A reverse phase ultra performance liquid chromatography (UPLC) method was developed for the rapid quantification of five anthraquinone derivatives (aloe-emodin, rhein, emodin, chrysophanol and physcion) in rhubarb using a Waters Acquity BEH C18, 50 mm x 2.1 mm, 1.7 microm column. The runtime was as short as 3 min. The influence of flow rate and column temperature on resolution was investigated. The method was validated according to the regulatory guidelines with respect to precision, accuracy, linearity and robustness. Comparison of system performance with conventional HPLC was made with respect to analysis time, efficiency and sensitivity. The proposed method was found to be reproducible and convenient for quantitative analysis of five anthraquinone derivatives in three species of rhubarb and related preparations.