Ation curve obtained by the F method (absorbance vs. caffeic acid concentration).In Table 4 the statistical parameters from the calibration curve are presented.Table 4. Statistical evaluation of calibration curve. Parameters Curve slope a Curve intercept b Limit of detection LOD (mg/L) Limit of quantification LOQ (mg/L) Coefficient of determination R2 Worth 0.147 0.004 0.0227 0.0096 0.13 0.31 99.Based on the parameters from the reference curve, the polyphenol content with regards to caffeic acid equivalent within the tested samples was calculated. The results are RIPGBM In Vivo presented in Table 5.Table five. Obtained results with statistical evaluation. Parameters/Samples Xmean SD. mg CAE/g Collagen Manage Xmean SD eight.22 1.9 Collagen/Meliss AXmean SD 9.39 1.CAE–caffeic acid equivalent; Xmean –average value; SD–standard deviation.3.five.two. Determination of Antioxidant Activity by FRAP Process The FRAP (ferric ion minimizing antioxidant parameter) system was proposed by Benzie et al. in 1996 to establish the antioxidant activity of plasma, and also a few years later, it was applied to study plant antioxidants . It truly is determined by the determination of AA by way of the capability to lower Fe3+ ions to Fe2+ ions under the influence of an antioxidant, and Fe(II) is complexed by TPTZ (two,4,6-tripyridyl-S-triazine) (Figure 8). The reduction reaction leads to the formation of a blue complicated (max = 595 nm) [55,57].Cosmetics 2021, 8,11 ofFigure 8. The schema of reaction in FRAP process .AA is determined by comparing the value from the transform in absorbance of your analyzed sample along with the normal option. The FRAP unit determines the ability to lessen 1 mole of Fe(III) to Fe(II). The transform within the absorbance worth is linear inside a wide array of concentrations, which is the benefit of this process . The optimum pH for this system, necessary to stabilize the iron ions, is 3.six, plus the redox possible on the samples should be reduced than 0.7 V since the redox potential of [Fe(TPTZ)2 ]3+ /[Fe(TPTZ)two ]2+ is 0.7 V. The FRAP system will not call for time-consuming sample preparation, is straightforward and swift to perform, and ensures repeatability with the obtained final results. FRAP has been employed in the determination from the antioxidant capacity of cells and tissues; nevertheless, it cannot measure the PF 05089771 Autophagy primary thiol antioxidant–glutathione. Moreover, Fe(II) ions are conveniently oxidized, creating an extremely damaging OHradical . The outcomes obtained for the reference curve have been shown in Table 6.Table 6. Data obtained for the calibration curve. Trolox volume (cm3 ) Concentration (mg/L) Absorbance 0.05 0.0125 0.153 0.ten 0.025 0.390 0.15 0.0375 0.643 0.20 0.0501 0.863 0.25 0.0626 1.Determined by the obtained final results, the dependence in the absorbance value on the concentration of Trolox was plotted (Figure 9). In Table 7, the statistical parameters on the calibration curve are presented.Figure 9. The calibration curve for FRAP method (absorbance vs. Trolox concentration).Cosmetics 2021, 8,12 ofTable 7. Statistical analysis of calibration curve. Parameter Curve slope a Curve intercept b Limit of detection LOD (mg/L) Limit of quantification LOQ (mg/L) Coefficient of determination R2 Worth 19.13 0.72 0.0853 0.0297 0.0024 0.0059 99.Based on the parameters from the calibration curve, the total antioxidant content with regards to Trolox equivalent in the tested samples have been calculated. The outcomes have been shown in Table eight.Table 8. Obtained benefits with statistical evaluation. Parameters/Samples Xmean SD. mg TE/g Collagen Control Xmean S.