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Raml, R; Ratzer, M; Obermayer-Pietsch, B; Mautner, A; Pieber, TR; Sinner, FM; Magnes, C.
Quantifying vitamin D and its metabolites by LC/Orbitrap MS
ANAL METHODS-UK. 2015; 7(20): 8961-8966. Doi: 10.1039/c5ay01583a
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Co-authors Med Uni Graz: Obermayer-Pietsch Barbara; Pieber Thomas; Sinner Frank Michael
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Abstract:: 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)D-2 and 25(OH)D-3. 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)D-3 concentrations. Thus, we show that quantitative HRMS is now within reach of clinical diagnostics for vitamin D and its metabolites.