Title: Development of a method for the quantification of 1alpha,25(OH)2-vitamin D3 in serum by liquid chromatography tandem mass spectometry without derivatization
Authors: Casetta, B ×
Jans, Ivo
Billen, Jaak
Vanderschueren, Dirk
Bouillon, Roger #
Issue Date: 2010
Publisher: IM Publications
Series Title: European Journal of Mass Spectrometry vol:16 issue:1 pages:81-89
Abstract: Quantification of 1alpha,25(OH)(2)-vitamin D(3) in serum is technically challenging because of its very low concentration. Even mass spectrometry faces a challenge due to the low sensitivity for this molecule, unless a specific derivatization is implemented. Therefore, there is still a need for a robust, simplified, sensitive and specific liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the quantification of 1alpha,25(OH)(2)-vitamin D(3). Thanks to tandem mass- spectrometry associated to an on-line sample extraction and clean-up, sample preparation has been reduced to a simple protein precipitation step and derivatization has been avoided. Specimen volume has been kept to a minimum. The innovative aspects of the hereby-presented method are the use of stable lithium adducts and a highly sensitive tandem mass spectrometer. The achieved limit of quantification is at a level of 15 pg mL(-1), with a % CV of 5-15% at physiological concentration levels. The linearity is good and spiking experiments yielded a recovery of 87-102%. Comparison of selected samples with an established reference method, using an extensive purification procedure (high-performance liquid chromatography and radioimmunoassay), has shown a good correlation.
ISSN: 1469-0667
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Laboratory of Clinical Bacteriology and Mycology
Clinical and Experimental Endocrinology
× corresponding author
# (joint) last author

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