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

Year 2021, Issue: 27, 972 - 984, 30.11.2021

Gizem Yıldırım Baştemur , Reyhan Akpınar Sabriye Perçin Özkorucuklu , Esengül Kır

https://doi.org/10.31590/ejosat.987628

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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