INTRODUCTION: The aims of the study were 1) to evaluate whether a multi-facetted simulated soccer game protocol, entitled the Copenhagen Soccer Test (CST), elicited a similar physiological loading as a competitive game, and 2) to determine muscle metabolites, blood variables and sprint performance in various phases of CST. METHODS: Twelve Danish Second and Third Division soccer players participated in the study. On separate days, heart rate (HR) measurements, frequent blood sampling and physical/technical tests were performed during 60- and 90-min versions of CST during which repeated m. vastus lateralis biopsies were collected. A competitive game (CG) was also played, where HR was recorded and pre-post muscle biopsies and blood samples were collected. RESULTS: No differences were observed between CST and CG in average HR (85±1 and 86±1%HRmax, P>0.05) or recovery plasma creatine kinase (24h: 312±57 and 324±76 U·L, P>0.05). Muscle glycogen decreased (P0.05). Rate of glycogen utilization was 4±1 mmol·kg d.w.·min during the warm-up and the first 15 min of CST and 1±1 mmol·kg d.w.·min (P