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Title

Quantitative analytical method to evaluate the metabolism of vitamin D

Summary A method for quantitative analysis of vitamin D (both D2 and D3) and its main metabolites - monohydroxylated vitamin D (25-hydroxyvitamin D2 and 25-hydroxyvitamin D3) and dihydroxylated metabolites (1,25- dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3) in human serum is here re- ported. The method is based on direct analysis of serum by an automated platform involving on-line coupling of a solid-phase extraction workstation to a liquid chromatograph–tandem mass spectrometer. Detection of the seven analytes was carried out by the selected reaction monitoring (SRM) mode, and quantitative analysis was supported on the use of stable isotopic labeled internal standards (SIL-Iss). The detection limits were between 0.3–75 pg/mL for the target compounds, while precision (expressed as relative standard deviation) was below 13.0% for between-day variability. The method was externally validated according to the vitamin D External Quality Assurance Scheme (DEQAS) through the analysis of ten serum samples provided by this organism. The analytical features of the method support its applicability in nutritional and clinical studies targeted at elucidating the role of vitamin D metabolism
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Matrix Human Serum
Market Clinical
Cartridge HySphere C8
Detection MS Agilent 6410 triple Quad
Compounds Vitamin D2, Vitamin D3, 25-Hydroxyvitamin D2 25-hydroxyvitamin D3, 1,25-dihydroxyvitain D2 and 1,25 dihydroxyvitamin D3
Compound Group Vitamin,
Sample_volume 200 µL
 
Contact A. Mena-Bravo; C. Ferreiro-Vera, F. Priego-Capote, M.A. Maestro, A. Mouriño, J.M. Quesada-Gómez, M.D. Luque de Castro
Account a Department of Analytical Chemistry, Annex Marie Curie Building, Campus of Rabanales, University of Córdoba, Córdoba, Spain b University of Córdoba Agroalimentary Excellence Campus, ceiA3, Córdoba, Spain c Maimónides Institute of Biomedical Research (IMIBIC), Reina Sofía University Hospital, University of Córdoba, Córdoba, Spain d Phytoplant Research S.L. Rabanales 21, Science and Technology Park of Córdoba, Astrónoma Cecilia Payne Building, Córdoba, Spain e Mineral Metabolism Unit, Reina So?a Hospital, University of Córdoba, Córdoba, Spain f Department of Fundamental Chemistry, Faculty of Sciences, University of La Coruña, La Coruña, Spain g Department of Organic Chemistry, Faculty of Chemistry, University of Santiago, Santiago de Compostela, Spain
Published Clinica Chimica Acta 442 (2015) 6 12
Date 4-8-2015
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Remarks bution of the latter as cutaneous synthesis always exists. The method is also very appropriate to determine the 1,25(OH)2D metabolite, the analysis of which is presently very demanded in patients with renal insuf?ciency, hypoparathyroidism and hyperparathyroidism, and also for screening of hypercalcemia, increase of extra renal synthe- sis, etc. Despite quantitation of the 24,25(OH)2D is not at present relevant in routine clinical analysis, it could be very useful for clinical re- search to de?ne the cut-off of vitamin D status, of great present controversy. The sensitivity of the proposed method could help in basic research on in vivo and in vitro models. Despite quantitation of vitamins D2 and D3 is not necessary from the clinical point of view owing to fast conversion into the corresponding 25(OH)D metabolite, methods to differentiate both forms are of interest in pharmacokinetics studies, and to know the absorption capacity of metabolites orally administrated. Also, the determination of both vitamin D2 and its 25(OH)D2 metabolite could help in the study of the capability of vitamin D absorption in malabsorption process as celiac disease, in?ammatory bowel disease or bariatric surgery. On the other hand, determination of vitamin D3 is useful to evaluate the ability of synthesis of this vitamin by ultraviolet irradiation. With these premises, the next step for assessment is the application of the method to an extended cohort recruited in clinical centers.

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