Simultaneous quantification of five anthraquinones in rat plasma by high‐performance liquid chromatography with fluorescence detection

A sensitive high-performance liquid chromatographic method with fluorescence detection (excitation 435 and emission 515 nm) was established and validated for quantification of five anthraquinones (aloe-emodin, rhein, emodin, chrysophanol and physcion) in rat plasma. Following a single-step liquid-liquid extraction, the analytes and internal standard (1,8-dihydroxyanthraquinone) were separated on a reversed-phase C(18) column with water-phosphoric acid-methanol as mobile phase at a flow rate of 1 mL/min. The linear ranges of the calibration curves were 6.5-1300 ng/mL for aloe-emodin, 20-4000 ng/mL for rhein, 40-8000 ng/mL for emodin, 15-3000 ng/mL for chrysophanol and 13-2600 ng/mL for physcion. The lower limit of quantification was 6.5 ng/mL for aloe-emodin, 20 ng/mL for rhein, 40 ng/mL for emodin, 15 ng/mL for chrysophanol and 13 ng/mL for physcion. The mean accuracy was 94.3-105.1% for aloe-emodin, 90.3-108.8% for rhein, 92.6-106.7% for emodin, 95.8-103.8% for chrysophanol and 98.7-101.2% for physcion. The within-batch and between-batch precisions were < or = 5.5% and < or = 13.4%, respectively. This method is suitable for determining the five anthraquinones in plasma simultaneously and thus investigating the pharmacokinetics of anthraquinones from Xiexin decoction in rats.