黄连解毒汤6种主要成分高效液相色谱检测方法的建立和应用  

Establishment and application of main components of Huang-Lian-Jie-Du decoction based on high performance liquid chromatography

作　　者：周芳芳 谷雪妹 王微[1] 林明[1] 王蕾[1] ZHOU Fangfang;GU Xuemei;WANG Wei;LIN Ming;WANG Lei(Department of Clinical Laboratory,Shanghai Eighth People's Hospital,Shanghai 200235,China)

机构地区：[1]上海市第八人民医院检验科,上海200235

出　　处：《检验医学》2024年第5期468-473,共6页Laboratory Medicine

基　　金：上海市卫生健康委员会上海市综合医院中西医结合专项(ZHYY-ZXYJHZX-202104);上海市徐汇区医学重点学科(SHXHZDXK202322)。

摘　　要：目的建立检测黄连解毒汤中6种主要成分的高效液相色谱(HPLC)方法。方法采用Zorbax Eclipse Plus C18色谱柱(250 mm×4.6 mm,5μm),以0.03 mg·mL-1磷酸二氢钾-乙腈(V∶V=7∶3)为流动相,等度洗脱,流速为1.0 mL·min-1,检测波长为345 nm,柱温为37℃,进样量为20μL,建立检测黄连解毒汤6种主要成分(京尼平苷、小檗碱、黄连碱、药根碱、黄芩苷、巴马汀)的HPLC方法。绘制HPLC检测6种成分的标准曲线,并进行方法学评价(线性、加样回收率、准确度和精密度、稳定性)。采用建立的HPLC方法检测黄连解毒汤样本。结果HPLC检测京尼平苷、小檗碱、黄连碱、药根碱、黄芩苷、巴马汀的线性范围分别为25.00~800.00、32.00~1024.00、20.00~640.00、20.30~650.00、13.13~420.00、25.00~800.00μg·mL-1,精密度和准确度均<2%,加样回收率分别为105.30%、102.88%、103.51%、107.76%、96.29%、99.34%。稳定性实验结果显示,室温放置24 h、反复冻融3次、2~8℃保存1周、-80℃冻存1个月,京尼平苷、小檗碱、黄连碱、药根碱、黄芩苷、巴马汀的峰面积相对标准偏差(RSD)均<2%,检测结果均稳定。采用HPLC检测黄连解毒汤的主要成分,浓度从高到低依次为药根碱(2799.00μg·mL-1)、小檗碱(2358.03μg·mL-1)、京尼平苷(1793.34μg·mL-1)、巴马汀(945.30μg·mL-1)、黄连碱(192.00μg·mL-1)、黄芩苷(171.00μg·mL-1)。结论建立的同时检测黄连解毒汤6种主要成分的HPLC方法快速、简便、重复性好、准确度高,可为后续黄连解毒汤的相关研究和临床应用奠定基础。Objective To establish a high performance liquid chromatography(HPLC)for the determination of 6 main components in Huang-Lian-Jie-Du decoction.Methods The chromatography was performed on a Zorbax Eclipse Plus C18 column(250 mm×4.6 mm,5μm)with 0.03 mg·mL^(-1) potassium dihydrogen phosphate acetonitrile(V∶V=7∶3)as the mobile phase.The flow rate was 1.0 mL·min^(-1),and the determination wavelength was 345 nm.The column temperature was 37℃,and the sample size was 20μL.The HPLC was established for the determination of the 6 main components of Huang-Lian-Jie-Du decoction(genipin,berberine,coptisine,jateorhizine,baicalin and palmatine).The standard curves of 6 components determined by HPLC were drawn,and the methodological evaluation(linear evaluation,recovery rate,accuracy and precision,stability)was carried out.The established HPLC was used to determine the samples of Huang-Lian-Jie-Du decoction.Results The linear ranges of genipin,berberine,coptisine,jateorhizine,baicalin and palmatine were 25.00-800.00,32.00-1024.00,20.00-640.00,20.30-650.00,13.13-420.00 and 25.00-800.00μg·mL^(-1),respectively.The precision and accuracy were<2%.The recoveries were 105.30%,102.88%,103.51%,107.76%,96.29% and 99.34%,respectively.The stability test results showed that the peak area relative standard derivations(RSD)of genipin,berberine,coptisine,jateorhizine,baicalin and palmatine were all<2% after 24 h at room temperature,3 times frozen and thawing,2-8℃ for 1 week,and -80℃ for 1 month,and the determination results were all stable.The main components of Huang-Lian-Jie-Du decoction were determined by HPLC.The concentrations from high to low were jateorhizine(2799.00μg·mL^(-1)),berberine(2358.03μg·mL^(-1)),genipin(1793.34μg·mL^(-1)),palmatine(945.30μg·mL^(-1)),coptisine(192.00μg·mL^(-1))and baicalin(171.00μg·mL^(-1)).Conclusions The established HPLC for the determination of the main components of Huang-Lian-Jie-Du decoction is rapid,simple,reproducible and accurate,which lays a foundation for the subsequent re

关 键 词：黄连解毒汤 药根碱 小檗碱 京尼平苷 巴马汀 黄连碱 黄芩苷 高效液相色谱 

分 类 号：R446.1[医药卫生—诊断学]