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

Abstract

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.

Citation

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, https://doi.org/10.1016/j.jchromb.2019.121884

• 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
• DOI: https://doi.org/10.1016/j.jchromb.2019.121884
• Keywords: B vitamins, amines, one-carbon metabolism, HILIC, mass spectrometry, sheep liver
• Public URL: https://nottingham-repository.worktribe.com/output/3456658
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S1570023219312693?via%3Dihub
• 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: https://doi.org/10.1016/j.jchromb.2019.121884; Content Type: article; Copyright: © 2019 The Authors. Published by Elsevier B.V.