Abstract Background and purpose Plyometric training and β-alanine supplementation are common among athletes, although its mixed-use had never been investigated in handball players. Thus, a randomized, double-blind, placebo-controlled trial was performed to investigate the impacts of a plyometric training regimen with or without β-alanine supplementation on anaerobic performance and carnosine levels in male handball players. Methods Eighteen male handball players were allocated to either a plyometric training group receiving placebo (placebo, n = 9; 24.33 ± 5.12 years) and a plyometric training group receiving β-alanine (β-alanine, n = 9; 18.89 ± 2.14 years) supplementation. Participants were evaluated for anaerobic power, serum level of carnosine, and blood lactate before and after a 4-week intervention. Results Average power and peak power were increased more after β-alanine ingestion than after placebo (10.4 and 9.1% versus − 1.7 and − 6.8%, respectively). Fatigue index was significantly decreased by 26.2% in the placebo group, with no significant change in the β-alanine group (+ 10.6%). The serum level of carnosine in both groups was significantly increased compared with pre-intervention. It was increased more after β-alanine supplementation than placebo (61.8 versus 43.6%). Conclusion It was concluded that β-alanine consumption during plyometric training might add further adaptive changes related to anaerobic power and carnosine levels.

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