## 3 Results and Discussion
### Physical Assay  
Hardness values obtained from sourdough bread often show mixed results. It has been seen on wheat bread that sourdough addition increases bread's hardness [28]. However, it has also been reported that sourdough addition lowers hardness values [29]. Often, high hardness values have been observed in GF products, generally due to the high amount of carbohydrates present in the food matrix [24]. Regarding GF bread, hardness results also often show mixed results. Addition of fermented _Lupinus angustifolius_ L. flour (3 g/100 g, 6 g/100 g) increased bread's hardness by 34% and 17%, respectively [30]. Fermented cassava flour has been correlated to increased bread hardness [31]. Hardness results obtained can be observed in Figure 5. All biscuits showed similar results, except for 2 h and 6 h, both being statistically different from each other. Surface response methodology has been used to identify trends in physical and rheological properties of breads. Bread hardness indicated that there is an optimum fermentation time window in which a maximum or minimum value is obtained [32]. Hardness values for 2 h and 6 h could be explained by a similar behavior, 2 h being the maximum value obtained and 6 h the minimum value. Larger cell size formation during baking would have a significant impact on bread's hardness. As seen in Table 1, carbohydrate content decreased while hardness values were maintained in control biscuit and fermented flour biscuit. This can be explained primarily by the baking temperature and moisture content in the food matrix. It has been reported that these two values have a positive correlation on bread crumb's hardness [33]. Both cell number and size changed considerably when comparing to the control biscuit, as can be seen in Figure 6. It has been seen that, with the use of sourdoughs, bread crumb gas cell size increases. However, a transfglutaminase (TG) was used to increase fermentation stability [34]. It is \begin{table}
\begin{tabular}{c c c c c c c} \hline \hline
**Fermentation Time** & **Crude Fiber (\%)** & **Fats (\%)** & **Carohydrates (\%)** & **Ash (\%)** & **Protein (\%)** & **Moisture (\%)** \\ \hline
0 h & 0.40 \(\pm\) 0.05 a & 8.66 \(\pm\) 1.66 b & 56.76 \(\pm\) 3.16 a & 0.08 \(\pm\) 0.03 a & 9.43 \(\pm\) 0.64 a & 24.6 \(\pm\) 2.31 a \\ \hline
2 h & 0.68 \(\pm\) 0.02 a & 14.3 \(\pm\) 1.08 a & 32.9 \(\pm\) 1.71 b & 4.63 \(\pm\) 0.09 b & 12.2 \(\pm\) 2.08 a & 35.1 \(\pm\) 1.01 b \\ \hline
4 h & 0.57 \(\pm\) 0.05 a & 14.2 \(\pm\) 0.45 a & 29.4 \(\pm\) 7.27 b & 4.82 \(\pm\) 0.14 b & 12.2 \(\pm\) 1.13 a & 38.6 \(\pm\) 6.43 b \\ \hline
6 h & 0.40 \(\pm\) 0.07 a & 13.2 \(\pm\) 0.84 a & 35.4 \(\pm\) 1.10 b & 4.79 \(\pm\) 0.10 b & 10.8 \(\pm\) 0.35 a & 35.3 \(\pm\) 1.5 b \\ \hline
8 h & 0.42 \(\pm\) 0.02 a & 12.4 \(\pm\) 0.16 a & 38.0 \(\pm\) 1.52 b & 4.76 \(\pm\) 0.13 b & 10.7 \(\pm\) 2.16 a & 33.5 \(\pm\) 0.40 b \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Results of chemical composition analyzes obtained from biscuits from flours fermented for different amounts of time. Data is mean of three replicates (\(\pm\)standard deviation). a, b different letters indicate significant differences within the same column (\(p<0.05\)). Figure 5: Hardness values obtained in biscuits made from fermented flour. Data is mean of three replicates (\(\pm\)standard deviation). a, b different letters indicate significant differences (\(p<0.05\)). thought that dough acidification could have a direct impact in gas cell formation and size by modifying the interaction between water and other molecules within the food matrix [35]. The decrease of \(\overline{\text{G}^{\prime}}\) could be the best explanation for this. It has been observed that the viscous portion of dough mass allows gas cells to expand [36]. In terms of color, there was no statistical difference in the parameters as can be seen in Table 1. However, there exists a browning effect with a statistical difference between 0 h (blank) and the rest of the biscuits.