Guo Sheng, Duan Jin-ao, Tang Yuping, Su Shulan, Shang Erxin, Ni Shumao, Qian Dawei
Jiangsu Key Laboratory for TCM Formulae Research, Nanjing University of Chinese Medicine, 138 Xianlin Road, Nanjing 210046, PR China.
J Pharm Biomed Anal. 2009 Jul 12;49(5):1296-302. doi: 10.1016/j.jpba.2009.03.006. Epub 2009 Mar 20.
A simple and sensitive HPLC-DAD method has been developed for the first time to simultaneously determine 10 triterpenoid acids (ceanothic acid, alphitolic acid, zizyberanal acid, zizyberanalic acid, epiceanothic acid, ceanothenic acid, betulinic acid, oleanolic acid, ursonic acid and zizyberenalic acid) in the dried fruit of Ziziphus jujuba (called Dazao) which has been widely used as one of the traditional Chinese medicines (TCMs). This HPLC assay was performed on a reversed-phase C(18) column (250 mm x 4.6mm, 5 microm) with the column temperature at 35 degrees C. The mobile phase was composed of A (acetonitrile) and B (0.05% aqueous phosphoric acid, v/v). The flow rate was 1.0 ml/min and the detection wave length was set at 205 nm for reference compounds 1-9 and 238 nm for reference compound 10. All calibration curves showed good linear regression (r(2)>0.9999) within the test range. The established method showed good precision and accuracy with overall intra-day and inter-day variations of 0.43-1.72% and 0.53-2.45%, respectively, and overall recoveries of 94.98-104.09% for the 10 compounds analyzed. The validated method was successfully applied for the simultaneous determination of the 10 triterpenoid acids in 42 batches of Dazao which contained 36 cultivars from 22 cultivation regions, and were investigated and authenticated as Z. jujuba. Hierarchical clustering analysis (HCA) and principal components analysis (PCA) were performed to differentiate and classify the samples based on the contents of the 10 triterpenoid acids. The presented HPLC-DAD method conjugated with chemometrics approach was demonstrated to be very helpful in using Dazao resources, and was possibly useful in chemotaxonomic characterization.
首次建立了一种简单灵敏的高效液相色谱 - 二极管阵列检测法（HPLC - DAD），用于同时测定酸枣（大枣）干燥果实中的10种三萜酸（ceanothic酸、alphitolic酸、zizyberanal酸、zizyberanalic酸、epiceanothic酸、ceanothenic酸、桦木酸、齐墩果酸、乌索酸和zizyberenalic酸）。酸枣作为传统中药之一已被广泛应用。该HPLC分析在反相C(18)柱（250 mm×4.6mm，5μm）上进行，柱温为35℃。流动相由A（乙腈）和B（0.05%磷酸水溶液，v/v）组成。流速为1.0 ml/min，参考化合物1 - 9的检测波长设定为205 nm，参考化合物10的检测波长设定为238 nm。所有校准曲线在测试范围内均显示出良好的线性回归（r(2)>0.9999）。所建立的方法具有良好的精密度和准确度，日内和日间总体变化分别为0.43 - 1.72%和0.53 - 2.45%，所分析的10种化合物的总体回收率为94.98 - 104.09%。该验证方法成功应用于同时测定42批大枣中的10种三萜酸，这些大枣包含来自22个种植区域的36个品种，并经调查和鉴定为酸枣。进行了层次聚类分析（HCA）和主成分分析（PCA），以根据10种三萜酸的含量对样品进行区分和分类。所提出的HPLC - DAD方法与化学计量学方法相结合，被证明对大枣资源的利用非常有帮助，并且可能有助于化学分类学表征。
