树型金银花与野生金银花HPLC指纹图谱对比研究

Comparative study on HPLC fingerprint of tree honeysuckle and wild honeysuckle

【目的】建立树型金银花与野生金银花的HPLC指纹图谱,为全面、客观评价金银花质量、快速鉴别金银花品种提供依据。【方法】从陕西洛南、丹凤和河南灵宝等地收集树型金银花样品13份、野生金银花样品12份进行HPLC分析,将数据导入“中药色谱指纹图谱相似度评价系统(2012年版)”,以绿原酸色谱峰为参照峰,采用中位数法进行多点校正,建立树型金银花、野生金银花指纹图谱;应用SPSS 13.0软件,采用离差平方和法,以相对峰面积为评价指标,对2种金银花分别进行聚类分析;以绿原酸、异绿原酸、木犀草苷标准品为对照,用HPLC仪测定各样品中的3种成分含量。【结果】野生金银花指纹图谱匹配共有峰28个,树型金银花指纹图谱匹配共有峰13个。12份野生金银花样品中有10份与其对照指纹图谱相似度达0.95以上,13份树型金银花样品中有12份与其对照指纹图谱相似度达0.95以上,有24份样品与对照指纹图谱相似度达0.90以上。根据聚类分析结果,野生金银花样品可分为4个分支,其中洛南石坡、巡检、灵口、景村4地金银花聚为一支,河南灵宝的2份样品为一支,丹凤县铁峪铺、武关、蔡川、留仙坪4地样品为一支,丹凤县花瓶子、竹林关2地样品为一支;树型金银花样品也可分成4个分支,其中陕南各地样品为一支;渭南蒲城、西安鄠邑、河南灵宝3地各为1支。野生金银花样品中的绿原酸、异绿原酸、木犀草苷3种成分平均含量分别为3.72%,0.39%,0.15%,树型金银花样品分别为3.42%,1.22%,0.06%。【结论】利用对照指纹图谱可快速鉴别树型金银花与野生金银花;野生金银花的化学物质较树型金银花丰富,但后者因是规范的人工栽培品种,品质更稳定,更符合药品质量稳定性、均一性的要求。

【Objective】The HPLC fingerprints of tree honeysuckle and wild honeysuckle were established to comprehensively and objectively evaluate quality of honeysuckle and to rapidly identify honeysuckle varieties.【Method】A total of 13 tree honeysuckle samples and 12 wild honeysuckle samples were collected from Shaanxi and Henan for HPLC analysis.The results were imported into a similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine with chlorogenic acid as the reference peak.The median method was used for multi point correction to establish the fingerprint of tree honeysuckle and wild honeysuckle.SPSS 13.0 software was used for cluster analysis based on sum of square deviations using relative peak area as evaluation index.Chlorogenic acid,isochlorogenic acid and luteolin were used as reference materials to determine the three components in each sample through HPLC method.【Result】There were 28 and 13 common peaks in the fingerprints of wild honeysuckle and tree honeysuckle,respectively.Ten out of the 12 wild honeysuckle samples had similarity of more than 0.95 to their control fingerprints,while 12 out of 13 the tree honeysuckle samples had the similarity of more than 0.95 similar to the control fingerprints and 24 samples had the similarity of more than 0.90.Wild honeysuckle samples were clustered into four clades according to SPSS analysis.Samples from Shipo,Xunjian,Lingkou and Jingcun were clustered into one clade,two samples from Lingbao,Henan were clustered into another clade,samples from Tieyupu,Wuguan,Caichuan and Liuxianping were clustered into the third clade,and samples from Huapingzi and Zhulinguan were clustered into the fourth clade.Tree honeysuckle samples were also clustered into four clades.Samples from southern Shaanxi were clustered into one clade,while samples from Pucheng,Huyi district and Lingbao,Henan were clustered into three different clades,respectively.The average contents of chlorogenic acid,isochlorogenic acid and luteolin in wild honeysuckle were 3.72%,0.39%and 0.15%and those in tree honeysuckle were 3.42%,1.22%and 0.06%,respectively.【Conclusion】The control fingerprint can be used to quickly identify tree honeysuckle and wild honeysuckle.Wild honeysuckle contains more chemical substances than tree honeysuckle,but fingerprints and effective component contents in tree honeysuckle are more stable to meet the requirements in quality stability and homogeneity of traditional Chinese medicine.