基于HPLC指纹图谱、多成分含量测定和化学计量学的二地消渴合剂质量评价研究  

Quality evaluation of Erdi Xiaoke mixture based on HPLC fingerprint,multi-component content determination and chemometrics

作　　者：孙丽芬 蔡依云 朱敏[1] 安晓飞[1] 吴磊[1] SUN Li-fen;CAI Yi-yun;ZHU Min;AN Xiao-fei;WU Lei(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Nanjing University of Chinese Medicine,Nanjing 210023,China;College of Traditional Chinese Medicine,China Pharmaceutical University,Nanjing 211198,China)

机构地区：[1]南京中医药大学附属医院,南京210029 [2]南京中医药大学,南京210023 [3]中国药科大学中药学院,南京211198

出　　处：《药物分析杂志》2024年第12期2171-2179,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金项目(82074359);南京药学会-常州四药医院药学科研基金项目(2023YX027)。

摘　　要：目的:建立二地消渴合剂的HPLC指纹图谱、多成分含量测定和化学计量学相结合的质量评价方法。方法:采用Acutfex PW-C_(18)(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.1%磷酸为流动相梯度洗脱,体积流量1.0 mL·min^(-1),柱温30℃,检测波长285 nm,建立二地消渴合剂的HPLC指纹图谱;对12批二地消渴合剂中的11个成分[5-羟甲基糠醛(5-HMF)、丹参素、绿原酸、咖啡酸、毛蕊异黄酮苷、毛蕊花糖苷、异绿原酸B、异绿原酸C、迷迭香酸、丹酚酸B、芒柄花黄素]进行含量测定,并结合相似度评价、聚类分析,主成分分析及正交偏最小二乘-判别分析对二地消渴合剂质量进行综合评价。结果:所测11个成分在各自浓度范围内线性关系良好(r≥0.9996),平均加样回收率为95.3%~101.5%,RSD为1.1%~2.3%。12批样品的HPLC指纹图谱中共确定18个共有峰,相似度均>0.960。12批样品中上述11个成分的含量的范围依次在43.40~65.70、219.43~274.70、119.00~164.78、11.87~18.17、42.80~56.86、10.01~20.71、8.21~14.73、40.15~57.39、43.29~60.29、755.22~1208.50、2.47~4.44μg·mL^(-1)。12批二地消渴合剂可聚为3类,不同批次间质量存在一定差异,5-HMF、丹酚酸B和绿原酸是影响样品质量的主要潜在标志物。结论:该方法稳定可靠,准确灵敏,可用于二地消渴合剂的质量控制和综合评价。Objective:To establish a quality evaluation method of Erdi Xiaoke mixture based on HPLC fingerprint,multi-component determination and chemometric analysis.Methods:The separation was performed on a 30℃thermostatic Acutfex PW-C_(18)(250 mm×4.6 mm,5μm)column,with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL·min^(-1)in a gradient elution manner,and the detection wavelength was set at 285 nm to establish HPLC fingerprint of Erdi Xiaoke mixture.Contents of 11 components[5-hydroxymethyl furfural(5-HMF),salvianic acid A,chlorogenic acid,caffeic acid,calycosin-7-glucoside,acteoside,isochlorogenic acid B,isochlorogenic acid C,rosmarinic acid,salvianolic acid B and formononetin]in 12 batches of Erdi Xiaoke mixture were determined,and similarity evaluation,cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were used to comprehensively evaluate the quality of Erdi Xiaoke mixture.Results:The 11 constituents showed good linear relationships within their respective ranges(r≥0.9996),with average recovery rates of 95.3%-101.5%,and the RSDs were in the range of 1.1%-2.3%.There were 18 common peaks in the fingerprints for 12 batches of samples with the similarities above 0.960.The range of the content of the above 11 components in the 12 batches of samples were as follows as 43.40-65.70,219.43-274.70,119.00-164.78,11.87-18.17,42.80-56.86,10.01-20.71,8.21-14.73,40.15-57.39,43.29-60.29,755.22-1208.50,2.47-4.44μg·mL^(-1).And 12 batches of Erdi Xiaoke mixture could be clustered into three categories with some differences in sample quality among different batches.In addition,5-HMF,isochlorogenic acid B and chlorogenic acid were the main potential markers affecting the quality of Erdi Xiaoke mixture.Conclusion:This method is stable,reliable,accurate and sensitive,and can be used to control and comprehensively evaluate the quality of Erdi Xiaoke mixture.

关 键 词：二地消渴合剂 指纹图谱 含量测定 化学计量学 质量评价 

分 类 号：R917[医药卫生—药物分析学]