RP-HPLC法测定糙叶五加不同部位中五加酸和贝壳烯酸含量

Determination of Acanthoic Acid and Kaurenoic Acid from Different Parts of Acanthopanax henryi (Oliv.) Harms by RP-HPLC

为了建立反相高效液相色谱法(RP-HPLC)测定糙叶五加不同部位中五加酸和贝壳烯酸的定量分析方法,本研究采用RP-HPLC外标法测定了五加酸和贝壳烯酸含量。所用色谱柱:ODS-C18(250mm×4.6mm,5μm);流动相:乙腈-水(0～60min时乙腈含量为10%～100%,60～70min时乙腈含量为100%);检测波长207nm;柱温30℃;流速1.0mL/min。结果表明:在上述色谱条件下,五加酸和贝壳烯酸获得良好分离,线性范围分别为:0.1070～0.5351g/L(相关系数r=0.9999)和0.0930～0.4649g/L(r=0.9998),加样回收率分别为97.93%(标准偏差为1.33%)和98.12%(标准偏差为1.16%)。糙叶五加根、茎、叶中五加酸的平均含量(n=3)分别为:39.90、19.72、49.19μg/g;贝壳烯酸的平均含量(n=3)分别为:21.01、21.70、25.88μg/g。由此可知,该方法准确性高,重现性好,为糙叶五加的质量控制提供了可借鉴的定量分析方法。

To establish a RP-HPLC method for quantitative analysis of acanthoic acid and kaurenoic acid in different parts of Acanthopanax henryi （Oliv.） Harms, an external standard method was adopped by HPLC with ODS-C18 column （250mm × 4.6mm, 5μm） at 30℃ as fixed phase and CH3CN-H2O （CH3CN 10% - 100% at 0 - 60 min, CH3CN 100% at 60 - 70 min） as mobile phase. The detection wavelength was 207nm and the flow rate was 1.0mL/min. The results showed that acanthoic acid and kaurenoic acid were well separated by this method. The liner range of acanthoic acid and kaurenoic acid are 0.1070 -0.5351 g/L and 0.0930 -0.4649 g/L, respectively, and give correlations （r） of 0.9999 and 0.9998. The recoveries of acanthoic acid and kaurenoic acid are 97.93% （standard deviation is 1.33%） and 98.12% （standard deviation is 1.16%）, respectively. The average contents （n=3） of acanthoic acid in the root, stem, leave of Acanthopanax henryi （Oliv.） Harms were 39.90, 19.72 and 49.19μg/g, and the average contents （n=3） of kaurenoic acid in that were 21. 01,21.70 and 25.88μg/g. In conclusion, this is an accurate and reproducible method and can be used for the quality control of Acanthopanax henrvi （Oliv.） Harms.