不同产地铁线蕨药材指纹图谱的建立、分析及差异成分的含量测定

Establishment and Analysis of the Fingerprints of Adiantum capillusveneris from Different Producing Areas and Content Determination of Its Differential Components

目的:建立不同产地铁线蕨药材的指纹图谱,并进行化学计量学分析和差异成分的含量测定,为铁线蕨药材质量控制提供参考。方法:采用高效液相色谱-二极管阵列检测器法(HPLC-DAD)检测并结合《中药色谱指纹图谱相似度评价系统》(2004A版)建立19批不同产地铁线蕨药材(S1~S19)的指纹图谱,同时进行共有峰指认和相似度评价。采用聚类分析、主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)等化学计量学分析方法对19批不同产地铁线蕨药材的质量进行评价,筛选差异成分,并对部分差异成分进行含量测定。结果:19批不同产地铁线蕨药材中,共确定22个共有峰,并指认了9号峰为绿原酸、17号峰为槲皮素-3-O-β-D-吡喃葡糖苷酸、20号峰为山柰酚-3-O-芸香糖苷;19批药材的相似度为0.530~0.962。经聚类分析发现,S7、S10聚为一类,S15、S18聚为一类,S1~S6、S8、S9、S11~S14、S16、S17、S19聚为一类;经PCA、OPLS-DA发现,S7、S10聚为一类,S15为一类,S18为一类,S1~S6、S8、S9、S11~S14、S16、S17、S19聚为一类;筛选出绿原酸、槲皮素-3-O-β-D-吡喃葡糖苷酸、山柰酚-3-O-芸香糖苷以及14号峰所代表的化学成分为该药材的差异成分。19批铁线蕨药材中,绿原酸、槲皮素-3-O-β-D-吡喃葡糖苷酸和山柰酚-3-O-芸香糖苷的平均含量分别为0.10~4.25、0.31~7.11、0.61~12.00mg/g。结论:本研究建立了不同产地铁线蕨药材的HPLC-DAD指纹图谱,指认了3个共有峰;筛选出了4个影响铁线蕨药材质量的差异成分,其中绿原酸、槲皮素-3-O-β-D-吡喃葡糖苷酸、山柰酚-3-O-芸香糖苷在不同产地铁线蕨药材中的含量差异明显。

OBJECTIVE:To establish the fingerprint of Adiantum capillusveneris from different producing areas,and to conduct chemometric analysis and content determination of differential components,so as to provide reference for quality control of A. capillusveneris. METHODS:HPLC-DAD combined with Similarity Evaluation System of TCM Chromatogramtic Fingerprint(2004 A edition)were used to establish fingerprint of 19 batches of A. capillusveneris from different producing areas(S1-S19).Common peaks were confirmed and their similarities were evaluated. Chemometric analysis methods such as cluster analysis,principle component analysis(PCA),orthogonal partial least squares discriminant analysis(OPLS-DA)were used to evaluate the quality of 19 batches of A. capillusveneris from different producing areas,screen the differential components,and determine the contents of some differential components. RESULTS:Among 19 batches of A. capillusveneris from different producing areas,22 common peaks were confirmed;peak 9 was chlorogenic acid,peak 17 was quercetin-3-O-β-D-glucopyranoside,peak 20 was kaempferol-3-O-rutoside;the similarity of 19 batches of sample were 0.677-0.962. Through cluster analysis,it was found that S7 and S10 were clustered into one category;S15 and S18 were clustered into one category;and S1-S6,S8,S9,S11-S14,S16,S17 and S19 were clustered into one category. PCA and OPLS-DA found that S7 and S10 were clustered into one category;S15 were clustered into one category;S18 were clustered into one category;S1-S6,S8,S9,S11-S14,S16,S17 and S19 were clustered into one category. Chlorogenic acid,quercetin-3-O-β-D-glucopyranoside,kaempferol-3-O-rutoside and chemical composition represented by peak 14 were the differential components of the medicinal material. Among 19 batches of A. capillusveneris,average contents of chlorogenic acid and quercetin-3-O-β-Dglucopyranoside and kaempferol-3-O-rutoside were 0.10-4.25,0.31-7.11,0.61-12.00 mg/g,respectively. CONCLUSIONS:HPLC-DAD fingerprints of A. capillusveneris from different producing areas are established in the study,and three common peaks are identified. Four differential components affecting the quality of A. capillusveneris are screened,and the contents of chlorogenic acid,quercetin-3-O-β-D-glucopyranoside and kaempferol-3-O-rutoside in A. capillusveneris from different producing areas were significantly different.