Abstract Background This study was developed to establish whether betaine was present in the sweat of females and to determine any correlations with other sweat components. Methods Sweat patches were placed on eight trained adolescent Highland dancers (age = 13.6 ± 2.3 yr), who then participated in a dance class for 2 hours. Patches were removed, and the sweat recovered via centrifugation. The sweat was subsequently analyzed for betaine, choline, sodium, potassium, chloride, lactate, glucose, urea and ammonia. Results Betaine was present in the sweat of all subjects (232 ± 84 μmol·L-1), which is higher than typically found in plasma. The concentration of several sweat components were correlated, in particular betaine with most other measured components. Conclusion Betaine, an osmoprotectant and methyl donor, is a component of sweat that may be lost from the body in significant amounts.