HPLC法同时测定墓头回中齐墩果酸和熊果酸的含量

Simultaneous determination of oleanolic acid and ursolic acid in P.heterophylla by HPLC

目的建立高效液相色谱法(high performance liquid chromatography,HPLC)同时测定墓头回药材中齐墩果酸和熊果酸含量的方法。方法采用HPLC色谱法,色谱柱为Sil Green C_(18)(5μm,4.6 mm×250 mm);以甲醇-0.2%乙酸铵水溶液(85∶15)为流动相,检测波长为210 nm,流速为1.0 ml/min,柱温为30℃。结果齐墩果酸和熊果酸分别在0.02510~1.0041mg/ml、0.02527~1.0109 mg/ml浓度范围内具有良好线性关系(r=0.99999),平均回收率分别为95.71%[相对标准偏差(relative standard deviation,RSD)=0.92%,n=6]、96.19%(RSD=1.16%,n=6)。结论 HPLC测定墓头回药材中齐墩果酸和熊果酸含量具有灵敏度高、快速、简便、专属性强、准确和重复性好等优点,可为墓头回药材质量控制标准的建立提供科学依据。

Objective The objective of this study was to establish a high performance liquid chromatography （HPLC） method for simultaneous measuring of the concentration of oleanolic acid and ursolic acid in P. heterophylla. Methods The analysis was performed on a SilGreen Cl8 （5μm,4.6 mm×250 mm） chromatographic column with mobile phase consisting of methanol - 0.2% ammonium acetate water （85 ： 15）. The detected wavelength was set at 210 nm. The flow rate was 1.0 ml/min and the column temperature was at 30℃. Results The linearity was good in the ranges of 0.02510-1.0041 mg/ml and 0.02527-1.0109 mg/ml respectively for oleanolic acid and ursolic acid （r=0.99999）： The average recovery rates of oleanolic acid and ursolic acid were 95.71% [relative standard deviation （RSD=0.92%, n=6] and 96.19% （RSD=1.16%, n=6） respectively. Conclusions The method of HPLC features high sensitivity, time efficiency, simplicity, strong specificity, high accuracy and good repeatability, which can be used for the quality control of P. heterophylla.