Full-scan mass spectral evidence for 3-epi-25-hydroxyvitamin D3 in serum of infants and adults

Autor:	An Vanderroost, Linda M. Thienpont, Hedwig Stepman, Dietmar Stöckl
Rok vydání:	2010
Předmět:	Chromatography Collision-induced dissociation Chemistry Metabolite Biochemistry (medical) Clinical Biochemistry General Medicine Fractionation Mass spectrometry High-performance liquid chromatography Mass spectrometric chemistry.chemical_compound Sephadex 3-epi-25-hydroxyvitamin D3
Zdroj:	cclm. 49:253-256
ISSN:	1437-4331 1434-6621
Popis:	Background: Since the introduction of liquid chromatography-mass spectrometry (LC/MS) for assessing vitamin D status, it has been shown that the C-3 epimer can account for a significant proportion of circulating 25-hydroxyvitamin D3 (25OHD3) concentrations in infants. However, some question whether monitoring a single MS transition at a chromatographic retention time typical for 3-epi-25OHD3 sufficiently warrants conclusions about the identity of the substance generating the signal. Therefore, we aimed to substantiate the evidence for 3-epi-25OHD3 in infants by collision induced dissociation (CID)-MS/MS product ion scans. A second objective was mass spectrometric investigation of the presence and prevalence of the 3-epi metabolite in serum from adults. Methods: Serum samples from six infants and 32 adults were studied using an ultra performance LC/tandem MS (UPLC/MS/MS) method designed to separate the 3-epi-25OHD3 from 25OHD3. Samples were submitted to liquid/liquid extraction and Sephadex LH-20 fractionation, prior to column-switching UPLC with MS/MS recording of CID product ion spectra of the [M+H]+ precursor ion. The respective standards were analyzed under identical UPLC/MS/MS conditions for comparison. Results: In the chromatograms of all samples, two peaks eluted with retention characteristics and spectra closely matching those observed for the 25OHD3 and the 3-epi standards. The percentage of the 3-epi metabolite relative to 25OHD3 in infants ranged from 15% to 41%, and in adults from 2.5% to 17%. Conclusions: This preliminary finding suggests that the prevalence of 3-epi-25OHD3 in serum of infants is considerable, and that even in adults the concentrations of this form should not be neglected.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::9cd38ac402e3e22932100a4a68b81356 https://doi.org/10.1515/cclm.2011.050 Zobrazit plný text záznamu