Reversed phase liquid chromatography (RPLC) is the gold standard analytical strategy in the pharmaceutical industry. In this contribution, we will describe two alternative chromatographic approaches, namely hydrophilic interaction chromatography (HILIC) and supercritical fluid chromatography (SFC), which are more and more commonly used in pharmaceutical analysis. Among their advantages, these two strategies offer better retention of polar substances, alternative selectivities and enhanced MS sensitivity, in comparison with RPLC.