Quantitative HPLC Determination and Extraction of Anthraquinones in Senna alata Leaves

Abstract

A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of four anthraquinones: rhein, aloe-emodin, emodin, and chrysophanol in Senna alata leaves. The method involves the use of a TSK-gel ODS-80Tm column (5 microm, 4.6 x 150 mm) at 25 degrees C with the mixture of methanol and 2% aqueous acetic acid (70:30, v/v) as the mobile phase and detection at 254 nm. The parameters of linearity, precision, accuracy, and specificity of the method were evaluated. The recovery of the method is 100.3-100.5%, and linearity (r(2) > 0.9998) was obtained for all anthraquinones. A high degree of specificity as well as repeatability and reproducibility (relative standard deviation values less than 5%) were also achieved. The solvent for extraction of anthraquinones from S. alata leaves was examined in order to increase the anthraquinone content of the leaf extract. It was found that a solution of 5% hydrochloric acid (v/v), 5% ferric chloride (w/v), and 15% water in methanol (v/v) was capable of increasing the anthraquinone content in the leaf extract up to 1.67% (w/w).