Quantification of cyclic AMP and cyclic GMP levels in Krebs-Henseleit solution by LC-MS/MS: Application in washed platelet aggregation samples.

Autor: Nash CES; Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), São Paulo, Campinas, Brazil., Antunes NJ; Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), São Paulo, Campinas, Brazil. Electronic address: nataliciaja@gmail.com., Coelho-Silva WC; Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), São Paulo, Campinas, Brazil., Campos R; Clinical Pharmacology Unit, Drug Research and Development Centre, Federal University of Ceará (UFC), Fortaleza, CE, Brazil; Superior Institute of Biomedical Sciences, Ceará State University (UECE), Fortaleza, Brazil., De Nucci G; Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), São Paulo, Campinas, Brazil; Department of Pharmacology, Institute of Biomedical Sciences, USP - University of São Paulo, São Paulo, Brazil; Clinical Pharmacology Unit, Drug Research and Development Centre, Federal University of Ceará (UFC), Fortaleza, CE, Brazil; Metropolitan University of Santos (UNIMES), Santos, Brazil.
Jazyk: angličtina
Zdroj: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2022 Nov 15; Vol. 1211, pp. 123472. Date of Electronic Publication: 2022 Sep 27.
DOI: 10.1016/j.jchromb.2022.123472
Abstrakt: Cyclic Nucleotides are important in regulating platelet function. Increases in 3'5'-cyclic adenosine monophosphate (cAMP) and 3'5'-cyclic guanosine monophosphate (cGMP) inhibit platelet aggregation and are pharmacological targets for antiplatelet therapy. Here we report an improved method for determining cAMP and cGMP concentrations and, for the first time, in washed platelet supernatants by combining high-performance liquid chromatography and tandem mass spectrometry (LC-MS/MS). Characteristic peaks of the substrates, cGMP or cAMP and their internal standards were identified in negative-ion electrospray ionisation using multiple reaction monitoring. Compared with previously reported methods, the method presented here shows high precision with the lowest lower limit of quantification (LLoQ) to date (10 pg/mL). The effect of a novel catecholamine, 6-nitrodopamine, on cyclic nucleotide levels was quantified. Our results showed that this new method was fast, sensitive, and highly reproducible.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2022 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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