This project investigates increasing the elution strength of mobile phases in HPLC without altering their composition, to retain the benefits of gradient analyses—such as refocusing, better detectability, broader applicability, and shorter cycle times—without the drawbacks of changing solvents. Conventional reversed-phase LC requires broad temperature ranges for effective use, so this study optimizes Temperature Responsive Liquid Chromatography (TRLC), which uses water as the mobile phase and offers sharper retention changes with temperature. Various gradient approaches will be tested: developing flow-field thermal gradients, exploiting longitudinal stationary phase density gradients, using mixed-flow thermal gradients, and studying pressure-enhanced gradients. The performance of these methods will also be combined into novel isocratic (pressure, stationary phase loading, temperature) gradient methods for enhanced TRLC.