Simultaneous determination of hydroxyanthraquinones in rhubarb and experimental animal bodies by high-performance liquid chromatography.

A simple and reliable high-performance liquid chromatographic (HPLC) method was developed for the simultaneous determination of five hydroxyanthraquinones (aloe-emodin, rhein, emodin, chrysophanol, and physcion) in Rhubarb and experimental animal bodies. A Zorbax SB-C18 column (250 mm x 4.6 mm i.d., 5 microm) and a methanol-0.5% acetic acid (85:15, v/v) mobile phase were used for the separation. The detection wavelength of a diode array detector (DAD) was set at 254 nm. Regression equations revealed a linear relationship (R2>0.9996) between the mass of hydroxyanthraquinones injected and the peak areas detected by DAD. The detection limits (S/N=3) ranged from 0.35 ng to 3.13 ng, and the recoveries ranged from 83% to 103% for different hydroxyanthraquinones. This method is simple, sensitive and suitable for the analysis of hydroxyanthraquinones in medicinal materials and pharmacological experiment samples.