ILOILO DOCTORS’ COLLEGE

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Experiment 3
CHEMISTRY OF CARBOHYDRATES

Acel Anne E. Habaradas

Group 3 / BSN 1 - C
CHM 103/ MC 2 – Biochemistry

ABSTRACT
The term carbohydrate or hydrates of carbon is derived from its basic elemental formula in which
carbon is joined to hydrogen and oxygen present in the same ratio as in water. The experiment
used various methods to determine the different chemical reactions of carbohydrates and how its
reaction relate to different reageants.

INTRODUCTION
Carbohydrates are polyhydroxy aldehyde (aldoses) or polyhydroxy ketone (ketoses) with the
general formula (CH2O) n, where n equals 3 or more. Carbohydrates are the most important
source of energy. They are widely distributed in plants and are even found in certain animal
tissues, such as the liver and muscles.

Carbohydrates exists as cyclic structures containing functional groups such as the hydroxyl
acetal, ketal, hemiacetal and hemiketal groups. The end which has a hemiacetal in its terminal is
referred to as the reducing agent while the end which does not contain a hemiacetal is the non-
reducing sugar end. In the oxidation process, reducing agents like carbohydrates are susceptible to
attack by oxidizing agents such as Barfoed’s, Fehling’s, and Benidict’s reagents. However, not all
disaccharides or polysaccharides contains reducing end. This experiment aims to; investigate the
reducing and non-reducing nature of carbohydrates, familiarize the different chemical reactions of
carbohydrates and understand and explain its reaction to different reagents.

Reactions with Different Reagents

Reagent Name Active Component End Color

Molisch’s reagent Alpha-Naphthol Purple/violet ring

Seliwanoff’s reagent Resorcinol-HCl Cherry red

Barfoed’s reagent Cupric acetate and glacial Yellow to green solution with
acetic acid brick red precipitate
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Fehling’s reagent Copper sulphate + NaOH + Brick red precipitate

SODIUM Potassium tartrate
Benedict’s reagent Sodium carbonate + sodium Brick red precipitate
citrate + CuSO4
Iodine solution Iodine Deep blue color

Tollen’s reagent Conc HCl + Phloroglucinol Silver mirror/black ppt

EXPERIMENTAL
Apparatus:

12 pcs - Tube tube (15 ml) Test tube rack Hot plate
Water bath Pipette (10 ml)

Chemical:

10 ml NaOH 20 ml Fehling’s reagent

10 ml Benedict’s reagent 10 ml Seliwanoff’s reagent
10 ml Barfoed’s reagent 10 ml Molisch’s reagent
10 ml Iodine solution 10 ml 5% Iodine solution
10 ml 5% Glucose solution 10 ml 5% Sucrose solution
10 ml 5% Fructose solution 10 ml 5% Lactose solution

Materials Needed:

Table sugar Corn starch Evaporated milk

Mango juice Honey or maple syrup

Procedure

Test for Reducing and Non-reducing sugars

In a one test tube, Fehling’s A and Fehling’s B were mixed and set aside to make a
solution. Six test tubes were then added and placed in each about 2 ml of the Fehling’s solution.

In test tube #1, a 10 drops of a one spoon of table sugar mixed with 1 ml tap water was
added to the 2 ml Fehling’s A and Fehling’s B solution. The mixture was mixed thoroughly and
boiled in water bath for 5 minutes. Observations were recorded.

In test tube #2, a 10 drops of 10 ml mango juice with equal part of water was added to
the 2 ml Fehling’s A and Fehling’s B solution. The mixture was mixed thoroughly and boiled in water
bath for 5 minutes. Observations were recorded.

In test tube #3, a 10 drops of 50 ml evaporated milk diluted with 100 ml of water was
added to thr 2ml Fehling’s A and Fehling’s B solution. The mixture was mixed thoroughly and boiled
in water bath for 5 minutes. Observations were recorded.
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5000 Philippines

In test tube #4, a 10 drops of one tablespoon honey diluted with 100 ml water was added
to the 2 ml Fehling”s A and Fehling’s B solution. The mixture was mixed thoroughly and boiled in
water bath for 5 minutes. Observations were recorded.

In test tube #5, a 10 drops of Iodine solution was added to the 2 ml Fehling’s A and
Fehling’s B solution. The mixture was mixed thoroughly and boiled in water bath for 5 minutes.
Observations were recorded.

In test tube #6, a 10 drops of starch solution was added to the 2 ml Fehling’s A and
Fehling’s B solution. The mixture was mixed thoroughly and boiled in water bath for 5 minutes.
Observations were recorded.

Test for Reactions using Different Reagents

In Barfoed’s test, a 1 ml Barfoed’s regeant was mixed to a 1 ml of 5% Glucose solution. It

was heated for 30 seconds in a boiling water bath and was allowed to stand for 15 minutes after.
The results were recorded and the procedure was repeated using 5% Sucrose, 5% Fructose, 5%
Lactose, and 5% Starch solution.

In Benedict’s test, a 1 ml of Benedict’s reagent was added with two drops of 5% Glucose
solution. It was boiled for two minutes and was allowed to cool. The results were recorded and the
procedure was repeated using 5% Sucrose, 5% Lactose, and 5% Starch solution.

