title: HPLC method for simultaneous determination of retinoids and tocopherols in human serum for monitoring of anticancer therapy.
abstract: A simple and rapid HPLC method requiring small volumes (250 microL) of human serum after C18 SPE sample preparation was developed using monolithic technology for simultaneous determination of all-trans-retinoic acid, 13-cis-retinoic acid, retinol, gamma- and alpha-tocopherol. The monolithic column, Chromolith Performance RP-18e (100x4.6 mm), was operated at ambient temperature. The mobile phase consisted of a mixture of acetonitrile (ACN) and 1% ammonium acetate in water (AMC) at pH 7.0. The mobile phase started at 98:2 (v/v) ACN/AMC (column pre-treatment) at a flow rate of 2 mL/min, then changed to 95:5 (v/v) ACN/AMC for 4 min at a flow rate of 1.5 mL/min and a further 3 min at a flow rate of 3.2 mL/min. Detection and identification were performed using a photodiode array detector. Retinol, 13-cis- and all-trans-retinoic acid were monitored at 325 nm. Both alpha- and gamma-tocopherol were detected at 295 nm. The total analysis time was 7.2 min. Tocol (synthesized tocopherol, not occurring in humans) was used as internal standard. The method was linear in the range of 0.125-10.00 micromol/L for all-trans-retinoic acid, 0.125-5.00 micromol/L for 13-cis-retinoic acid, 0.25-10.00 micromol/L for retinol, 0.5-50.00 micromol/L for gamma-tocopherol, and 0.5-50.00 micromol/L for alpha-tocopherol. The present method may be useful for monitoring of retinoids and tocopherols in clinical studies.
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Hradec Králové, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Krcmová
authors.forename: Lubor
authors.initials: L
authors.lastname: Urbánek
authors.forename: Dagmar
authors.initials: D
authors.lastname: Solichová
authors.forename: Markéta
authors.initials: M
authors.lastname: Kasparová
authors.forename: Hana
authors.initials: H
authors.lastname: Vlcková
authors.forename: Bohuslav
authors.initials: B
authors.lastname: Melichar
authors.forename: Lubos
authors.initials: L
authors.lastname: Sobotka
authors.forename: Petr
authors.initials: P
authors.lastname: Solich
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D015394:Molecular Structure; D009369:Neoplasms / Q000097:blood / Q000628:therapy; D015203:Reproducibility of Results; D012176:Retinoids / Q000097:blood* / Q000737:chemistry; D012680:Sensitivity and Specificity; D024505:Tocopherols / Q000097:blood* / Q000737:chemistry
chemical_list: D012176:Retinoids; D024505:Tocopherols