Portulaca oleracea is a fleshy annual herb which is distributed throughout the temperate and tropical areas of the world. The chromatographic fraction 2 of this plant has been reported to have deleterious effects on reproductive parameters in male rats. Air-dried specimen of Portulaca oleracea (3.2 kg) was cold-extracted in methanol for 72 hours. The resulting methanol extract was then subjected to open column chromatography on silica gel for fractionation. Out of the 5 fractions obtained, fraction 2 was then subjected to male rats’ reproductive bioassays. Twenty male rats (120-150 g) were divided into control (distilled water) and fraction 2 (1, 2, 3 mg/kg) treated groups (5 per group) for hormonal assay and andrological studies. The animals were orally treated on daily basis for 50 days and allowed a recovery (withdrawal) period of 50 days after which plasma testosterone level was assayed using Enzyme-Linked Immunosorbent Assay (ELISA) and semen analysis was done microscopically. There were significant (p<0.05) decrease in sperm motility of fraction 2 (1 mg/kg, 2 mg/kg, 3 mg/kg) recovery groups relative to control. Also, there were significant (p<0.05) decrease in sperm counts of fraction 2 (1 mg/kg, 3 mg/kg) recovery groups relative to the control. It can therefore be concluded that the deleterious effects induced by chromatographic fraction 2 of Portulaca oleracea on the reproductive parameters in male rats were probably not totally reversible.