IA # 2

DATE: October 19, 2016

TITLE: BIOLOGICAL MOLECULES- Carbohydrates

AIM:

To Determine the Relative Concentration of Reducing Sugars present in a Sample of

Orange juice, Grape fruit juice and Lime juice

APPARATUS:

Bunsen burner, tripod stand, gauze, 4 % glucose solution, six test tubes, distilled water,
5 cm3 syringes, benedicts solution, samples of lime juice, orange juice and grapefruit
juice, beaker and tongs.

METHOD:

Using a syringe, 1 cm3 of 4 % glucose solution was placed into a test tube and this test
tube was labelled 4%. Again, using a syringe, 5 cm3 of 4% glucose solution was placed
into another test tube and 5cm3 of distilled water was added to the solution. This yielded 2
% glucose solution. From this new formed solution, 1 cm3 was placed into a test tube and
this test tube was labelled 2%. To the remaining 9 cm3 of 2 % glucose solution, 9 cm3 of
distilled water was added. This was gently swirled and from this now 1 % solution, 1 cm3
was removed using a syringe and placed in a test tube labelled 1 %. 1 cm3 of orange juice,
grape fruit juice and lime juice was placed in three test tubes and labelled respectively.
Again, using a syringe, 5 cm3 of benedicts solution was added to each test tube making
the total volume in each test tube 6 cm3. A water bath was prepared and the test tubes
were all placed in the bath. Upon observation of colour changes, the test tubes were
removed from the bath using tongs and placed on a test tube rack to make comparisons.

Brittany Pitt
L6H1
OBSERVATIONS:

TABLE SHOWING THE COLOUR CHANGES OF THE STANDARD GLUCOSE

SOLUTIONS UPON CARRYING OUT THE BENEDICTS TEST

CONCENTRATION COLOUR CHANGES

OF STANDARD
SOLUTION
This was the first of the standard solutions to change and the bottom of the
test tube began changing before it entered the water bath. Upon heating, the
solutions initial blue colour (due to the colourless glucose solution gaining
4% the blue colour of the benedicts solution) changed to green then to yellow
then to a brick red colour. When the test tube was gently swirled, numerous
fine specks stuck to the test tube glass indicating that a large amount
precipitate was formed.
This solution changed shortly upon entering the water bath. Upon heating,
2% the solutions initial blue colour (due to the colourless glucose solution
gaining the blue colour of the benedicts solution) changed to green then to
yellow and finally a red-orange colour was observed. When the test tube
was gently swirled, a small amount of fine specks stuck to the test tube
glass indicating that a precipitate was formed.
This solution was the last of the standard solutions to begin changing
1% colour. Upon heating, the initial blue coloured solution (due to the
colourless glucose solution gaining the blue colour of the benedicts
solution) developed a green colour which finally changed to a yellow-green
colour. When the test tube was gently swirled, fine specks stuck to the test
tube glass indicating that a precipitate was formed however the amount of
small specks observed was less than that formed by both the 4% and 2%
glucose solutions.

Brittany Pitt
L6H1
TABLE SHOWING THE COLOUR CHANGES OF VARIOUS JUICE SAMPLES UPON
CARRYING OUT THE BENEDICTS TEST

JUICE COLOUR CHANGES

SAMPLE
Grapefruit The grapefruit juice was initially light grey in colour however upon mixing with
the benedicts solution gained its blue colour. Upon heating, this was the first
juice sample to begin its colour change. It changed from blue to green to yellow
and finally a red orange colour was observed. When the test tube was gently
swirled, numerous fine specks stuck to the test tube glass indicating that a
precipitate was formed.
Orange The orange juice was initially pale yellow in colour however upon mixing with
the benedicts solution gained its blue colour. Upon heating, this was the second
sample to begin changing colour. It changed from blue to green to yellow and
finally a yellow-orange colour was observed. When the test tube was gently
swirled, a small amount of fine specks stuck to the test tubes glass indicating
that a precipitate was formed
Lime The lime juice was initially light grey in colour however upon mixing with the
benedicts solution gained its blue colour. Upon heating, this was the last sample
to begin its colour change. It changed from blue to green then to a yellow-green
colour. When the test tube was gently swirled, fine specks stuck to the test tube
glass indicating that a precipitate was formed however the amount of small
specks observed was less than that of both the orange and grapefruit solutions.

