高效液相色谱-电雾式检测器测定牛黄解毒片中的6种胆汁酸  

Determination of Six Kinds of Bile Acids in Niuhuang Jiedu Tablets Using High Performance Liquid Chromatography Coupled with Charged Aerosol Detector

作　　者：林楚慧 柴瑞平 张璐 付志博 章弘扬 张敏 胡坪 LIN Chu-Hui;CHAI Rui-Ping;ZHANG Lu;FU Zhi-Bo;ZHANG Hong-Yang;ZHANG Min;HU Ping(Shanghai Key Laboratory of Functional Materials Chemistry,School of Chemistry and Molecular Engineering,East China University of Science and Technology,Shanghai 200237,China;Thermo Fisher Scientific China Co.Ltd.,Shanghai 200000,China;Shanghai Key Laboratory of New Drug Design,School of Pharmacy,East China University of Science and Technology,Shanghai 200237,China)

机构地区：[1]华东理工大学化学与分子工程学院,上海市功能性材料化学重点实验室,上海200237 [2]赛默飞世尔科技中国有限公司,上海200000 [3]华东理工大学药学院,上海市新药设计重点实验室,上海200237

出　　处：《分析化学》2022年第5期764-771,共8页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(No.81973285)资助。

摘　　要：建立了牛黄解毒片中6种胆汁酸类物质(甘氨胆酸、猪脱氧胆酸、胆酸、甘氨脱氧胆酸、鹅脱氧胆酸与脱氧胆酸)的高效液相色谱-电雾式检测器(HPLC-CAD)测定方法。样品经70%(V/V)甲醇超声提取,氮吹浓缩后,以Agilent Poroshell 120 EC C_(18)色谱柱(100 mm×4.6 mm,2.7μm)分离,采用甲醇和5 mmol/L乙酸铵-0.1%甲酸溶液梯度洗脱,设置电雾式检测器的雾化温度为55℃,幂律参数(PFV)为1.35。甘氨胆酸、猪脱氧胆酸、胆酸、甘氨脱氧胆酸、鹅脱氧胆酸和脱氧胆酸分别在2.10~972μg/mL、2.09~1046μg/mL、2.05~978μg/mL、2.56~1022μg/mL、1.25~658μg/mL和2.05~1025μg/mL浓度范围内线性良好,相关系数(R^(2))大于0.9991,检出限在0.86~1.53μg/mL之间,定量限在1.20~2.56μg/mL之间,平均回收率在94.2%~105.3%之间,相对标准偏差(RSD)小于1.6%。将本方法应用于实际牛黄解毒片样品的测定,结果表明,不同厂家生产的牛黄解毒片中胆汁酸的含量差异显著。建立的6种胆汁酸的HPLC-CAD含量测定方法灵敏、准确、可靠,可用于中药复方牛黄解毒片质量评价。A high performance liquid chromatography-charge aerosol detector(HPLC-CAD)method for determination of six kinds of bile acids(Glycocholicacid,hyodeoxycholic acid,cholic acid,glycodeoxycholic acid,chenodeoxycholic acid and deoxycholic acid)in Niuhuang Jiedu tablets was developed.The tablet samples were extracted by ultrasonication in 70%methanol and concentrated under nitrogen flow.The separation of bile acids was achieved on an Agilent Poroshell 120 EC C_(18) column(4.6 mm×100 mm,2.7μm)with gradient elution using methanol and aqueous solution containing 0.1%formic acid and 5 mmol/L ammonium formate.CAD evaporation temperature was set at 55℃and the power function value(PFV)was 1.35.The linearity ranges were 2.10-972μg/mL for glycocholic acid,2.09-1046μg/mL for hyodeoxycholic acid,2.05-978μg/mL for cholic acid,2.56-1022μg/mL for glycodeoxycholic acid,1.25-658μg/mL for chenodeoxycholic acid and 2.05-1025μg/mL for deoxycholic acid,respectively.All relative coefficients(R 2)were higher than 0.9991.The limits of detection and the limits of quantitation ranged from 0.86 to 1.53μg/mL and 1.20 to 2.56μg/mL,respectively.The average recoveries of six kinds of bile acids were in the range of 94.2%-105.3%with RSD≤1.63%.The method was used to determine the real tablet samples,and the results indicated that the contents of six bile acids had an obvious difference among the Niuhuang Jiedu tablets of different manufacturers.The HPLC-CAD method for the determination of six kinds of bile acids was sensitive,accurate and reliable,which could be applied to evaluate the quality of the Niuhuang Jiedu tablets.

关 键 词：牛黄解毒片 高效液相色谱 电雾式检测器 胆汁酸 

分 类 号：O657.72[理学—分析化学] R286.0[理学—化学]