Development of microemulsion of phytosteryl glycoside in DMSO and Tween20 for increase antiamoebic activity

Abstract

© 2016 Trans Tech Publications, Switzerland. Peperomia pellucida is the plant in Piperaceae family; Thai name is "Phak Kra Sang". It was the medicinal plant eaten for protecting glaucoma, eye inflammation. Recently, the dried crude methanol of P. pellucida was killed Acanthamoeba spp. Acanthamoeba spp is protozoa, free-living in the environment and feed on bacteria. It was the causative agents of acanthamoeba keratitis. The study, dried crude methanol of P. pellucida was isolated phytosteryl glycoside (PG) by column chromatography (CC) using ethyl acetate and methanol (7:3). The developing of microemulsion (ME) of PG was dissolved in normal saline or aqueous solution.ME-PG was treated in Acanthamoeba cysts for antiamoebic test. Preparing of ME-PG by using the ratio of Smix, PG and H2O. Smix was the mixture of DMSO and Tween20 (co-surfactant and surfactant) in the ratio of 1:1(Smix1), 2:1(Smix2), 3:1(Smix3) and 4:1(Smix4). PG was concentrated in 200μg/ml in ethanol. The pseudo-ternary phase diagram for identifying ME-PG region and they were plotted by the percentage of Smix, PG and water. The stability of ME-PG was formed in condition of Smix3 and Smix4. The formulation of ME-PG was characterized by zeta potential, which was found to 121-397 nm, and conductivity showed in the range of -7.13-9.38 mV. ME-PG was treated and was absorbed in Acanthamoeb cysts.The dead cysts were stained with toluidine blue, were detected under the light microscope. Inhibited of concentration (IC50) showed 0.88±0.02 μg/ml for antiamoebic activity.