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Introduction: The identification of modifiable health span-promoting factors is a public health priority. Objective: To explore the socio-environmental, lifestyle, behavioural, and psychological determinants of a clinical phenotypic measure of biological ageing in the Singapore Longitudinal Ageing Study (SLAS) cohort. Methods: Using cross-sectional data on 2,844 SLAS-2 adults with a chronological age (CA) ≥55 years, we estimated biological age (BA) using a validated panel of clinical, biochemical, physiological, and functional indicators (8 in men and 10 in women) and calculated the difference between BA and CA (BA – CA in years). Potential determinants included education, housing status, loss of a spouse, living alone, lifestyle and health activity, smoking, alcohol consumption, nutritional risks, consumption of milk, soy, fruit, vegetables, coffee and tea, sleep parameters, and life satisfaction. Results: The mean CA was 67.0 (standard deviation [SD] 7.9; range 55–94) years. The estimated BA varied more widely (SD 8.9 years; range 47.5–119.9 years), and BA – CA ranged from –11.3 to 30.0 years. In stepwise selection regression analyses, multiple significant independent determinants in a final model were larger for private housing, being single/divorced/widowed, productivity, cognitive and leisure time activity scores, 10 h/week of moderate-to-vigorous physical activity, unintended loss of weight, life satisfaction, and daily consumption of fruits 1–2 or ≥3 servings and Chinese tea 1–2 or ≥3 cups daily, together explaining 16% of BA – CA variance in men and 14% in women. Associated BA – CA estimates were highest in men with high-end housing status (–1.8 years, effect size 0.015) and unintended weight loss (1.5 years, effect size 0.017). Conclusion: We identified determinants of biological ageing which can promote health span.