DEVELOPMENT OF ETHYL ACETATE FRACTION OF GREEN TEA (CAMELLIA SINENSIS (L.) KUNTZE) ETHANOL EXTRACT USING NANOSUSPENSION FORMULA AS A TYROSINASE AND ANTIOXIDANT ENZYME INHIBITOR AND ITS EFFECT ON ACTIVITIES

• Main Author: Akhsani, Amalia
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/73967
• Institution: Institut Teknologi Bandung

Tyrosinase and antioxidants play an essential role in the condition of hyperpigmentation. Green tea is known to have activity as a tyrosinase enzyme inhibitor and a good antioxidant. Research shows the formation of nanoparticles can increase pharmacological activity. This study aims to develop the ethyl acetate fraction from the ethanolic extract of green tea using the nanosuspension formula as an inhibitor of the tyrosinase enzyme and antioxidant and its effect on both activities. Extraction was carried out by maceration with ethanol solvent. Then fractionation was continued by the liquid-liquid extraction method with n-hexane, ethyl acetate, and water as solvents. The sonication technique used a bath or probe to reduce the particle size. The nanosuspension was developed with Tween 20 and sodium lauryl sulfate (SLS) as stabilizers at a concentration of 0.1% (w/v). The ability to inhibit the tyrosinase enzyme is determined by the IC50 value. The IC50 values of the kojic acid, extract, ethyl acetate fraction (F2), water fraction (F3), NST-F2 (F2 nanosuspension with Tween 20 as stabilizer), and NSS-F2 (F2 nanosuspension with SLS as stabilizer) were respectively 6.37±0.15; 362.47±20.59; 131.73±7.72; 710.13±32.52; 199.91±20.79; 95,77±2,81 µg/mL. Activity as an antioxidant is determined by the IC50 value of DPPH reduction. The IC50 values of the ascorbic acid, extract, ethyl acetate fraction (F2), water fraction (F3), NST-F2, and NSS-F2 were respectively 5,38±0,09; 4.68±0.08 µg/mL; 4.94±0.06 µg/mL; 16.42±0.09 µg/mL; 5.58±0.02 µg/mL; 4.68±0.16 µg/mL. The F2 nanosuspension formula with Tween 20 stabilizer decreased the activity of inhibiting tyrosinase and antioxidant enzymes. While the F2 nanosuspension formula with SLS stabilizer experienced an increase in tyrosinase enzyme inhibition and antioxidant activity. The development of an ethyl acetate fraction nanosuspension formula from green tea ethanol extract (NS-F2) with an F2 of 1% and Tween 20 0.1% (NST-F2) has good stability after storage for 28 days at 4°C, 25°C, and 40°C. The formula that gives increased activity results is the use of sodium lauryl sulfate stabilizer at 0.1% (NSS-F2). The results of the NSS-F2 activity test showed that the inhibition of the tyrosinase enzyme increased to 137% and the antioxidant activity to 105% compared to the activity of the ethyl acetate fraction without nanosuspension development.