OUR LADY OF FATIMA UNIVERSITY

1118, 1 Esperanza, Novaliches, Lungsod Quezon, 1118 Kalakhang Maynila

LABORATORY REPORT:

QUALITATIVE TEST FOR CARBOHYDRATES

Name: Quimno, Ivy N.

Course: BSN 1 YB-22
Subject: BIOCHEMESTRY
I. Introduction
Carbohydrates are carbon, hydrogen, and oxygen-containing compounds. Monosaccharide’s,
disaccharides, and polysaccharides are the three types of carbohydrates. Monosaccharaides are simple
sugars with one oxygen atom and two hydrogen atoms for every carbon atom. Disaccharides are
polymers made up of many monosaccharide molecules (more than two) linked together by
condensation processes, whereas Polysaccharides are polymers made up of many monosaccharide
molecules (more than two). The Barfoed test is a chemical procedure for detecting monosaccharaides. It
was invented by Danish chemist Christen Thomsen Barfoed. The trial, as mentioned earlier,’s primary
goal is to detect reducing carbs and distinguish between reducing monosaccharaides and disaccharides.

Carbohydrate-rich foods are an essential part of a balanced diet. Carbohydrates supply glucose
to the body, which is transformed into energy for biological functions and physical activities. However,
carbohydrate quality matters; some carbohydrate-rich diets are superior to others. Carbohydrates are
assessed on the Molisch Test. Hans Molisch, an Austrian botanist, was the inspiration for this test. The
purple ring test is another name for it. The experiment's primary goal is to determine whether or not a
given substance contains carbohydrates. The most practical application of this test is that if a food
product is labeled as sugar-free, you can quickly test it using Molisch's test to see if it contains
carbohydrate (or sugar).

Ketose and aldose are monosaccharaides distinguished by the group in which they are found.
An aldose is a monosaccharide with an aldehyde group in its carbon backbone. They can only be found
in plants. Ketose is a monosaccharide with a ketone group on its carbon backbone. They can only
isomerize to aldose in the presence of reducing sugar. They're commonly found in processed foods.
Ribulose, fructose, and other sugars are examples of ketoses. To identify between aldose and ketose, we
can use Seliwanoff's Test. The color of aldose changes to light pink after this test, while the color of
ketose changes to deep cherry red. Chemist Theodor Seliwanoff introduced the Selwanoff Test.

Fehling's test is one of the most widely used methods for estimating or detecting reducing and
non-reducing sugars. H.C., a German chemist, devised the test. Von Fehling also distinguishes between
water-soluble carbohydrates and ketone functional groups. Benedict's test determines whether or not a
substance contains simple carbohydrates. Benedict's test detects reducing sugars with a free ketone or
aldehyde active group (monosaccharaides and some disaccharides). The presence of glucose in the urine
can be detected using Benedict's solution. Tollens' test is a chemical method for distinguishing between
reducing and non-reducing carbohydrates. Because of the result, this test is also known as the silver
mirror test. The primary goal of this assessment is to distinguish between reducing and non-reducing
sugars, identify aldehyde-containing carbs and distinguish them from ketone-containing carbohydrates.
II. Materials • Boiling water bath

D. Barfoed’s Test

A. Molisch’s Test • Test tubes

• Test tubes • Test tube rack

• Test tube rack • Dropper

• Dropper • Marker and paper tape (for label)

• Marker and paper tape (for label) • Sugar solutions (glucose, fructose,
lactose, sucrose)
• Carbohydrate solutions (glucose,
fructose, lactose, sucrose) • Barfoed’s reagent

• Molisch’s reagent • Boiling water bath

• Concentrated sulfuric acid

B. Benedict’s Test E. Seliwanoff’s Test

• Test tubes • Test tubes

• Test tube rack • Test tube rack

• Dropper • Dropper

• Marker and paper tape (for label) • Marker and paper tape (for label)

• Carbohydrate solutions (glucose, • Sugar solutions (glucose, fructose,

fructose, lactose, sucrose) lactose, sucrose)

• Benedict’s reagent • Seliwanoff’s reagent

• Boiling water bath • Boiling water bath

C. Fehling’s Test F. Iodine Test

• Test tubes • Test tubes

• Test tube rack • Test tube rack

• Dropper • Dropper

• Marker and paper tape (for label) • Marker and paper tape (for label)

• Sugar solutions (glucose, fructose, • Iodine solution

lactose, sucrose) • Starch solution
• Fehling’s reagent A and B • Boiling water bath
• Water
III. Method
Molisch’s Test

Procedures:

1. Sulfuric acid used in this test.

2. Add 2 ml of each carbohydrate Solution

3. Add 10 drops of moles reagent to each carbohydrate sample and mix well.

4. Carefully, add 15 to 20 drops of concentrated sulfuric acid.

5. After that, Color formed at the zone of the two liquids.

Benedict’s Test

Procedures:

1. Add 2 ml Benedict Solution

2. Add 4 empty tubes and placed into a water bath for 30 seconds.

3. If, the solution remains clear blue, we can continue the test which means the reagent is not
contaminated.

4. Need, 4 test tube need to cool down until they warm.

5. After that place the four test tubes, contain benedict solution

and carbohydrates samples boiling into heater bath for 2 mins.

