Publication Date

Summer 2016

Degree Type


Degree Name

Master of Science (MS)




Joseph Pesek


Chromatography, Diol, HPLC, Silica Hydride, Stationary phase, UPLC

Subject Areas

Analytical chemistry; Chemistry


The impact of chromatography across many scientific fields and applications is limitless. It is a vital everyday separation, characterization, and purification tool for many scientists worldwide. Innovations in high performance liquid chromatography (HPLC) stationary phases have led to more diverse separations essential to many fields including the pharmaceutical industry and research. Silica hydride based stationary phases have been shown to display both reverse phase (RP) and aqueous normal phase (ANP) chromatographic behavior. This is a result of both the silica hydride surface and the bonded phase. The goal of this work was to characterize silica hydride based diol stationary phases for HPLC. A wide range of compounds with varying polarities were analyzed. Retention was observed under ANP and RP conditions. Two representative silica hydride based diol stationary phases were compared to demonstrate the effect that the length of the bonded phase has on the separation capabilities of the column. The diol bonded phase with a longer carbon chain retained analytes with more hydrophobic (or non-polar) characteristics longer than analytes with more hydrophilic properties. As part of a larger study the effect of buffer concentration on the ANP retention of model compounds was investigated. Retention dramatically decreased when the concentration of some buffers was increased. This trend is opposite of what has been observed in hydrophilic interaction liquid chromatography (HILLIC), indicating a clear distinction in the retention mecahism for HILLIC and ANP.