This project aims to compare three distinct workflows using different sample collection/preparation plate supports (EVO96 oil-free, LF48 oil-based and LF48 oil-free, a streamlined automated sample resuspension and direct injection protocol). Then, digestion efficiency using a range of protein/enzyme ratios (1:1; 10:1; 20:1; 50:1) to maximize peptide recovery was evaluated. Finally, the chromatographic setup was refined to determine the best compromise between throughput and robustness.