Noradrenaline hydrochloride

The following drugs were used: acetylcholine chloride (ACh), sodium nitroprusside (SNP), noradrenaline hydrochloride (NA) (Sigma-Aldrich, St. Louise, MO, USA); potassium chloride (KCl) (Chempur, Piekary Slaskie, Poland); pinacidil and U-46619 (Cayman Chemical, Ann Arbor, MI, USA). The stock solutions (10 mM) of drugs were prepared in distilled water, except for Noradrenaline (NA) which was dissolved in a NaCl (0.9%) + ascorbic acid (0.01% w/v) solution; pinacidil was dissolved in DMSO, and U-46619 in ethanol. These solutions were kept at −20 °C, and appropriate dilutions were made in KHS (in mM: NaCl 115; CaCl₂ 2.5; KCl 4.6; KH₂PO₄ 1.2; MgSO₄ 1.2; NaHCO₃ 25; glucose 11.1) on the day of the experiment. At a concentration of 0.01%, the solvents did not alter the reactivity of isolated aortic rings.
Myo-inositol, trichloroacetic acid (TCA), thiobarbituric acid (TBA), sodium dodecyl sulfate (SDS), ethylenediaminetetraacetic acid (EDTA), 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB), 1-(trimethylsilyl) imidazole, pyridine and hydrogen peroxide (H₂O₂) were purchased from Sigma-Aldrich (St. Louis, MO, USA); while ethanol was purchased from Stanlab (Lublin, Poland). Other reagents were of analytical grade and were provided by commercial suppliers, including POCh (Gliwice, Poland), and Chempur (Piekary Slaskie, Poland).

ATP and its metabolic degradation products as sodium salts as well as clodronate disodium, Evans blue (EB), 4,4′-diisothiocyanatostilbene-2,2′-disulfonate (DIDS), noradrenaline hydrochloride (NA), 3,4-dihydroxybenzylamine hydrobromide, reserpine chlorhydrate, quercetin, cyclodextrin adenine, and adenosine deaminase (ADA) from calf intestinal mucosa type II crude powder were purchased from Sigma-Aldrich Chemicals (Saint Louis, MO, United States). N⁶-etheno-purine standards for chromatographic column calibration or the development of specific experimental protocols (eATP and eADO) were previously synthetized in our laboratory. Chemicals used to prepare buffers and mobile chromatography phases were of analytical grade and purchased from Merck Chemicals (Darmstadt, Germany).

Acetylcholine chloride, sodium nitroprusside, and noradrenaline hydrochloride were obtained from Sigma-Aldrich (St. Louise, MO, USA); potassium chloride from Chempur (Piekary Śląskie, Poland); pinacidil, NS1619, and U-46619 from Cayman Chemical (Ann Arbor, MI, USA). Stock solutions (10 mM) of these drugs were prepared in distilled water, except for noradrenaline, which was dissolved in NaCl (0.9%) + ascorbic acid (0.01% w/v) solution; pinacidil, NS-1619 were dissolved in DMSO, and U-46619 in ethanol.
These solutions were stored at −20 °C, and appropriate dilutions were made in Krebs-Henseleit solution (KH in mmol/L: NaCl 115; CaCl₂ 2.5; KCl 4.6; KH₂PO₄ 1.2; MgSO₄ 1.2; NaHCO₃ 25; glucose 11.1) on the day of the experiment. The maximal solvent concentration in the medium was less than 0.01% (vol/vol). At these concentrations, solvents did not alter the reactivity of the studied arteries.
Hemp seeds were purchased from Ekogram (Zielonki, Poland), and unrefined, cold-pressed hemp seed oil was obtained from Ol’Vita (Panków, Poland).

(R)-7-OH-DPAT: Cedarlane (Axon Medchem # 1013), (S)-7-OH-DPAT: Cedarlane (Axon Medchem # 1014), (R)-5-OH-DPAT: Cedarlane (Axon Medchem # 1007), (S)-5-OH-DPAT: Cedarlane (Axon Medchem # 1008), hordenine: Sigma # 04476, m-tyramine hydrochloride: Sigma # D017, Lot: 063K4620, p-tyramine hydrochloride: Sigma # T90344, noradrenaline hydrochloride: Sigma # 74480, serotonin hydrochloride: Sigma # H9523, dopamine hydrochloride: Sigma # H8502.