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SHR Neuro Cancer Cardio Lipid Metab Microb
Herrmann, M; Harwood, T; Gaston-Parry, O; Kouzios, D; Wong, T; Lih, A; Jimenez, M; Janu, M; Seibel, MJ.
A new quantitative LC tandem mass spectrometry assay for serum 25-hydroxy vitamin D.
Steroids. 2010; 75(13-14):1106-1112
Doi: 10.1016/j.steroids.2010.07.006
Web of Science
PubMed
FullText
FullText_MUG
- Leading authors Med Uni Graz
- Herrmann Markus
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- Abstract:
- The accurate measurement of 25-hydoxy vitamin D (25OH-D) in serum has been a challenge for many years. We developed a liquid chromatography tandem mass spectrometry (LC Tandem MS) assay for the quantitative determination of 25OH-D(2) and 25OH-D(3) in serum. The new method was compared with two widely used commercially available immunoassays. Sample preparation involved protein precipitation with acetonitrile containing deuterated forms of the target species as internal standards. An API 5000 mass spectrometer coupled with a photoionization source was used for quantitation. The performance of the new LC Tandem MS assay was compared with a radioimmunoassay (RIA, Diasorin) and a chemiluminescence immunoassay (ECLIA, Roche Diagnostics), analysing serum obtained from 152 individuals. Using 100 μl of serum, the LC Tandem MS assay had a limit of quantitation of 1.3 nmol/L for both 25OH-D(2) and 25OH-D(3) with a linear response between 1.3 and 625 nmol/L and accuracy of between 95 and 124%. Intra- and inter-assay precision were ≤7% and ≤4%, respectively. Measurement of 25OH-D levels in 152 serum samples gave run averages of 71, 56 and 62 nmol/L for LC Tandem MS, ECLIA and RIA, respectively. Correlations between the various methods were: LC Tandem MS vs. RIA: r=0.931; LC Tandem MS vs. ECLIA: r=0.784; RIA vs. ECLIA: r=0.787. The LC Tandem MS method had a positive proportional bias of 26% over the RIA, whereas the ECLIA showed variable differences. The new LC Tandem MS assay is accurate and precise at physiologically relevant 25OH-D concentrations, and compares favourably with the RIA. In contrast, the ECLIA shows variable bias with the other assays tested. Copyright © 2010 Elsevier Inc. All rights reserved.
- Find related publications in this database (using NLM MeSH Indexing)
- 25-Hydroxyvitamin D 2 - blood
- Blood Chemical Analysis - methods
- Calcifediol - blood
- Chromatography, Liquid - methods
- Female -
- Humans -
- Male -
- Middle Aged -
- Tandem Mass Spectrometry - methods
- Time Factors -
- Find related publications in this database (Keywords)
- Vitamin D
- Serum
- LC Tandem MS
- Immunoassay