5 amino 2 3 dihydro 1 4 phthalazinedione luminol

Taxifolin, quercetin, catechin, eriodictyol, hesperetin, naringenin, horseradish peroxidase (150 U/mg), 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine-4,40-disulfonic acid monosodium salt hydrate (ferrozine), bathocuproinedisulfonic acid (BCDS), copper (II) sulfate, ascorbic acid, 1-butanol, 1,1,3,3 tetraethoxypropane, and 5-amino-2,3-dihydro-1,4-phthalazinedione (luminol) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Acetic acid, orthophosphoric acid (H₃PO₄), sodium acetate, ferrous sulfate (FeSO4·7H₂O), ferric chloride (FeCl₃·6H₂O), and hydrogen peroxide (H₂O₂) were purchased from Reakhim (Moscow, Russia). Phosphate buffered saline (PBS) was purchased from Paneco (Moscow, Russia), thiobarbituric acid (TBA) from Dia-m (Moscow, Russia), and lecithin from AppliChem (Darmstadt, Germany). The product of the condensation of Taxifolin with glyoxylic acid (DfTf) was synthesized as described previously [31 (link)]. All reagents were of analytical grade purity. Water used for the preparation of the solutions was purified by a Milli-Q system (Millipore, Burlington, MA, USA).

Paracetamol (PCM, 4-Acetaminophenol, purity ≥98%) was purchased from ACROS Organics and used as received. For the iodide dosimetry experiments, potassium iodide (KI, purity ≥99.5%) and ammonium molybdate tetrahydrate ((NH₄)₆Mo₇O_24, purity ≥99%) were obtained from Fisher Chemicals and Sigma-Aldrich, respectively. The sodium hydroxide (NaOH, purity ≥98%), and luminol (5-Amino-2,3-dihydro-1,4-phthalazinedione, purity ≥97%) used for sonochemiluminescence (SCL) measurements were obtained from Sigma-Aldrich. All solutions were prepared using Milli-Q water (18.2 MΩ).

We evaluated the effect of SPLP ingestion on blood ROS activity with a chemiluminescence (CL) analyzing system (CLD-110, Tohoku Electronic Industrial, Sendai, Japan), as described previously [19 (link)]. The system contains a photon detector (Model CLD-110), chemiluminescence counter (Model CLC-10), water circulator (Model CH-200), and personal computer system. A cooler circulator is connected to the Model CLD-110 photon detector to keep the temperature at 5 °C. The CLD-110 model is sensitive enough to detect as little as 10⁻¹⁵ W of radiant energy. The chemiluminescence was measured in an absolutely dark chamber of the CL analyzing system. We demonstrated that using the CL-emitting substance luminol (5-amino-2,3-dihydro-1,4-phthalazinedione, Sigma, St. Louis, MO, USA) for H₂O₂ or HOCl (hydrogen peroxide, sodium hypochlorite, Sigma, St. Louis, MO, USA) to enhance the CL counts provided similar data to those reported in our previous study [19 (link)]. The CL in the tested sample was continuously measured for a total of 600 s. The assay was performed in duplicate for each sample, and the results are expressed as CL counts (10 s)⁻¹. The total amount of CL in 600 s was calculated by integrating the area under the curve. The mean ± SD CL level for each sample was calculated.

Potassium iodide (KI, purity ≥ 99.5 %) was obtained from Fisher Chemicals. Ammonium molybdate tetrahydrate ((NH₄)₆Mo₇O_24, purity ≥ 99 %), Sodium Hydroxide (NaOH, purity ≥ 98 %), and Luminol (5-Amino-2,3-dihydro-1,4-phthalazinedione, purity ≥ 97 %) were obtained from Sigma-Aldrich. Paracetamol (PCM, 4-Acetaminophenol, purity ≥ 98 %) was purchased from ACROS Organics and used as received. All solutions were prepared using Milli-Q water (18.2 MΩ). Methanol, acetonitrile, water, and formic acid Optima^TM grade (purity ≥ 99.9 %) from Fisher Chemicals, were used as the mobile phase for liquid chromatography/mass spectrometry (LCMS).