RP-HPLC-PAD法同时测定枸骨中熊果酸和齐墩果酸含量

Simultaneous determination of ursolic acid and oleanolic acid in Ilex cornuta Lindl. ex Paxt. by RP-HPLC with Photodiode Array Detector

目的结合超声技术提取枸骨中熊果酸和齐墩果酸,建立RP-HPLC-PAD法同时测定其含量。方法采用Kromasil C18色谱柱(250mm×4.6mm,5μm),流动相为甲醇-0.2%磷酸水溶液(88∶12),光电二极管阵列检测器(PAD),检测波长210nm,流速0.8mL/min,柱温30℃。以保留时间和紫外光谱对分离出的组分予以定性确证,用峰面积进行定量。结果组分的质量浓度与其峰面积在一定的范围内呈现良好的线性关系,相关系数均为0.9999,熊果酸进样量在0.443～7.088μg、齐墩果酸进样量在0.247～3.952μg时呈现良好的线性关系,平均加样回收率分别为99.12%、98.76%,RSD分别为1.2%、1.7%(n=6)。结论本方法灵敏、准确,重现性好,可用于枸骨中熊果酸和齐墩果酸含量的测定。

Objective To extract ursolic acid and oleanolic acid from Ilex cornuta Lindl. ex Paxt. by supersonic technology and develop a RP-HPLC-PAD method to determine the content of ursolic acid and oleanolic acid simultaneously. Method Ursolic acid and oleanolic acid were separated and analyzed on Kromasil C18 column （250 mm× 4.6 mm, 5 μm） using methanol-0.2% aqueous solution of phosphoric acid （88： 12） as mobile phase at flow rate of 0. 8 mL/min. UV detection was at 210 nm and the column temperature was 30 ℃. The compounds were identified by retention time data and spectral characteristics and quantified by peak area. Results There were beth good linear correlations between the concentrations and peak areas of the components with the correlation coefficients of 0. 999 9. An average recovery of 99.12% was obtained with a RSD of 1.2% （n = 6） in the range of 0. 443 - 7. 088 μg for ursolic acid, and an average recovery of 98.76% was obtained with a RSD of 1.7% （n =6） in the range of 0.247 -3.952 μg for oleanohc acid. Conclusion RP-HPLC-PAD method was proved to be sensitive, fast and accurate. It can be used for the determination of ursolic acid and oleanolic acid in Ilex cornuta Lindl. ex Paxt.