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目的:测定四味消扶协定方不同浓度乙醇提取液中熊果酸及齐墩果酸含量。方法:分别以500mL,300mL的0%～80%乙醇溶液分2次水浴回流提取四味消扶协定方中熊果酸及齐墩果酸,提取液经预处理后采用RP-HPLC法同时测定它们的含量。色谱条件:色谱柱为Lichrospher C18柱(250mm×4.6mm,5μm,江苏汉邦科技有限公司);检测波长为215nm;柱温为20℃;流速为1.0mL.min-1;进样量为25μL;流动相为pH7.5的0.1%磷酸溶液与乙腈(v/v:25:75)。结果:齐墩果酸与熊果酸各自在1.54～308.00μg.mL-1与6.54～1308.00μg.mL-1浓度范围内,峰面积与其浓度具有良好的线性关系,平均加样回收率为99.63%～111.54%。提取液中齐墩果酸和熊果酸含量与加入的乙醇浓度成正比。结论:本方法简便、快速、准确,可同时测定四味消扶协定方乙醇提取液中齐墩果酸和熊果酸的含量。
Objective: To determine the contents of ursolic acid and oleanolic acid in ethanol extracts with different concentrations of Siweixiaozhe protocol. METHODS: Ursolic acid and oleanolic acid were extracted with 500 mL and 300 mL of 0%-80% ethanol solution in two water baths at reflux. The extracts were pretreated and their RP-HPLC was used to determine their contents. content. Chromatographic conditions: The column was a Lichrospher C18 column (250mm×4.6mm, 5μm, Jiangsu Hanbang Technology Co., Ltd.); detection wavelength was 215nm; column temperature was 20°C; flow rate was 1.0mL.min-1; sample injection volume was 25μL. The mobile phase was a pH 7.5 solution of 0.1% phosphoric acid and acetonitrile (v/v: 25:75). Results: Oleanolic acid and ursolic acid were in the range of 1.54～308.00μg.mL-1 and 6.54～1308.00μg.mL-1 respectively, and the peak area and concentration had a good linear relationship. The average recovery rate was 99.63. % ~ 111.54%. The content of oleanolic acid and ursolic acid in the extract is directly proportional to the ethanol concentration added. Conclusion: This method is simple, rapid and accurate. It can simultaneously determine the contents of oleanolic acid and ursolic acid in the ethanol extract of the four-way dietary supplement protocol.