Silica Hydride-Based Chromatography of Lc–Ms Response-Altering Compounds Native to Human Plasma

Abstract

An investigation was carried out into the chromatographic behavior, on a silica hydride-based phase and a comparator silica-based phase, of an important group of lipids endogenous to human plasma, which are associated with matrix effect and in the context of quantitative peptide analysis. The propensity for aqueous normal phase (ANP) retention on the silica hydride-based phase was strong and extensive in comparison with the silica-based comparator, and the lipophilic interferences in question were readily eluted using the ANP mode, a contrast to over-retention issues with accompanying implications for method ruggedness typically found with silica-based phases. The silica hydride-based phase, with ANP operation, offered selectivity conducive to rapid lipophilic interferent elimination and the bimodal retention involved in suitable gradient elution was appropriate for general peptide analytical application.