6. And see the results, after 2 mins.

Fehling’s Test

Procedures:

1. Each of the tubes, we are going to mix 1 ml fehling solution A and 1 ml fehling solution B.

2. Add 3 ml of water.

3. After that, the 4 test tubes are placed in boiling water bath in 1 minutes.

4. If the solution remains color blue or clear blue, add 8 drops of sugar solution of each test tubes.

5. Repeat again for 4 test tubes with a sugar solution for another two minutes.

6. See the results, after 2 mins.

Barfoed’s Test

Procedures:

1. Each test tubes, add 2 ml of barfoed’s reagent

2. Then, add 10 drops of each of sugar solution

3. Place the test tubes in boiling water bath for five minutes

4. After 5 minutes, remove the test tubes from boiling water bath and let it cool

5. Observe the first 15 minutes

Seliwanoff’s Test

Procedures:

1. Add 2 ml of the sugar solution to 3 ml of seliwanoff’s reagent

2. Immerse the solutions in boiling water bath

3. Observe the color changes during the first 10 minutes of boiling

Iodine Test

Procedures:

1. Place 2 ml of plant starch solution

2. Then, add a drop of iodine solution

3. Heat the solution and note the change

4. After heating the solution and cool it down

IV. Data

V. Analysis

Carbohydrates are polyhydroxy aldehydes or ketones that are the most abundant bio macromolecules
on the earth. All carbohydrates are hydrates of carbon and they contain C, H, O, while some also contain
nitrogen, phosphorus, and sulfur. In the laboratory activity, carbohydrates underwent some tests to see
how they will react to it.

The data gathered is to characterize carbohydrates present in an unknown solution on the basis of
various chemical assays.

First, the Molisch's Test. Molisch's test is a general test of carbohydrates to determine if they are
present in a certain substance; hence, all the substances that were tested were positive for
carbohydrates as they appear to produce a purplish color that indicates a positive test.

Second, the Benedict's Test. Benedict's test is used to determine the concentration of reducing sugar as
the substance is being heated; hence, glucose, fructose, and lactose have the presence of reducing
sugars, wherein glucose has the highest concentration of it, while sucrose and starch resulted in
negative results.
Third, the Fehling's Test. Just like Benedict's test, Fehling's test also indicates the presence of reducing
sugars in a substance by manifesting an appearance of a reddish-brown precipitate; hence, glucose,
fructose, and lactose tested positive while sucrose and starch were negative.

Fourth, the Barfoed's Test. Barfoed's test determines if there is a carbohydrate present in a substance,
or even if the carbohydrate is a monosaccharide or a disaccharide; hence, glucose and fructose were
monosaccharides because a red precipitate appeared much faster than the red precipitate of lactose
and sucrose.

Fifth, the Seliwanoff's test. Seliwanoff's test is a times color reaction test used to differentiate between
carbohydrates that have ketose and those that have aldose; hence, fructose and sucrose have ketose for
they manifested a reddish-brown color for a couple of minutes, while glucose and lactose have aldose
which is they do not manifest the same color.

Lastly, the Iodine Test. Iodine test is used to determine the presence of starch in a substance; hence,
above all the substances tested, only the starch tested positive as it is a starch itself by manifesting a
blue-colored appearance while the others were green.

VI. Conclusion

It is concluded that the experiment was successful in using benedict test and iodine test. Benedict
test is used to detect the presence of reducing sugar such as glucose, fructose and lactose. All
monosaccharaides are reducing sugar, they all have a free reactive carbonyl group. Some disaccharides
have expose carbonyl group and are also reducing sugar.

Lactose which is disaccharides also called reducing sugar as it exposed carbonyl group. Other
disaccharides such as sucrose and starch are non - reducing sugar and will not react with Benedict test.
Next, we use iodine test to test for the presence of starch. Starch is a type of polysaccharides
carbohydrates which is made up of amylose and amylopectin. Starch gave positive result in iodine test
as the color of the solution change from yellow to dark blue.

After that, we try to minimize as low error as we can which is the uses of volumetric glassware. We
ensure that we make the most accurate measured volumes as possible. It is the proper way and will
gave an accurate reading of the volumes. In future labs, we would emphasize the importance of keeping
all chemicals in closed containers, and the proper set-up of data collection systems, in order to ensure
accurate results. We would also increase the number of trials for this lab, in order to achieve more
plentiful and accurate data.
VII. References

Aryal, S. (2020, February 24). Carbohydrates – Monosaccharides, Disaccharides, Polysaccharides.

