Result From the Total Reducing Sugar Based on DNS Method and Hplc Method

By comparing the result from the total reducing sugar based on DNS method and HPLC method, it can be confirmed that HPLC is more accurate and precise than DNS method. HPLC have a better absorptivity onto either in the mobile phase and stationary phase. HPLC method can separate pure compound from the substances while the DNS method is analytical. Tihomirova et al., (2016) proved that the sugar concentration from HPLC data are more than two times higher than the DNS data. According to Xu et al.

, (2015) , authors such as Nejib et al., (2011) and Sims (1995) have reported that HPLC method as an established and preferred method for the determination of the individual sugars in carbohydrate mixtures, for its accuracy and simplicity.

Fourier-transform infrared spectroscopy is a routine analytical technique. It can used to determine chemical compositions of food being quick and environmental friendly. Absorption bands in the infrared region are characteristic linkages and functional groups of a molecule. These absorption bands characterize a molecule state, including stretching or bending, so that the whole spectrum is a fingerprint of a specific compound or product (Mureşan et al.

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, 2015).From the graph above, it is noticed that the spectra showed strong and broad absorption at peak 3380〖cm〗^(-1). According to Patty et al., (2017), for regions 3300 – 3450 cm-1, the functional group is alcohols, hydrogen bonded O-H and the peak intensity is strong and broad. A very broad peak in the region between 3100 and 3600 cm-1 indicates the presence of exchangeable protons, typically from alcohol, amine, amide or carboxylic acid group.

Spectra showed strong absorption bands, characteristic to water molecules between 2800 – 3700 〖cm〗^(-1), indicating an apparent similarity to all solutions (Mureşan et al., 2015).

Based on the graph, the small peak on range 1048 to 1216.50〖cm〗^(-1) can be determined as presence of functional group of sucrose, which is ether (C-O-C). According to Mureşan et al., (2015), characteristic absorption bands of sucrose are on peak 995, 1055, 1113 and 1138〖cm〗^(-1). According to Coates (2006), the peak on (1150 – 1050〖cm〗^(-1)) represents the ether, C-O-C stretch. The particular frequency ranges from (1320–1000 〖cm〗^(-1)) is C-O stretching, which is the presence of alcohols, carboxylic acids, esters, ethers (Venkatesan et al., 2015). For the control which is without enzyme, there is no peak in this range which proved that the enzyme did hydrolyze the sucrose from the soya bean waste sample. Based on the graph, the spectra showed the absorption band at 1641〖cm〗^(-1).

According to the infrared spectrum table, the peak at range (1740-1660〖cm〗^(-1) ) show the presence of a simple carbonyl compound such as a carboxylic acid or aldehyde (Coates, 2006). According to Czegledy (2014) and Sharba, Al-Bayati, Aouad, & Rezki (2005), absorption band at 1640〖cm〗^(-1) is aldehyde functional group. So, it can deduce that the hydrolyzate of soya bean waste contains the functional group of aldehyde which is the presence of arabinose.