Chlorophytum alismifolium is widely utilized as remedy for pain, inflammation and diabetes mellitus. Toxicological evaluation of the plant demonstrated an appreciable degree of safety following sub-acute administration. This study was performed to evaluate the effect of sub-chronic (13 weeks) administration of C. alismifolium in Wistar rats. The tubers of C. alismifolium were obtained, processed and extracted with methanol to get the methanol extract of C. alismifolium (MECA). MECA (150, 300, and 600 mg/kg) was administered orally and daily to rats for 13 weeks. Thereafter, toxicological parameters were evaluated. Administration of MECA for 13 weeks did not significantly alter the feed intake, water consumption and body weights of the animals. Hematological analysis showed a significant (p < 0.05) rise in granulocyte levels in male rats at 600 mg/kg. In the biochemical investigations, alanine aminotransferase was significantly (p < 0.05) raised in the female rats at 600 mg/kg relative to the control. A significant (p < 0.05) reduction in glucose level was also observed in the male rats at 300 mg/kg. Histological investigations showed normal features in some organs as no major distortions in the histo-architecture of tissues were observed. However, slight to moderate alterations in the hepatic and renal tissues were observed at 300 and 600 mg/kg of the extract. The findings acquired following 13 weeks of administration of MECA indicate that MECA is comparatively not toxic in rats. However, prolonged administration of the extract at larger doses may predispose the liver to toxic effect.