Journal article
Determination of four sulfated vitamin D compounds in human biological fluids by liquid chromatography-tandem mass spectrometry
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, Vol.1009-1010, pp.80-86
2016
PMID: 26708628
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Source: InCites
Abstract
The determination of both the water-soluble and lipid-soluble vitamin D compounds in human biological fluids is necessary to illuminate potentially significant biochemical mechanisms. The lack of analytical methods to quantify the water-soluble forms precludes studies on their role and biological functions; currently available liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods are able to determine only a single sulfated form of Vitamin D. We describe here a highly sensitive and specific LC-MS/MS method for the quantification of four sulfated forms of vitamin D: vitamins D2- and D3-sulfate (D2-S and D3-S) and 25-hydroxyvitamin D2- and D3-sulfate (25(OH)D2-S and 25(OH)D3-S). A comparative evaluation showed that the ionization efficiencies of underivatized forms in negative ion mode electrospray ionisation (ESI) are superior to those of the derivatized (using 4-phenyl-l,2,4-triazoline-3,5-dione (PTAD)) forms in positive ion mode ESI. Separation was optimised to minimise co-elution with endogenous matrix compounds, thereby reducing ion suppression/enhancement effects. Isotopically labelled analogues of each compound were used as internal standards to correct for ion suppression/enhancement effects. The method was validated and then applied for the analysis of breastmilk and human serum. The detection limits, repeatability standard deviations, and recoveries ranged from 0.20 to 0.28fmol, 2.8 to 10.2%, and 81.1 to 102%, respectively.
Details
- Title
- Determination of four sulfated vitamin D compounds in human biological fluids by liquid chromatography-tandem mass spectrometry
- Creators
- Fabio P Gomes - University of QueenslandP Nicholas Shaw - University of QueenslandAmitha K Hewavitharana (Corresponding Author) - University of Queensland
- Publication Details
- Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, Vol.1009-1010, pp.80-86
- Publisher
- Netherlands
- Identifiers
- 991012902100002368
- Academic Unit
- Faculty of Science and Engineering; Southern Cross Plant Science; Science
- Language
- English
- Resource Type
- Journal article