A voucher specimen is deposited at the KKU Herb arium, Division of Biology, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand. Chemical compounds and almost all of the pure requirements of phenolic acids have been bought from Sigma Aldrich Corporation. The pure requirements of m hydroxybenzaldehyde and p hydroxybenzoic acid have been purchased from Fluka and Acros Organics. respectively. Crude ethanolic extraction 5 grams of air dried ground rhizome had been macerated and periodically stirred in 50 ml of absolute ethanol for 48 hrs. The suspension was filtered by way of Whatman No. four filter paper and centrifuged at five,000 rpm for 15 minutes. The supernatant was air dried to yield an ethanolic crude extract. The residue was reconstituted in dimethyl sulfoxide or ethanol before testing along with the solvent was utilized as a adverse handle. Fractionated solvent extraction 5 grams of air dried ground rhizome have been macerated and periodically stirred in 50 ml of hexane for 48 hrs.
The suspension selleck inhibitor was filtered as a result of the filter paper and centrifuged at 5,000 rpm for 15 minutes. The super natant was air dried to acquire the hexane soluble frac tion. The precipitate remaining from hexane extraction was dispersed, macerated and periodically stirred in 50 mL of ethyl acetate for 48 hrs. The ethyl acetate sus pension was filtered by means of the filter paper, centrifuged at 5,000 rpm for 15 minutes, and air dried to obtain the ethyl acetate soluble fraction. The precipitate remaining from ethyl acetate extraction was dispersed, macerated and periodically stirred in 50 ml of methanol for 48 hours. The methanol suspension was filtered by means of the filter paper, centrifuged at five,000 rpm for 15 minutes, and air dried to get the methanol soluble fraction.
Just about every solvent fraction was reconstituted in an appropri ate car, DMSO or ethanol, ahead of testing. Phenolic extraction Phenolic extraction was carried out through the use of acidic hy drolysis system with some modifications. Briefly, two hundred milliliters of 70% methanol GSK2118436 manufacturer had been extra to a beaker containing ten grams of ground rhizome. The mixture was stirred for 2 hours at room temperature and after that filtered via the filter paper. The filtrate was evaporated to 60 ml by a rotary evaporator. The remaining filtrate was additional with 50 ml of two M NaOH and stirred constantly for 12 hours at space tempera ture. The mixture was centrifuged at one,700 g for twenty mi nutes and then filtered by means of the filter paper. The supernatant was repeatedly extracted three times with 80 ml of diethyl ether, in which the aqueous phase was collected along with the diethyl ether phase was discarded. The aqueous phase was adjusted to pH one.