Microbe Notes.

Sapkota, A. (2020, November 19). Barfoed’s Test- Definition, Principle, Procedure, Result, Uses. Microbe
Notes.

Wikipedia contributors. (2021, May 12). Barfoed’s test. Wikipedia.

Carbohydrates. (2019, May 22). The Nutrition Source.

Vedantu. (2022, February 3). Molisch’s Test.

Wilson, M. (2019, October 24). What is the importance of Molisch test? – Restaurantnorman.com.
Restaurant Norman

Biology Online. (2022, January 27). Iodine test - Definition and Examples - Biology Online Dictionary.
Biology Articles, Tutorials & Dictionary Online.

A. (2021, March 22). General Data Protection Regulation (GDPR) Guidelines BYJU’S. BYJUS

Aryal, S. (2019, August 15). Benedict’s Test- Principle, Composition, Preparation, Procedure and Result
Interpretation. Microbiology Info.Com.

Sapkota, A. (2020b, November 19). Tollens’ Test- Definition, Principle, Procedure, Result, Uses. Microbe
Notes.
 OUR LADY OF FATIMA UNIVERSITY
1118, 1 Esperanza, Novaliches, Lungsod Quezon, 1118 Kalakhang Maynila

LABORATORY REPORT:
QUALITATIVE ANALYSIS OF
CARBOHYDRATES

Name: Quimno, Ivy N.

Course: BSN 1 YB-22

Subject: BIOCHEMESTRY
I. Introduction

Carbohydrates are the primary energy source for all living species, and they are divided into three types:
monosaccharaides, oligosaccharides, and polysaccharides. Simple crystalline sugars that are readily
soluble in water are known as monosaccharaides. Low molecular weight sugars are found in
oligosaccharides, a polymer of monosaccharaides covalently bonded via glycoside bonds.

Carbohydrates are essential for good health and can be found in both natural and processed diets. A
polysaccharide is a starch. Cereals (wheat, maize, and rice), potatoes, and processed foods containing
cereal flour, such as bread, pizza, and pasta, are all high in it. Table sugar (sucrose, produced from
sugarcane or sugar beets), lactose (found in milk), glucose, and fructose (found in honey, many fruits,
and certain vegetables) are the most common sugars in the human diet. Drinks and many prepared
dishes, such as jam, biscuits, and cakes, commonly contain table sugar, milk, or honey.

Carbohydrates play a variety of roles in living things. Polysaccharides are energy storage molecules (e.g.,
starch and glycogen) and structural elements (e.g., cellulose in plants and chitin in arthropods). Ribose, a
5-carbon monosaccharide, is a crucial component of coenzymes (such as ATP, FAD, and NAD) and the
backbone of the genetic material RNA. Deoxyribose, which is related, is a component of DNA. Many
other critical macromolecules are made up of saccharides and their derivatives, and they play essential
roles in the immune system, fertilization, pathogenesis prevention, blood coagulation, and
development.

II. Materials

A. Molisch’s Test

• Test tubes

• Test tube rack

• Dropper

• Marker and paper tape (for label)

• Carbohydrate solutions (glucose, fructose, lactose, sucrose)

• Starch

• Distilled Water

B. Benedict’s Test

• Test tubes

• Test tube rack

• Dropper

• Marker and paper tape (for label)

• Carbohydrate solutions (glucose, fructose, lactose, sucrose)

• Starch

• Distilled Water

C. Fehling’s Test

• Test tubes

• Test tube rack

• Dropper

• Marker and paper tape (for label)

• Sugar solutions (glucose, fructose, lactose, sucrose)

• Starch

• Distilled Water

D. Barfoed’s Test

• Test tubes

• Test tube rack

• Dropper

• Marker and paper tape (for label)

• Sugar solutions (glucose, fructose, lactose, sucrose)

• Starch

• Distilled Water

E. Seliwanoff’s Test

• Test tubes

• Test tube rack

• Dropper

• Marker and paper tape (for label)

• Sugar solutions (glucose, fructose, lactose, sucrose)

• Starch

• Distilled Water

F. Iodine Test

• Test tubes

• Test tube rack

• Dropper

• Marker and paper tape (for label)

• Iodine solution

• Starch

• Distilled Water

III. Methods

Molisch’s Test

Procedures:

1. Take a sample and add water to make 2 ml of solution.

2. Add 2 drops of Molisch’s reagent

3. Then, pour 5 ml of concentrated H2SO4 from the side of the inclined test tube.

4. Observe the colour change in the tube.

Benedict ’s test

Procedures:

