Background/Objectives: Data from human studies that have investigated the association between vitamin D status and cognitive function in elderly adults are conflicting. The objective of this study was to assess vitamin D status (reflected by serum 25-hydroxyvitamin D (25(OH)D)) in older European subjects (n ¼ 387; aged 55-87 years) and examine its association with measures of cognitive function. Subjects/Methods: Serum 25(OH)D was assessed using enzyme-linked immunosorbent assay, whereas measures of cognitive function were assessed using a comprehensive Cambridge Neuropsychological Testing Automated Battery (CANTAB). Results: In all, 12, 36 and 64% of subjects had serum 25(OH)D concentrations o30, o50 and o80 nmol/l, respectively, throughout the year. Serum 25(OH)D was significantly and inversely correlated with four assessments within the spatial working memory (SWM) test parameter (SWM between errors (r ¼ À0.166; P ¼ 0.003); SWM between errors 8 boxes (r ¼ À0.134; P ¼ 0.038); SWM strategy (r ¼ À0.246; Po0.0001); and SWM total errors (r ¼ À0.174; Po0.003)). When subjects were stratified on the basis of tertiles (T) of serum 25(OH)D (o47.6 (T 1 ); 47.6-85.8 (T 2 ); and 485.8 (T 3 ) nmol/l), fewer errors in SWM test scores occurred in subjects in the third T when compared with the first T (Po0.05-0.084). Stratification by sex showed that these differences between tertiles strengthened (Po0.001-0.043) in the females, but the differences were not significant (P40.6) in males. Conclusions: Vitamin D insufficiency, but not deficiency, is widespread in the older population of several European countries. Low vitamin D status was associated with a reduced capacity for SWM, particularly in women.