Antityrosinase activity and LC-MS/MS analysis of optimized ultrasound-assisted condition extracts and fractions from Strawberry tree (Arbutus unedo L.)

Serdar Demir^a,*, Halil Koyu^b, Mustafa Abdullah Yilmaz^c,d, Abbas Tarhan^d,

Sura Baykan Ozturk^a

^a Department of Pharmaceutical Botany, Faculty of Pharmacy, Ege University, 35040, Bornova-Izmir, Turkey

^b Department of Pharmaceutical Botany, Faculty of Pharmacy, Izmir Katip Celebi University, 35620, Cigli-Izmir, Turkey

^c Department of Analytical Chemistry, Faculty of Pharmacy, Dicle University, 21280, Sur-Diyarbakir, Turkey

^d Dicle University Science and Technology Research and Application Center, 21280, Sur-Diyarbakir, Turkey

Investigation of utilization possibilities of natural sources has been an important area for research. Tyrosinase inhibitory activity plays a key role in food and medicine industry. Strawberry tree (Arbutus unedo), a widely distributed plant among Mediterranean countries, possess fruits and leaves with rich bioactive phytochemicals, especially polyphenolic compounds. In this study, we aimed to investigate the antityrosinase activity of the fruit and leaf extracts of the plant, and to determine the phenolic compounds that contribute to the antityrosinase activity. In this regard, we evaluated the effect of solvent composition on the extraction of phenolic compounds from A. unedo and on its antityrosinase activity using a simplex centroid design approach, and used chromatographic and LC-MS/MS techniques. The leaf extracts prepared using EtOH:water (50:50) provided higher TPC (456.39 mg GAE/g extract) and acetone:EtOH:water (33:33:33) provided higher TFC (56.15 mg QE/g extract) values than of fruit extracts. LC-MS/MS analysis revealed 23 phenolic/flavonoid compounds in leaf extracts (L1-8), and major metabolites were detected as quercitrin, quinic acid, catechin, tannic acid, isoquercitrin, gallic acid, and ellagic acid. Among the leaf extracts, L3 (aceton:water, 50:50) exhibited 72.01% tyrosinase inhibition at 500 μg/mL. After fractionation studies guided by antityrosinase activity, its subfraction L3-Fr2 exhibited 40.06% inhibition at 50 μg/mL concentration (IC₅₀: 146±7.75 μg/ml), and catechin (113.19 mg/g), tannic acid (53.14 mg/g), ellagic acid (22.14 mg/g), gallic acid (10.27 mg/g), and epicatechin gallate (8.65 mg/g) were determined as major metabolites. Its subfraction L3-Fr2-sub7 exhibited better antityrosinase activity (IC₅₀: 206.23±9.87μg/mL), and quantitative analysis results revealed the presence of tannic acid (127.40 mg/g), gallic acid (13.96 mg/g), ellagic acid (7.66 mg/g), quercetin-3-O-glucuronide (5.06 mg/g), and quinic acid (3.2 mg/g) as major metabolites, and correlation analysis showed that ellagic acid and quinic acid were positively correlated with antityrosinase activity.