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建立了HPLC-DAD法测定血满草中熊果酸和齐墩果酸含量,并进行方法学考察。采用HPLC-DAD进行分析,fusion-RP C18柱(4.6 mm×250 mm,4μm),甲醇-0.2%磷酸水溶液(90∶10)为流动相,检测波长210 nm,体积流量1.0 mL/min。同时采用微波辅助提取、回流提取、索氏提取、冷浸提取、超声提取五种方法对血满草中熊果酸和齐墩果酸含量进行测定并比较不同方法所得结果的差异,还比较了血满草不同部位中熊果酸和齐墩果酸的含量差异。测定结果表明熊果酸进样量在3.6～8.4μg范围内,齐墩果酸进样量在3.2～16μg范围内,呈良好线性关系。血满草中熊果酸和齐墩果酸平均回收率分别为98.3%和101.4%(n=5),相对标准偏差分别为1.13%和0.72%(n=5)。五种方法比较得出索氏提取得熊果酸和齐墩果酸含量最高;血满草花中熊果酸和齐墩果酸含量最高,而根中含量最低。该方法使血满草中熊果酸和齐墩果酸达到基线分离,操作简便,结果稳定可靠。
A HPLC-DAD method was established for the determination of ursolic acid and oleanolic acid in S. korshinskii and the methodological study. The mobile phase consisted of fusion-RP C18 column (4.6 mm × 250 mm, 4 μm) and methanol-0.2% phosphoric acid (90:10) with a detection wavelength of 210 nm at a volumetric flow rate of 1.0 mL / min using HPLC-DAD. At the same time, the contents of ursolic acid and oleanolic acid were determined by microwave-assisted extraction, reflux extraction, Soxhlet extraction, cold soak extraction and ultrasonic extraction. The differences of the results obtained by different methods were also compared Differences in content of ursolic acid and oleanolic acid in different parts of Satsuma mandarinus. The results show that the injection volume of ursolic acid in the range of 3.6 ~ 8.4μg, oleanolic acid injection volume in the range of 3.2 ~ 16μg, showed a good linear relationship. The average recoveries of ursolic acid and oleanolic acid in S. purpurea were 98.3% and 101.4%, respectively (n = 5), with relative standard deviations of 1.13% and 0.72%, respectively (n = 5). Compared with the five methods, the content of ursolic acid and oleanolic acid was the highest in Soxhlet extraction; the content of ursolic acid and oleanolic acid in the flower of Sominia gracilis was the highest, while the content in the root was the lowest. The method separates the ursolic acid and the oleanolic acid in the blood serum to baseline separation, has the advantages of simple operation and stable and reliable results.