The aim of the current study was to examine the effects of very long chain fatty acids (VLCFA) alone at 2 dietary levels, or in combination of VLCFA at the lower level with lecithin (LT) or phytosterols (PS), on lipid profiles and cholesterol biosynthesis in hamsters. Seventy-five male Golden Syrian hamsters, weighing 100 to 120 g, were fed a regular rodent chow for 2 weeks before being randomly assigned into 5 groups of 15 animals each fed semisynthetic diets for 4 weeks. Group 1 was given a control diet that contained 0.25% cholesterol and 5% fat with a polyunsaturated to saturated fatty acids ratio of 0.4. Groups 2 to 5 were fed the control diet and given 25 mg/kg BW per day of VLCFA (Licowax) (VLCFA25), 50 mg/kg BW per day of VLCFA (VLCFA50), 25 mg/kg BW per day of VLCFA+1000 mg/kg BW per day of LT (VLCFA25/LT), and 25 mg/kg BW per day of VLCFA+1000 mg/kg BW per day of PS (Cholestatin, VLCFA25/PS), respectively. Results showed that HDL-cholesterol (HDL-C) levels were not changed by VLCFA25, although increased by VLCFA50 (P<.05) relative to control. Total cholesterol (T-C) and non-HDL-C levels were not affected by VLCFA25 and VLCFA50 as compared with control. VLCFA25/LT had higher (P<.02) T-C and HDL-C levels than any other treatments and increased (P<.05) liver weight relative to control. In contrast, VLCFA25/PS reduced T-C (P=.0004) and non-HDL-C (P=.007) without effect on HDL-C levels compared with control. Triglyceride levels were not affected by any treatment. Cholesterol biosynthesis rate was higher (P<.05) in animals fed VLCFA25 and VLCFA50 than those fed control or VLCFA25/LT or VLCFA25/PS. Results suggest that PSs can decrease total and non-HDL-C cholesterol, whereas VLCFA may increase HDL-C in hamsters.