# Antioxidant Properties of Wafers with Added Pumpkin Seed Flour Subjected to In Vitro Digestion
## 4 Discussion  
Recently, food enriched with bioactive ingredients has been increasingly available on the food market. This trend is aimed at delivering products with desirable health-promoting properties. Bakery products (e.g., biscuits, muffins, cakes, wafers) are good materials for enrichment with bioactive ingredients [15; 16; 18; 29]. Wafers are low in some bioactive compounds, as they are made only from wheat flour, sugar, and fat. One of the possibilities of increasing their nutritional potential and functional attractiveness is to add other ingredients. Thus, in this study, pumpkin flour was added to wafers in the following amounts: 1 wt. %, 2 wt. %, 3 wt. %, and 4 wt. %. The compounds with antioxidant activity were characterized after the in vitro digestion process. This addition was intended to enhance the health benefits of wafers, because pumpkin flour contains many valuable biologically active compounds. For such wafers to be attractive to consumers, they should meet certain criteria related to appearance and textural features. The color of wafers is one of the most significant characteristics of their appearance, as it has a decisive impact on the attractiveness of the product. In this study, the total color change increased with the increasing pumpkin seed flour added to the wafers. This may be the result of Maillard reaction products formed during the baking of wafers. These results correspond well with those reported by [PERSON] et al. [18], where the L parameter, indicating brightness, was reduced as a result of the increasing percentage of almond peel added to the wafer recipe. Moreover, a darker color of the wafers was more attractive to potential consumers. Considering the acceptance of wafers as the sum of various quality characteristics (appearance, color, taste, crispiness, and texture) can be used to assess their potential. The overall acceptance score of wafers varied from 3.42 \(\pm\) 1.06 for P4 to 4.45 \(\pm\) 0.81 for C. In [PERSON] et al. 's [30] study, the overall acceptance score of cookies with pumpkin flour varied from 6.66 to 8.15 on the nine-point hedonic scale. The authors successfully partially replaced wheat flour to produce wholesome biscuits with an improved nutritional \begin{table}
\begin{tabular}{c c c c c} \hline \multirow{2}{*}{\(\leq\)**3.5 kDa Fraction**} & \multicolumn{4}{c}{**Antioxidant Properties**} \\ \cline{2-5}  & **ABTS*+** & **DPPH* & **Fe2+ Chelation** & **Reducing Power** \\
**Samples** & **[IC\({}_{50}\) mg/mL]** & **[IC\({}_{50}\) mg/mL]** & **[IC\({}_{50}\) mg/mL]** & **[mmol Trolox/mL]** \\ \hline Cf & 0.523 \(\pm\) 0.018 a & 5.583 \(\pm\) 0.846 b & 0.245 \(\pm\) 0.041 c & 0.030 \(\pm\) 0.002 a \\ Pf1 & 0.513 \(\pm\) 0.021 a & 5.986 \(\pm\) 0.907 b & 0.157 \(\pm\) 0.028 ab & 0.029 \(\pm\) 0.001 a \\ Pf2 & 0.512 \(\pm\) 0.056 a & 5.123 \(\pm\) 0.388 b & 0.192 \(\pm\) 0.037 b & 0.028 \(\pm\) 0.002 a \\ Pf3 & 0.507 \(\pm\) 0.036 a & 4.118 \(\pm\) 0.281 ab & 0.190 \(\pm\) 0.028 b & 0.028 \(\pm\) 0.002 a \\ Pf4 & 0.491 \(\pm\) 0.037 a & 3.843 \(\pm\) 0.291 a & 0.114 \(\pm\) 0.015 ab & 0.028 \(\pm\) 0.001 a \\ \hline \end{tabular}
\begin{tabular}{c c c c} \hline Cf, Pf1 to Pf4—fractions \(\leq\) 3.5 kDa after hydrolysis of control wafers and wafers with the addition of pumpkin flour. Different letters indicate statistically significant differences between samples (\(p\leq\) 0.05, Tukey’s test).  
\end{table} 
Table 5: Antioxidant properties of fractions with \(\leq\)3.5 kDa molecules. value without disturbing their overall acceptability. The analysis of texture in the study of [PERSON] (2020) seemed to reveal a significantly improved hardness, which resulted due to a higher Newton force in cookies prepared from composite pumpkin seed powder when compared with the control cookies [31]. In our study, the hardness and fracturability were investigated.