# Enhancing the Content of Hesperidin and Nobiletin in Citrus Wines through Multi-Strain Fermentation
## 2 Materials and Methods
### Physicochemical Analysis
\begin{table}
\begin{tabular}{c c c c} \hline \hline \multirow{2}{*}{**Control**} & **Multi-Strain Combination** & **Multi-Strain Combination** & **Multi-Strain Combination** \\  & **Fermentation** & **Fermentation** & **Fermentation** \\  & **(Sequential Fermentation)** & **(1:1 Mixed Fermentation)** & **(1:100 Mixed Fermentation)** \\ \hline \multirow{4}{*}{Sc} & (Ce.Hg)-10-Sc & Sc(Ce.Hg)-1:1 & Sc(Ce.Hg)-1:100 \\  & (Ce 0.5 \(\times\) 10\({}^{7}\) CFU/mL, Hg 0.5 \(\times\) 10\({}^{7}\) & (Ce 0.5 \(\times\) 10\({}^{7}\) CFU/mL, Hg 0.5 \(\times\) 10\({}^{7}\) & (Ce 0.5 \(\times\) 10\({}^{7}\) CFU/mL, Hg 0.5 \(\times\) 10\({}^{7}\) \\  & CFU/mL, Sc 1 \(\times\) 10\({}^{7}\) CFU/mL) & CFU/mL, Sc 1 \(\times\) 10\({}^{7}\) CFU/mL) & CFU/mL, Sc 1 \(\times\) 10\({}^{7}\) CFU/mL) \\  & (Ce.Ht-10-Sc & Sc(Ce.Ht)-1:1 & Sc(Ce.Ht)-1:100 \\ \multirow{4}{*}{Hg-10-Se} & (Ce 0.5 \(\times\) 10\({}^{7}\) CFU/mL, Ht 0.5 \(\times\) 10\({}^{7}\) & (Ce 0.5 \(\times\) 10\({}^{7}\) CFU/mL, Ht 0.5 \(\times\) 10\({}^{7}\) & (Ce 0.5 \(\times\) 10\({}^{7}\) CFU/mL, Ht 0.5 \(\times\) 10\({}^{7}\) \\  & CFU/mL, Sc 1 \(\times\) 10\({}^{7}\) CFU/mL) & CFU/mL, Sc 1 \(\times\) 10\({}^{7}\) CFU/mL) & CFU/mL, Sc 1 \(\times\) 10\({}^{8}\) CFU/mL) \\  & (Hg.Ht-10-Sc & Sc(Hg.Ht)-1:1 & Sc(Hg.Ht)-1:10 \\  & (Hg.0 \(\times\) 10\({}^{7}\) CFU/mL, Ht 0.5 \(\times\) 10\({}^{7}\) & (Hg.0 \(\times\) 10\({}^{7}\) CFU/mL, Ht 0.5 \(\times\) 10\({}^{7}\) & (Hg.0 \(\times\) 10\({}^{7}\) CFU/mL, Ht 0.5 \(\times\) 10\({}^{7}\) \\  & CFU/mL, Sc 1 \(\times\) 10\({}^{7}\) CFU/mL) & CFU/mL, Sc 1 \(\times\) 10\({}^{7}\) CFU/mL) & CFU/mL, Sc 1 \(\times\) 10\({}^{8}\) CFU/mL) \\  & (Ce.Hg.Ht)-10-Sc & Sc(Ce.Hg.Ht)-1:1 & Sc(Ce.Hg.Ht)-1:10 \\  & (Ce.3 \(\times\) 10\({}^{7}\) CFU/mL, Hg 0.3 \(\times\) 10\({}^{7}\) & (Ce 0.3 \(\times\) 10\({}^{7}\) CFU/mL, Hg 0.3 \(\times\) 10\({}^{7}\) & (Ce 0.3 \(\times\) 10\({}^{7}\) CFU/mL, Hg 0.3 \(\times\) 10\({}^{7}\) \\  & CFU/mL, Ht 0.3 \(\times\) 10\({}^{7}\) CFU/mL, Sc & CFU/mL, Ht 0.3 \(\times\) 10\({}^{7}\) CFU/mL, Sc & CFU/mL, Ht 0.3 \(\times\) 10\({}^{7}\) CFU/mL, Sc \\  & \(1\times\) 10\({}^{7}\) CFU/mL) & \(1\times\) 10\({}^{7}\) CFU/mL) & \(1\times\) 10\({}^{3}\) CFU/mL) \\ \hline \hline \end{tabular}
\end{table} 
Table 1: The fermentation groups. 3.8 mL of 0.1 mM DPPH (Sinopharm Group Chemical Reagent, Beijing, China) solution was prepared. Following a 30-min incubation in darkness, the absorbance was recorded at a wavelength of 517 nm. Every sample underwent testing three times. To determine the percentage of the DPPH radical scavenging rate, the following formula was used:  
DPPH radical scavenging rate (%) = (1 - AS/Ab) \(\times\) 100%  
AS represents the absorbance of the sample, while Ab represents the absorbance of the blank. The determination of the ABTS radical scavenging method was carried out with slight modifications to the methods as described previously [20]. A mixture of ABTS solution (7.0 mM, 10 mL, Sinopharm Group Chemical Reagent, Beijing, China) and potassium persulfate (140 mM, 176 \(\upmu\)L, Sinopharm Group Chemical Reagent, Beijing, China) was left to incubate at room temperature in the dark for 14 h. An appropriate quantity of ethanol was added to the ABTS solution to reach an absorbance of 0.70 \(\pm\) 0.02 at 734 nm. After diluting the samples with anhydrous ethanol to the desired concentration, 200 \(\upmu\)L of the diluted sample was mixed with 3.8 mL of the ABTS+-solution. Following a 6-min incubation period at ambient temperature, the absorption was recorded at a wavelength of 734 nm. Every sample underwent testing three times. To determine the rate of ABTS+-scavenging, use the following formula:  
ABTS radical scavenging rate (%) = (1 - AS/Ab) \(\times\) 100%  
AS represents the absorbance of the sample, while Ab represents the absorbance of the blank.