{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T03:28:26Z","timestamp":1790479706506,"version":"4.1.0"},"reference-count":17,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2021,10,19]],"date-time":"2021-10-19T00:00:00Z","timestamp":1634601600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>A new HPLC method for the simultaneous quantitative analysis of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) was developed and validated. ATP, ADP, and AMP were extracted from human bronchial epithelial cells with a rapid extraction procedure and separated with a C18 column (3 \u00d7 150 mm, 2.7 \u00b5m) using isocratic elution with a mobile phase consisting of 50 mM of potassium hydrogen phosphate (pH 6.80). The absorbance was monitored at 254 nm. The calibration curves were linear in 0.2 to 10 \u00b5M, selective, precise, and accurate. This method allowed us to quantify the nucleotides from two cell models: differentiated NHBE primary cells grown at the air\u2013liquid interface (ALI) and BEAS-2B cell line. Our study highlighted the development of a sensitive, simple, and green analytical method that is faster and less expensive than other existing methods to measure ATP, ADP, and AMP and can be carried out on 2D and 3D cell models.<\/jats:p>","DOI":"10.3390\/molecules26206324","type":"journal-article","created":{"date-parts":[[2021,10,20]],"date-time":"2021-10-20T03:01:13Z","timestamp":1634698873000},"page":"6324","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4314-5501","authenticated-orcid":false,"given":"Ana Teresa","family":"Juarez-Facio","sequence":"first","affiliation":[{"name":"Normandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Violaine","family":"Martin de Lagarde","sequence":"additional","affiliation":[{"name":"Normandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4981-0800","authenticated-orcid":false,"given":"Christelle","family":"Monteil","sequence":"additional","affiliation":[{"name":"Normandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8296-672X","authenticated-orcid":false,"given":"Jean-Marie","family":"Vaugeois","sequence":"additional","affiliation":[{"name":"Normandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C\u00e9cile","family":"Corbiere","sequence":"additional","affiliation":[{"name":"Normandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1558-9810","authenticated-orcid":false,"given":"Tiphaine","family":"Rogez-Florent","sequence":"additional","affiliation":[{"name":"Normandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,19]]},"reference":[{"key":"ref_1","first-page":"2","article-title":"ATP and Brain Function","volume":"9","author":"Silver","year":"1989","journal-title":"Br. J. Pharmacol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Fan, T., Sun, G., Sun, X., Zhao, L., Zhong, R., and Peng, Y. (2019). Tumor Energy Metabolism and Potential of 3-Bromopyruvate as an Inhibitor of Aerobic Glycolysis: Implications in Tumor Treatment. Cancers, 11.","DOI":"10.3390\/cancers11030317"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"4680","DOI":"10.1074\/jbc.272.8.4680","article-title":"Energy Metabolism during Apoptosis","volume":"272","author":"Garland","year":"1997","journal-title":"J. Biol. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"12653","DOI":"10.1074\/jbc.M117.793018","article-title":"Deconvoluting AMP-activated protein kinase (AMPK) adenine nucleotide binding and sensing","volume":"292","author":"Gu","year":"2017","journal-title":"J. Biol. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1373\/clinchem.2006.076364","article-title":"Human Plasma ATP Concentration","volume":"53","author":"Gorman","year":"2007","journal-title":"Clin. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1016\/j.bbamcr.2008.01.024","article-title":"Nucleotide- and Nucleoside-Converting Ectoenzymes: Important Modulators of Purinergic Signalling Cascade","volume":"1783","author":"Yegutkin","year":"2008","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1134\/S1061934809070028","article-title":"Methods for the determination of adenosine triphosphate and other adenine nucleotides","volume":"64","author":"Khlyntseva","year":"2009","journal-title":"J. Anal. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"100245","DOI":"10.1016\/j.xpro.2020.100245","article-title":"Measurement of mitochondrial respiration in adherent cells by Seahorse XF96 Cell Mito Stress Test","volume":"2","author":"Gu","year":"2020","journal-title":"STAR Protoc."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1007\/978-1-4939-9018-4_19","article-title":"Determination of ATP, ADP, and AMP Levels by Reversed-Phase High-Performance Liquid Chromatography in Cultured Cells","volume":"1925","author":"Menegollo","year":"2019","journal-title":"Calcium Signal."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1016\/j.tiv.2017.04.025","article-title":"Impact of after-treatment devices and biofuels on diesel exhausts genotoxicity in A549 cells exposed at air-liquid interface","volume":"45","author":"Barraud","year":"2017","journal-title":"Toxicol. In Vitro"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/j.lwt.2015.11.047","article-title":"Analysis of 10 nucleotides and related compounds in Litopenaeus vannamei during chilled storage by HPLC-DAD","volume":"67","author":"Qiu","year":"2016","journal-title":"LWT"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.jchromb.2015.07.005","article-title":"Development of an ATP measurement method suitable for xenobiotic treatment activated sludge biomass","volume":"1000","author":"Nguyen","year":"2015","journal-title":"J. Chromatogr. B"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1556\/1326.2017.00344","article-title":"HPLC\u2013UV method for simultaneous determination of adenosine triphosphate and its metabolites inMycobacterium smegmatis","volume":"31","author":"Akhova","year":"2019","journal-title":"Acta Chromatogr."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1007\/978-1-4939-7598-3_15","article-title":"Determination of Adenine Nucleotide Concentrations in Cells and Tissues by High-Performance Liquid Chromatography","volume":"1732","author":"Guigas","year":"2018","journal-title":"Methods Mol. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"122110","DOI":"10.1016\/j.jchromb.2020.122110","article-title":"Determination of adenosine phosphates in mouse myocardium tissue by HPLC with UV detection and using porous graphite carbon column","volume":"1145","author":"Feng","year":"2020","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_16","first-page":"104","article-title":"Development of a standardized in vitro approach to evaluate microphysical, chemical, and toxicological properties of combustion-derived fine and ultrafine particles","volume":"113","author":"Castilla","year":"2021","journal-title":"J. Environ. Sci."},{"key":"ref_17","unstructured":"(, January November). ICH Harmonized Tripartite Guideline: Validation of Analytical Procedures: Text and Methodology Q2(R1). Proceedings of the International Conference of Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use, Geneva, Switzerland. Available online: https:\/\/www.ema.europea.eu\/en\/documents\/scientific-guideline\/ich-q-2-r1-validation-analytical-procedures-text-methodology-step-5_en.pdf."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/20\/6324\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:18:25Z","timestamp":1760167105000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/20\/6324"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,19]]},"references-count":17,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2021,10]]}},"alternative-id":["molecules26206324"],"URL":"https:\/\/doi.org\/10.3390\/molecules26206324","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,19]]}}}