Pleurotus tuber reguim is a mushroom widely used as food, and in traditional medicine for weight loss and treatment of headache, fever and stomach pain among other ailments. This present study is aimed at the evaluation of antihyperlipidemic activities of chromatography fractions obtained from an earlier reported anti-hyperlipidemic chloroform extract of Pleurotus tuber reguim using triton X-100 induced hyperlipidemic rat model as well as investigate the phytochemical constituent(s) from the active chromatography fraction(s). The pulverized sclerotia sample was extracted exhaustively by cold maceration with chloroform and the chloroform extract (6.42g) fractionated on a chromatography column(silica gel G, 200 -400 mesh-stationary phase) eluting with a gradient mixtures of n-hexane: chloroform: methanol (4:0:0; 3:1:0; 1:3:0; 0:1:0; 0:3:1; 0:2:2; 0:1:3; 0:4- v/v/v). Fractions with similar chromatography fingerprint on thin layer chromatography examination were pooled. Phytochemical screening was done using standard methods. The chloroform extract afforded four pooled chromatography fractions (F1, F2, F3 and F4). The trend: Total cholesterol [atovastatin (47.5%) > F1 (42.3%) > F2 (37.6%) > F4 (29.9%) > F3 (29.8%)]; Triglyceride [F4 (47.6%) > F1 (44.8%) > Atorvastatin (35.6%) > F3 (32.9%) > F2 (4.3%)]; High density lipoprotein-cholesterol [F2 (101.3%) > F1 (91.2%) > atovastatin (53.7%) > F3 (17.7%) > F4 (14.9%)];Very low density lipoprotein-cholesterol [F4 (47.6%) > F1 (44.8%) > Atorvastatin (35.7%) > F3 (32.9%) > F2 (4.3%)]; Low density lipoprotein-cholesterol [Atorvastatin (12.5%) > F2 (10.0%) > F1 (7.6%) > F3 (3.5%) > F4 (-4.9%)]. Fractions F1 and F2 where found to contain triterpenoids and fatty acids while fractions F3 and F4 were found to contained amino acids or peptide derivatives. Pleurotus tuber reguim exhibited hyperlipidemic activity and the presence of triterpenoids and amino acids or peptide derviatives could be responsible for the observed trend in antihyperlipidemic activity.