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Comprehensive and quantitative profiling of B vitamins and related compounds in the mammalian liver

Xu, Juan; Clare, Constance E; Brassington, Amey H; Sinclair, Kevin; Barrett, David


Juan Xu

Constance E Clare

Amey H Brassington

Professor of Developmental Biology

David Barrett


A method for the simultaneous quantification of B vitamins and related amines in one-carbon (1C) metabolism would benefit the study of diet and genetic/epigenetic regulation of mammalian development and health. We present a validated method for the simultaneous quantitative analysis of 13 B vitamers and four related 1C-pathway amine intermediates in liver using hydrophilic interaction chromatography (HILIC) coupled to electrospray ionization tandem mass spectrometry. Frozen sheep liver samples (50 mg) were homogenized in cold 50% acetonitrile containing 1% acetic acid with the addition of two isotope labelled internal standards. Hot acid hydrolysis was applied to release the protein-bound forms. The separation of 17 analytes was achieved using a pHILIC column with a total run time of 13 min. Detection was achieved in electrospray positive ionisation mode. Limits of detection for the majority of analytes were within the range of 0.4-3.2 pmol/g. The method was applied to 266 sheep liver samples and revealed that adenosylcobalamin, methylcobalamin, pyridoxic acid, flavin adenine dinucleotide and thiamine were the major forms of the B vitamers present with pyridoxal 5’-phosphate and thiamine pyrophosphate being detected at lower concentrations. Trimethylglycine and methylglycine were the predominant 1C-related amines measured. As anticipated, the B vitamin status of individuals varied considerably, reflecting dietary and genetic variation in our chosen outbred model species. This method offers a simple sample extraction procedure and provides comprehensive coverage of B vitamins coupled with good sensitivity and reliability.


Xu, J., Clare, C. E., Brassington, A. H., Sinclair, K., & Barrett, D. (2020). Comprehensive and quantitative profiling of B vitamins and related compounds in the mammalian liver. Journal of Chromatography B, 1136,

Journal Article Type Article
Acceptance Date Nov 13, 2019
Online Publication Date Nov 27, 2019
Publication Date Jan 1, 2020
Deposit Date Jan 30, 2020
Journal Journal of Chromotography B
Print ISSN 1570-0232
Electronic ISSN 1873-376X
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 1136
Article Number 121884
Keywords B vitamins, amines, one-carbon metabolism, HILIC, mass spectrometry, sheep liver
Public URL
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Additional Information This article is maintained by: Elsevier; Article Title: Comprehensive and quantitative profiling of B vitamins and related compounds in the mammalian liver; Journal Title: Journal of Chromatography B; CrossRef DOI link to publisher maintained version:; Content Type: article; Copyright: © 2019 The Authors. Published by Elsevier B.V.


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