HPLC-UV Method for Determing Flavonoids in Hawthorn Flowers and Leaves

Abstract

An HPLC technique with UV spectrophotometric detection was developed for qualitative and quantitative determination of vitexin, rutin, hyperoside, and quercetin in Crataegus sanguinea hawthorn herbs (flowers and leaves). The separation used a Luna C18(2) column (4.6 × 150 mm, 5 μm) and a stepwise gradient. Calibration curves for the four flavonoids exhibited good linearity (r² > 0.995) in the studied ranges. The analytical recoveries over one day/week were 95.9-104.3%/95.6 – 102.9% for vitexin; 94.0 – 102.2%/93.5 – 101.5% for rutin; 98.0 – 102.2%/97.5 – 102.9% for hyperoside; and 96.0 – 109.8%/97.2 – 111.0% for quercetin. The intra- and interday coefficients of variation (RSD, %) were <3%. The relative error (ε, %) of a single determination at confidence level 95% was <3% for rutin, hyperoside, and vitexin and <4% for quercetin.