Simultaneous determination of nine flavonoids and qualitative evaluation of Herba Epimedii by high performance liquid chromatography with ultraviolet detection

A simple high performance liquid chromatographic method has been developed for the determination of nine flavonoids in Herba Epimedii, including quercetin-3-O-glucoside, epimedin A, hexandraside A, epimedin B, epimedin C, icariin, icariside 1, icariside 11, and icaritin. The chromatographic separation was performed on a Diamonsil C-18 column (5 mu m, 250 x 4.6 mm) with gradient elution by acetonitrile-0.5% acetic buffer (adjusted to pH 5.00 with triethylamine). Methodological validation gave acceptable linearities (r(2) >0.9992) and recoveries (ranging from 98.9 to 102.4%). The limits of detection of these flavonoids ranged from 16.3 to 83.3 ng. The results indicated that the contents of flavonoids in Herba Epimedii varied significantly from habitat to habitat with contents ranging from 0.05 to 39.0 mg/g. Twenty five Herba Epimedii samples prepared from five different botanical materials were investigated by the established method, and the results showed the influence of the botanical material and the habitat on the quality of Herba Epimedii. The proposed method is simple, effective, and suitable for the evaluation of this traditional Chinese medicine.