The effect of glutamine on hepatic steatosis and serum amino acid pattern was studied in rats receiving total parenteral nutrition (TPN) with different levels of caloric intake. Rats were divided into four groups; a control group (n = 10) was fed a chow diet and infused with saline only. Three experimental groups (n = 8 to 10) received TPN solutions at energy levels of 25 kcal, 30 kcal, and 35 kcal/100 g body weight, respectively. The experimental groups were maintained with TPN for a period of 6 days. Each experimental group was divided into two subgroups, one of which was supplemented with glutamine, replacing 40% of the total amino acid nitrogen. All of the basal TPN solutions were isonitrogenous and identical in nutrient composition, except for the difference in energy level, which was adjusted with glucose. The results demonstrated that liver fat increased in accordance with the increase of glucose supply, and this increase was mainly due to triglyceride accumulation. Very‐low‐density lipoprotein‐triglyceride and serum free fatty acid were significantly higher in the 30‐kcal groups. There were no differences in hepatic lipid content, very‐low‐density lipoprotein‐triglyceride secretion, or hepatic uptake of fatty acids between subgroups with and without glutamine supplementation. It was concluded that glutamine enrichment of a TPN solution did not have any effect on hepatic steatosis in normal rats. (Journal of Parenteral and Enteral Nutrition18:40–44, 1994)