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Background: Vitamin B12 and folate function as co-factors in pathways used during physical activity. Physical activity may therefore increase vitamin requirements, leading to a risk of deficient plasma concentrations. We aimed to investigate the relationship between intake and plasma concentrations of vitamin B12 and folate in physically active adults, as well as identify other determinants of vitamin B12 and folate plasma concentrations., Methods: The study population consisted of 873 adults (528 men and 345 women), aged 19-78 years, who participated in a 4-day walking event. The relationship between intake and plasma concentrations of vitamin B12 and folate was assessed using correlation and linear regression analyses. In addition, potential other determinants (sex, age, body mass index, energy intake and physical activity) of vitamin plasma concentrations were investigated., Results: Significant positive correlations were observed between intake and plasma concentrations of vitamin B12 [Pearson's correlation coefficient = 0.15; 95% confidence interval (CI) = 0.08-0.21] and folate (Pearson's correlation coefficient = 0.18; 95% CI = 0.12-0.25). In addition to vitamin intake, sex, age and energy intake were also determinants of both vitamin B12 and folate plasma concentrations in multivariable regression models., Conclusions: The results suggest a positive association between intake and plasma concentrations for both vitamin B12 and folate in physically active people. By contrast to our hypothesis, physical activity was not a determinant of vitamin B12 and folate plasma concentrations. However, sex, age and energy intake were found to be determinants. Thus, when studying the relationship between intake and plasma concentrations of vitamin B12 or folate, these factors should be taken into account., (© 2020 The Authors. Journal of Human Nutrition and Dietetics published by John Wiley & Sons Ltd on behalf of British Dietetic Association.)