Collagen, a critical biomaterial with wide applications in pharmaceuticals, cosmetics, and medical industries, can be sourced sustainably from fish scales. This study optimizes the extraction of col-lagen using Tris-Glycine buffer from fish scales via the Taguchi method. Various extraction pa-rameters—buffer concentration, temperature, pH, and time—were evaluated to identify optimal conditions. Under optimal conditions (0.5 of acetic acids, volume of acids of 100mL, soaking time of 120 min, and Tris-Glycine buffer of 10mL), the results demonstrate that temperature and buffer concentration significantly influence collagen yield, with a collagen purity of 17.14 ± 0.05 mg/g. R2 value of 73.84% was obtained for the mathematical model). FTIR analysis confirmed the pres-ence of characteristic collagen peaks at 1611 cm⁻¹ (amide I), 1523 cm⁻¹ (amide II), and 1300 cm⁻¹ (amide III), indicating the successful extraction of Type I collagen. SDS-PAGE analysis revealed a protein banding pattern consistent with the molecular weight of collagen, and amino acid analysis shows high percentages of glycine (20.98%), proline (15.43%), and hydroxyproline (11.51%), im-plying fibrous collagen structures. The finding suggests that the Taguchi method offers an efficient and sustainable approach for collagen extraction, reducing waste from fish processing industries.