Objective: To explore the association between vitamin B 12 (B 12 )-containing supplement use, low B 12 concentrations and biochemically defined B 12 deficiency in US adults. Design: A cross-sectional study with adjustment for survey design. Prevalence ratios for two age groups (18-50 and .50 years) were estimated using unconditional logistic models. Outcome measures included prevalence of low serum B 12 concentration (,148 pmol/l) and biochemical B 12 deficiency (serum B 12 , 148 pmol/l with concomitant homocysteine . 10 mmol/l). Setting: A population survey of health and nutritional measures. Subjects: Subjects were non-institutionalized adults, aged 18 years and older, who participated in Phase 2 of NHANES III (Third National Health and Nutrition Examination Survey). Results: Low B 12 concentrations were less prevalent among persons consuming B 12 -containing supplements (P 5 0?001) with an adjusted prevalence ratio of 0?6 (95 % CI 0?3, 1?0). Biochemical B 12 deficiency showed a similar trend (P 5 0?0002), with an adjusted prevalence ratio of 0?3 (95 % CI 0?1, 0?8). Prevalence ratios were similar in adults .50 years of age, although the prevalence of low B 12 and biochemical deficiency was proportionally higher. Conclusions: Consumption of B 12 -containing supplements was associated with at least 50 % lower prevalence of both low serum B 12 and biochemical B 12 deficiency in a nationally representative sample of US adults, suggesting increased consumption of B 12 from supplements or from fortified foods may reduce the prevalence of B 12 deficiency. Additionally, the current Recommended Daily Allowance for B 12 of 2?4 mg may be insufficient for those aged .50 years. The Institute of Medicine (IOM) extensively reviewed the available data on vitamin B 12 (B 12 ) deficiency and requirements, and has identified biochemical B 12 deficiency and effective methods to reduce risk of B 12 deficiency as high priorities for research (1) . The Recommended Daily Allowance (RDA) was set at 120 % of the daily requirement and is expected to protect 97-98 % of healthy persons from deficiency. Based on the lower bioavailability of food-bound B 12 in older individuals, the IOM recommended adults over the age of 50 years meet the RDA for B 12 of 2?4 mg by consuming synthetic vitamin B 12 in B 12 -fortified foods or B 12 supplements (1) . Previously, three population-based studies examined biochemical indicators of B 12 deficiency (serum B 12 and/or homocysteine and methylmalonic acid (MMA) concentrations) in relation to vitamin supplement status and concluded that B 12 -containing supplements may protect against B 12 deficiency