ISOLATION OF TYROSINASE ENZYME INHIBITOR COMPOUND FROM ETHANOLIC EXTRACT OF MALAPARI SEED DREGS (PONGAMIA PINNATA (L.) PIERRE)

• Main Author: Prabasari Dewi, Maryam
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/85289
• Institution: Institut Teknologi Bandung

Hyperpigmentation is a dermatological condition characterized by the production of excessive melanin in specific areas of the skin, leading to pigmentation that is darker than the surrounding tissue. The enzyme tyrosinase acts as a catalyst in the melanin synthesis pathway by hydroxylating tyrosine and oxidizing L-DOPA to DOPA-quinone. Inhibiting the tyrosinase enzyme presents a potential approach to preventing hyperpigmentation. Traditional uses of the malapari plant (Pongamia pinnata (L.) Pierre) have suggested various benefits for skin health. This research focuses on utilizing the seed dregs of the malapari plant to assess the efficacy of its ethanol extract and its isolated compounds as inhibitors of the tyrosinase enzyme while also characterizing the isolates obtained. The extraction process for the malapari seed dregs employed the maceration technique using 96% ethanol, followed by fractionation through a liquid-liquid extraction method utilizing n-hexane, ethyl acetate, and water. To quantify the tyrosinase enzyme inhibitory activity, a microplate assay was conducted, measuring absorbance at 475 nm. The IC50 values obtained for the extracts and the positive control, kojic acid, were 1306,96 ± 164,79 g/mL and 7,92 ± 0,47 g/mL, µ µ respectively. A qualitative activity assessment was performed using bioautography thin-layer chromatography (TLC), where the most intense white spots indicated the strongest potential for tyrosinase inhibition in the n-hexane fraction. This fraction was further subfractionated using classical column chromatography with a gradient elution method. Subsequent stage II subfractionation was executed utilizing the radial chromatography method and further purification via preparative thin layer chromatography to isolate compound Y. The purity of isolate Y was assessed using single development chromatography across three mobile phases and two-dimensional chromatography. Characterization of isolate Y, through specific spot appearance and thin layer chromatography-densitometry, confirmed it as a flavonoid compound from the flavone subgroup. Isolate Y was evaluated for its quantitative activity, demonstrating a tyrosinase enzyme inhibition rate of 5.833% at a concentration of 100 ?g/mL.