Hypercarb Columns for Urinary Metabolite Separation

Separations of polar compounds can be challenging due to inadequate retention on common stationary phases. Selected urinary metabolites will be examined on Hypercarb porous graphitic carbon columns. These columns are more effective in separating polar compounds than traditional C18 columns due to complex retention mechanisms. Retention is important in determining efficiency of separation and peak capacity. Retention of these metabolites will first be characterized on Hypercarb and C18 columns using HPLC. Various concentrations of water and methanol will be used to compare retention at various mobile phase conditions. The metabolites will then be characterized using stop flow experiments on HPLC columns. Stop flow rates are useful in calculating the diffusion of compounds on the stationary phase. Diffusion also plays a role in separation efficiency and peak broadening. These experiments will inform further studies on UHPLC-MS for metabolomics of human urine samples.Bachelor of Art