Development of an RP-HPLC method for the analysis of phenolic compounds in Achillea millefolium L.

Several major phenolic constituents present in yarrow (Achillea millefolium L. s.l.) were determined for a homogenized plant sample. In order to optimize the conditions for sample preparation of the botanical matrix two different solvent extraction methods (maceration and ultrasonic agitation) were assayed. The preliminary maceration studies were performed to determine the influence of extracting solvents on the recovery of phenolics by using a different concentration of aqueous ethanol (40-96%, v/v) as extractant. On the basis of these results, sonication extractions were carried out at different time intervals (5, 10, 20, 30, and 60min) using optimum extractant concentration. Levels of seven constituents (chlorogenic acid, vicenin-2, luteolin-7-O-glucoside, rutin, apigenin-7-O-glucoside, luteolin, and apigenin) were measured by means of high performance liquid chromatography (HPLC) with ultraviolet and photodiode array detection systems. The main validation steps of the HPLC method were evaluated to demonstrate its selectivity, linearity, and precision. In addition, the method was applied to characterization of the flavonoid and phenolcarbonic acid complex in different samples of yarrow.