Casein Supplementation Timing and Exercise Performance in Soccer Players: Pre-Sleep vs. Post-Exercise Intake—A Randomized Controlled Trial

Background: Nutrient timing strategies may influence recovery and performance in athletes; however, the effects of ingesting casein protein before sleep versus immediately after exercise remain unclear. This study compared the acute effects of pre-sleep and post-exercise ingestion of casein on high-intensity anaerobic performance in highly trained soccer players. Methods: Twenty-four highly trained male soccer players (mean age: 20.6 ± 1.8 years) were randomly assigned to three groups: post-exercise casein ingestion group (PECIG; n = 8), pre-sleep casein ingestion group (PSCIG; n = 8), and control group (CG; n = 8). Following a standardized high-intensity resistance training protocol, participants consumed 30 g of micellar casein dissolved in 300 mL of water either immediately after exercise or 30–60 min before bedtime. Performance was assessed using the Countermovement Jump (CMJ), the Illinois Agility Test (IAT), and the Running-based Anaerobic Sprint Test (RAST), which were conducted both before and 24 h after the training session. Results: Repeated-measures ANOVA revealed significant group-by-time effects for CMJ (F = 8.21, p = 0.002, η²p = 0.36) and RAST performance variables, including peak power (F = 6.94, p = 0.003, η²p = 0.32), mean power (F = 7.42, p = 0.002, η²p = 0.34), and fatigue index (F = 5.87, p = 0.008, η²p = 0.28). Post hoc analyses showed that PSCIG significantly improved CMJ (Δ = +1.45 cm; p < 0.001, d = 2.04) and RAST mean power (Δ = +125.14 W; p = 0.002, d = 1.55) compared with the control condition. PECIG significantly enhanced RAST peak power (Δ = +205.79 W; p = 0.001, d = 1.64), mean power (Δ = +128.43 W; p = 0.013, d = 1.24), and fatigue index (Δ = +4.12 W/s; p = 0.010, d = 1.29) relative to CG. No performance differences were observed between PSCIG and PECIG timing conditions (all p > 0.05). Conclusions: Both pre-sleep and post-exercise casein ingestion enhanced anaerobic performance in highly trained soccer players, with each timing method favoring different performance outcomes. However, neither strategy demonstrated superiority over the other, suggesting that casein timing may be flexibly applied according to athletes’ preferences and training contexts.