Introduction: Short and long sleep durations are associated with heightened risk for cardiovascular disease and vascular risk factors. Elevated homocysteine is also associated with greater risk for cardiovascular disease; however, studies have yet to investigate the relationship between sleep duration and homocysteine. Hypothesis: We hypothesized that short and long sleep duration would be associated with clinical levels of homocysteine. Methods: Adults (n=2,469; ≥20y) from the 2005-2006 National Health and Nutrition Examination Survey (NHANES) were assessed for habitual sleep duration (coded as <5, 5, 6, 7, 8, 9, and ≥10hrs) and fasting plasma homocysteine levels (<10 [normative], 10 to <15 [pre-clinical] and ≥15 [clinical] μmol/L). Participants were excluded if pregnant, lactating, missing data on the primary variables, or if they had a history of cardiovascular disease, cancer, diabetes, kidney disease, or diagnosed sleep disorder. Population weighted, multinomial logistic regression analyses assessed the relationship between sleep duration and homocysteine after adjustment for age, sex, race/ethnicity, marital and menopausal status, shift work, dietary folate, alcohol intake, cotinine levels, reported physical activity, hypertension, and reported frequency of cessation of breathing at night. Results: Pre-clinical and clinical levels of homocysteine were present in 13.7% and 2.5% of the sample, respectively. The mean sleep duration was 6.9 ± 1.4 hours. In adjusted analyses, sleep duration was significantly related to homocysteine ( p < 0.001). See Table. Very short sleepers (<5hrs) were more likely to have clinical levels of homocysteine (OR: 3.01, 95%CI: 1.38, 6.57) compared to 7-hr sleepers. Conclusions: In a U.S. representative sample of adults without cardiovascular disease or other major conditions, short sleepers were at greater odds for clinical levels of homocysteine Findings suggest that homocysteine may be one mechanism linking short sleep duration to cardiovascular disease.