Abstract

A simple, rapid and specific liquid chromatographic–diode array detector (DAD) method was developed and fully validated for simultaneous quantification of five major active ingredients (markers) from Apocynum venetum L. Samples were ultrasonically extracted with 70% ethanol. Then the foreign materials were removed in solid phase extraction column. The chromatographic separation was performed on a Zorbax SB-C₁₈ column (250mm×4.6mm i.d., 5μm) with a gradient of acetonitrile and water containing 0.1% glacial acetic acid, at a flow rate of 1.0 ml/min, detected at 360 nm. Five regression equations showed good linear relationships (r² > 0.999) between the peak areas of each marker and concentrations. The assay was reproducible with overall intra- and inter-day variation of less than 6.0%. The recoveries, measured at three concentration levels, varied from 97.8% to 102.5%. This assay was successfully applied to the determination of the 5 bioactive compounds in 10 samples. The results indicated that the developed assay method was rapid, accurate, reliable and could be readily utilized as a quality control method for A. Venetum L.

Key words: HPLC, Apocynum venetum L., chlorogenic acid, hyperoside, isoquercitrin, quercitroside, quercetin.