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

Campos, Rafael; Pinheiro, David Halen Araújo; Britto-Júnior, José; Castro, Heleson Alves de; Mendes, Gustavo Duarte; Moraes, Manoel Odoríco de; Moraes, Maria Elisabete Amaral de; Lopes-Martins, Rodrigo Álvaro Brandão; Antunes, Natalícia J.; Nucci, Gilberto De

doi:10.1016/j.jchromb.2021.122668

Cited by 16 publications

(17 citation statements)

References 11 publications

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“…The method employed for 6‐ND quantification 18 was modified to allow the measurement of the four catecholamines in a single chromatographic run. Briefly, the extraction of the catecholamines from KHS (1 ml) was performed by solid phase extraction.…”

Section: Methodsmentioning

confidence: 99%

“…The dissolved residues were injected by a SIL‐30AC autoinjector, at a temperature of 8°C. The transitions monitored by electrospray multiple reaction monitoring, injection volume, run‐time, and limit of quantitation were described elsewhere 18 . The linearity was given in a range of 0.1–20 ng/ml, and the method validation was carried following the Food and Drug Adminisration (FDA) guidelines for bioanalytical methods 19 …”

Section: Methodsmentioning

confidence: 99%

“…To 1 ml of KHS was added 50 μl (100 ng/ml) of the deuterated catecholamines used as internal standards, and the samples were homogenized for 10 s. multiple reaction monitoring, injection volume, run-time, and limit of quantitation were described elsewhere. 18 The linearity was given in a range of 0.1-20 ng/ml, and the method validation was carried following the Food and Drug Adminisration (FDA) guidelines for bioanalytical methods. 19…”

Section: Determination Of Catecholamine Concentrations By Mass Spectr...mentioning

confidence: 99%

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6‐nitrodopamine is a major endogenous modulator of human vas deferens contractility

et al. 2022

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Background Rat isolated vas deferens releases 6‐nitrodopamine (6‐ND), and the spasmogenic activity of this novel catecholamine is significantly reduced by tricyclic compounds such as amitriptyline, desipramine, and carbamazepine and by antagonists of the α1‐adrenergic receptors such as doxazosin, tamsulosin, and prazosin. Objectives To investigate the liberation of 6‐ND by human epididymal vas deferens (HEVDs) and its pharmacological actions. Methods The in vitro liberation of 6‐ND, dopamine, noradrenaline, and adrenaline from human vas deferens was evaluated by LC‐MS/MS. The contractile effect of the catecholamines in HEVDs was investigated in vitro. The action of tricyclic antidepressants was evaluated on the spasmogenic activity ellicited by the catecholamines and by the electric‐field stimulation (EFS). The tissue was also incubated with the inhibitor of nitric oxide (NO) synthase L‐NAME and the release of catecholamines and the contractile response to EFS were assessed. Results 6‐ND is the major catecholamine released from human vas deferens and its synthesis/release is inhibited by NO inhibition. The spasmogenic activity elicited by EFS in the human vas deferens was blocked by tricyclic antidepressants only at concentrations that selectively antagonize 6‐ND induced contractions of the human vas deferens, without affecting the spasmogenic activity induced by dopamine, noradrenaline, and adrenaline in this tissue. Incubation of the vas deferens with L‐NAME reduced both the 6‐ND release and the contractions induced by EFS. Discussion and conclusion 6‐ND should be considered a major endogenous modulator of human vas deferens contractility and possibly plays a pivotal role in the emission process of ejaculation. It offers a novel and shared mechanism of action for tricyclic antidepressants and α1‐adrenergic receptor antagonists.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Determination Of Catecholamine Concentrations By Mass Spectr...mentioning

confidence: 99%

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6‐nitrodopamine is a major endogenous modulator of human vas deferens contractility

et al. 2022

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“…The release of 6‐ND, and the classical catecholamines dopamine, noradrenaline and adrenaline, in Krebs–Henseleit's solution, can be quantified simultaneously by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS; Campos et al, 2021). Here it is described the method and the full validation for quantification in Krebs–Henseleit’s solution, not only 6‐ND, but also other NO‐related catecholamines, such as 6‐nitrodopa and 6‐nitroadrenaline.…”

Section: Introductionmentioning

confidence: 99%

Measurement of 6‐cyanodopamine, 6‐nitrodopa, 6‐nitrodopamine and 6‐nitroadrenaline by LC–MS/MS in Krebs–Henseleit solution. Assessment of basal release from rabbit isolated right atrium and ventricles

