Sequential injection chromatographic system for the expeditious molecular weight screening in hydrolysates

Introduction and objective: According to European Union regulations, the use of additives in animal feed must be rigorously evaluated to ensure safety, efficacy, and quality. The guidelines highlight the importance of reliable and efficient analytical tools for characterizing bioactive compounds in feed. Currently, protein evaluation is predominantly performed using High-Performance Liquid Chromatography (HPLC). Despite its robustness, presents disadvantages. The main objective is to develop a low-pressure chromatographic flow-based spectrophotometric method for the molecular weight screening of peptides used in animal feed. Method: This work presents a low-pressure chromatographic method based on a sequential injection analysis (SIA) system. A laboratory-made chromatographic column was coupled to the flow system, enabling a rapid molecular weight-based separation and quantification. This method approach provides an expeditious, cost-effective, miniaturized, and automated solution with strong potential for real- time control in industrial production environments. On going work: Optimization of the chromatographic conditions aiming for a higher chromatogram resolution and improved sensitivity through the evaluation of different column sizes, sample volume and flow-rate.