Reverse phase high performance liquid chromatography with photodiode array detection method for the identification and detection of fat-soluble vitamins in plasma

The purpose of this study was to establish a method for simultaneous evaluation and quantification of fat-soluble vitamins in human blood samples using reverse-phase high performance liquid chromatography coupled with photodiode array detection. The biochemical forms of fat-soluble vitamins analyzed include retinol, cholecalciferol, calcidiol, α-tocopherol, γ-tocopherol, and phylloquinone. This method is novel in the use of prostaglandin B1 as an internal standard for quantification and utilizes a simple method of extraction without derivatization or saponification. The method had a high degree of reproducibility as evidenced by coefficients of variation 1.7% and 2.5% of the mean for retinal and cholecalciferol respectively.