藏药西藏凹乳芹及近似种HPLC指纹图谱对比研究

Comparative study on HPLC fingerprints of Vicatia thibetica de Boiss.and its similar species

目的:建立藏药西藏凹乳芹的HPLC指纹图谱方法,并与其近似种进行对比分析,寻找各品种间成分的共性和差异性。方法:收集主产区具有代表性的26批西藏凹乳芹样品,建立其HPLC指纹图谱方法。采用《中药色谱指纹图谱相似度评价系统》(2012.130723版)对西藏凹乳芹及其近似种(迷果芹、西藏棱子芹、刺果峨参、羽苞藁本、凹乳芹、峨参、东当归、松潘棱子芹)的指纹图谱进行相似度评价分析。同时以共有峰面积量化数据为变量,采用SIMCA软件对临床常用的3个加哇基原药材(西藏凹乳芹、迷果芹、西藏棱子芹)进行主成分分析(PCA)和偏最小二乘法判别分析(PLS-DA)。结果:HPLC指纹图谱共标定了西藏凹乳芹17个共有峰,并指认了L-色氨酸、绿原酸、阿魏酸、异绿原酸A、异绿原酸C、洋川芎内酯I及藁本内酯7个共有峰。通过相似度比较分析,西藏凹乳芹与近似种具有明显差异。PCA及PLS-DA结果显示西藏凹乳芹、迷果芹及西藏棱子芹具有显著差异性,并筛选出6个种间差异的标志物。结论:建立的HPLC指纹图谱结合多元统计学分析可鉴别西藏凹乳芹及其近似种,各品种间成分上存在着较大的差异性,所指认的7个共有峰均为其主要有效成分,为其质量标准的制定打下基础。

Objective:To establish the HPLC fingerprint of Vicatia thibetica de Boiss.and to compare its similar species to find out the commonalities and differences of the components among the species.Methods:The HPLC fingerprint method was established by collecting 26 representative batches of Vicatia thibetica de Boiss.samples from the main production area.The chromatography fingerprint similarity evaluation software(2012.130723 version)was used to compare Vicatia thibetica de Boiss.and its similar species,including 14 batches of Sphallerocarpus gracilis(Bess.)K.-Pol.,4 batches of Pleurospermum hookeri var.thomsonii C.B.Clarke,1 batch of Anthriscus.nemorosa(M.Bie.)Spreng.,6 batches of Ligusticum daucoides(Franch.)Franch.,1 batch of Vicatia coniifolia(Wall.)DC.,2 batchs of Anthriscus sylvestris(L.)Hoffm.,1 batch of Angelica acutiloba(Sieb.et Zucc.)Kitagawa,and 5 batchs of Pleurospermum franchetianum Hemsl.Meanwhile,principal component analysis(PCA)and partial least squares discriminant analysis(PLS-DA)were performed on the three clinically commonly used Jiawa species,using SIMCA software with the quantified data of shared peak areas as variables.Results:Seventeen shared peaks were comfirmed,and seven shared peaks of L-tryptophan,chlorogenic acid,ferulic acid,isochlorogenic acid A,isochlorogenic acid C,chuanxiong lactone I and ligustrolactone were identified.The results of similarity evaluation and PCA and PLS-DA showed that there were significant differences among species,and six markers of inter-species differences were screened.Conclusion:The established HPLC fingerprint combined with multivariate statistical analysis can identify Vicatia thibetica de Boiss.and its similar species,and there is a large variability in the composition among the species.The seven shared peaks were identified as the main active ingredients,laying the foundation for the development of its quality standard.