Expt #10

Date: 21st February, 2019.

Title: Quantitative test for reducing sugars

Aim: To determine the concentration of reducing sugar in soda samples

Materials:

10% glucose solution, distilled water, pipette, regular soda sample, diet soda sample,
spectrophotometer, Benedict’s solution, test tubes, water bath, stop watch, filter paper, cuvette

Procedure:

1. 10% glucose solution and distilled water was used to prepare a series of standard glucose
solutions of different concentrations:

TABLE SHOWING THE DILUTION SERIES OF GLUCOSE

Dilution 10% glucose 5% glucose 2% glucose 1% glucose 0.5% 0% glucose

glucose
Volume of 10 5 2 1 0.5 0
10% glucose
solution
(cm3)
Volume of 0 5 8 9 9.5 10
water (cm3)
2. Identical Benedict’s tests were carried out on the standard glucose solutions prepared above
and the soda samples.
 - 2 cm3 of each of the standard glucose solutions or soda sample was pipetted into labelled test
tubes.
 6 cm3 of Benedict’s solution was added.
 The solution was mixed by shaking.
 The tubes were placed in a water bath at 80°C for 5 mins.
 The tubes were removed from the water bath
 Any observations were recorded.
3. The tubes were left to stand for about 10 mins to allow settling and then the samples were
filtered to remove any precipitate formed when the reducing sugar in the samples reacted with
the Benedict’s solution. The filter papers were discarded. The filtrates were kept. Any
observations were recorded.
4. Each filtrate was placed in a cuvette and the spectrophotometer was used to measure the
absorbance at 735nm. Distilled water was used as the blank to zero the spectrophotometer.
5. A calibration curve was plotted to show the absorbance of the standard glucose solutions.
6. The calibration curve was used to determine the concentration of reducing sugars in the soda
samples.

Observations: Place all observations from steps #2 and #3 in this section. Also include a table showing
the absorbance values for each solution. Place the calibration curve in this section.
Discussion: Explain your observations. Determine the concentration of reducing sugars in each of the
soda samples.

The test for reducing sugars requires the use of the Benedict’s solution. This solution contains Copper (II)
Sulfate. During the test for reducing sugars, the available carbonyl carbons reduce the blue cu 2+ ions
present in the solution to the insoluble red-brown copper oxide which contains Cu+ ions. Since copper
oxide is insoluble it was expected to form a red-brown precipitate.

Solutions that remained blue, or had traces of blue, like the solution which had 0% glucose, had no
reducing sugars. Solutions that slight colour change, such blue to green, had minimal amounts of
reducing sugars. Examples of solutions that contained little amounts of reducing sugars are the solutions
that contained 0.5%, 1%, and 2% of glucose. Solutions that turned brick red contained a high
concentration of reducing sugars. Examples of such solutions are: the solutions containing 5% and 10%
glucose.

The wavelength used for the spectrophotometer was to determine the value of red that is present in
each sample, after the precipitate has been filtered out. The closer the absorbance value is to zero the
greater red wave lengths is being read from it. This means that there’s a high concentration of reducing
sugars in that sample. As determined by the graph and the results collected, the regular soda has ____%
of reducing sugars while the diet soda has less than 0% of reducing sugars.

Conclusion: A sentence or two that clearly summarise the finding of your experiment as it relates to your
aim

The amount of reducing sugars present in the regular soda sample was found to be ____ whereas the
amount of reducing sugars present in the diet soda sample was found to be ____...