This population-based cross-sectional study examined the prevalence and risk factors of suboptimal vitamin D levels (assessed using circulating 25-hydroxycholecalciferol (25(OH)D)) in a multi-ethnic sample of Asian adults. Plasma 25(OH)D concentration of 1139 Chinese, Malay and Indians (40–80 years) were stratified into normal (≥30 ng/mL), and suboptimal (including insufficiency and deficiency, <30 ng/mL) based on the 2011 Endocrine Society Clinical Practice Guidelines. Logistic regression models were used to assess the associations of demographic, lifestyle and clinical risk factors with the outcome. Of the 1139 participants, 25(OH)D concentration was suboptimal in 76.1%. In multivariable models, age ≤65 years (compared to age >65 years), Malay and Indian ethnicities (compared to Chinese ethnicity), and higher body mass index, HbA1c, education and income levels were associated with suboptimal 25(OH)D concentration (p < 0.05). In a population-based sample of Asian adults, approximately 75% had suboptimal 25(OH)D concentration. Targeted interventions and stricter reinforcements of existing guidelines for vitamin D supplementation are needed for groups at risk of vitamin D insufficiency/deficiency.