Júnior,

Britto‐Júnior,

Magalhaes

et al. 2023

Biomedical Chromatography

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This study presents the validation of a sensitive method for the determination of 6‐nitrodopa, 6‐nitrodopamine, 6‐nitroadrenaline and 6‐cyanodopamine in Krebs–Henseleit solution by LC–MS/MS with ESI+. HRMS was used to precisely characterize the structures of the fragment ions. The method was applied to investigate the catecholamine basal release from rabbit isolated atria and ventricles. The atria and ventricles were suspended separately in a 5 ml organ bath containing Krebs–Henseleit solution with ascorbic acid (3 mM), gassed (95%O2/5%CO2) at 37°C for 30 min. Strata‐X 33 μm SPE cartridges were employed for the extraction of the catecholamines and the internal standard 6‐nitrodopamine‐d4. The catecholamines were separated employing a 150 × 3 mm Shim‐pack GIST C18‐AQ (3 mm particle size), placed in an oven at 40°C and perfused by 65% of mobile phase A (MeCN/H2O; 90/10, v/v) + 0.4% CH3COOH and 35% mobile phase B (deionized H2O) + 0.2% CH2O2 at 320 μl/min in isocratic mode. The method was linear at 0.1–20 ng/ml. The method was used to identify for the first‐time basal release of the three nitrocatecholamines mentioned above and a member of a novel class of catecholamines, the cyanocatecholamines.

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“…The method employed for 6-ND quantification ( 14 ) was modified to allow the measurement of the four catecholamines in a single chromatographic run. Briefly, the extraction of the catecholamines from KHS (1 mL) was performed by solid phase extraction.…”

Section: Methodsmentioning

confidence: 99%

Relaxation of thoracic aorta and pulmonary artery rings of marmosets (Callithrix spp.) by endothelium-derived 6-nitrodopamine

Britto

Lima

Santos-Xavier

et al. 2023

Braz J Med Biol Res

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6-Nitrodopamine is a novel catecholamine released by vascular tissues, heart, and vas deferens. The aim of this study was to investigate whether 6-nitrodopamine is released from the thoracic aorta and pulmonary artery rings of marmosets ( Callithrix spp. ) and to evaluate the relaxing and anti-contractile actions of this catecholamine. Release of 6-nitrodopamine, dopamine, noradrenaline, and adrenaline was assessed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The relaxations induced by 6-nitrodopamine and by the selective dopamine D2 receptor antagonist L-741,626 were evaluated on U-46619 (3 nM)-pre-contracted vessels. The effects of 6-nitrodopamine and L-741,626 on the contractions induced by electric-field stimulation (EFS), dopamine, noradrenaline, and adrenaline were also investigated. Both aorta and pulmonary artery rings exhibited endothelium-dependent release of 6-nitrodopamine, which was significantly reduced by the NO synthesis inhibitor L-NAME. Addition of 6-nitrodopamine or L-741,626 caused concentration-dependent relaxations of both vascular tissues, which were almost abolished by endothelium removal, whereas L-NAME and the soluble guanylate cyclase inhibitor ODQ had no effect on 6-nitrodopamine-induced relaxations. Additionally, pre-incubation with 6-nitrodopamine antagonized the dopamine-induced contractions, without affecting the noradrenaline- and adrenaline-induced contractions. Pre-incubation with L-741,626 antagonized the contractions induced by all catecholamines. The EFS-induced contractions were significantly increased by L-NAME, but unaffected by ODQ. Immunohistochemical assays showed no immunostaining of the neural tissue markers S-100 and calretinin in either vascular tissue. The results indicated that 6-nitrodopamine is the major catecholamine released by marmoset vascular tissues, and it acts as a potent and selective antagonist of dopamine D 2 -like receptors. 6-nitrodopamine release may be the major mechanism by which NO causes vasodilatation.

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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

Cited by 16 publications

References 11 publications

6‐nitrodopamine is a major endogenous modulator of human vas deferens contractility

6‐nitrodopamine is a major endogenous modulator of human vas deferens contractility

Measurement of 6‐cyanodopamine, 6‐nitrodopa, 6‐nitrodopamine and 6‐nitroadrenaline by LC–MS/MS in Krebs–Henseleit solution. Assessment of basal release from rabbit isolated right atrium and ventricles

Relaxation of thoracic aorta and pulmonary artery rings of marmosets (Callithrix spp.) by endothelium-derived 6-nitrodopamine

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