1. Prepare 1 ml of a solution by adding a test sample and water.

2. Add 2 ml of Benedict’s solution in the test tube.

3. Heat the solution in the water bath for 3 minutes.

4. Observe the test tube for any colour change.

Fehling’s Test

Procedures:

1. Add 2 ml of the test sample in a test tube.

2. Then, add Fehling’s A and B solution in equal proportion to the above sample.

3. After that, place the test tube in a hot water bath for 3 minutes.

4. At last, observe the colour change in a test tube.

Barfoed’s Test

Procedures:

1. Add 1 ml of the test sample in a test tube.

2. Then add 2 ml of Barfoed’s solution in the test tube.

3. Place the tube in a water bath for boiling up to 1 min.

4. At last, observe the colour change in a test tube.

Bial’s Test

Procedures:

1. Take 2 ml of the test solution in the test tube.

2. Then, add 5 ml of Bial’s reagent.

3. Heat the solution in a water bath for 1 min.

4. Cool and observe the solution for any colour change.

Seliwanoff’s Test

Procedures:

1. Take 1 ml of test solution in the test tube.

2. Then, add 3 ml of Seliwanoff’s reagent.

3. Heat the solution in a water bath for 1 min.

4. Cool and observe the solution for the colour change.

Iodine Test

Procedures:

1. Take 2 ml of the test solution in the test tube.

2. Then, add 2 drops of iodine solution.

3. Observe the solution for the colour change.

IV. Data

DATA MOLISCH’S BENEDICTS FEHLING’S BARFOED’S IODINE’S TEST

TEST TEST TEST TEST
Glucose It looks weird Dark Orange Dark Red Silver Looks like
because of the orange urine
purple ring
interface

Lactose Purple with Orange Dark Red Looks like Orange

touch of green, metal
yellow color

Sucrose Dark blue, with Dark Blue Royal Blue Some touch of Orange
a red dots black
 Starch Orange color Clear green Royal blue Black Black
and purple
after the white
V. Analysis

Carbohydrates act as the major energy source for all the living organisms, and they are primarily of three
kinds, namely mono, oligo and polysaccharides. Monosaccharaides include simple crystalline sugars that
are readily soluble in water. Oligosaccharides include low molecular weight sugars and exist as a
polymer of monosaccharides covalently linked through the glycosidic linkages.

Molisch’s test detects the carbohydrate presence, which principle is based upon the dehydration
reaction. The carbohydrates in the sample get dehydrated into aldehyde by the addition of concentrated
H2SO4.

The reducing sugar will reduce into the enediols by reacting with alkaline reagent, i.e. Benedict’s
solution. The reducing sugar gives green to brick red colour precipitate depending upon the sugar
concentration. The colour change is due to the reduction reaction of copper (II) to copper (I) in the
solution that develops a red coloured precipitate.

In the Fehling’s test, a reduction reaction occurs between the aldehyde or keto groups of the reducing
sugar and the alkaline cupric hydroxide that later reduces into cuprous oxide. This cuprous oxide gives a
brick red coloured precipitate in the solution.

In Barfoed’s test, the reducing monosaccharide is oxidized by the copper ion in the solution to form a
carboxylic acid and copper (I) oxide, which results in the formation of a red coloured precipitate.

In the Iodine test, an iodine solution reacts with the starch (contains α-amylose and amylopectin
polymers). This reaction between starch and iodine results in a starch-iodine complex, which gives a
blue-black colour to the solution.

VI. Conclusion

It was concluded that carbohydrates can qualitative analyze using different test. The positive result for
Molisch test is the presence of purple liquid layer. In the Orcinol’s test the positive result for pentose is
blue or green color and hexoses is yellow or brown color. For Seliwanoff’s test, the positive result is red
product. Lastly, for Barfoed’s and Benedicts test were the presence of brick red precipitate.

Reference:

https://biologyreader.com/qualitative-analysis-of-carbohydrate.html

https://vlab.amrita.edu/?sub=3&brch=63&sim=631&cnt=1

amrita.olabs.edu.in,. (2015). Qualitative Analysis of Carbohydrates. Retrieved 23 February 2022, from

amrita.olabs.edu.in/?sub=73&brch=8&sim=209&cnt=582

Al-Ghamdi, Sharifa & Al- Shaibi, Huda. Carbohydrates. Retrieved from

www.kau.edu.sa/Files/0007306/Subjects/3)carbohydrates.doc

Cox, M. & Nelson, D. Lehninger Principles of Biochemistry. 4th edition

Experiment 1- Qualitative Test for Carbohydrates. Retrieved from: http://www.chem.boun.edu.tr/wp-
content/uploads/2014/04/Chem-415-Experiment-1.pdf

Ferrier, D. & Harvey, R. Biochemistry (2011). Lippincott Williams & Wilkins, a Wolters Kluwer business