Analysis of nine rhubarb anthraquinones and bianthrones by micellar electrokinetic chromatography using experimental design

An efficient micellar electrokinetic chromatography (MEKC) method has been developed for the analysis of nine anthraquinones and bianthrones in rhubarb. A chemometric approach was used to search for the optimum conditions of separation. Those factors which were found to be significant with a screening design were further optimized with a central composite face-centered (CCF) design. Acetonitrile concentration was found to be the most influential, not only in resolution, but also in analysis time and peak asymmetry. With the optimized conditions: 15 mM sodium tetraborate/15 mM sodium dihydrogenphosphate buffer, 30 mM sodium deoxycholate, pH 8.6, 17 vol.% acetonitrile and 28 kV, nine tested analytes were baseline-separated within 14 min. The method was validated to analyze the rhubarb material. Solid-phase extraction (SPE) was manipulated to remove interfering substances. Five anthraquinones and two glycosidic bianthrones were detected and quantificated. The method should be suitable for determining these major active principles in rhubarb crude drugs.