Liquid chromatography-tandem mass spectrometry for monitoring Vitamin D hydroxymetabolites in human aqueous humor

Liquids; Mass spectrometry; Metabolites; Vitamins; Average recovery; Cataract operations; Human aqueous humor; Limits of quantitations; Liquid chromatography tandem mass spectrometry (LC MS/MS); Liquid chromatography-tandem mass spectrometry; Sample preparation; Liquid chromatography

Abstract :

[en] A stable isotope dilution liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) and 25-hydroxyvitamin D3/D2 (25(OH)D3/D2) in human aqueous humor samples from the Eye-D study. Optimum sample preparation is based on simple liquid-liquid extraction (LLE) followed by derivatization with an Amplifex reagent prior to LC-MS/MS in order to enhance analyte sensitivity. The average recoveries for 24,25(OH)2D3 (0.02, 0.05, 0.4 μg L-1), 25(OH)D3 (0.2, 0.5,3.75 μg L-1) and 25(OH)D2 (0.15, 0.4, 3 μg L-1) ranged from 92 to 112 with a coefficient of variance (CV) lower than 15%. The limits of quantitation (LOQs) were from 0.02 μg L-1 (24,25(OH)2D3) to 0.2 μg L-1 (25(OH)D3) using a sample volume of 50 μL. We demonstrated that the present method provides adequate sensitivity, selectivity, accuracy, and robustness to screen for vitamin D metabolites in aqueous humor samples by analyzing five samples which were withdrawn during cataract operation. © 2019 The Royal Society of Chemistry.

Disciplines :

Laboratory medicine & medical technology

Author, co-author :

Fabregat Cabello, Neus ; Department of Clinical Chemistry, University of Liège, CHU Sart-Tilman, Liège, Belgium, Hematology and Hemotherapy Group, Health Research Institute Hospital la Fe (IIS la Fe), Valencia, Spain

DARIMONT, Pierre ; Centre Hospitalier Universitaire de Liège - CHU > Département de pharmacie hospitalière > Service administratif, dispositifs médicaux et stérilisation

Huyghebaert, Loreen ; Centre Hospitalier Universitaire de Liège - CHU > Unilab > Laboratoire techniques séparatives et stress oxydant

Reynier, Pascal; Department of Biochemistry and Genetics, CHU d'Angers, Angers, France

Annweiler, Cédric; Department of Geriatric Medicine, Angers University Hospital, Research Center on Autonomy and Longevity, University Memory Clinic of Angers, UPRES EA 4638, University of Angers, Angers, France, Robarts Research Institute, Department of Medical Biophysics, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada

Milea, Dan; Department of Ophthalmology, Angers University Hospital, Angers, France, Singapore Eye Research Institute, Singapore National Eye Centre, Duke-NUS, Neuroscience and Behavioral Diseases, Singapore

Le Goff, Caroline ; Université de Liège - ULiège > Département des sciences de la santé publique > Département des sciences de la santé publique

Cavalier, Etienne ; Université de Liège - ULiège > Département de pharmacie > Chimie médicale

Language :

English

Title :

Liquid chromatography-tandem mass spectrometry for monitoring Vitamin D hydroxymetabolites in human aqueous humor

Publication date :

2019

Journal title :

Analytical Methods

ISSN :

1759-9660

eISSN :

1759-9679

Publisher :

Royal Society of Chemistry

Volume :

Issue :

Pages :

5282-5288

Peer reviewed :

Peer Reviewed verified by ORBi

Funders :

UA - Université d'Angers

Available on ORBi :

since 09 April 2020

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