Simultaneous Measurement of Three Tocopherols, All-trans-retinol, and Eight Carotenoids in Human Plasma by Isocratic Liquid Chromatography

Abstract

We describe a simplified isocratic HPLC method for the simultaneous determination of all-trans-retinol, 3 tocopherols (α-, δ-, and γ- tocopherol), and 8 carotenoids (lutein, zeaxanthin, canthaxanthin, β-cryptoxanthin, all-trans-lycopene, 5-cis-lycopne, α-carotene, and β-carotene) in human plasma, which uses a single C18 reversed-phase column (4.6 × 250 mm, 3 µm). A photodiode array detector was used to measure the UV-vis wavelength absorbance of retinol and the eight carotenoids, and fluorescence detection was used for the tocopherols. The linear ranges of the calibration curves for the calibration solutions injected into the HPLC column are 0.02–6.0 µg/mL for all-trans-retinol, β-cryptoxanthin and α-carotene; 0.01–3.0 µg/mL for δ-tocopherol, lutein, and lycopene; 0.08–24.0 µg/mL for γ-tocopherol; 0.3–90.0 µg/mL for α-tocopherol; 0.005-1.5 µg/mL for zeaxanthin and canthaxanthin; and 0.04–12.0 µg/mL for β-carotene.