title: A sensitive and specific method for measurement of multiple retinoids in human serum with UHPLC-MS/MS.
abstract: Retinol (vitamin A) circulates at 1-4 μM concentration and is easily measured in serum. However, retinol is biologically inactive. Its metabolite, retinoic acid (RA), is believed to be responsible for biological effects of vitamin A, and hence the measurement of retinol concentrations is of limited value. A UHPLC-MS/MS method using isotope-labeled internal standards was developed and validated for quantitative analysis of endogenous RA isomers and metabolites. The method was used to measure retinoids in serum samples from 20 healthy men. In the fed state, the measured concentrations were 3.1 ± 0.2 nM for atRA, 0.1 ± 0.02 nM for 9-cisRA, 5.3 ± 1.3 nM for 13-cisRA, 0.4 ± 0.4 nM for 9,13-dicisRA, and 17.2 ± 6.8 nM for 4oxo-13-cisRA. The concentrations of the retinoids were not significantly different when measured after an overnight fast (3.0 ± 0.1 nM for atRA, 0.09 ± 0.01 nM for 9-cisRA, 3.9 ± 0.2 nM for 13-cisRA, 0.3 ± 0.1 nM for 9,13-dicisRA, and 11.9 ± 1.6 nM for 4oxo-13-cisRA). 11-cisRA and 4OH-RA were not detected in human serum. The high sensitivity of the MS/MS method combined with the UHPLC separation power allowed detection of endogenous 9-cisRA and 4oxo-atRA for the first time in human serum.
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, University of Washington, Seattle, WA.
authors.forename: Samuel L M
authors.initials: SLM
authors.lastname: Arnold
authors.affiliation: Department of Internal Medicine, School of Medicine, University of Washington, Seattle, WA.
authors.forename: John K
authors.initials: JK
authors.lastname: Amory
authors.affiliation: Department of Urology, School of Medicine, University of Washington, Seattle, WA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Walsh
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, University of Washington, Seattle, WA. Electronic address: ni2@u.washington.edu.
authors.forename: Nina
authors.initials: N
authors.lastname: Isoherranen
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D012176:Retinoids / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014212:Tretinoin / Q000097:blood; D055815:Young Adult
chemical_list: D012176:Retinoids; D014212:Tretinoin