# Enzymatic Degumming of Soybean Oil for Raw Material Preparation in BioFuel Production
## 3 Results
### Analysis of the Transesterification Product by GC-MS  
To confirm the composition of the transesterification products of soybean oil, gas chromatography-mass spectrometry (GC-MS) analysis was performed. The study was performed on an Agilent 8890 gas chromatograph equipped with a DB-5 ms capillary column (30 m \(\times\) 0.32 mm) coupled to an Agilent 5977C mass detector. The mass scan range was 45.00-450.00 \(m/z\), and methanol was used as the solvent. The main objective was to determine the content of fatty acid methyl esters (FAME) in the 240-320 \({}^{\circ}\)C fraction of the biofuel. The results of the study showed that the biofuel contains the following main fractions, the main ones of which were observed at retention times of about 8.193; 8.675; 8.814; 8.882; 8.973, and 9.196 min.  
\begin{table}
\begin{tabular}{c c c} \hline \hline
**Enzymatic Degumming** & **Phosphorous Content** & **FFA (Acid Value,** \\
**Time** & **(ppm)** & **mg KOH/g)** \\ \hline
30 & 178.2 & 0.58 \\
60 & 59.2 & 0.65 \\
90 & 25.3 & 0.73 \\
120 & 9.2 & 0.84 \\
150 & 9.0 & 0.95 \\
180 & 8.8 & 1.18 \\
210 & 8.5 & 1.47 \\
240 & 8.4 & 1.95 \\
270 & 8.1 & 2.49 \\
300 & 8.0 & 3.18 \\ \hline \hline \end{tabular}
\end{table}
Table 6: Influence of the enzymatic degumming time on degumming efficiency.  
\begin{table}
\begin{tabular}{c c c} \hline \hline
**Dosage of Water (\%)** & **Phosphorous Content** & **FFA (Acid Value,** \\  & **(ppm)** & **mg KOH/g)** \\ \hline
0.50 & 254.2 & 0.59 \\
1.00 & 171.9 & 0.67 \\
1.50 & 56.2 & 1.07 \\
2.00 & 10.0 & 0.93 \\
2.50 & 9.1 & 0.87 \\
3.00 & 9.0 & 0.85 \\
3.50 & 8.8 & 0.81 \\
4.00 & 8.8 & 0.78 \\
4.50 & 8.7 & 0.74 \\
5.00 & 8.6 & 0.73 \\ \hline \hline \end{tabular}
\end{table}
Table 5: Effect of water dosage on degumming efficiency.  
Analysis of GC-MS results confirmed the presence of fatty acid methyl esters as the main components in the studied biofuel fraction (Table 7).  
FTIR Spectral Analysis of Crude Soybean Oil, Enzymatically Desummed Soybean Oil, and the Resulting Biofuel  
The Fourier-transform infrared (FTIR) spectra of samples were recorded on a Spectrum Two spectrometer PerkinElmer. The FTIR spectra of crude soybean oil and degummed soybean oil were nearly similar, and there were no significant differences, as shown in Table 8. The main differences were observed in the spectrum of biofuel, which is explained by the transesterification reaction. The identification of functional groups and bands corresponding to different stretching and bending vibrations clearly indicates the corresponding characteristic bands of the expected functional groups.  
\begin{table}
\begin{tabular}{c c c c} \hline
**FAME** & **R.T. (min)** & **Contents (\% of Total)** & **Characteristic Fragments with Masses \(m/z\)** \\ \hline Palmitic acid, methyl & & & \\ ester (C\({}_{17}\)H\({}_{34}\)O\({}_{2}\)) & 8.193 & 9.808 & 74; 87; 43; 55; 41; 143; 75; 57; 69; 227 \\ Linolenic acid, methyl & & & \\ ester (C\({}_{19}\)H\({}_{32}\)O\({}_{2}\)) & & & \\ linoleic acid, methyl & & & \\ ester (C\({}_{19}\)H\({}_{34}\)O\({}_{2}\)) & & & \\ Oleic acid, methyl & & & \\ ester (C\({}_{19}\)H\({}_{36}\)O\({}_{2}\)) & & & \\ Stearic acid, methyl & & & \\ ester (C\({}_{19}\)H\({}_{38}\)O\({}_{2}\)) & & & \\ Isolinoleic acid, methyl & & & \\ ester (C\({}_{19}\)H\({}_{34}\)O\({}_{2}\)) & & & \\ Isolinoleic acid, methyl & & & \\ ester (C\({}_{19}\)H\({}_{34}\)O\({}_{2}\)) & & & \\ Isolinoleic acid, methyl & & & \\ ester (C\({}_{19}\)H\({}_{34}\)O\({}_{2}\)) & & & \\ \hline \end{tabular}
\end{table}
Table 7: GC-MS analysis of the fatty acid methyl ester composition of the transesterification product of degummed soybean oil.  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Observation** & \multicolumn{3}{c}{**Frequency, cm\({}^{-1}\)**} \\ \cline{2-4}  & **Crude Soybean Oil** & **Degummed Soybean Oil** & **BioFuel** \\ \hline -OH & 3475.00 & 3475.46 & 3464.00 \\ -C=C–H & 3008.93 & 3008.93 & 3009.00 \\ -CH\({}_{2}\) & 2922.00 & 2922.81 & 2923.49 \\ -CH\({}_{2}\) & 2853.00 & 2853.27 & 2853.70 \\ C=O & 1741.30 &