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

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-C₁₈ chromatographic column (250 mm×4.6 mm, 5 μm) 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 (3S/N) of 0.04, 0.03 mg·L^-1, and lower limits of determination (10S/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.