Quantifying vitamin D and its metabolites by LC/Orbitrap MS. Issue 20 (28th August 2015)

• Record Type: Journal Article
• Authors: Raml, Reingard; Ratzer, Maria; Obermayer-Pietsch, Barbara; Mautner, Anton; Pieber, Thomas R.; Sinner, Frank M.; Magnes, Christoph
• Is Part Of: Analytical methods. Volume 7:Issue 20(2015)
• Page Start: 8961
• Page End: 8966
• Publication Date: 2015-08-28
• Subjects: Chemistry, Analytic -- Periodicals; Analytical biochemistry -- Periodicals; Chemical laboratories -- Standards -- Periodicals; 543.1905
• DOI: 10.1039/c5ay01583a
• Languages: English
• ISSNs: 1759-9660

Abstract

Abstract : We developed a HPLC-HRMS method for the determination of 25(OH)D2, 25(OH)D3, epi-25(OH)D3, and vitamins D2 and D3 as well as 24, 25(OH)2 D3 . 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)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.