Simultaneous Separation and Detection of Ten Common FAT-Soluble Vitamins in Milk

Abstract

A method for the separation of ten fat-soluble vitamins on a reversed phase C₁₈ column with UV detection is described. The choice of a suitable organic solvent in conjunction with an aqueous phase composition is important in achieving a good separation of the ten common fat-soluble vitamins which include different forms of vitamins A, D, K and α-tocopherol (E). Two different mobile phases were developed for the isocratic separation of the vitamins. It was found that the separation of the ten vitamins is very sensitive to a small change in the composition of the mobile phase. The method was applied to determine the vitamins in milk sample, after sample treatment which includes saponification, drying, and reconstitution of the vitamins with the mobile phase.