## 3 Materials and Methods
### Analysis of Bioactive Properties of HOC and FHOC  
Determination of ABTS\({}^{+}\), DPPH, and hydroxyl free radical binding capacity: The free radical binding capacities of the samples were determined spectrophotometrically [130]. The reaction of 0.1 mL of sample solution with 1.9 mL of 7 mmol/L ABTS radical solution (prepared in 2.5 mmol/L potassium persulfate) was determined kinetically by measuring the change in absorbance at 734 nm for 6 min. For hydroxyl radical binding capacity, 100 \(\upmu\)L of sample and 3 mL of Smirnoff reagent were incubated at 37 \({}^{\circ}\)C for 30 min. Absorbance measurements were then performed at 510 nm. The results were expressed as trolox equivalents. Determination of iron chelating activity: The iron ion chelating capacity of the samples was determined by spectrophotometric method [130]. A volume of 2 mL of appropriately diluted sample solution was mixed with 0.1 mL of 1 mmol/L FeCl\({}_{2}\)-4H\({}_{2}\)O solution and incubated for 30 min. Then, 0.1 mL of 5 mmol/L ferrozine was added, mixed, and incubated for another 10 min. The resulting color was measured spectrophotometrically at 562 nm and the results were expressed as EDTA conjugate. Determination of Angiotensin Converting Enzyme (ACE) inhibition activity: A volume of 10 \(\upmu\)L of 0.25 units/mL ACE prepared in 0.01 mol/L saline phosphate buffer (pH: 7.0, NaCl concentration: 0.5 mol/L) was mixed with 10 \(\upmu\)L sample solution. The mixture was incubated at 37 \({}^{\circ}\)C for 15 min, then the reaction was initiated by adding 150 \(\upmu\)L of 1.75 mmol/L FAPGG substrate solution (37 \({}^{\circ}\)C) prepared in saline phosphate buffer. The absorbance of the reaction was kinetically measured at 340 nm at 37 \({}^{\circ}\)C for 30 min. ACE activity was determined using the slope of the initial linear region of the absorbance-time curve and captopril was used as control [2]. Determination of \(\upalpha\)-amylase and \(\upalpha\)-glucosidase inhibition activity: \(\upalpha\)-glucosidase and \(\upalpha\)-amylase inhibition assays were performed according to the methods described in [PERSON] et al. (2022) [110]. A volume of 100 \(\upmu\)L of \(\upalpha\)-glucosidase enzyme (1 unit/mL) prepared in 0.1 mol/L sodium phosphate buffer (pH 6.8) was pre-incubated with 50 \(\upmu\)L sample at 37 \({}^{\circ}\)C for 10 min. Then, the enzymatic reaction was initiated by adding 50 \(\upmu\)L of 10 \(\upmu\)M (4-Nitrophenyl \(\upbeta\)-D-Galactopyranoside) as substrate and the reaction at 37 \({}^{\circ}\)C for 30 min was terminated by adding 1 mL of 0.1 mol/L Na\({}_{2}\)CO\({}_{3}\). Enzyme activity was determined by measuring the p-nitrophenyl formed at 400 nm. For the other assay, 100 \(\upmu\)L of \(\alpha\)-amylase enzyme (1 unit/mL) prepared in 0.1 mol/L sodium phosphate buffer (pH 6.8) was incubated with 100 \(\upmu\)L of sample for 5 min at 37 \({}^{\circ}\)C and 250 \(\upmu\)L of 1% (mg/mL) starch prepared in sodium phosphate buffer (pH 6.8) was added to initiate the reaction. The reaction was carried out at 37 \({}^{\circ}\)C for 5 min and 200 \(\upmu\)L of DNS reagent (1% 3,5-dinitrosalicylic acid and 12% sodium potassium tartrate in 0.4 mol/L NaOH) was added. The reaction was carried out by heating at 95 \({}^{\circ}\)C for 10 min. It was then diluted with 1 mL of distilled water in an ice bath. \(\alpha\)-amylase activity was determined by measuring absorbance at 540 nm. Sample-free solution as control, substrate-free solution as blind, and enzyme-free solution as enzyme-blind were used (for each sample). Determination of acetylcholine esterase (AChE) inhibition activity: The inhibition activity of the samples was carried out as described in [PERSON] et al. (2007) [131].