高效液相色谱法同时测定刺三加叶中芦丁和没食子酸含量

Simultaneous Determination of Rutin and Gallic Acid in Acanthopanax Trifoliatus Leaves by HPLC

刺三加叶样品先后经沸水和石油醚浸泡,所得滤渣再经60%(体积分数)乙醇溶液超声提取,提取液经高效液相色谱法测定其中芦丁和没食子酸含量。以ZORBAXSB-C_(18)色谱柱(250 mm×4.6 mm,5μm)为固定相,以体积比为44:55.6:0.4的CH2 OH-H2O-CH3COOH混合溶液为流动相,以260 nm和272 nm分别作为芦丁和没食子酸的检测波长。芦丁和没食子酸的质量浓度分别在0.55~11.00,0.26~5.18 mg·L^(-1)内与其对应的峰面积呈线性关系,芦丁和没食子酸的检出限(3S/N)分别为0.04,0.03 mg·L^(-1);芦丁和没食子酸的测定下限(10S/N)分别为0.11,0.08 mg·L^(-1)。芦丁和没食子酸的的平均加标回收率分别为99.7%,99.4%,相对标准偏差(n=6)分别为0.81%,0.97%。

The sample of Acanthopanax trifoliatus leaves was successively soaked by boiling water and petroleum ether,and then the residue obtained was extracted ultrasonically with 60%(φ)ethanol solution.The contents of rutin and gallic acid in the extract were determined by HPLC with ZORBAXSB-C_(18)chromatographic column(250 mm×4.6 mm,5μm)as stationary phase,the mixture of CH3OH-H2O-CH3COOH(the volume ratio of 44:55.6:0.4)as mobile phase,and 260 nm,272 nm as the detection wavelengths of rutin and gallic acid,respectively.Linearity relationships between peak areas and mass concentrations of rutin and gallic acid were kept in the ranges of 0.55-11.00 mg·L^(-1)and 0.26-5.18 mg·L^(-1),with detection limits(3 S/N)of 0.04,0.03 mg·L^(-1),and lower limits of determination(10 S/N)of 0.11,0.08 mg·L^(-1),respectively.Values of average recovery of rutin and gallic acid obtained by added standard were found 99.7%and 99.4%,with RSDs(n=6)of measured values of 0.81%and 0.97%,respectively.