A review of the background, operating parameters and applications of microemulsion liquid chromatography (MELC)

Microemulsions are dispersions of nanometre‐sized droplets of an immiscible liquid within another liquid. Droplet formation is facilitated by the addition of surfactants and often also cosurfactants. Microemulsions are classified as either oil‐in‐water (O/W) (oil droplets such as octane dispersed throughout aqueous buffer) or water‐in‐oil (W/O) (aqueous droplets in oil such as hexane). Both microemulsion types have been used as mobile phases for separation in microemulsion HPLC (MELC). There has been a recent increase of interest in this area with new applications and developments such as gradient elution and optimisation of methods using experimental design. O/W microemulsions have been employed as eluents for RP‐HPLC while W/O microemulsions have been used for normal phase chromatography. Separations can have superior speed and efficiency to conventional HPLC modes while offering a unique selectivity with excellent resolution. The capability for quantitative and stability‐indicating analysis has also been demonstrated. Specific advantages include the ability to operate at low UV wavelengths and elimination of the need for an equilibration rinse between gradients. Operational issues associated with the use of MELC have been identified including the need to add salt to the gradient eluent, relatively high back‐pressures and increased need for equipment cleaning compared to conventional RP eluent. This report details the different microemulsion types and compositions used and their reported applications. The use of gradient and isocratic elution is described. The effects on separations of varying operating parameters such as temperature, oil type and concentration, surfactant type and concentration, sample solvent, column type, and organic solvent addition will be discussed and illustrated.