Brittany Pitt
L6H1
CALCULATIONS:
Density of water = 1gcm-3 or 1000mgcm-3

Considering that only 1 cm3 of each solution

4% Glucose
was used
4
1000
100

= 40 mgcm-3

After performing the Benedicts test, the grapefruit juice was found to produce a colour similar to that
produced by the 4% glucose and therefore is deduced to have a concentration of 40 mgcm-3 of reducing
sugar.

2% Glucose

2
1000
100

= 20 mgcm-3

After performing the Benedicts test, the orange juice was found to produce a colour similar to that
produced by the 2% glucose and therefore is deduced to have a concentration of 20 mgcm-3 of reducing
sugar.

1% Glucose

1
1000
100

= 10 mgcm-3

After performing the Benedicts test, the lime juice was found to produce a colour similar to that
produced by the 1% glucose and therefore is deduced to have a concentration of 10 mgcm-3 of reducing
sugar.

Brittany Pitt
L6H1
DISCUSSION:

Qualitative analysis deals with the identification of substances present in a sample while
quantitative analysis deals with the measurement of the quantities of constituents present
in a substance. Benedicts solution is a chemical reagent that changes colour in the
presence of reducing agents (the category to which reducing sugars belong). Benedicts
solution is a mixture of sodium or potassium citrate, sodium carbonate and copper
sulphate. How Benedicts solution works is that the Cu2+ ions from the copper sulphate
are blue in colour however when reduced to copper (i) the ions are precipitated as red
copper (i) oxide which is insoluble in water, accounting for the precipitates observed.

The more reducing sugar present, the more Cu 2+ ions will be reduced to form the red
copper (i) oxide and in turn a redder colour change will be observed. It is therefore safe to
say that the qualitative analysis may be linked to the quantitative analysis as any form of
change will indicate the presence of a reducing sugar (qualitative) however the
degree/intensity of the colour change will indicate how much reducing sugar is present
(quantitative).

Quantitative analysis involves the comparison of an unknown concentration of a

substance to a known concentration of a substance (referred to as a standard solution). In
the experiment conducted, the 4 % glucose was that of known concentration. Serial
dilution is the repeated dilution of a solution to amplify the dilution factor. To broaden
the spectrum of colours to compare the juice solutions to after carrying out the Benedicts
test, this 4% glucose solution was serially diluted to yield 2% and 1% glucose solution.

PRECAUTION:

A syringe was used to measure the solutions instead of a measuring cylinder to

increase the experiments precision.

Brittany Pitt
L6H1
LIMITATION:

Only solutions of concentrations 4% 2% and 1% by mass were available for comparison.

As a result, it is unknown whether or not the juice solutions possibly contained
concentrations of reducing sugar not include in the standard solutions used for
comparison ( 5%, 3% or 1.5% reducing sugar concentration by mass). The experiment
can be improved by increasing the amount of standard solutions of varying
concentrations present for the comparison. Also, the solutions were not stirred to evenly
mix with the benedicts solution. This resulted in most of the colour change being
concentrated to the bottom of the test tube. Also, one test tube began changing at the
bottom before it was placed into the water bath due to the heat present in the lab. This can
be eliminated by the stirring of each solution with the Benedicts reagent before applying
heat as well as keeping the test tubes at room temperature until it is ready to be placed in
the water bath.

CONCLUSION:

From the experiments conducted it was found that grapefruit juice had the highest
concentration of reducing sugar which was approximately 40mgcm-3 followed by orange
juice which was approximately 20 mgcm-3 . Lime juice was found to contain the least
amount of reducing sugar which was approximately 10 mgcm-3.

Brittany Pitt
L6H1