DISCUSSION

DNS reagent is used to determine the reducing sugar. Reducing sugar contains free
carboxyl group and reduces DNS reagent. Different reducing sugar yields different
colour intensity and absorbance. Amount of absorbance is directly related to the
amount of reducing sugar. When there are no reducing ends present, the final colour
is yellow. Formation of a yellow-orange colour indicates a positive result.
First of all, a known concentration of carbohydrates solution and its serial dilution
need to be done to get a standard curve preparation. In order to get the standard
curves, all of four carbohydrates absorbance need to be calculated based on their
concentration. After the standard curves were prepared, they were used to estimate
the unknown concentration of carbohydrates which are sample A, B, C, and D. In
order to identify the type of carbohydrate among the sample, the colour comparison
needs to be done.
From the result obtained, glucose and maltose showed the positive result, which
turned the mixture of sugars and DNS reagent to yellow-orange colour after heated.
Despite both of the reducing sugar give the same final colour among the five
different concentrations; they gave slightly different absorbance result. The results
were tabulated and the graphs were plotted. For glucose, the slope is 0.0046 and
maltose is 0.057. Based on the unknown samples that were given, the conclusion
that can be made was sample A is maltose and D is glucose after the colour
comparison. However, the average reading of absorbance showed a few amount of
reducing sugar. That means both absorbance reading of sample A and D differ from
analytical value based on late experiment. Amount of absorbance directly related to
amount of reducing sugar. So, meaning that reducing sugar may have higher
absorbance. After the calculations were performed, the concentration of sample A is
0.0322mg/ml and sample D is -0.07mg/ml. On one hand, the concentration of
sample A is acceptable although it too small, but on the other hand, sample D cannot
be make sense because of the concentration is negative.
Otherwise, sucrose and starch proved that they are non-reducing sugars which
turned the mixture into yellow in colour. Since there are no free carbonyl groups in
sucrose, it is therefore non-reducing sugar (Kong Lee, 2007). For sucrose, the slope
of the graph is 0.0093 and for starch is 0.2423. It is easier to conclude these sugars
are non-reducing because their absorbance not so obviously showed the significant.
That means the absorbance readings were not high. So, the results of these two
graphs correspond with analytical data experiment. It is because non-reducing sugar
does not have the properties to reduce many of the reagents; in this case, DNS
reagent. Since the absorbance is proportional to the amount of reagent reduced, so
these sugars do not have higher reading of it. Sample B gave the same colour
characteristic with starch while sample C with sucrose. The sample A and B gave the
concentration of 0.027mg/ml and 0.057mg/ml respectively.

As the experiment was carried out, there are many sources of error that influenced
the results. As mentioned in the results, the fluctuation of reading from
spectrophotometer gave the significant inaccuracies especially reading for
absorbance of glucose, maltose, sample A and D. So, one of the error that have
been figured out was the cuvettes compartment arent cleaned up thoroughly. This
precaution need to be handled proper to avoid any spills inside the cuvettes.
Therefore, if there are any spills inside, the light that was ejected from
spectrophotometer through them had been blocked.

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