Retinol, α-tocopherol, lycopene, and α- and β-carotene simultaneously determined in plasma by isocratic liquid chromatography

Abstract

Retinol, α-tocopherol, lycopene, and α- and β-carotene can be simultaneously determined in human plasma by reversed-phase liquid chromatography. Plasma - 0.5 mL plus added internal standard, retinyl acetate - is deproteinized with 0.5 mL of ethanol, then extracted with 1.0 mL of petroleum ether. The organic layer is removed and evaporated, the residue is redissolved in 0.25 mL of ethanol, and 8-μL samples are injected into a 60 x 4.6 mm column of Hypersil ODS 3-μm particles at 35°C. An isocratic methanol mobile phase, flow rate 0.9 mL/min, is used for the 9-min run. Retinol and retinyl acetate are monitored at 305 nm, the tocopherols at 292 nm, and the carotenoids at 460 nm. Between-run CVs were 3.1, 6.9, 6.1, and 6.5% for retinol, α-tocopherol, lycopene, and β-carotene, respectively. Small sample requirement, simplicity of extraction, short run time, and good reproducibility make this procedure ideal for clinical or research use.