## 2 Materials and Methods
### Extraction of Polyphenols from Green Tea Leaves and Determination of the Polyphenol Content  
The tea leaves were removed from their bags and pre-washed with Milli-Q water. They were then dried in an oven at 60 \({}^{\circ}\)C until constant weighing. To extract the polyphenols, 5 g of the dry tea sample was weighed and transferred to a three-necked flask. A total of 100 mL of Milli-Q water was added as a solvent and reflux extraction was carried out for 20 min, as previously described in the literature [59]. At the end of the process, the tea extract was separated from the solids by triple vacuum filtration. To avoid oxidative degradation of polyphenols, the resulting tea extract was stored in a freezer at \(-22\)\({}^{\circ}\)C until use [60]. The composition of the natural extract was qualitatively analyzed using the following procedure. Initially, the samples were filtered through 0.45 \(\upmu\)m nylon membrane filters. Subsequently, an HPLC system (Agilent Technologies HPLC 1260 Series, Santa Clara, CA, USA) equipped with a UV/Vis-Photo Diode Array Detector (DAD) and a Luna C18 column (3 \(\upmu\)m, 150 mm \(\times\) 2 mm, 100 A, Phenomenex, Torrance, CA, USA) was utilized to separate and identify the polyphenols present in the sample. Two different mobile phases were employed. The first one (A) consisted of 0.1% formic acid in water (\(v/v\)), while the second one (B) was composed of 0.1% formic acid in methanol. The experiment involved the use of various gradients: 0-3 min, 10% B; 3-30 min, 50% B; 30-40 min, 60% B; 40-45 min, 60% B; 45-50 min, 100% B; 50-60 min, 10% B. A consistent flow rate of 0.25 mL-min\({}^{-1}\) was maintained. The column temperature was held constant at 30 \({}^{\circ}\)C, and the injection volume was 20 \(\upmu\)L. The DAD operated at \(\uplambda\) = 280 nm. For the sake of clarity, determination of the total polyphenol content in the leaf extract was preferred due to its composition as a mixture of various polyphenic compounds. Quantification was conducted by UV-VIS spectrophotometry (Thermo Fisher Scientific, Waltham, MA, USA) using the Folin-Ciocalteu method [61]. This method relies on the reaction between phenolic compounds and Folin's reagent under alkaline conditions, achieved by the addition of Na\({}_{2}\)CO\({}_{3}\). Folin's reagent in this assay consists of a solution of sodium tungstate and sodium tungstate in phosphoric acid. When this is reduced by phenolic groups, the yellow phosphomolybdotungscit acid (formed by both salts in an acidic medium) produces an intense blue complex, whose absorbance was determined spectrophotometrically using a Shimadzu UV-1800 Ultraviolet/Visible spectrophotometer (Cole-Parmer, Vernon Hills, IL, USA). It is important to note that the quantification was performed based on a gallic acid standard line. Consequently, the concentration of polyphenols was determined as gallic acid equivalents. The procedure for establishing the standard line was as follows: Solutions with varying concentrations (ranging from 2.3 to 7.9 mg/L) of gallic acid standards were prepared in 25 mL flasks using a 196 mg/L stock solution. Subsequently, 0.5 mL of [PERSON]'s reagent and 10 mL of 7.5% Na\({}_{2}\)CO\({}_{3}\) were added to these solutions, followed by the addition of ultrapure water to reach the desired volume. The flasks were kept in the dark for one hour, after which the absorbance was measured at a fixed wavelength of 760 nm against a blank prepared in the same manner but without gallic acid. Following the establishment of the calibration line, the quantification of polyphenols in the green tea extract was conducted. Various volumes of the extract (specifically, 50, 75, and 100 \(\upmu\)L) were measured. To each volume, 0.5 mL of [PERSON]'s reagent and 10 mL of Na\({}_{2}\)CO\({}_{3}\) were added, and the solution was brought to a final volume of 25 mL by adding ultrapure water. After allowing the solution to stand for one hour in the absence of light, the absorbance was measured at 760 nm.