Abstract The Maillard reaction (MR) of tilapia byproduct protein hydrolysates was investigated for the use of byproduct protein as a food ingredient and to mask its fishy odor and bitter flavor. The flavor differences in tilapia byproduct hydrolysates before and after the MR were analyzed to explore the key flavor precursor peptides and amino acids involved in MR. The results suggested that eight key volatile substances, including 2,5‐dimethylpyrazine, 2‐pentylfuran, hexanal, octanal, nonanal, ( E )‐2‐decenal, decanal, and 1‐octen‐3‐ol contributed most to the MR products group (ROAV > 1). Ten volatile compounds, including 1‐octen‐3‐ol, hexanal, 2‐pentylfuran, 2,5‐dimethylpyrazine, methyl decanoate, and 2‐octylfuran, were the flavor markers that distinguished the different samples (VIP > 1). The four most consumed peptides were VAPEEHPTL, GPIGPRGPAG, KSADDIKKAF, and VWEGQNIVK. Umami peptides and bitter free amino acids (FAAs) were the key flavor precursor peptide and FAAs, respectively. Overall, the hydrolysates of tilapia byproducts with flavor improved by MR are a promising strategy for the production of flavorings.