ISOLATION AND STUDY OF BIOLOGICAL ACTIVITY OF METABOLITE OF IDAT (CRATOXYLUM GLAUCUM KORTH.)

• Main Author: Juanda, Dadang
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/56595
• Institution: Institut Teknologi Bandung

Superoxide radicals such as reactive oxygen species (ROS) and reactive nitrogen species (RNS) have been known involved in the pathogenesis of various diseases. One source of free radicals is from the oxidation process of xanthine or hypoxanthine to uric acid by xanthine oxidase. Hyperuricemia and gout are metabolism disorders that cause accumulation of uric acid in the joint and kidney. Allopurinol is used for the treatment of hyperuricemia and gout clinically. But it has an undesirable effect including, hepatitis, nephropathy, hypersensitivity and skin rash. The level of superoxide radicals that are inadequate with the antioxidants system can cause oxidative stress. Increased uric acid levels and oxidative stress can increase metabolic syndrome complications, so it is necessary to discover a component that has xanthine oxidase inhibitory activity along with antioxidant capabilities. One group that has antioxidant activity and xanthine oxidase inhibitory activities is polyphenols. The main source of polyphenols comes from plants. The genus Cratoxylum has been traditionally used in food poisoning, internal bleeding, tonic, treating stomach pain, diuretic, preventing obesity, reducing blood lipid and lowering blood pressure. The well-known pharmacological activities of the genus Cratoxylum include antioxidants, cytotoxic activity, antibacterial, anti-malarial, anti-ulcer, tyrosine phosphatase inhibition and ?-glucosidase inhibition. One of the Cratoxylum species that has not been evaluated related to its activity and active compound information is idat (Cratoxylym glaucum Korth.). This research was conducted to isolate and to evaluate the antioxidant and xanthine oxidase inhibitory activities of idat (Cratoxylum glaucum Korth.). The research went through several main steps: crude drug standardization, phytochemical screening, extraction, determination of total phenolic and total flavonoid content, determination of antioxidant activity, in vitro xanthine oxidase inhibitory evaluation and evaluation by thin layer chromatography, isolation, biological activity evaluating of isolated compounds and characterization-identification. iv Leaf, twig and cortex of idat crude drug contained flavonoids, quinones, phenols, saponins and steroids/triterpenoids. The results of the determination of total ash content, ethanol soluble content and watersoluble content were: leaves crude drug 3.36; 12.26; 13.03%; twig crude drug 1.73; 5.28; 4.78%; cortex crude drug 1.65; 10.03; 7.09%; respectively. Extraction was carried out by reflux using n-hexane, ethyl acetate and ethanol as the solvents. The yields were: leaf extract 4.61; 7.63; 16.12%; twig extract 3.29; 3.26; 3.18%; cortex extract 2.33; 4.25; 12.84%; respectively. The extracts had density in range 0.62-0.91 (extract 1% in solvent of extraction). The results of measured of total phenolic and total flavonoid content of leaves, twigs, cortex extract varied from 6.62 to 48.77 (g GAE/100 g extract), 1.54 to 25.96 (g QE/100 g extract) respectively. The antioxidant activity test using DPPH (2,2-diphenyl-1-picrylhydrazyl) method, was obtained AAI (Antioxidant Activity Index) values varied from 0.33 - 10.50 meanwhile ascorbic acid and quercetin as standard with AAI values were 9.46 and 14.81, respectively. The inhibitory XO activity was determined by in vitro using microplate reader, IC50 values varied from 36.64 to 96.96 µg/mL and allopurinol with IC50 5.02 µg/mL. Isolate DE2-A and DE2-C have been successfully isolated from the ethyl acetate leaves extract of idat. A Qualitative test by TLC exposed that both isolates had antioxidant and XO inhibitory activities. The quantitative result presented that DE2-A and DE2-C had antioxidant activities with AAI values were 8.14 and 14.97, also XO inhibitory activities with IC50 39.84 and 61.24 µg/mL, respectively. Based on characterization and identification by Nuclear Magnetic Resonance (NMR) was known that DE2-A was protocatechuic acid and DE2-C was quercitrin, which had antioxidant and XO inhibitory activities. Idat leaves had the potential to be developed as source of antioxidant and XO inhibitor agent.