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Objective Emerging evidence indicates that vitamin D has anti-oxidative properties. The present study investigates whether serum 25‑hydroxyvitamin D [25(OH)D] is associated with biomarkers of anti-oxidative status in community-dwelling older adults using cross-sectional and longitudinal data. Methods A total of 302 subjects aged 62 to 92 years from Germany (50.6°N) were analysed via cross-sectional approach. For longitudinal analysis, data of 153 subjects were available. Fasting blood samples from 2004 and 2012 were analysed for 25(OH)D concentrations, total anti-oxidative status (TAOS) as well as anti-oxidative enzymes, such as catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD). Multiple regression analyses were performed to examine the associations between 25(OH)D and parameters of anti-oxidative status. Results In cross-sectional analyses, 25(OH)D was a significant predictor of CAT ( β  = −0.166; P  = 0.010), lg 10 GPx ( β  = 0.136; P  = 0.037) and TAOS ( β  = 0.121; P  = 0.048) after adjusting for age, sex, percentage total body fat (TBF), month of blood sampling, smoking behaviour and use of vitamin D supplements. Longitudinal change in 25(OH)D concentration positively predicted change in TAOS ( β  = 0.224; P  = 0.006) after adjusting for sex, baseline TAOS, age, smoking behaviour, use of vitamin D supplements and change in TBF, physical activity level, current time spent outdoors and dietary vitamin D intake. Conclusion The maintenance of an adequate vitamin D status may have a beneficial impact on the anti-oxidative defence system in older adults on a long-term perspective. [ABSTRACT FROM AUTHOR]