The chemical constituent investigation on the root bark of Ailanthus altissima leads to the isolation of a new β-carboline alkaloid, 14(S),15-dihydroxy-6-methoxy-β-carboline (1), along with nine known alkaloids. The structure of new compound was elucidated on basis of extensive spectroscopic analysis, especially two-dimensional (2D) NMR techniques and the absolute configuration of C-14 was determined by ECD calculation. The neuroprotective effect of the isolated compounds on PC12 cells against the serum deprivation injury was evaluated by MTT method. As a result, compound 7 revealed protective effect on PC12 cells and the cell survival rate was significantly increased. © 2022 Informa UK Limited, trading as Taylor & Francis Group.