Asian Journal
Of Traditional Medicines

Abstract: A high-performance liquid chromatographic method was established for the simultaneous determination of three major components, i.e. limonin, evodiamine, and rutaecarpine, of Fructus Evodiae. These three compounds, limonin a limonoid, and evodiamine and rutaecarpine, alkaloids, were simultaneously separated on a Lichrospher-rp18e analytical HPLC column (4.6 mm×250 mm, 5 μm; Merck, Germany) at a column temperature of 25 ºC. Elution was carried out under isocratic conditions using acetonitrile-water (46:54, v/v) and the flow rate was 1 ml/min. The detection wavelength was set at 225 nm. All calibration curves showed good linear regression (r >0.9999) within the concentration ranges of 5.45-545 μg/ml for limonin, 0.34-79.8 μg/ml for evodiamine, and 0.36-46.4 μg/ml for rutaecarpine, respectively. The established method showed a good precision and accuracy with overall intra-day and inter-day variations of (0.98-2.29) % and (0.77-2.63) %, respectively, and the detection limits were 2.18, 0.10 and 0.23 μg/ml for limonin, evodiamine and rutaecarpine, respectively. Extraction of samples ultrasonically with 80 % ethanol at room temperature allowed a good analyte recovery, and the recovery rates were 101.59 % (RSD 1.05 %), 98.11 % (RSD 0.99 %) and 100.47 % (RSD 0.72 %) for limonin, evodiamine and rutaecarpine, respectively. The developed method has been successfully applied to simultaneously evaluate the intrinsic quality of different species of Fructus Evodiae. The results indicate that there are statistically significant differences in the contents of the three active components among the samples which were collected from different places and from species to species.

Key words: HPLC, quantitative determination, limonin, evodiamine, rutaecarpine, Fructus Evodiae