Methods for focusing analyte peaks in liquid chromatography using a spatial temperature gradient are provided. Also provided are methods for focusing analyte peaks and improving resolution using a trap column upstream of a separation column. Further, methods are provided in which the trap column placed upstream of the separation column is packed with a temperature-sensitive polymer/copolymer, and a spatial temperature gradient is applied along the trap column for obtaining improved retentivity by trap column stationary phase, and overall improved resolution of analyte peaks.