Optimization of a Reverse-Phase High Performance Liquid Chromatography (RP-HPLC) Method for Simultaneous Separation of Aloe-Emodin, Rhein, Emodin, Chrysophanol and Physcion

Abstract

In this study, a reverse-phase high performance liquid chromatography (RP-HPLC) method with high separation efficiency, high detection sensitivity and excellent selectivity was successfully developed for the simultaneous quantitative determination of the aloe-emodin, rhein, emodin, chrysophanol, and physcion compounds. These optimum method conditions were determined by studying various columns, mobile phases and compositions, flow rates and column temperatures. The successful separation was carried out with a Supelcosil LC-18 column (250 × 4.6 mm, 5 µm) and a gradient program. The procedure was carried out at 20 °C with the flow rate of 1.0 mL/min and the injection volume of 20 μL utilizing an RP-HPLC method with DAD detector at 225 nm. Deionized water containing 0.5% orthophosphoric acid and methanol was used as the mobile phases A and B. This study showed an optimized analytical method can be effectively utilized to identify and measure anthraquinone compounds.

Keywords

• Reverse Phase High Performance Liquid Chromatography
• Anthraquinon
• Secondary Metabolites
• Method Optimization

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