Previous research suggests that n-3 PUFA may play a role in bone health. The present analysis aimed to investigate the impact of n-3 PUFA supplementation on bone resorption in adult men and women. Serum samples from 113 mild-moderately depressed individuals (twenty-six males and eighty-seven females, aged 18-67 years) randomised to receive 1·48 g EPA+DHA/d (n 53) or placebo (n 60) for 12 weeks as part of a large recent randomised controlled trial were assayed for n-3 PUFA status and a bone resorption marker, C-terminal cross-linking telopeptide of type 1 collagen (β-CTX). Regression analyses revealed that n-3 PUFA status following supplementation was associated with randomisation (placebo/n-3 PUFA) (B = 3·25, 95 % CI 2·60, 3·91, P < 0·01). However, β-CTX status following supplementation was not associated with randomisation (B = - 0·01, 95 % CI - 0·03, 0·04). Change in β-CTX status was also not associated with change in n-3 PUFA status (B = - 0·002, 95 % CI - 0·01, 0·01). These findings provide no evidence for an association between n-3 PUFA supplementation (1·48 g EPA+DHA/d) for 12 weeks and bone resorption in humans assessed by β-CTX, and suggest that n-3 PUFA supplementation may be unlikely to be of benefit in preventing bone loss.