RP-HPLC法同时测定毛冬青根中4种环烯醚萜类化合物的含量

Simultaneous determination of the contents of four iridoids in roots of Ilex pubescens Hook. et Arn. by RP-HPLC

目的建立RP-HPLC法同时测定毛冬青根中女贞子苷、橄榄苦苷、oleoacteoside和ligstroside共4种环烯醚萜类化合物的含量。方法采用Kromasil C18柱(250 mm×4.60 mm,5μm),以乙腈-质量分数为0.1%的磷酸水溶液为流动相梯度洗脱,流速为1.0 mL·min^(-1),柱温为30℃,检测波长为230 nm,进样量为10μL。结果女贞子苷、橄榄苦苷、oleoacteoside和ligstroside与其邻近的色谱峰均能良好的分离,进样量分别在0.061 35~1.227 0μg(r=0.999 6)、0.294 75~5.895 0μg(r=0.999 5)、0.064 00~1.280 0μg(r=0.999 5)和0.049 76~0.995 2μg(r=0.999 7)内与峰面积呈良好的线性关系,平均加样回收率分别为99.0%(RSD=1.8%)、99.6%(RSD=1.8%)、98.9%(RSD=1.2%)和100.3%(RSD=1.8%)。结论该法准确、可靠,为毛冬青根中环烯醚萜类成分的开发和应用提供参考。

Objective To establish an RP-HPLC method for the simultaneous determination of nuezhenide,oleuropein,o leoacteoside and ligstroside in roots of Ilex pubescens Hook.et Arn..Methods A Kromasil C18column（250 mm × 4.60 mm,5 μm） was adopted with a mobile phase of acetonitrile -0.1% phosphoric acid aqueous by gradient elution at the rate of 1.0 mL·min-1.The column temperature was 30 ℃,the detection wavelength was set at 230 nm,and the injection volume was 10 μL.Results Nuezhenide,oleuropein,o leoacteoside and ligstroside was well separated with their near peaks.The method had a good linearity in the range of 0.061 35-1.227 0 μg（r=0.999 6） for nuezhenide,0.294 75-5.895 0 μg（r=0.999 5） for oleuropein,0.064 00-1.280 0 μg（r=0.999 5） for oleoacteoside,0.049 76-0.995 2 μg（r=0.999 7） for ligstroside,respectively.The average recoveries were 99.0%（RSD=1.8%）,99.6%（RSD=1.8%）,98.9%（RSD=1.2%） and 100.3%（RSD=1.8%）.Conclusions This method is accurate and reliable for the development and application of the iridoids in roots of Ilex pubescens Hook.et Arn.to provide the reference.