The current study was designed to investigate the physiochemical properties, protein and metabolite profiles of muscle exudate obtained from chicken breast fillets affected by wooden breast (WB) myopathy. Twenty-four fillets were categorized into varying degrees of WB condition including normal, moderate and severe. Results indicated that exudate loss, free hemoglobin concentration, protein and lipid oxidation were affected by WB myopathy. Electrophoresis analysis showed eight distinct protein bands of differential relative abundance in WB samples compared with the normal, and the identified proteins were mostly involved in carbohydrate metabolic process. H nuclear magnetic resonance-based metabolomics identified eleven metabolites including amino acids, nucleotides and organic acid as the most influential metabolites affected by WB myopathy. Overall, this study shows differential molecular profiles of myopathic chicken muscle exudate, and provides a valuable resource for further recognition of WB myopathy.