A method for simultaneous determination of puerarin,ferulic acid,hesperidin,osthole and isoimperatorin in Gubiling Tablets by high performance liquid chromatography was established.CAPCELL PAK C18 column(250 mm×4.6 mm,5 μm)was used for gradient elution with acetonitrile-0.1%formic acid solution as the mobile phase at a flow rate of 1.0 mL/min.The column temperature was 30℃.The detection wavelengths were set at 250 nm for puerarin,284 nm for hesperidin,322 nm for ferulic acid,osthole and isoimperatorin,respectively.The injection volume was 10 μL.The mass concentrations of the 5 constituents had a good linear relationships with the corresponding chromatographic peak areas in their respective ranges,and the correlation coefficients were all 0.999 9.The detection limits were 0.04-0.14 μg/mL,and the quantification limits were 0.13-0.46 μg/mL.The average recoveries of sample spiked were 96.0%-103.1%.and the relative standard deviations of the determination results were 0.8%-1.7%(n=6).This method is simple,reliable,and has good reproducibility,which can be used for quality control of Gubiling Tablets.
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