Ionic liquids (ILs) have drawn considerable interest for their use in various analytical techniques including chromatography, extractions, and mass spectrometry. This is largely due to the flexibility in tuning the physicochemical properties of ILs. There has been a significant increase in the number of publications over the last decade in which ILs have been employed as a chromatographic stationary phase, extraction solvent, or sorbent material in various preconcentration techniques. This review article highlights the recent advancements in the use of ILs in separation science including gas chromatography, high-performance liquid chromatography, capillary electrophoresis, extraction and microextraction techniques as well as mass spectrometry.