Materials and Methods: Dried root of P. glabra was extracted under reflux with methyl alcohol, fractionated through the vacuum liquid chromatography technique, and evaporated and then purified the compounds using column chromatography and preparative thin-layer chromatography. THP-1 cells were treated with amentoflavone, 5,7,4'-hydroxyflavonoid, and stigmasterol with various concentrations (0-30 µg/mL) and then incubated with MTS reagent for 2h. Treatment was done for 24, 48, and 72h. Then, effects of these compounds were also tested on PGE2, TNF-α, and IL-6 expression in human THP-1-derived macrophage cells for 24h.
Results: Three new compounds such as amentoflavone, 5,7,4'-hydroxyflavonoid, and stigmasterol were isolated. After 24h of incubation, a significant decrease in cell viability was reported with IC50 values of amentoflavone, 5,7,4'- hydroxyflavonoid, and stigmasterol (21 µg/mL ≡ 38 M), (18 µg/mL ≡ 66 M) and (20 µg/mL ≡ 48.5 M), respectively. Whereas for 48 and 72h treatment showed a less decreased cell viability compared with 24h treatment. These compounds also showed a significant reduction in the production of TNF-α, IL-6, and PGE2 in a dose-dependent manner.
Conclusions: The isolated new compounds showed significant cytotoxicity and anti-inflammatory effects.
Aims and Objectives: The aim of this study was to identify the optimum method to obtain one of the chemical compounds in the water fraction and to identify the hypothesized chemical isolates in the water fraction katuk leave's ethanol extract.
Materials and Methods: The methods used in this study included the collection and determination of the katuk plant, the processing of the katuk, phytochemical filtrating, extracting with ethanol 96%, and fractionation using the liquid-liquid extraction method with n-hexane, ethyl acetate, and water solvents The water fraction of katuk leaves was analyzed by its components by thin-layer chromatography using the stationary phase of silica gel 60 F254, developer of n-butanol:acetic acid:water (4:1:5), and detection under ultraviolet (UV) light at a wavelength of 366 and 254nm, as well as with vanillin-sulfuric acid reagent. To isolate the compounds from water fraction of katuk leaves, it was then eluted with a vacuum column chromatography by eluent with a level polarity that would get 11 subfractions. Each subfraction was checked by two-dimensional thin-layer chromatography to see subfraction purity characterized by the appearance of a spot on the chromatogram plate. The isolate was analyzed using spot test, ultraviolet-visible spectrophotometer, infrared spectrophotometer, and liquid chromatography-mass spectrometry.
Results: The isolate was an alkaloid compound with a molecular mass of 406.3131 m/z with the molecular formula C21H39N6O2 as S, S-5, 5'-amino-4,4'-dihexyl-propyldihydropyrazol-3, 3-one.
Conclusion: One of the chemical compounds contained in the water fraction of the ethanol extract of the katuk leaf was an alkaloid group.