This study was aimed to establish an analytical method to determine spinosin, jujuboside A, jujuboside B and betulinic acid content of Ziziphi Spinosae Semen quickly and accurately. The UPLC determination of Ziziphi Spinosae Semen method was established on an HSS T3 Column (2.1 mm × 100 mm, 1.8 μm) eluted with the mobile phase consisted of water containing 0.03% phosphoric acid and acetonitrile in gradient mode with the flow rate of 0.5 mL?min-1 and the detection wavelength was set at 204 nm. The standard curves of spinosin, jujuboside A, jujuboside B, betulinic acid showed a good linearity in 29.70 - 594.0 μg?mL-1, 10.68 - 213.6 μg?mL-1, 6.175 - 123.5 μg?mL-1, 22.00 - 440.0 μg?mL-1, respectively. The mean recoveries (n = 9) of the four components were between 98 . 3% and 99 . 4%, RSD < 1 . 4%. This method which is quick and accurate can be used to determine spinosin, jujuboside A, jujuboside B and betulinic acid in Ziziphi Spinosae Semen.