High-performance liquid chromatographic determination of vitamin D3 in foods with particular reference to eggs

Abstract

A high-performance liquid chromatographic (HPLC) method for the determination of vitamin D₃ in foods is described. The method involves addition of vitamin D₂ to the sample as an internal standard followed by saponification and extraction of the unsaponifiable matter. Sterols are removed by precipitation and other interfering compounds by a thin-layer chromatographic clean-up procedure. Vitamins D₂ and D₃ are then separated using reversed-phase HPLC. Details of the accuracy and precision of the method are presented. This method has been applied successfully to the determination of vitamin D₃ in eggs, butter, milk and cheese.