Simple and fast HPLC method for simultaneous determination of retinol, tocopherols, coenzyme Q10 and carotenoids in complex samples

The effects of fat-soluble vitamins (such as vitamins A and E) and lipid microconstituents (such as carotenoids) on human health are now well established. However, high-performance liquid chromatography (HPLC) methods able to detect these molecules in simultaneous runs are often difficult to set up. We report here a 35-min reversed-phase HPLC method using a single C30 column kept at 35 °C with a gradient system of methanol, methyl-tert-butyl ether and water at a flow-rate of 1 mL/min. This method resolves 11 carotenoids, retinol, α- and γ-tocopherol from complex matrixes such as food samples, human plasma and human adipose tissue within 35 min. The method is also able to separate coenzyme Q10. The intra-day and inter-day coefficients of variation are suitable for routine clinical and scientific applications for the determination of lipid micronutrients from various sample types.