A liquid chromatography-tandem mass spectrometry based method for the quantification of adenosine nucleotides and NAD precursors and products in various biological samples

Frontiers in Immunology

Immunology

Johanna Hiefner¹  Anna Worthmann¹  Johann Rische²  Madeleine J. Bunders³ 

[1] Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany;III. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany;III. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany;Research Department of Virus Immunology, Leibniz Institute of Virology, Hamburg, Germany;Hamburg Center of Translational Immunology, Hamburg, Germany;

关键词: metabolomics;    adenine nucleotides;    quantification;    LC-MS/MS;    HILIC;    purinergic signaling;   

DOI  :  10.3389/fimmu.2023.1250762

 received in 2023-06-30, accepted in 2023-08-18,  发布年份 2023

来源: Frontiers

【 摘 要 】

Adenine nucleotides (AN) are ubiquitous metabolites that regulate cellular energy metabolism and modulate cell communication and inflammation. To understand how disturbances in AN balance arise and affect cellular function, robust quantification techniques for these metabolites are crucial. However, due to their hydrophilicity, simultaneous quantification of AN across various biological samples has been challenging. Here we present a hydrophilic interaction high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) based method for the quantification of 26 adenosine nucleotides and precursors as well as metabolic products of nicotinamide adenine dinucleotide (NAD) in plasma, liver, and adipose tissue samples as well as cell culture supernatants and cells. Method validation was performed with regard to linearity, accuracy, precision, matrix effects, and carryover. Finally, analysis of cell culture supernatants derived from intestinal organoids and RAW 264.7 cells illustrates that the here described method is a reliable and easy-to-use tool to quantify AN and opens up new avenues to understand the role of AN generation and breakdown for cellular functions.

【 授权许可】

Unknown   
Copyright © 2023 Hiefner, Rische, Bunders and Worthmann