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Quantification of 6-nitrodopamine in Krebs-Henseleit's solution by LC-MS/MS for the assessment of its basal release from Chelonoidis carbonaria aortae in vitro

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Author(s):
Campos, Rafael ; Araujo Pinheiro, David Halen ; Britto-Junior, Jose ; de Castro, Heleson Alves ; Mendes, Gustavo Duarte ; Moraes, Manoel Odorico ; Moraes, Maria Elisabete A. ; Brandao Lopes-Martins, Rodrigo Alvaro ; Antunes, Natalicia J. ; De Nucci, Gilberto
Total Authors: 10
Document type: Journal article
Source: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES; v. 1173, p. 6-pg., 2021-04-02.
Abstract

In this study, the development and validation of a method for quantification of 6-nitrodopamine in Krebs-Henseleit's solution by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) with positive ion electrospray ionization is described. Aortic rings taken from tortoise were either denuded or left with endothelium intact (15 mm, N = 6) and were incubated for 30 min in 5 mL Krebs-Henseleit's solution in an organ bath. Solid phase extraction (SPE) was performed for aliquots of 1 mL of the supernatant. The separation of 6-nitrodopamine was obtained on a 150 mm x 3 mm Shim-pack GIST-HP C18 column, using 75% of mobile phase A consisted of deionized water with 0.1% formic acid (v/v) and 25% of mobile phase B consisted of acetonitrile/deionized water (50/50, v/v) + 0.1% formic acid at a flow rate of 350 mu L/min in an isocratic mode. The method was linear over the concentration range of 0.1-20 ng/mL. The method was sensitive, precise and accurate for the assessment of the basal release of 6-nitrodopamine from Chelonoidis carbonaria aortae in vitro. The mean +/- SEM concentrations of 6-nitrodopamine released from endothelium-intact and endothelium-denuded aortae were 0.44 +/- 0.06 ng/mL and 0.18 +/- 0.05 ng/mL, respectively. These results indicate that tortoise's aortae display a basal endothelium-derived 6-nitrodopamine release. (AU)

FAPESP's process: 16/22506-1 - Metabolism of dapaconazole
Grantee:Natalícia de Jesus Antunes
Support Opportunities: Scholarships in Brazil - Post-Doctoral