Abstract: A comprehensive analytical method based on UPLC-MS/MS was developed for the simultaneous determination of fourteen components include three stilbenes (stilbeneglucoside, polydatin, resveratrol), five anthraquinones (emodin, physcion, emodin-8-β-D-glucopyranoside, aloe-emodin, rhein), five flavonoids (epicatechin, quercetin, rutin, astragalin, hyperoside), and one phenolic acid (gallic acid) in Polygoni Multi-flori Radix. The samples was extracted with 70% methanol water solutions and assisted by ultrasonic machine. The extraction conditions which were optimized by experiment as following: the solid liquid ratio was 1∶25, the extraction time was 45 min at room -temperature, the extractant phase was separated by centrifugation at 12 000 r/min for 10 min, then filtered through 0.22 μm nylon membranes. Under the optimized chromatographic conditions, good separation for fourteen target compounds were obtained on a Waters BEH C18 （100 mm×2.1 mm×1.7 μm） eluted by a mobile phase of water (0.1% acetic acid, A) acetonitrile (B) at a flow rate of 0.25 mL/min. The target compounds were analyzed by multiple reaction monitoring (MRM) mode. The results show that fourteen components have good linearity, the linearity of the detected components is excellent more than 0.991 5, and the limits of detection are all satisfied. The average recoveries of standard addition for the compounds are between 95.85% and 102.13%, and the relative standard deviations are less than 5%. The established method is simplify, sensitive and accurate, which is suitable for determination of fourteen components in Polygoni Multiflori Radix, and also can provide a reliable and effective technique for the quality control of Polygoni Multiflori Radix.