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.
RESULTS: Pineapple juice vinegar, which had the highest total phenolic acid content, also exhibited the greatest in vitro antioxidant capacity compared to coconut juice and nipah juice vinegars. Following acute and sub-chronic in vivo toxicity evaluation, no toxicity and mortality were evident and there were no significant differences in the serum biochemical profiles between mice administered the vinegars versus the control group. In the sub-chronic toxicity evaluation, the highest liver antioxidant levels were found in mice fed with pineapple juice vinegar, followed by coconut juice and nipah juice vinegars. However, compared to the pineapple juice and nipah juice vinegars, the mice fed with coconut juice vinegar, exhibited a higher population of CD4+ and CD8+ T-lymphocytes in the spleen, which was associated with greater levels of serum interleukin-2 and interferon-γ cytokines.
CONCLUSIONS: Overall, the data suggested that not all vinegar samples cause acute and sub-chronic toxicity in vivo. Moreover, the in vivo immunity and organ antioxidant levels were enhanced, to varying extents, by the phenolic acids present in the vinegars. The results obtained in this study provide appropriate guidelines for further in vivo bioactivity studies and pre-clinical assessments of vinegar consumption. © 2017 Society of Chemical Industry.