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Introduction: Textile industry needs more bio centered solutions - the industry needs to adopt circular economy principles to repurpose its by-products; There is a need to re-utilize leather by-products and to develop biobased products1,2; SE-HPLC aids in the selection and production of peptide-rich solutions, these solutions are intended for use as functional coatings in textiles. This methodology helps optimize the formulation of coatings for desired functional properties. Objectives: Utilize SE-HPLC to gather fundamental data on the hydrolysis of protein-rich byproducts. Select and produce peptide-rich solutions suitable for functional textiles coatings. Evaluate different enzymes and hydrolysis conditions using SE-HPLC. Repurpose protein-rich leather by-products effectively using SE-HPLC guided strategies. Conclusions: Alcalase 10% 6 hours had higher hydrolysis degree (70.12%) than all the other enzymes percentage/time of hydrolysis Between time 0 and pre-final, molecules MW lower than 0.2 kDa consistently have a higher area. Conducting an enzyme inactivation (2 hours/90ºC) might result in molecule aggregation, leading to an increase in molecules with MW exceeding 50 kDa. Utilizing this technique facilitates the identification of the optimal time/enzyme concentration ratio. It enables industries to repurpose waste effectively while developing new bio-based products through hydrolysis. Specifically, it aids in the production of functional coatings, offering a sustainable solution for waste utilization and product innovation.