title: New approach for the clinical monitoring of 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 by ultra high performance liquid chromatography with MS/MS based on the standard reference material 972.
abstract: Biomarkers, 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 , are important indicators of the vitamin D general status and are monitored in several pathophysiological disorders, such as osteoporosis, diabetes, heart disease, etc. A novel ultra-HPLC with MS/MS methodology for the analysis of 25-hydroxyvitamin D derivatives coupled with a very simple and highly rapid sample preparation step was developed. Analytical parameters obtained showed linearity (R(2) ) above 0.999 for both vitamins with accuracies between 95.8 and 102%. The LODs were as low as 0.22 and 0.67 nmol/L for 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 , respectively. Intra-assay precision (%RSD) was lower than 4.5%, and inter-assay precision (%RSD) was lower than 6.5%. The feasibility of the developed methodology to be applied in clinical routine analysis has been proved by its application in blood samples from non-agenarian patients, patients with familial hypercholesterolemia and patients suffering from age-related macular degeneration.
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského, Hradec Králové, Czech Republic; III. Internal Gerontometabolic Clinic, University Hospital, Sokolská, Hradec Králové, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Plíšek
authors.forename: Lenka Kujovská
authors.initials: LK
authors.lastname: Krčmová
authors.forename: Jana
authors.initials: J
authors.lastname: Aufartová
authors.forename: Tanausú V
authors.initials: TV
authors.lastname: Morales
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Esponda
authors.forename: Roman
authors.initials: R
authors.lastname: Oros
authors.forename: Eva
authors.initials: E
authors.lastname: Kasalová
authors.forename: Jose J
authors.initials: JJ
authors.lastname: Santana-Rodriguez
authors.forename: Luboš
authors.initials: L
authors.lastname: Sobotka
authors.forename: Petr
authors.initials: P
authors.lastname: Solich
authors.forename: Dagmar
authors.initials: D
authors.lastname: Solichová
mesh_terms: D015652:25-Hydroxyvitamin D 2 / Q000097:blood*; D001774:Blood Chemical Analysis / Q000379:methods*; D002112:Calcifediol / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D057230:Limit of Detection; D012015:Reference Standards; D053719:Tandem Mass Spectrometry; D013997:Time Factors
chemical_list: D015652:25-Hydroxyvitamin D 2; D002112:Calcifediol
keywords: Clinical research; Mass spectrometry; Standard reference material 972; Vitamin D