ISOLASI SENYAWA AKTIF INHIBITOR ENZIM TIROSINASE DARI FRAKSI N-HEKSANA KULIT BATANG KAYU MANIS (CINNAMOMUM BURMANNII (NEES & T. NEES) BLUME)

• Main Author: Aini Sa'ah Dini, Nur
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/78167
• Institution: Institut Teknologi Bandung

Hyperpigmentation is described as one of skin disorder showed by the increase in melanin production. The tyrosinase enzyme acts as a catalyst during melanogenesis. Previous studies have shown that there were polyphenolic compounds in essential oils and triterpenoids in the n-hexane fraction of cinnamon bark (Cinnamomum burmannii (Nees & T. Nees) Blume) which inhibited the tyrosinase. This study aims to determine the potency of extracts and fractions of cinnamon bark (C. burmannii) as inhibitors of the tyrosinase enzyme, to isolate the active compounds as inhibitors of the tyrosinase enzyme in the n-hexane fraction of cinnamon bark, and followed by the isolate characterization and identification. Crud drug was extracted by maceration with 96% ethanol. Fractionation was carried out using the liquid-liquid extraction method using n-hexane, ethyl acetate, and ethanol-water as solvents. Extracts and fractions were monitored by TLC using universal and specific spray reagent. The inhibitory effect was determined qualitatively using the bioautographic TLC method and quantitatively by calculating the IC50 value. The chromatogram of bioautographic showed tyrosinase inhibition in the extract, n-hexane fraction, ethyl acetate fraction, ethanol-water fraction, and kojic acid as a comparison. The ethanol extract, n-hexane fraction, ethyl acetate fraction, ethanol-water fraction, and kojic acid had IC50 values of 6,496.83 ± 267.44; 637.88 ± 37.11; 3,612.28 ± 73.53; >12,000 (37.23% inhibition at a concentration of 12,000 µg/mL); 8.40 ± 1.24 µg/mL respectively. Subfractionation and purification was carried out using consecutive classical column chromatography method. The purity was checked using a single development TLC method with three mobile phases and two-dimensional TLC. The results of the purity test obtained isolate X which was tested for its activity qualitatively. Isolate X was characterized by specific single spot appearance, IR spectrophotometry, and TLC densitometry. The n-hexane fraction showed the strongest tyrosinase enzyme inhibitory activity, followed by the ethyl acetate fraction and the ethanol extract. Meanwhile, the ethanol-water fraction only showed inhibition below 50% at a 12,000 µg/mL. The Isolate X from n-hexane fraction of cinnamon bark showed inhibition activity against tyrosinase qualitatively and identified as a flavonoid compound of the isoflavone subgroup.