Antioxidant vitamins have been reported to be associated with an improvement in blood lipid profiles, but results are not consistent. The present study was designed to determine whether long-term vitamin C supplementation could alter serum lipid concentrations in subjects who completed a 5-year population-based double-blind intervention trial. A total of 439 Japanese subjects with atrophic gastritis initially participated in the trial using vitamin C and β-carotene to prevent gastric cancer. Before and upon early termination of β-carotene supplementation, 134 subjects dropped out of the trial; finally, 161 subjects assigned to the high-dose group (500 mg vitamin C/d) and 144 subjects assigned to the low-dose group (50 mg vitamin C/d) were studied. No favourable effect of vitamin C supplementation on serum concentrations of total cholesterol, HDL- and LDL-cholesterol, and triacylglycerol was observed, although high-dose vitamin C supplementation increased serum vitamin C concentrations substantially. Among women, the mean change in serum triacylglycerol decreased (−0·12 mmol/l, 95 % CI −0·32, 0·09) in the high-dose group, but increased (+0·12 mmol/l, 95 % CI 0·03, 0·22) in the low-dose group. In addition, the mean change in serum triacylglycerol among women with hypertriacylglycerolaemia was statistically significant (−1·21, 95 % CI −2·38, −0·05) after high-dose vitamin C supplementation. The 5-year vitamin C supplementation had no markedly favourable effects on the serum lipid and lipoprotein profile. However, our present results do not preclude the possibility that vitamin C supplementation may decrease triacylglycerol concentrations among women with hypertriacylglycerolaemia.