Separation and Determination of Active Components in Evodiae fructus and its Pharmaceutical Preparations by Non-Aqueous Micellar Electrokinetic Chromatography

Abstract

In this paper, an easy, economical and effective nonaqueous micellar electrokinetic capillary chromatography (NAMECC) method for the simultaneous determination of two bioactive components (evodiamine and rutaecarpine) in Evodiae Fructus and its pharmaceutical preparations was developed for the first time. After optimization of the separation conditions and concentration conditions, the two alkaloids can be separated by using a fused-silica capillary column (57 × 75 μm I.D.) with a running buffer of 140 mM sodium cholate (SC) in methanol and detected with a diode-array detector (228nm). The second-order derivative electropherograms were applied for resolving overlapping peaks in complicated preparations. Regression equations revealed good linear relationships (correlation coefficients 0.9969-0.9999) between peak heights in second-order derivative electropherograms and concentrations of the two analytes. The relative standard deviations (RSD) of the migration times and the peak height of the two constituents were less than 0.49% and 1.6%. The recoveries of two constituents ranged from 93.5 to 105.6%. The results indicated that baseline separation of the analytes was hard to be achieved in real samples sometimes and second-order derivative electropherograms was applicable for the resolving and analysis of overlapping peaks.