4,4’-dimethoxy-5,6,5’,6’-bis(methylenedioxy)-2’-morpholine methylbiphenyl-2-methyl formate methanesulfonate(IMH),a new drug for the treatment of fatty liver disease.Methods:The NP-HPLC method was carried out using a Phenomenex Luna■Silica column(250 mm×4.6 mm,5μm)maintained at 35℃.The mobile phase was a mixture of n-hexane and isopropanol in a constant proportion of 95∶5 at a flow rate of 0.8 mL·min^(-1).The detection wavelength was 226 nm,and the injection volume was set at 50μL.Chloride was quantified by external standard method.Results:Chloride that was easy to decompose in aqueous phase could be effectively separated under the condition of NP-HPLC,and the specificity was good.The linear relationship of chloride was good in the concentration ranges of 0.03-3.0μg·mL^(-1);,and the detection limit and quantitative limit were 0.15 ng and 0.31 ng,respectively.The average recovery was 99%-103%,and the precision and solution stability were good.The chlorides in 3 batches of IMH samples were below the detection limit,thereby indicating that the subsequent process could effectively remove chloride in IMH.Conclusion:The chromatographic method is rapid,accurate and sensitive,which can accurately analyze chloride in IMH,and effectively control the drug safety.At the same time,it can provide a reference method for the quality control of other drugs containing haloalkane genotoxic impurities.

关 键 词：抗脂肪肝药物 氯代物 4 4’-二甲氧基-5 6 5’ 6’-双(亚甲二氧基)-2’-吗啉亚甲基联苯-2-甲酸甲酯甲磺酸盐 4 4’-二甲氧基-5 6 5’ 6’-双(亚甲二氧基)-2’-氯甲基-2-甲酸甲酯 质量控制 正相高效液相色谱法 “警示结构”片段 潜在遗传毒性杂质 

分 类 号：R917[医药卫生—药物分析学]