Analysis of Four Flavonoids in Hawthorn Fruit by HPLC
A HPLC method was developed to determine four flavonoids in hawthorn fruit,including rutin,vitexin,hypericin and quercetin.An Agilent 5 TC-C18 column(250 mm×4.6 mm,5 μm)was used with the mobile phases consisting of acetonitrile(A)and 0.2％acetic acid(B)at the flow rate of 1 mL/min.The injection volume was 10 μL,while the detection wavelength was set at 355 nm and the column temperature was 40℃.The linear gradient elution procedure was 20％～30％A in 0～15 min,and 30％～60％A in 15～30 min.Methodological results showed that the four components have a good linear relationship.The intraday precision retention time RSD was＜0.5％,and the peak area RSD was＜2.5％.The RSD of interday precision retention time was＜0.5％and the RSD of peak area was＜5.0％.The average recoveries ranged from 103.4％to 106.7％,and the RSD was＜2.5％.The optimal extraction solvent was 60％ethanol.The contents of rutin and hypericin in 6 batches of hawthorn samples were in the range of 51.8～264.5 μg/g and 148.0～441.6 μg/g,respectively.Quercetin was detected in 2 batches of hawthorn samples,with contents of 11.0 and 12.7 μg/g,and vitexin was undetectable in all samples.
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