Optimization of the extraction conditions and simultaneous quantification by RP-LC of six alkaloids in evodiae fructus

Abstract

A reversed phase liquid chromatographic (LC) method coupled with DAD (250 nm) has been developed and validated for simultaneous quantification of six alkaloids, dehydroevodiamine (1), wuzhuyuamide-I (2), 5-hydroxyrutaecarpine (3), 14-formyldihydrorutaecarpine (4), evodiamine (5) and rutaecarpine (6), in 12 batches evodiae fructus [the dried, unripe fruits of Evodia rutaecarpa (Juss.) Benth. or E. rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang, E. rutaecarpa (Juss.) Benth. var. bodinieri (Dode) Huang] as a traditional Chinese medicine. The method was carried out by a C18 column (250 × 4.6 mm) with a gradient mobile phase of methanol, acetonitrile, and phosphoric acid-triethylamine-buffer solution. The contents of 1-6 in the evodiae fructus could easily be determined within 70 min. The experimental results were satisfactory for the intra-day and inter-day precision and accuracy of the method for simultaneous determination. The linear calibration ranges of 1-6 were 40-1,000, 20-500, 1-100, 10-500, 40-1,000 and 80-1,000 μg mL-1. The recoveries of 1-6 were 97.43-103.73% with RSDs from 0.21 to 1.99%. The limits of detection for 1-6 were 2.0, 2.0, 0.1, 1.0, 5.0 and 5.0 μg mL -1, and the limits of quantification were 6.6, 6.6, 0.3, 3.3, 16.5 and 16.5 μg mL-1. The method was successfully applied to the quantification of six alkaloids in the evodiae fructus. © 2008 Friedr. Vieweg & Sohn Verlag/GWV Fachverlage GmbH.