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Herrmann, M., Harwood, T., Gaston-Parry, O., Kouzios, D., Wong, T., Lih, A., Jimenez, M., Janu, M. & Seibel, M. J. (2010) A new quantitative LC tandem mass spectrometry assay for serum 25-hydroxy vitamin D. Steroids, 75 1106–1112. 
Added by: Sarina (02 Aug 2022 15:40:41 UTC)   Last edited by: Sarina (26 Sep 2022 12:38:06 UTC)
Resource type: Journal Article
DOI: https://doi.org/10.1016/j.steroids.2010.07.006
ID no. (ISBN etc.): 0039-128X
BibTeX citation key: Herrmann2010
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Categories: Englisch = English
Keywords: Vitamin D = Vitamin D
Creators: Gaston-Parry, Harwood, Herrmann, Janu, Jimenez, Kouzios, Lih, Seibel, Wong
Collection: Steroids
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Abstract
Background 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-D2 and 25OH-D3 in serum. The new method was compared with two widely used commercially available immunoassays. Methods 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. Results Using 100μl of serum, the LC Tandem MS assay had a limit of quantitation of 1.3nmol/L for both 25OH-D2 and 25OH-D3 with a linear response between 1.3 and 625nmol/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 62nmol/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. Conclusion 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.
Added by: Sarina  
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