Abstract: Using the branches and leaves of Taxus cuspidata Siebold et Zucc. as raw materials and petroleum ether as extraction solvent, the taxinine was extracted by the ultrasonic-assisted method. The extraction process was optimized by single factor test and orthogonal method, and taxinine was obtained by separation and purification. In addition, the anti-tumor activity test was conducted on different separation components of petroleum ether and taxinine. The optimal conditions for ultrasonic-assisted method of 10 g taxinine were ultrasonic power of 168 W, extraction time of 50 min, material to liquid ratio of 1∶8(g∶mL) and extraction temperature of 50 ℃. The petroleum ether extract was separated and purified by silica gel column and C18 reversed-phase column to obtain taxanes. The compound was identified by nuclear magnetic resonance(NMR) and single crystal X-ray diffraction structure analysis. The results of anti-tumor activity of Fr.1-Fr.7 fraction and taxinine obtained from Fr.4 fraction showed that Fr.1, Fr.3, Fr.4 and Fr.5 had good inhibitory activity on human cervical cancer cells, with half inhibitory mass concentration(IC₅₀) values of 32.3, 31.9, 22.3 and 38.4 mg/L, respectively. Taxinine had a good inhibitory effect on human breast cancer cells, human non-small cell lung cancer cells and human cervical cancer cells. Among them, the inhibitory effect on non-small cell lung cancer was relatively best, with IC₅₀ value of 13.1 mg/L, followed by the inhibitory effect on breast cancer cells and IC₅₀ value was 17.9 mg/L. It also had a certain inhibitory effect on cervical cancer cells, with IC₅₀ value of 35.5 mg/L.