Studies on the retention mechanism of solutes in hydrophilic interaction chromatography using stoichiometric displacement theory II. HILIC/RPLC dual-retention mechanism of solutes in hydrophilic interaction chromatography over the entire range of water concentration in mobile phase

This paper is a continuation of research into the retention behavior and mechanism of solutes in hydrophilic interaction chromatography (HILIC) using stoichiometric displacement theory (SDT). A HILIC/reversed-phase liquid chromatography (RPLC) dual-retention mechanism was studied in detail using a β -CD HILIC column. The retention behaviors of three groups of solutes with varying polarities were studied over the entire range of water concentrations in the mobile phase on the β -CD column, resulting in the formation of “U-shape” curves when lg k ’ was plotted against lg[H 2 O]. Additionally, the effect of hydrophobic distribution coefficient lg P O/W on the retention behaviors of solutes in HILIC and RPLC modes was also examined. A four-parameter equation derived from the SDT-R was found to accurately describe the “U-shaped” curves of solutes with RPLC/HILIC dual-retention mechanisms on β -CD column. The theoretical lg k ’ values of solutes calculated using the equation were found to be in agreement with their experimental values, with correlation coefficients greater than 0.99. This indicates that the four-parameter equation derived from SDT-R can effectively describe the retention behaviors of solutes over the entire range of water concentrations in the mobile phase in HILIC. As such, SDT can be used as a theoretical guide for the development of HILIC, including the exploration of new dual-function stationary phases to enhance separation efficiency.