Simultaneous determination of fat-soluble vitamins and provitamins in dairy products by liquid chromatography with a narrow-bore column

Abstract

A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of vitamins A, D₂, D₃, E and K₁, retinyl acetate, retinyl palmitate, tocopherol acetate, ergosterol and 7-dehydrocholesterol in milk and butter. Narrow-bore columns are recommended because this alternative provides a good separation and efficiency, plus greater economy and sensitivity. Detection limits for individual vitamins range from 0.14 to 6.9 ng. All vitamins are separated in less than 33 min. For the simultaneous determination of these vitamins and provitamins we use two sample pre-treatment methods, a liquid–liquid extraction with hexane or a solid-phase extraction with a C₁₈ cartridge. Recovery studies show good results for all solutes (84–108% and 85-108% for milk and butter, respectively) and the intra-day coefficients of variations range from 1.6 to 4.5%. These methods permit the simple determination of fat-soluble vitamins using a small sample volume.