Electric eel ache type 6 s

Extracts were investigated for their in vitro cholinesterase inhibitory activity at 200 μg/mL using ELISA microplate reader. AChE and BChE inhibitory activity was measured by slightly modified spectrophotometric method of Ellman et al. (Ellman et al., 1961) . Electric eel AChE (Type-VI-S; EC 3.1.1.7, Sigma, St. Louis, MO, USA) and horse serum BChE (EC 3.1.1.8, Sigma, St. Louis, MO, USA) were the enzyme sources used, while acetylthiocholine iodide and butyrylthiocholine chloride (Sigma, St. Louis, MO, USA) were employed as the substrates of the reaction. 5,5'-Dithiobis(2-nitrobenzoic)acid (DTNB; Sigma, St. Louis, MO, USA) was used for the measurement of the anticholinesterase activity. All reagents and conditions were same as described in our previous publication (Sevim et al., Galanthamine (Sigma, St. Louis, MO, USA), the anticholinesterase alkaloid-type of drug obtained from the bulbs of Galanthus sp., was used as the reference. The measurements and calculations were evaluated by using Softmax PRO 4.3.2.LS software (Sunnyvale, CA, USA). Experiments were run in triplicate and the results were expressed as average values with S.E.M.

The measurements and calculations of the activity results were evaluated by using bioactivity measurements, they were carried out on a 96-well microplate reader, Perkin Elmer Multimode Plate Reader EnSpire at Center of Biotechnology Research. 1,1-diphenyl-2-picrylhydrazyl (DPPH), butylatedhydroxylanisole (BHA), butylatedhydroxyltoluene (BHT), α-Tocopherol, Folin-ciocalteu's reagent (FCR), 3-(2-Pyridyl)-5,6-di(2-furyl)-1,2,4-triazine-5′,5′′-disulfonic acid disodium salt (Ferrene), Ethylenediaminetetraacetic acid (EDTA), Neocuproine, 2,2'-azino-bis(3-ethylbenzothiazoline-6sulfonicacid)diammoniumsalt (ABTS), p-nitrophenyl-α-D-glucopyranoside, 5,50-dithiobis (2-nitrobenzoic) acid (DTNB), acetylthiocholine iodide, butyrylthiocholine chloride, and galantamine were obtained from Sigma Chemical Co (Sigma-Aldrich GmbH, Stern-heim, Germany), horse serum butyrylcholinesterase (BChE, EC 3.1.1.8, 11. 4 U/mg) and electric eel AChE (Type-VI-S, EC 3.1.1.7, 425.84 U/mg) were obtained from Sigma, St. Louis, MO, Copper (II) chloride, Potassium persulfate, Sodium Carbonate, Aluminum Nitrate, Iron (II) chloride, Potassium acetate, Ammonium acetate were obtained from Biochem Chemopharma. All other chemicals and solvents were of analytical grade.