In Molisch’s test, a 1 ml of 5% Glucose solution was placed in a test tube. The solution was
added with a one drop of Molisch’s reagent and was mixed thoroughly. After that, the test tube was
inclined and a 1 ml of concentrated Sulfiric Acid was allowed to flow in the side of the test tube. The
Sulfiric Acid forms a layer at the bottom of the test tube. The results produced in color at the
junction of the two liquids were observed. The procedure was repeated using 5% Sucrose, 5%
Fructose, 5% Lactose, and 5% Starch solution.

In Phloroglucinol-HCI (Tollen’s Test), a 0.5 ml of 5% Glucose solution was placed in a test

tube. It was added with a 0.5 ml concentrated HCI and 5 drops of Phloroglunicol. After adding the
chemicalos, it was heated in a boiling water bath. The results were observed and repated with the
same procedure using Sucrose, Fructose, Lactose, and Starch solution.

In Seliwanoff’s Test (Resorcinol – HCI Reaction), a 1 ml of Seliwanoff’s reagent was placed

in a test tube. It was added a 3 drops of 5% Glucose solution and boiled in a water bath. The colord
produced was observed and the time required for the development of pink color (if there was any)
was recorded. Repeat the procedure using Sucrose, Fructose, Lactose, and Starch solution.

In Iodine test, a 1 ml of 5% Glucose solution in a test tube. It was added with a one drop
of Iodine solution and heated to note the changes in color. The solution was set aside to cool and
observed. The procedure was repeated using Sucrose, Fructose, Lactose, and Starch solution.

Hydrolysis of Carbohydrates

In test tube #1, a 1 ml of diluted HCI and a 5 ml 5% Sucrose solution were stirred
and heated in boiling water bath for 20 minutes. The solution was set aside to cool and itv
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was added with diluted NaOH dropwise until the solution turns neutral on litmus paper.
The 10 drops of this solution was transferred to another test tube then was added 2 ml of
Fehling’s A and B solution to the other test tube and was added 2 drops of Iodine solution.
The results were observed and recorded.

In test tube #2, the procedure in test tube #1 was repeated but a 5% Glucose was
used instead of 5% Sucrose.

In test tube #3, a 2 ml of 5% Starch solution and a 2 ml diluted HCI solution were
stirred and heated in a boiling water bath for 20 minutes. The solution was set aside to
cool and was added with diluted NaOH dropwise until the solution turns neutral on litmus
paper. The solution was divided into 2 test tubes, in one tube a 2 ml Fehling’s A and B was
added. On the other hand, to the other test tube, a 2 drops of Iodine solution. The results
were observe and recorded.

Results and Discussions

Table 1. Test for Reducing and Non-reducing sugar

Reducing sugars are the most abundant organic molecules found in nature that
contain free aldehyde and ketone groups. They can donate electrons to other compounds
and cause the reduction of other compounds. However, reducing sugars contain free
anomeric carbon. The anomeric carbon is that which is derived from the carbonyl group
and contains two oxygen substituents.

Non-reducing sugars are carbohydrate which does not contains free aldehyde and
ketone group is known as non-reducing sugars. They do not contain anomeric carbon
attached to the hydroxyl (-OH) group.

Substance Observation Results

Table sugar

Mango Juice

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Evaporated Milk

Honey

Iodine Solution

Starch Solution

Table 2. Test for Reactions Using Different Reagents

The word ''reagent'' appears in various chemistry terms, such as ''limiting

reagent'' and ''analytical reagent”. A reagent is a chemical that is added to other chemicals
in order to cause them to react, or to detect a certain category of substance, such as a
sugar or an amine. The laboratory uses of reagents are often for the purpose of
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synthesizing a desired molecule. In other words, they induce a chemical change in another
molecule for the purpose of producing the product.

Testing Reagents

Sample

Barfoed Benidict Molisch Seliwanoff Tollen Iodine

Glucose

Sucrose

Fructose

Lactose

Iodine

Table 3. Hydrolysis of Carbohydrates

Hydrolysis is any chemical reaction in which a molecule of water breaks one or

more chemical bonds that exists between a particular substance. The term is widely in use
for substitution, elimination, and solvation reactions in which water is the nucleophile.
Biological hydrolysis is the breaking of biomolecules, where a water molecule is consumed

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to affect the separation of a larger molecule into components. Carbohydrate is broken into
a component of sugar molecules by hydrolysis.

Samples Observations Result

Sucrose solution

Glucose solution

Starch solution

Conclusions

(Make a generalizations on the objectives of this experiment.)

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REFERENCES

Sajid, M. A. (2023, February 7). Reducing vs. Non Reducing Sugars: Comparison and Examples.
PSIBERG.

https://psiberg.com/reducing-vs-non-reducing-sugars/#google_vignette

Admin. (2022c). List of Reagent- Examples of reagents with FAQs. BYJUS.

https://byjus.com/chemistry/list-of-reagent/

What is Hydrolysis: Definition, Types of Hydrolysis , Examples. (2021, May 28). Toppr-guides.

https://www.toppr.com/guides/chemistry/biomolecule/hydrolysis/

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West Avenue, Molo, Iloilo City
5000 Philippines

MODE OF REFERENCE (MOV)