Application of a new validated HPLC-DAD method for simultaneous determination of ten active components in Xiedu San

In this research, a sensitive high-performance liquid chromatography-diode array detector (HPLC-DAD) method was established and validated for simultaneous detection of ten active constituents (gallic acid, catechin, berberine, palmatine, baicalin, baicalein, wogonin, rhein, emodin and chrysophanol) in Xiedu San, a traditional Chinese medicine compound preparation with effects of clearing away heat and toxic materials. The analysis was achieved on Agilent ZORBAX SB-C₁₈ column (5μm, 250mm×4.6mm) with the temperature of 30°C. Gradient elution was applied using methanol (A)-0.1% phosphoric acid solution (B) as mobile phase at the flow rate of 1.0mL• min^-1. The determination was performed at the wavelength of 225, 245, 278 and 348 nm along with the sample volume of 5μL. The tested constituents demonstrated good linear relationships within their respective determination ranges (r>0.9995). Average recoveries varied from 99.97% to 101.12% with RSDs of 0.71% to 1.92%. The contents of tested constituents ranged from 0.970 to 24.602 mg•g^-1. The developed method was proved to be simple, accurate and sensitive, which can provide a quantitative analysis method for the quality evaluation and quality control of Xiedu San.