Abstract: To explore the differences in the muscle nutritional components and quality among Hemibagrus guttatus, H. macropterus, and H. pluriradiatus, we applied the biochemical analysis methods to determine and evaluate the contents of general nutritional components, as well as the composition and contents of amino acids, fatty acids, and mineral elements in the dorsal muscles of the three fish species. The results show that the water content of H. guttatus was significantly higher than that of H. macropterus and H. pluriradiatus (p<0.05); the protein contents of H. pluriradiatus and H. macropterus were significantly higher than that of H. guttatus (p<0.05). The contents of crude fat and total sugar decreased following a descending order of H. pluriradiatus>H. macropterus>H. guttatus, with significant differences (p<0.05), while there was no significant differences in the ash content (p>0.05). Sixteen common amino acids had been detected in the three fish species. Glutamic acid had the highest content among them, and lysine had the highest content among essential amino acids (EAA). The total amino acid content (TAA) and essential amino acid index (EAAI) were the lowest in H. guttatus but the highest in H. pluriradiatus. Amino acid score (AAS) and chemical score (CS) indicate that the first limiting amino acid of the three fish species was methionine+cysteine. Twenty-four fatty acids had been detected in the three fish species. In terms of the contents of polyunsaturated fatty acids, H. guttatus had the highest content, and C18:2n6c (linoleic acid) of the n-6 series was the main one in the three fish species. The atherogenic index (AI=0.47) and thrombogenic index (TI=1.21) of H. guttatus were the lowest. The muscles of the three fish species were rich in various inorganic elements. The content of the major element potassium (K) was the highest, showing the characteristic of "high potassium and low sodium". The contents of trace elements iron (Fe), zinc (Zn), and boron (B) were relatively high. Selenium (Se) was only detected in H. pluriradiatus, and manganese (Mn) was only detected in H. macropterus. The study indicates that the protein, crude fat, total sugar, total amino acid content, and essential amino acid index of H. pluriradiatus are higher than H. guttatus and H. macropterus. And the water content and polyunsaturated fatty acid content of H. guttatus are higher than H. pluriradiatus and H. macropterus.