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
•A novel method for analysis of 6-nitrodopamine in Krebs-Henseleit’s solution is presented.•The method was linear in the range of 0.1–20.0 ng/mL.•Tortoise’s aortae display a basal endothelium-derived 6-nitrodopamine release. In this study, the development and validation of a method for quantificatio...
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Veröffentlicht in: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2021-05, Vol.1173, p.122668-122668, Article 122668 |
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Zusammenfassung: | •A novel method for analysis of 6-nitrodopamine in Krebs-Henseleit’s solution is presented.•The method was linear in the range of 0.1–20.0 ng/mL.•Tortoise’s aortae display a basal endothelium-derived 6-nitrodopamine release.
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 × 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 μL/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. |
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ISSN: | 1570-0232 1873-376X |
DOI: | 10.1016/j.jchromb.2021.122668 |