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Issue 20, 2015
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Quantifying vitamin D and its metabolites by LC/Orbitrap MS

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Establishing accurate and precise analytical methods for vitamin D species is of rising importance as awareness of the vitamin D deficiency problem grows. Here, we report an analytical method for quantifying vitamin D and relevant vitamin D metabolites by high-resolution mass spectrometry (HRMS). We combine a straightforward time saving sample preparation and chromatographic separation, employing gradients and a column-switching technique, with high-resolution MS detection, extending its applicability to a broader range of relevant vitamin D metabolites. We calibrated the method to NIST standards and validated it for 25(OH)D2 and 25(OH)D3. We also present short- and long-term sample-stability data and proof-of-concept by applying the method to serum samples from a healthy patient library that were previously measured by immunoassay. Compared to the immunoassay data, our method showed a good correlation of 25(OH)D3 concentrations. Thus, we show that quantitative HRMS is now within reach of clinical diagnostics for vitamin D and its metabolites.

Graphical abstract: Quantifying vitamin D and its metabolites by LC/Orbitrap MS

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Supplementary files

Article information

19 Jun 2015
09 Aug 2015
First published
28 Aug 2015

Anal. Methods, 2015,7, 8961-8966
Article type
Technical Note
Author version available

Quantifying vitamin D and its metabolites by LC/Orbitrap MS

R. Raml, M. Ratzer, B. Obermayer-Pietsch, A. Mautner, T. R. Pieber, F. M. Sinner and C. Magnes, Anal. Methods, 2015, 7, 8961
DOI: 10.1039/C5AY01583A

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