Diosmin 3 5 7 trihydroxy 4 methoxyflavone 7 rutinoside

Diosmin (3′,5,7-Trihydroxy-4′-methoxyflavone 7-rutinoside), escin (β-D-Glucopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-(22α-(acetyloxy)-16α,24,28-trihydroxy-21β-{[(2Z)-2-methylbut-2-enoyl]oxy}olean-12-en-3β-yl β-D-glucopyranosiduronic acid), and bromelain from a pineapple stem were purchased from Sigma-Aldrich (St. Louis, MO, USA). All the investigated compounds were dissolved according to the suggestions of the manufacturers. Diosmin was dissolved in dimethyl sulfoxide (DMSO); the final concentration of the solvent at the highest concentration did not exceed 0.02%. Escin was dissolved in methanol, and similarly the final concentration of the solvent at its highest did not exceed 0.02%. An additional control for solvents at 0.02% concentration was performed. bromelain was dissolved in PBS. Resazurin, 1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene (TMA-DPH), 11-(Dansylamino)undecanoic acid (DAUDA), 2′,7′-dichlorofluorescin diacetate (H₂DCFDA), dihydroethidium (DHEt), and diaminofluorescein-FM (DAF-FM) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Sodium salts and other reagents were of the highest available purity and were purchased from POCH (Gliwice, Poland).

The following chemicals were purchased from Sigma Chemical Co. (St. Louis, MO, USA): 4-Amino-TEMPO (tempamine), 4-Maleimido-TEMPO (MSL), 4-(2-Iodoacetamido)-TEMPO (ISL), 5-doxyl-stearic acid (5-DS), 12-doxyl-stearic acid (12-DS), 16-doxylstearic acid (16-DS), o-phthalaldehyde (OPA), 4,4-dithiodipyridine, 2,4,6-trinitrobenzene sulfonic acid (TNBS), 2,4-dinitrophenylhydrazine (DNPH), and 2,4,6-tripyridyl-S-triazine (TPTZ). All other reagents of analytical purity were obtained from POCH S.A. (Gliwice, Poland). The investigated compounds, diosmin (3′,5,7-Trihydroxy-4′-methoxyflavone 7-rutinoside) and bromelain from the pineapple stem, were purchased from Sigma-Aldrich (St. Louis, MO, USA). Both compounds were dissolved according to the manufacturers’ suggestions: diosmin was dissolved in DMSO, and bromelain was dissolved in PBS.