基于HPLC指纹图谱、多指标成分含量测定及化学计量学的湿热痹片质量评价  

Quality evaluation of Shirebi Tablets based on combinative methods of HPLCfingerprint,quantitative analysis of multi-components and chemometricsanalysis

作　　者：高森[1] 王苹 唐铖 白雪 文柳静 李正翔[1] GAO Sen;WANG Ping;TANG Cheng;BAI Xue;WEN Liu-jing;LI Zheng-xiang(Department of Pharmacy,General Hospital,Tianjin Medical University,Tianjin 300052,China;Tianjin Modern Innovation TCM Technology Co.,Ltd.,Tianjin 300392,China;Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics(Theranostics),College of Pharmacy,Tianjin Medical University,Tianjin 300070,China;State Key Laboratory of Chinese Medicine Key Technology Innovation,Tasly Holding Group Academy of Traditional Chinese Medicine Development Center,Tianjin 300402,China;Tianjin Key Laboratory of Cancer Prevention and Treatment,State Cancer Clinical Medical Research Center,Tianjin Medical University Cancer Hospital,Tianjin 300060,China)

机构地区：[1]天津医科大学总医院药剂科,天津300052 [2]天津现代创新中药科技有限公司,天津300392 [3]天津医科大学药学院,天津市临床药物关键技术重点实验室,天津300070 [4]天士力控股集团研究院中药开发中心,创新中药关键技术国家重点实验室,天津300402 [5]天津医科大学肿瘤医院药学部,国家肿瘤临床医学研究中心,天津市“肿瘤防治”重点实验室,天津300060

出　　处：《中草药》2020年第21期5454-5461,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(81872236);天津市自然科学基金项目(18JCYBJC95100)。

摘　　要：目的建立指纹图谱和多指标定量与化学计量学相结合的湿热痹片质量评价方法。方法采用Waters Symmetry C18色谱柱(250 mm×4.6 mm,5μm),柱温30℃;检测波长分别为303 nm(检测桑皮苷A、桑皮苷F、桑辛素M)和270 nm(检测连翘酯苷B、连翘酯苷A、连翘苷、苍术素醇、白术内酯Ⅱ、苍术素);流动相为乙腈-0.2%磷酸水溶液,梯度洗脱,体积流量1.0 mL/min。利用中药色谱指纹图谱相似度评价系统(2012.130723版)建立湿热痹片的HPLC指纹图谱,确定共有峰并进行相似度评价;并对桑皮苷A、桑皮苷F、桑辛素M、连翘酯苷B、连翘酯苷A、连翘苷、苍术素醇、白术内酯Ⅱ、苍术素的含量测定方法进行方法学验证;基于指纹图谱共有峰峰面积测定结果,采用聚类分析和主成分分析等化学计量学方法对不同批次的湿热痹片进行质量评价。结果湿热痹片HPLC指纹图谱确认了16个共有峰,指认9个共有峰,10批湿热痹片样品相似度均大于0.95,相似度良好;9种成分在各自的质量浓度范围内线性关系良好(r2≥0.999 1),平均加样回收率分别为98.87%、97.44%、97.94%、98.39%、100.13%、99.06%、96.80%、98.44%、99.15%,RSD分别为1.42%、1.17%、1.30%、0.91%、0.86%、1.23%、1.08%、1.37%、0.79%;10批次样品中桑皮苷A、桑皮苷F、桑辛素M、连翘酯苷B、连翘酯苷A、连翘苷、苍术素醇、白术内酯Ⅱ、苍术素的质量浓度分别在0.192~0.289、0.057~0.095、0.113~0.158、0.309~0.375、1.537~1.916、0.478~0.596、0.049~0.072、0.279~0.354、0.629~0.759 mg/g。10批湿热痹片聚为2类;主成分1~6是影响湿热痹片质量评价的主要因子。结论所建立的方法操作便捷、结果准确、重复性好,可用于湿热痹片的质量控制和评价。Objective To establish a quality evaluation method of Shirebi Tablet based on HPLC fingerprints, quantitative analysis of multi-components and chemometrics analysis. Methods The chromatographic column was Waters Symmetry C18 column(250 mm × 4.6 mm, 5 μm), and the column temperature was set at 30 ℃. The detection wavelength was set at 303 nm(for mulberroside A, mulberroside F and moracin M) and 270 nm(for forsythoside B, forsythoside A, forsythin, atractylodinol, atractylenolide Ⅱ, and atractylodin). The mobile phase was composed of acetonitrile-0.2% phosphate acid solution in gradient elution manner at a flow rate of 1.0 mL/min. The HPLC fingerprint of Shirebi Tablet was established, the common peaks were determined by similarity evaluation system for chromatographic fingerprint of TCM(Version 2012.130723), and the similarity was calculated. The content determination methods for mulberroside A, mulberroside F, moracin M, forsythoside B, forsythoside A, forsythin, atractylodinol, atractylenolide Ⅱ, and atractylodin were validated. The chemometrics methods such as cluster analysis and principal component analysis were used to evaluate the quality of Shirebi Tablet from different batches based on the results of fingerprint common peak area. Results The fingerprint of Shirebi Tablet was established. Sixteen common peaks were identified. The similarity of fingerprints of 10 batches of Shirebi Tablet was more than 0.95. Nine components had good linear relationship in the range of mass concentration(r2 ≥ 0.999 1), and the average recoveries were 98.87%, 97.44%, 97.94%, 98.39%, 100.13%, 99.06%, 96.80%, 98.44% and 99.15% with the RSDs of 1.42%, 1.17%, 1.30%, 0.91%, 0.86%, 1.23%, 1.08%, 1.37% and 0.79%, respectively. The concentrations of mulberroside A, mulberroside F, moracin M, forsythoside B, forsythoside A, forsythin, atractylodinol, atractylenolide Ⅱ, and atractylodin in 10 batches were 0.192-0.289, 0.057-0.095, 0.113-0.158, 0.309-0.375, 1.537-1.916, 0.478-0.596, 0.049-0.072, 0.279-0.354, and 0.629-0.759

关 键 词：湿热痹片 HPLC 指纹图谱 化学计量学 聚类分析 主成分分析 桑皮苷A 桑皮苷F 桑辛素M 连翘酯苷B 连翘酯苷A 连翘苷 苍术素醇 白术内酯Ⅱ 苍术素 质量评价 

分 类 号：R286.02[医药卫生—中药学]