An ultra-sensitive and high-throughput trapping-micro-LC-MS method for quantification of circulating vitamin D metabolites and application in multiple sclerosis patients

Quantitative analysis of the biologically-active metabolites of vitamin D (VitD), which are crucial in regulating various physiological and pathological processes, is important for clinical investigations. Liquid chromatography-tandem mass spectrometry (LC-MS) has been widely used for this purpose b...

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Vydáno v:Scientific reports Ročník 14; číslo 1; s. 5545 - 14
Hlavní autoři: Qu, Flora, Zhang, Ming, Weinstock-Guttman, Bianca, Zivadinov, Robert, Qu, Jun, Zhu, Xiaoyu, Ramanathan, Murali
Médium: Journal Article
Jazyk:angličtina
Vydáno: London Nature Publishing Group UK 06.03.2024
Nature Publishing Group
Nature Portfolio
Témata:
631/1647/2196
631/1647/2196/1380
631/1647/296
Calcitriol
Chromatography, Liquid
Humanities and Social Sciences
Humans
Liquid chromatography
Liquid Chromatography-Mass Spectrometry
Mass spectrometry
Mass spectroscopy
Metabolites
multidisciplinary
Multiple Sclerosis
Science
Science (multidisciplinary)
Tandem Mass Spectrometry
Trapping
Vitamin D
ISSN:2045-2322, 2045-2322
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Popis
Shrnutí:Quantitative analysis of the biologically-active metabolites of vitamin D (VitD), which are crucial in regulating various physiological and pathological processes, is important for clinical investigations. Liquid chromatography-tandem mass spectrometry (LC-MS) has been widely used for this purpose but existing LC-MS methods face challenges in achieving highly sensitive and accurate quantification of low-abundance VitD metabolites while maintaining high throughput and robustness. Here we developed a novel pipeline that combines a trapping-micro-LC-(T-µLC) with narrow-window-isolation selected-reaction monitoring MS(NWI-SRM) for ultra-sensitive, robust and high-throughput quantification of VitD metabolites in serum samples after derivatization. The selective-trapping and delivery approach efficiently removes matrix components, enabling high-capacity sample loading and enhancing sensitivity, throughput, and robustness. The NWI-SRM further improves the sensitivity by providing high selectivity. The lower limits of quantification (LOQs) achieved were markedly lower than any existing LC-MS methods: 1.0 pg/mL for 1,25(OH) 2 D3, 5.0 pg/mL for 24,25(OH) 2 D3, 30 pg/mL for both 25(OH)D2 and 25(OH)D3, all within a 9-min cycle. The method is applied to quantify VitD metabolites from 218 patients with multiple sclerosis. This study revealed negative correlations(r=− 0.44 to − 0.51) between the levels of 25(OH)D2 and all the three D3 metabolites in multiple sclerosis patients.
Bibliografie:ObjectType-Article-1
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ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-024-55939-0