Back-exchange and peptide carry-over are crucial parameters to control in HDX-MS, which can be negatively influenced by an extended flow path and/or an additional column matrix. To investigate the impact of the integrated delipidation system, we conducted PhosB measurements on both configurations and compared back-exchange and carry-over on a large ensemble of peptides (Figure ). For the backexchange control, we pre-digested PhosB and maximally deuterated the generated peptides with 100% D 2 O.