HPLC快速测定沙棘叶中的苯丙素与黄酮类成分

Rapid determination of phenylpropanoids and flavonoids components in the leaves of Hippophae rhamnoides by HPLC

目的 采用HPLC法快速测定沙棘叶中苯丙素类(没食子酸、咖啡酸、对香豆酸、阿魏酸等)和黄酮类(原儿茶酸、儿茶素、山柰酚-3-O-芸香糖苷、异鼠李素-3-O-葡萄糖苷-7-O-鼠李糖苷等)化合物的含量。方法 采用Kromasil C18色谱柱(250 mm×4.6 mm, 5μm),0.1%甲酸水溶液-乙腈为流动相,梯度洗脱,流速1.0 mL·min-1,柱温30℃,检测波长为280 nm,进样量10μL。结果 没食子酸、原儿茶酸、儿茶素、咖啡酸、对香豆酸、阿魏酸、山柰酚-3-O-芸香糖苷、异鼠李素-3-O-葡萄糖苷-7-O-鼠李糖苷的线性范围分别为0.012~0.870、0.696~3.913、0.203~4.174、0.203~3.043、0.035~2.609、0.023~3.478、0.565~8.478、0.522~7.826μg(r2≥0.9992),平均回收率为93.58%~108.11%,RSD≤1.81%。采用该方法测定不同产地的沙棘叶中均含上述8种酚酸类化合物,但含量有差异。结论 所用方法分离效果好、操作简便、重复性和稳定性较好,可控制沙棘叶中酚酸类成分的质量。

OBJECTIVE To rapidly determine the contents of phenylpropanoids and flavonoids including gallic acid,protocatechuic acid,catechin,caffeic acid,p-coumaric acid,ferulic acid,kaempferol-3-O-rutinoside and isorhamnetin 3-O-glucoside 7-O-rhamnoside in the leaves of Hippophae rhamnoides L.by HPLC.METHODS The separation was performed on a Kromasil C18 column(250 mm×4.6 mm,5μm).The mobile phase consisted of acetonitrile and 0.1%formic acid water.The elution procedure was gradient elution.The flow rate was set at 1.0 mL·min-1.The detection wavelength was 280 nm.The injection volume was 10μL.The column temperature was 30℃.RESULTS The calibration curves of all eight standards showed good linearity(r 2≥0.9992)in a relatively wide concentration range.The linear ranges of gallic acid,protocatechuic acid;catechin,caffeic acid,p-coumaric acid,ferulic acid,kaempferol-3-O-rutinoside,isorhamnetin-3-O-glucoside-7-O-rhamnoside were 0.012-0.870,0.696-3.913,0.203-4.174,0.203-3.043,0.035-2.609,0.023-3.478,0.565-8.478,0.522-7.826μg,respectively.The mean recoveries of the eight compounds were in the range of 93.58%-108.11%with RSD≤1.81%.However,there existed differences in the content of phenolic compounds from different regions.CONCLUSION The method is simple with excellent separation effect,repeatability and stability,which can be used for quality control of phenolic acids from the leaves of Hippophae rhamnoides L.