Differences in Flavour Compounds and Key Metabolic Markers in High-Quality Broiler Rooster Breast Muscle Based on Broad-Target Metabolomics and Volatile Metabolomics

Flavor is a pivotal indicator influencing the meat quality and palatability of premium broiler chickens, shaped by multiple factors. The flavor differences among broiler chicken breeds/lines stem from the specificity of their metabolite profiles and volatile flavor compounds. This study aims to identify key metabolites and pathways that regulate flavor variations in high-quality broilers, providing data support and theoretical references for breeding superior broiler lines and developing technologies to enhance flavor quality. Breast Muscle tissue from 15-week-old roosters of the S3 and H lines (<i>n</i> = 6) was used as experimental material. Broad-targeted metabolomics and volatile metabolomics technologies were employed to identify key metabolites and volatile organic compounds (VOCs) influencing the flavor of breast meat in these two high-quality broiler lines. Broad-target metabolomics identified 167 differentially expressed metabolites (VIP > 1, <i>p</i> < 0.05) between the two strains, including 141 upregulated and 26 downregulated metabolites. These metabolites were primarily amino acids and their derivatives, and were significantly enriched in metabolic pathways such as ABC transporters (<i>p</i> < 0.05). Leu-Tyr, Ile-Tyr, Val-Leu, Val-Ile, and Tyr-Ala were identified as key metabolites influencing the flavor formation of breast meat from both high-quality broiler lines. Volatile metabolomics results identified 33 downregulated VOCs (VIP > 1 and <i>p</i> < 0.05). The flavor differences between the two strains primarily involved fatty and grassy flavor. Key flavor markers included 2-Nonanone, 2-Nonanone, 3-hydroxymethyl, 2-Methylheptanoic acid, and Hexanoic acid, butyl ester as the primary flavor markers. These significantly downregulated volatiles are formed through lipid oxidation and amino acid degradation pathways, respectively, collectively shaping the more pronounced fatty and grassy aromas in the S3 strain. Correlation analysis revealed a significant negative correlation between Met-Asn and Hexanoic acid, butyl ester, suggesting it may represent a key regulatory pathway influencing green flavor formation. In summary, this study elucidates key metabolites and pathways governing flavor differences in high-quality broiler rooster breast meat, providing a scientific foundation for poultry breeding, optimization of farming practices, and flavor regulation in meat products.