Malathion is a chemical pesticide, commonly used by Egyptian farmers; however, its contamination of water sources has become an important issue. It is suggested that the role of vitamin C in alteration of enzymes responsible for energy metabolism was induced by administration of Malathion. The effect of malathion and malathion with vitamin C on some enzymes in the rat were recorded in the present study. Significant increases in the level of hepatic enzymes (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma glutamyl transpeptidase) were recorded. Furthermore, renal markers such as urea, cholesterol and creatinine were increased in rats treated with malathion. Additionally, serum triglyceride was significantly decreased. No significant changes were observed in total protein, albumin and bilirubin in all treated groups as compared to the control. The present results showed that a significant (p < 0.001) decrease in glycolytic enzymes (hexokinase, pyruvatekinase, glucose phosphate isomerase and phosphofructokinase) were observed in treated groups, and lactate dehydrogenase enzyme activity also showed significant (p < 0.01) reduction. Coadministration of vitamin C to the groups restored all the parameters cited above to near-normal values. Therefore, the present investigation revealed that vitamin C appeared to be a promising agent for protection against malathion-induced toxicity.