ObjectiveTo investigate the extraction, isolation, and structure identification of chemical constituents from petroleum ether fractions of Gnaphalium affine D. Don. MethodsIsolation and purification were carried out by silica gel column and Sephadex LH-20 column chromatography. The structures of the purified compounds were identified by spectral and physicochemical methods. ResultsTen compounds were identified from petroleum ether fractions of Gnaphalium affine D. Don: taraxasteryl acetate(1),α-amyrin(2),β-amyrin (3), betulinic acid (4), physcioin(5), emodin(6),(20R)-cholest-4-en-3-one (7),3β-hydroxy- stigmast-5, 22-dien-7-one(8),β-sitosterol(9),and n-hexacosanic acid(10). ConclusionCompounds 1-8 have been isolated from affine genus for the first time.