title: Study of blood collection and sample preparation for analysis of vitamin D and its metabolites by liquid chromatography-tandem mass spectrometry.
abstract: The analysis of vitamin D status, with special emphasis on 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D, is gaining interest in clinical studies due to the classical and non-classical effects attributed to this prohormone. In this research, the influence of the two steps preceding determination (viz. sample collection and preparation) on the quantitative analysis of vitamin D and its more important metabolites has been studied. Two preparation approaches, deproteination and solid-phase extraction (SPE), have been evaluated in terms of sensitivity to delimit their application, thus establishing that detection of 1,25-dihydroxyvitamin D cannot be addressed by protein precipitation. Concerning sample collection, serum and plasma reported high accuracy (above 83.3%) for vitamin D and metabolites, while precision, expressed as relative standard deviation, was below 12.9% for all analytes in both samples. Statistical analysis revealed that serum and plasma provided similar physiological levels for vitamin D3, 24,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3, while significantly different levels were obtained for 1,25-dihydroxyvitamin D3, always higher in plasma than in serum. Sample collection and treatment have proved to be significant in the analysis of vitamin D and its relevant metabolites.
authors.affiliation: Department of Analytical Chemistry, Annex Marie Curie Building, Campus of Rabanales, University of Córdoba, Córdoba, Spain; Agroalimentary Excellence Campus, ceiA3, University of Córdoba, Córdoba, Spain; Maimónides Institute of Biomedical Research (IMIBIC), Reina Sofía University Hospital, University of Córdoba, Córdoba, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Mena-Bravo
authors.affiliation: Department of Analytical Chemistry, Annex Marie Curie Building, Campus of Rabanales, University of Córdoba, Córdoba, Spain; Agroalimentary Excellence Campus, ceiA3, University of Córdoba, Córdoba, Spain; Maimónides Institute of Biomedical Research (IMIBIC), Reina Sofía University Hospital, University of Córdoba, Córdoba, Spain. Electronic address: q72prcaf@uco.es.
authors.forename: F
authors.initials: F
authors.lastname: Priego-Capote
authors.affiliation: Department of Analytical Chemistry, Annex Marie Curie Building, Campus of Rabanales, University of Córdoba, Córdoba, Spain; Agroalimentary Excellence Campus, ceiA3, University of Córdoba, Córdoba, Spain; Maimónides Institute of Biomedical Research (IMIBIC), Reina Sofía University Hospital, University of Córdoba, Córdoba, Spain. Electronic address: qa1lucam@uco.es.
authors.forename: M D
authors.initials: MD
authors.lastname: Luque de Castro
mesh_terms: D001800:Blood Specimen Collection / Q000379:methods*; D002117:Calcitriol / Q000097:blood* / Q000378:metabolism; D002762:Cholecalciferol / Q000097:blood / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D005260:Female; D006801:Humans; D057230:Limit of Detection; D008297:Male; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014807:Vitamin D / Q000031:analogs & derivatives / Q000097:blood* / Q000378:metabolism
chemical_list: C118756:dihydroxy-vitamin D3; D014807:Vitamin D; D002762:Cholecalciferol; D002117:Calcitriol
keywords: Blood; Hydroxymetabolites; Plasma; Selected reaction monitoring; Serum; Vitamin D