高效液相法同时测定土茯苓中五种多酚成分

Quantitative Analysis of Five Polyphenols in Rhizoma smilacis Glabrae by HPLC

摘要:

本文建立了土茯苓中五种多酚成分：5-O-咖啡酰莽草酸、落新妇苷、花旗松素、黄杞苷及白藜芦醇的高效液相色谱分析方法。色谱柱为Agilent Zorbax SB C18柱（250 mm×4.6 mm i.d., 5 μm），流动相为乙腈（A）及0.1%醋酸水溶液（B），线性梯度洗脱程序为0~15 min，16~21% A；15~40 min，21~40% A。五种成分线性良好（r>0.998），保留时间相对标准偏差介于0.22~0.76%，峰面积相对标准偏差介于2.67~4.64%，加标回收率介于96.5~103.7%。对18批土茯苓样品分析结果表明，落新妇苷是土茯苓中最主要黄酮成分，含量范围为5.48~25.75 mg/g；花旗松素、黄杞苷、异黄杞苷、落新妇苷及其三种异构体(以落新妇苷计)等七种黄酮总量介于6.72~35.3 mg/g，平均含量为18±8.5 mg/g；5-O-咖啡酰莽草酸含量也较高，含量范围为0.97~6.29 mg/g，该成分在土茯苓中的含量测定为首次报道。但花旗松素和白藜芦醇含量较低，在一些样品中未检测到。

Abstract:

A HPLC quantitative method for simultaneous determination of 5-O-caffeoylshikimic acid, astilbin, taxifolin, engeletin and trans-resveratrol in Rhizoma Smilacis Glabrae (RSG) was developed. An Agilent Zorbax SB C18 column (250 mm×4.6 mm i.d., 5 μm) was used. The mobile phase consisted of acetonitrile (A) and 0.1% acetic acid aqueous solution (B) with linear gradient program of 16~21% (A) in 0~15 min, 21~40% (A) in 15~40 min. The calibration curve of each analyte was constructed under six concentrations. The correlation coefficients were all >0.998, and the RSD values of tR and the peak area ranged from 0.22 to 0.76% and from 2.67 to 4.64%, respectively. Among 18 RSG samples, astilbin was the most dominant constituent with content ranged from 5.48 to 25.75 mg/g. The total amount of seven flavonols including taxifolin, engeletin, isoengeletin, astilbin and its three isomers (quantified as astilbin) was in the range of 6.72~35.3 mg/g, with average of 18±8.5 mg/g. 5-O-caffeoylshikimic acid was the second dominant component with content ranged from 0.97 to 6.29 mg/g, while the contents of taxifolin and trans-resveratrol were relatively low, and in some samples was even undetected.