Two new steroidal alkaloids, neoverapatuline (1) and (1 beta,3 alpha,5 beta)-1,3 Phi-dihydroxyjervanin-12-en-11one (2), together with the four known compounds, veratramine (3), rubijervine (4), veratrosine (5). and veratroylzygadenine (6), were isolated from the roots and rhizomes of Veratrion nigrum L. Their structures were established through combined analyses of physicochemical properties and spectroscopic evidence. All compounds 1- 6 were tested for their cytotoxicities in vitro against the human glioma cell line SF 188.