Tupistra chinensis(Liliaceae), mainly distributed in southwestern China, is a rich source of steroidal sapogenins, saponins and flavans. These components exhibit diverse biological activities such as anti-inflammatory, anti-fungal and anti-tumor properties. As a part of a continuing study for the discovery of novel cytotoxic agents, eight compounds were isolated from the fresh rhizomes of Tupistra chinensis, and their structures were established on the basis of chemical methods and spectral data(MS, IR, NMR), including tupistroside J(1), (25S)-26-O-β-D-glucopyranosyl-furost-1β,3β,22α,26-tetraol-3-O-β-D-glucoside(2), digitoxigenin-3-O-α-L-fucopyranose(3), watti-genin B(4), epiruscogenin(5), epiruscogenin(6), orobol(7), (+)-catechin(8). Compound 1 was a new compound, and compounds 3, 4, 7 and 8 were isolated from Tupistra Ker-Gawl for the first time. All compounds were tested in vitro for their cytotoxic activities against the Human LoVo and BGC-823 cell lines, and compounds 5 and 6 were found to possess significant cytotoxicity against the tested tumor cell lines with half maximal inhibitory concentration(IC50) values of 0.532 and 0.757 μmol/L, respectively.