The discovery of taste compounds in foodstuffs is challenged by traditional one-dimensional liquid chromatographic techniques that have inadequate resolving power to facilitate identification of target analytes. The purpose of this study was to investigate the development and application of an off-line two-dimensional high pressure liquid chromatographic (2D HPLC) system for taste analysis. A simple geometric approach was applied to characterize the practical orthogonality of the analytical system for column selection. This mathematical prediction of resolving power correlated well with the experimental findings. A highly orthogonal 2D HPLC system was developed for the analysis of bitter compounds in whole wheat bread.