Phosphorus-specific, liquid chromatography inductively coupled plasma mass-spectrometry for analysis of inositol phosphate and inositol pyrophosphate metabolism

Sprigg, Colleen, Whitfield, Hayley L., Leftwich, Philip T., Kuo, Hui-Fen, Chiou, Tzyy-Jen, Saiardi, Adolfo, Shipton, Megan L., Riley, Andrew M., Potter, Barry V. L., Scholey, Dawn, Burton, Emily, Bedford, Mike R. and Brearley, Charles A. (2025) Phosphorus-specific, liquid chromatography inductively coupled plasma mass-spectrometry for analysis of inositol phosphate and inositol pyrophosphate metabolism. Biochemical Journal. ISSN 0264-6021

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Abstract

Inositol phosphate (InsP) and diphosphoinositol phosphate (PP-InsP) analysis in tissues is plagued by multiple difficulties of sensitivity, regioisomer resolution and the need for radiolabeling with metabolic precursors. We describe a liquid chromatography (LC) inductively coupled plasma (ICP) mass spectrometry (MS) method (LC-ICP-MS) that addresses all such issues and use LC-ICP-MS to analyse InsPs in avian tissues. The highly sensitive technique tolerates complex matrices and, by powerful chromatography, resolves in a single run multiple non-enantiomeric myo-inositol tetrakisphosphates, myo-inositol pentakisphosphates and all inositol hexakisphosphates, including myo-inositol 1,2,3,4,5,6-hexakisphosphate (phytate), known in nature. It also separates and quantifies diphospho myo-inositol pentakisphosphate (PP-InsP5) isomers from their biological precursors and from 1,5-bis-diphospho myo-inositol 2,3,4,6 tetrakisphosphate (1,5-[PP]2-InsP4). Gut tissue inositol phosphates, belonging to a non-canonical, lipid-independent pathway, are shown to differ from phytate digestion products and to be responsive to diet.

Item Type:	Article
Additional Information:	Data availability statement: Data associated with Figure 6 are available 469 at https://research-portal.uea.ac.uk/en/persons/charlesbrearley/datasets/ Funding information: This study was funded by award of a BBSRC Norwich Research Park Doctoral Training Studentship (Ref. BB/M011216/1) to C.S. with contribution from AB Vista and a NERC grant NE/W000350/1 to C.A.B. and H.W. Feeding trials described in this study were designed by AB Vista and commissioned at Nottingham Trent University by AB Vista. BVLP is a Wellcome Trust Senior Investigator (grant 101010). AS is supported by the Medical Research Council grant MR/T028904/1.
Faculty \ School:	Faculty of Science > School of Biological Sciences
UEA Research Groups:	Faculty of Science > Research Groups > Biosciences Teaching and Education Research Faculty of Science > Research Groups > Plant Sciences Faculty of Science > Research Groups > Molecular Microbiology
Depositing User:	LivePure Connector
Date Deposited:	07 Oct 2025 16:31
Last Modified:	08 Oct 2025 13:32
URI:	https://ueaeprints.uea.ac.uk/id/eprint/100673
DOI:	10.1042/BCJ20253151

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