DETERMINATION OF INTACT MOLECULAR-SPECIES OF BOVINE-MILK 1, 2-DIACYL-SN-GLYCERO-3-PHOSPHOCHOLINE AND 1, 2-DIACYL-SN-GLYCERO-3-PHOSPHOETHANOLAMINE BY REVERSED-PHASE HPLC, A MULTIVARIATE OPTIMIZATION

The molecular species profile, representing the fatty acid combinations on the glycerol backbone, is the most fundamental description of an intact glycerophospholipid class and provides a key to understanding its functional characteristics. We have developed a powerful RP-HPLC method for the separation of molecular species, including so-called ''critical pairs'', of natural mixtures of 1, 2-diacyl-sn-glycero-3-phosphocholine and 1, 2-diacyl-sn-glycero-3-phosphoethanolamine from bovine milk and other natural sources. We have used a novel multivariate development and optimization strategy to locate the region of optimum separation conditions within a pre-chosen domain of the chromatographic parameter space.