基于HPLC指纹图谱和一测多评多成分定量分析的连翘质量评价研究

Quality Evaluation of Forsythiae Fructus Based on HPLC Fingerprint and QAMS Multi-Component Content Determination

目的 建立连翘药材的高效液相色谱(HPLC)法指纹图谱及基于“一测多评”(QAMS)技术的多成分定量测定方法。方法 采用HPLC法建立河南、山东、山西、陕西4个省份47批连翘药材的指纹图谱,进行相似度评价,结合聚类分析(HCA)、正交偏最小二乘法-判别分析(OPLS-DA)对连翘进行化学模式识别分析。利用QAMS技术,以连翘酯苷A为内参物,同时对连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷和连翘脂素6种活性成分进行含量测定,分析不同产地样品差异。结果 建立了47批连翘药材的指纹图谱,相似度为0.785~0.989,共标定30个共有峰。聚类分析将47批样品聚为3类,OPLS-DA分析能有效将几个产区连翘药材进行区分(Q^(2)>0.5)。采用QAMS法与外标法(ESM)测定47批样品中6种活性成分的含量结果无明显差异(P>0.05),不同产地样品中活性成分含量存在明显差异(P<0.01)。连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素6种成分在各省样品中的平均含量分别为:河南(4.50、20.58、0.52、2.39、7.46、1.76 mg·g^(-1))、山东(0.90、9.26、0.36、1.39、3.84、3.87 mg·g^(-1))、山西(0.82、8.74、0.29、1.16、2.64、1.75 mg·g^(-1))、陕西(5.18、55.10、0.88、3.36、13.11、9.25 mg·g^(-1))。结论 不同来源的连翘药材样品中化学成分近似,但产地因素对有效成分含量有较大的影响,来自陕西的连翘样品中各检测成分含量最高。建立的连翘药材HPLC指纹图谱及多成分含量测定方法稳定、可靠,结合化学识别模式可为连翘药材产地判别及质量评价提供依据。

OBJECTIVE To establish the HPLC fingerprint of Forsythiae Fructus and develop a multi-component quantitative analysis method based on quantitative analysis of multi-components by single-marker(QAMS).METHODS The fingerprints of 47 Forsythiae Fructus samples from four provinces,including Henan,Shandong,Shanxi and Shaanxi,were established by high performance liquid chromatography(HPLC),while their similarity was evaluated.Chemical pattern recognition of Forsythiae Fructus was studied by hierarchical cluster analysis(HCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA).The content of forsythoside B,forsythoside A,hyperin,arctiin,phillyrin and phillygenin were determined by using QAMS,and the differences of samples from different producing areas were analyzed.RESULTS The fingerprints of 47 Forsythiae Fructus samples were established with similarities ranged from 0.785 to 0.989,and a total of 30 common peaks were identified.According to HCA,the samples were divided into three groups.OPLS-DA analysis can effectively distinguish Forsythiae Fructus from several producing areas(Q~2>0.5).There was no significant difference in the content of 6 components in 47 batches of samples by QAMS and external standard method(ESM)(P>0.05),but there was significant difference in the content of active components in samples from different origin(P<0.01).The average contents of forsythoside B,forsythoside A,hyperin,arctiin,phillyrin and phillygenin in samples from different provinces were,Henan(4.50、20.58、0.52、2.39、7.46、1.76 mg·g~(-1)),Shandong(0.90,9.26、0.36、1.39、3.84、3.87 mg·g~(-1)),Shanxi(0.82、8.74、0.29、1.16、2.64、1.75 mg·g~(-1)),Shaanxi(5.18、55.10、0.88、3.36、13.11、9.25 mg·g~(-1)).CONCLUSION The chemical composition of Forsythiae Fructus samples from different locations was similar,but the origin factors had a significant effect on the content of active ingredients,and the samples from Shaanxi province had the highest content.The HPLC fingerprint combined with the simultaneous determination of multi-component content analysis method established in this study is stable and reliable.The fingerprint combined with the chemical pattern recognition can provide a basis for the identification of origins and quality evaluation of Forsythiae Fructus.