A reverse-phase high-performance liquid chromatographic method for analyzing complex mixtures of triglycerides. Application to the fat fraction of an aged cheese

Analysis of complex mixtures of triglycerides that span a broad range of partition numbers can entail a number of difficulties. The present study was therefore intended to develop a method of reverse-phase high-performance liquid chromatography using a light scattering detector for analyzing such mixtures. Stationary phase type, mobile phase gradient, column temperature, and injection solvent were the factors used to develop the method. The method improved the selectivity of critical pairs of milk fat triglycerides, providing good resolution for 115 peaks with partition numbers of between 22 and 53. The use of tristearin as internal standard and the relative response factors for different partition number ranges made it possible to more accurately estimate the weight of the sample triglycerides.