Practical No.
– 02
SCT 222 – 2
BIOCHEMISTRY
CARBOHYDRATES
UWU/SCT/20/035
Rameesh Ishak
2022.10.01
Date – 2022.09.26
Practical no. – 02
Practical Title – Carbohydrates
Objective –
To identify carbohydrates according to there reducing and non-reducing properties and effects
of different catalyst on the hydrolysis of carbohydrates.
Introduction -
Carbohydrates are polyhydroxy aldehydes, ketones or compounds that are transformed into
one of a large group of organic compounds that occur in foods and living tissues, including
sugars, starch, and cellulose. Due to the presence of aldehyde or ketone group in the
carbohydrates they can be categorized according to the reducing and non-reducing factor.
Reducing and non-reducing properties of carbohydrates –
Aldose
Due to the existence of free aldehyde functional groups, all aldoses are reducing sugars.
Mild oxidizing agents are used to oxidize the aldehydes, generating the appropriate
carboxylates.
Ketose
All ketoses are reducing sugars due to containing a functional group next to an alcohol.
Hemiacetal functional groups
Because they may readily generate aldehydes, carbohydrates with hemiacetal functional
groups can decrease weak oxidizing agents like Benedict's reagent.
Materials –
Glucose Fehling’s B
Fructose 3M Sulfuric acid
Lactose Sodium hydroxide
2% sucrose Potassium hydroxide
2% starch Iodine solution
Fehling’s A
Methodology –
a) Reducing or non-reducing carbohydrates
5ml of Fehling’s A solution and 5ml of Fehling’s B solution were taken to make the Fehling’s
solution. Five boiling tubes were taken and for each 2ml of the made Fehling’s solution were
added. These boiling tubes were labeled as Glucose, Lactose, Fructose, Sucrose and Starch. For
each of these boiling tubes, 10 drops from the labeled solution were added. Then a water bath
was prepared, and it was boiled. In the boiling water bath, the boiling tubes were placed inside.
After 5 minutes the boiling tubes were taken out and observation was recorded.
b) Hydrolysis of starch (enzymes vs acid catalysts)
Two boiling tubes were taken and 2ml of 2% starch were added to each boiling tube. For one
boiling tube 2ml of your own saliva was added and 2ml 3M of sulfuric acid was added to the
other boiling tube. Then a water bath was taken, and it was boiled to a temperature of 45 o C.
Then the two boiling tubes were placed in the water bath for thirty minutes and during this
time period the temperature was maintained at 45oC throughout. Then few drops from the two
boiling tubes were taken to two separate boiling tubes and they were labeled accordingly. Two
drops of Iodine solution were added to the two separate solutions and the change was
observed and it was recorded.
c) Hydrolysis of sucrose (Moore’s test)
Two boiling tubes were taken and 3ml of 3% sucrose were added to each boiling tube. 3ml of
water was added to the two boiling tubes and for one boiling tube 3 drops of 3M sulfuric acid
was added and for the other boiling tube 3 drops of 3M sodium hydroxide was added and these
two boiling tubes were labeled as acid and base. The two boiling tubes were immersed in a hot
water bath for 5 minutes. Then the two boiling tubes were taken out and to the ‘acid’ labeled
boiling tube sodium hydroxide was added until the red litmus paper turned blue. Then the two
solutions were taken, and 3-5 drops of Fehling’s solution was added. Then the change occurred
was recorded.
Observations –
a) Reducing or non-reducing carbohydrates
When the carbohydrates were added to the 5 boiling tubes with Fehling’s solutions the fructose
labeled boiling tube showed a part of the solution turned its’ color to orange from blue as
shown in the figure 1.1.
Figure 1.1
When the boiling tubes were immersed in the hot bath, we saw the gradual change of color of
the glucose, fructose and lactose as shown in the figure 1.2.
Figure 1.2
After the experiment on the reducing factor of carbohydrates were finished, the following
observations could be seen.
Carbohydrate Observation
Glucose Orange precipitate
Fructose Orange precipitate
Lactose Orange precipitate
Sucrose No color change (blue color)
Starch No color change (blue color)
Figure 1.3
b) Hydrolysis of starch (enzymes vs acid catalysts)
The only change we observed were the color change of the two solutions after the iodine
solution was added. The two solutions were turned to yellow color from colorless solutions.
This can be seen in the figure 2.2.
Solution with
Acid
After adding Iodine
Figure 2.1 Figure 2.2
Solution with
Saliva
c) Hydrolysis of sucrose
No observable change was observed after the experiment and the two solutions were heated
again in a hot bath and in the boiling tube with 3M sulfuric acid the formation of orange color
precipitate was observed.
Figure 3
Discussion –
To the aqueous solution of carbohydrate Fehling’s solution is added and heated in water bath.
The formation of red precipitate confirms the presence of reducing sugars. The copper ions
present in Fehling’s solution in +3 state is reduced to +2 oxidation state and it is precipitated as
red cuprous oxide.
In the hydrolysis of starch, saliva acts as a catalyst due to the presence of amylase which is an
enzyme. This enzyme can breakdown starch into maltose and this maltose reacts with iodine
test and change the color from yellow to purple. For this occurrence the proteins in the saliva
must not get destroyed, since the reason for the temperature being maintained at 45 o C. But
even if we maintained the temperature at 42o C – 45o C the solution did not show any color
change to the iodine solution. We concluded the reason for this might had been the time we
kept the solutions in the water bath might be greater than 30 minutes.
In the presence of sulfuric acid and high temperature sucrose acts as a reducing agent and react
with the Fehling’s solution to give the orange precipitate.
Conclusion –
The carbohydrates can be identified by the reducing and non-reducing property which caused
by the structure of carbohydrates (the presence of aldose or ketose). The non-reducing
carbohydrates also can act as reducing agent under certain conditions (the presence of catalyst,
presence of acidic medium and high temperature) which were identified in the hydrolysis of
sucrose experiment and the hydrolysis of starch experiments.
References –
“Tests of Carbohydrates - Chemistry Practicals Class 12.” BYJUS,
byjus.com/chemistry/tests-of-carbohydrates/.
