## 4 Discussion  
Among the widely used methods, the FRAP and DPPH radical scavenging assays are prominent. Although both assays aim to evaluate antioxidant potential, they operate via distinct mechanisms. This study compares the fundamental principles, reaction mechanisms, and applications of these two assays. The FRAP assay is based on the principle of electron transfer, where antioxidants reduce ferric ion (Fe\({}^{3+}\)) to ferrous ion (Fe\({}^{2+}\)). The reaction involves the ferrocyanide, which, upon reduction, forms a Prussian blue complex. This assay is particularly effective in assessing reducing power rather than radical scavenging capacity. The reducing power of pinocembrin dihydrohalcone was found to be lower than that of ascorbic acid, with EC\({}_{50}\) values of 0.113 mg mL\({}^{-1}\) for pinocembrin dihydrohalcone and 0.028 mg mL\({}^{-1}\) for ascorbic acid. The observed linear increase in antioxidant activity with concentration suggests that pinocembrin dihydrohalcone could be an even stronger antioxidant at higher concentrations. First of all, it may be expected that pinocembrin dihydrohalcone will exhibit strong antioxidant activity when used as a fungicide at the recommended dose of 0.3-0.6 mg mL\({}^{-1}\). These findings are consistent with previous studies, such as those by [PERSON] et al. [35], who reported comparable reducing power between phloridzin (a dihydrohalcone) and ascorbic acid within a concentration range of 0.1-0.3 mg mL\({}^{-1}\). [PERSON] and [PERSON] [36] tested the reducing power of some synthetic chalcones and stated that the EC50 ranged from 0.048 to 0.156 mg ml\({}^{-1}\) depending on the structure, compared to the EC50 for ascorbic acid which is 0.053 mg ml\({}^{-1}\). The highest activity exhibited chalcones containing a p-dimethylamino group, but the authors also stated that reducing power strictly depended on the position of the hydroxyl group on ring A. Moreover, electron-donating substituents in ring B also increased the antioxidant activity. A similar relationship was also observed in the analysis of DPPH free radical scavenging activity. In turn, the DPPH assay evaluates the ability of antioxidants to scavenge free radicals by donating hydrogen or electrons. Unlike the FRAP assay, the DPPH assay operates in a neutral  
Figure 6: The reducing power of dihydrohalcone in comparison to ascorbic acid. or slightly acidic organic solvent (typically methanol or ethanol). It is widely used for assessing both electron-donating and hydrogen-donating antioxidants. In this matter, regarding the structural activity relationship (SAR), the pinocembrin dihydrochalcone possess antioxidant activity thanks to three hydroxyl groups, which allow electron transfer (ET), hydrogen atom transfer (HAT), and radical adduct formation (RAF) during the contact with free radicals [14]. The number of phenolic hydroxyl groups within the pinocembrin dihydrochalcone structure may enhance antioxidant activity, while glycosylation could reduce it [14; 37]. Moreover, the antioxidant activity is also enhanced by the formation of the intramolecular hydrogen bond (IHB) between 2\({}^{\prime}\)-OH (or 6\({}^{\prime}\)-OH) and the adjacent keto group. In general, the FRAP and DPPH assays provided a comprehensive evaluation of the antioxidant properties of pinocembrin dihydrochalcone. Our results showed differences in the direction of activity of the tested compound. It should be emphasized that the choice of assay depends on the nature of the antioxidants being tested and the desired analytical outcome. Therefore, in many studies, both assays are used together to provide a complex insight into the mechanisms of action [16; 38], while others have successfully incorporated only one method for the antioxidative assessment of chalcones and phenolic compounds [39]. The dissipation study, which aimed to determine the degradation rates of pinocembrin dihydrochalcone, revealed that it follows a single first-order (SFO) kinetic model.