A liquid chromatography-tandem mass spectrometry based method for the quantification of adenosine nucleotides and NAD precursors and products in various biological samples
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,...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-09-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1250762/full |
| _version_ | 1797679220275544064 |
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| author | Johanna Hiefner Johann Rische Madeleine J. Bunders Madeleine J. Bunders Madeleine J. Bunders Anna Worthmann |
| author_facet | Johanna Hiefner Johann Rische Madeleine J. Bunders Madeleine J. Bunders Madeleine J. Bunders Anna Worthmann |
| author_sort | Johanna Hiefner |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-11T23:11:21Z |
| format | Article |
| id | doaj.art-3a7a919376cd4767a0c9bc6c9005a0f3 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-11T23:11:21Z |
| publishDate | 2023-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-3a7a919376cd4767a0c9bc6c9005a0f32023-09-21T08:13:18ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-09-011410.3389/fimmu.2023.12507621250762A liquid chromatography-tandem mass spectrometry based method for the quantification of adenosine nucleotides and NAD precursors and products in various biological samplesJohanna Hiefner0Johann Rische1Madeleine J. Bunders2Madeleine J. Bunders3Madeleine J. Bunders4Anna Worthmann5Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyIII. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyIII. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyResearch Department of Virus Immunology, Leibniz Institute of Virology, Hamburg, GermanyHamburg Center of Translational Immunology, Hamburg, GermanyDepartment of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyAdenine 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.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1250762/fullmetabolomicsadenine nucleotidesquantificationLC-MS/MSHILICpurinergic signaling |
| spellingShingle | Johanna Hiefner Johann Rische Madeleine J. Bunders Madeleine J. Bunders Madeleine J. Bunders Anna Worthmann 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 metabolomics adenine nucleotides quantification LC-MS/MS HILIC purinergic signaling |
| title | A liquid chromatography-tandem mass spectrometry based method for the quantification of adenosine nucleotides and NAD precursors and products in various biological samples |
| title_full | A liquid chromatography-tandem mass spectrometry based method for the quantification of adenosine nucleotides and NAD precursors and products in various biological samples |
| title_fullStr | A liquid chromatography-tandem mass spectrometry based method for the quantification of adenosine nucleotides and NAD precursors and products in various biological samples |
| title_full_unstemmed | A liquid chromatography-tandem mass spectrometry based method for the quantification of adenosine nucleotides and NAD precursors and products in various biological samples |
| title_short | A liquid chromatography-tandem mass spectrometry based method for the quantification of adenosine nucleotides and NAD precursors and products in various biological samples |
| title_sort | liquid chromatography tandem mass spectrometry based method for the quantification of adenosine nucleotides and nad precursors and products in various biological samples |
| topic | metabolomics adenine nucleotides quantification LC-MS/MS HILIC purinergic signaling |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1250762/full |
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