Butyrylcholinesterase from equine

Quercetin, morin, rutin, pyrogallol, mandelic acid, hydroxy benzoic acid, phloroglucinol and chlorogenic acid, Follin-ciocalteu reagent, sodium carbonate, 2,2-diphenyle-1 picrylhydrazyle [DPPH], aluminum chloride, sodium nitrate, sodium hydroxide, ethanol, methanol, ascorbic acid, distilled water, 5,5-dithio-bis-nitrobenzoic acid (DTNB), 2,2′-azino-bis-3 ethylbenzothiazoline-6-sulfonic acid (ABTS) were all purchased from Sigma Aldrich. Butyrylthiocholine iodide, acetylthiocholine iodide, acetyl cholinesterase from Electric eel (type-VI-S) and butyrylcholinesterase from Equine were obtained from Sigma-Aldrich USA and were used in enzyme inhibition assays. All solvents used in the assays were of HPLC grades purchased from Dae-Jung Korea.

The antioxidant standards: quercetin, morin, rutin, pyrogallol, mandalic acid, hydroxy benzoic acid, phloroglucinol and chlorogenic acid were obtained from Sigma-Aldrich France. From the same firm galanthamine (from Lycoris Sp.) was also obtained. Follin-ciocalteu reagent, sodium carbonate, 5,5-dithio-bis-nitrobenzoic acid (DTNB) and 2,2-Diphenyle-1 picrylhydrazyle (DPPH) were purchased from Sigma Aldrich CHEMIE GmbH USA. Aluminum chloride, sodium nitrite, sodium hydroxide, ethanol, methanol, ascorbic acid, 2,2′-azino-bis-3 ethylbenzothiazoline-6-sulfonic acid (ABTS) were bought from sigma Aldrich Germany. Phosphate buffer (pH 8), butyryl thiocholine iodide, acetylthiocholine iodide, acetyl cholinesterase from Electric eel (type-VI-S) and butyryl cholinesterase from equine used in enzyme inhibition assays were obtained from Sigma-Aldrich USA. All the chemicals used were of analytical grade except the HPLC solvents which were HPLC grade and were purchased from DaeJung Korea. They were used as such without any further purification. Distilled water used for HPLC analysis and antioxidant assays were prepared in biochemistry lab using automatic water still (Daihen labtech, china). The HPLC solvents were subjected to sonication (Elmasonic, Model E 30 H) to remove air bubbles before use.