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We have recently developed a reliable method to pack an HPLC column with two phases particles in a variety of ratios, such as AQ-C 18/Amide or AQ-C 18/SCX.We have conducted a number of studies of the behavior of the columns and developed a number of practical applications based on their very unique features.In this presentation, we will demonstrate that the columns can be used in 4 types of unique applications: 1-Improving the separation by using the unique selectivity of the mix-phases 2-Analyzing a sample having both the very polar and non-polar substances in one run, especially under isocratic conditions.3-A novel systematic approach to optimize the separation of a complex mixture containing multiple critical pairs 4-A convenient solution for LC-MS analysis of highly water soluble and ionizable substance.