This study was performed to investigate the relationship between serum L-ascorbic acid, vitamin C intake, and diabetes in a nested case-control study. A cross-sectional survey of diet and health was conducted in 2,048 adults with an age of 30 y or older in Yonchon County, Korea. An oral glucose tolerance test was administered to all participants. One hundred cases of newly diagnosed diabetes were identified. Two healthy controls for each case matched with age, gender, drinking status, and smoking status were selected among the survey participants. L-Ascorbic acid levels were analyzed in fasting serum samples and one 24-h dietary recall was performed. Dietary vitamin C intake of persons with diabetes was 50.1±47.6 mg/d and that of controls was 55.1±41.1 mg/d. People with diabetes (22.3±16.8 µmol/L) have lower serum ascorbic acid levels than their controls (26.3±17.0 µmol/L) and the difference was significant by paired t-test (p<0.01). The association between diabetes and serum ascorbic acid level was still significant in non-smokers (24.2±17.8 µmol/L for the diabetes group and 29.5±16.7 µmol/L for the control group, p<0.01) but not in smokers (19.4±15.7 µmol/L for the diabetes group and 21.2±16.0 µmol/L for the control group). Our results suggest that diabetes and smoking interactively affect serum ascorbic acid levels. Since this population had poor nutritional status of vitamin C, further investigation of association between serum ascorbic acid level and diabetes and smoking by the level of vitamin C consumption is warranted.