Confectionery fat analysis with high performance liquid chromatography

Natural triglyceride mixtures have been analyzed by a variety of techniques. Earlier methods, such as crystallization, were unsuitable because they were very time-consuming, required large amounts of material and were not reproducible. Later advances in chromatography gave successful separations of triglycerides but required a combination of techniques to give suitable separations. High performance liquid chromatography (HPLC) with very efficient columns has been successfully applied to triglyceride separations on a routine basis. In the present work, triglycerides of cocoa butter as well as several replacement fats were separated by HPLC. The system consisted of a highly efficient column packed with octadecylbonded spherical silica (5 µ) and a mobile phase composed of acetone and acetonitrile. Peaks corresponding to eluted components were collected and their composition determined by gas chromatography (GC) of the corresponding methyl esters to allow unambiguous assignment of triglyceride structure to the components. A profile of the triglycerides present in such fats can be obtained within 30 min.