Abstract
Background: In this study, type B gelatin was extracted from Oreochromis niloticus scales under hydrothermal conditions at 60 °C to evaluate the effect of ultrasound-assisted pretreatment on its structural, physicochemical, thermal, and functional properties. Methods: Gelatin obtained with and without ultrasound pretreatment was systematically characterized through molecular weight analysis, proteomic profiling, size determination, surface morphology, proximate composition, thermal behavior, and gelation-related functional properties in order to assess the influence of the extraction method on gelation performance. Results: Ultrasound pretreatment slightly increased gelatin yield from 1.46 to 1.70%, indicating enhanced collagen solubilization. Proteomic analysis confirmed the predominance of fibrillar collagen proteins in both samples, although differences in protein distribution were observed. Furthermore, weight-average molecular weight analysis revealed a reduction from 212.3 ± 11.8 to 170.9 ± 13.2 kDa in the ultrasound-treated sample, suggesting partial fragmentation of collagen chains induced by cavitation effects. Structural modifications were also reflected in increased porosity and surface changes, contributing to improved colloidal stability. However, these changes significantly affect the functional behavior of the gelatin. Ultrasound-treated sample exhibited limited gel-forming capacity and failed to form stable gels at the evaluated concentration, despite complete dissolution. In contrast, gelatin extracted without ultrasound treatment retained higher-molecular-weight fractions and formed stable gels at both 5 and 10% (w/w). Thermal and spectroscopic analyses suggested that the fundamental collagen structure was preserved in both samples, although differences were observed in thermal degradation behavior. Conclusions: These results highlight the importance of controlling ultrasound-assisted extraction conditions to balance collagen recovery with the preservation of molecular integrity required for gelation, providing insights for the development of sustainable fish-derived biomaterials for pharmaceuticals and biomedical applications.