title: Suitability of ultra-high performance liquid chromatography for the determination of fat-soluble nutritional status (vitamins A, E, D, and individual carotenoids).
abstract: Our aim was to assess the suitability of ultra-high performance liquid chromatography (UHPLC) for the simultaneous determination of biomarkers of vitamins A (retinol, retinyl esters), E (alpha- and gamma-tocopherol), D (25-OH-vitamin D), and the major carotenoids in human serum to be used in clinical practice. UHPLC analysis was performed on HSS T3 column (2.1 x 100 mm; 1.8 microm) using gradient elution and UV-VIS detection. The system allows the simultaneous determination of retinol, retinyl palmitate, 25-OH-vitamin D, alpha- and gamma-tocopherol, lutein plus zeaxanthin, alpha-carotene, beta-carotene, alpha- and beta-cryptoxanthin and lycopene. The method showed a good linearity over the physiological range with an adequate accuracy in samples from quality control programs. Suitability of the method in clinical practice was tested by analyzing samples (n = 286) from patients. In conclusion, UHPLC constitutes a reliable approach for nutrient/biomarker profiling allowing the rapid, simultaneous and low-cost determination of vitamins A, E, and D (including vitamers and ester forms) and the major carotenoids in clinical practice.
authors.affiliation: Unidad de Vitaminas. Bioquímica Clínica, Edificio laboratorios (Peine 7) Planta 1a, Hospital Universitario Puerta de Hierro-Majadahonda, c/ Maestro Joaquin Rodrigo, 2, 28222 Madrid, Spain. fgranado.hpth@salud.madrid.org
authors.forename: F
authors.initials: F
authors.lastname: Granado-Lorencio
authors.forename: C
authors.initials: C
authors.lastname: Herrero-Barbudo
authors.forename: I
authors.initials: I
authors.lastname: Blanco-Navarro
authors.forename: B
authors.initials: B
authors.lastname: Pérez-Sacristán
mesh_terms: D002338:Carotenoids / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D016014:Linear Models; D012680:Sensitivity and Specificity; D014801:Vitamin A / Q000097:blood*; D014807:Vitamin D / Q000097:blood*; D014810:Vitamin E / Q000097:blood*; D014815:Vitamins / Q000097:blood
chemical_list: D014815:Vitamins; D014801:Vitamin A; D014807:Vitamin D; D014810:Vitamin E; D002338:Carotenoids