麝香保心丸中蟾酥的质量控制标准提升  

Improvement of Quality Control Standard of Bufonis Venenum in Shexiang Baoxin Pills

作　　者：石莉尧 房蕴歌 陈两绵[2] 王瑞 王智民[2] 高慧敏[2] 姜鹏 詹常森 SHI Li-yao;FANG Yun-ge;CHEN Liang-mian;WANG Rui;WANG Zhi-min;GAO Hui-min;JIANG Peng;ZHAN Chang-sen(Shanxi University of Chinese Medicine,Jinzhong 030619,China;National Engineering Laboratory for Quality Control Technology of Chinese Herbal Medicines,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Shanghai Hutchison Pharmaceuticals Co.Ltd.,Shanghai 200201,China)

机构地区：[1]山西中医药大学,山西晋中030619 [2]中国中医科学院中药研究所中药质量控制技术国家工程实验室,北京100700 [3]上海和黄药业有限公司,上海200201

出　　处：《中国实验方剂学杂志》2022年第2期159-165,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81773897);中央级公益性科研院所基本科研业务费专项(QZPT004)。

摘　　要：目的:建立麝香保心丸中蟾毒灵、华蟾酥毒基和脂蟾毒配基含量测定的一测多评法,为完善麝香保心丸国家标准提供检测方法。方法:采用高效液相色谱法(HPLC)建立麝香保心丸中3种蟾毒配基含量的同步测定方法,并进行方法学验证,色谱条件为Nucleosil 100-5 C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相乙腈-0.1%磷酸二氢钾水溶液(磷酸调节pH 3.2)(48∶52),流速0.6 mL·min^(-1),检测波长296 nm,柱温35℃;以华蟾酥毒基为内参物,建立其对蟾毒灵和脂蟾毒配基的相对校正因子,并对相对校正因子的关键影响因素进行考察,采用相对保留时间进行待测色谱峰定位,结合在线紫外光谱和超高效液相色谱-四级杆飞行时间质谱法(UPLC-QTOF/MS)提供的精确相对分子质量辅助识别待测色谱峰;采用外标法验证一测多评法测定结果的准确性。结果:建立了麝香保心丸中3种蟾毒配基的一测多评质量评价模式,华蟾酥毒基对蟾毒灵和脂蟾毒配基的相对校正因子分别为0.922和1.01;11批样品中三者总量范围33.7~36.0μg/丸,外标法和一测多评法测定结果无显著性差异。结论:建立的方法可用于麝香保心丸中蟾毒灵、华蟾酥毒基和脂蟾毒配基的质量控制,建议麝香保心丸国家标准含量测定项下蟾酥药材的质量控制新增蟾毒灵,并以这3种成分总量制定含量限度值。Objective:To develop a quantitative analysis of multi-components by single marker(QAMS)for determination of bufalin,cinobufagin and resibufogenin in Shexiang Baoxin pills,and to provide a method for improving the national standard of the pills.Method:High performance liquid chromatography(HPLC)was developed for simultaneous determination of bufalin,cinobufagin and resibufogenin in Shexiang Baoxin pills and the methodology validation was carried out.The chromatographic separation was performed on a Nucleosil 100-5 C^(18) column(4.6 mm×250 mm,5μm)with the mobile phase of acetonitrile-0.1%potassium dihydrogen phosphate aqueous solution(pH adjusted to 3.2 with phosphoric acid)(48∶52),and the flow rate was 0.6 mL·min^(-1),the detection wavelength was set at 296 nm and the column temperature was 35℃.Taking cinobufagin as the internal standard,the relative correction factors(RCFs)of bufalin and resibufogenin were calculated,and the key influencing factors of RCFs were investigated.Relative retention time was used for the chromatographic peak location of the analyte,combining with the on-line ultraviolet spectroscopy and accurate relative molecular weight obtained by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS).The external standard method was used to verify the contents of three components obtained by QAMS.Result:QAMS was established for the determination of bufalin,cinobufagin and resibufogenin in the samples,and RCFs of cinobufagin to bufalin and resibufogenin were 0.922 and 1.01,respectively.The total content of the three marker compounds in 11 batches of Shexiang Baoxin pills was 33.7-36.0μg per pill.There was no significant difference between the quantitative results of QAMS and external standard method.Conclusion:The established method can be used for the quality control of bufalin,cinobufagin and resibufogenin in Shexiang Baoxin pills.It is suggested that bufalin should be considered as one of three marker compounds,and the sum of bufalin,cinobufagin an

关 键 词：蟾酥 麝香保心丸 华蟾酥毒基 蟾毒灵 脂蟾毒配基 一测多评法 高效液相色谱法(HPLC) 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学]