High-temperature micro liquid chromatography for lipid molecular species analysis with evaporative light scattering detection

The need for a rapid and sensitive chromatographic technique for analyzing lipid molecular species, has led to the development of an high-temperature micro liquid chromatographic system (HTLC) coupled to an evaporative light scattering detector. The increased diffusion coefficients and reduced viscosity at higher temperatures allowed lipids to be analyzed rapidly with solvents differing from those classically used in lipids chemistry. Hypercarb, a reverse phase material, was used for its different properties including heat resistance in high temperature micro HPLC. We have investigated the temperature effect on kinetic performances in HTLC, established pure solvents eluent strength at high temperature and studied different classes of lipids with seven pure solvents. We found that it was possible to use alcohols solvents in the mobile phase to elute lipids without the use of chlorinated solvents. A quick and simple method was developed to analyze a complex lipid simple, ceramide type III and type IV.