BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022

Far Eastern University – Manila 2nd semester

BIOCHEMISTRY LABORATORY Qualitative Analysis for Carbohydrates Data

Sheet

Objective
• to demonstrate how some qualitative tests for carbohydrates are performed and
discuss your observations in this virtual lab.

Simulation
• You can access the simulation for the qualitative analysis of carbohydrates via this
link:
http://amrita.olabs.edu.in/?sub=73&brch=8&sim=209&cnt=4

Concept Overview
Carbohydrates are the most abundant biomolecules on Earth. They have a wide range
of functions, including providing a significant fraction of the dietary calories for most
organisms, acting as a storage form of energy in the body, and serving as cell
membrane components that mediate some forms of intercellular communication
(Seager & Slabaugh, 2013) . Carbohydrate polymers (also called glycans) serve as
structural and protective elements in the cell walls of bacteria and plants and in the
connective tissues of animals.

Carbohydrates are polyhydroxy aldehydes or ketones, or substances that yield such

compounds on hydrolysis. Many, but not all, carbohydrates have the empirical formula
(CH2O)n; some also contain nitrogen, phosphorus, or sulfur. They can be classified
according to the size of the molecules. Monosaccharides consist of a single
polyhydroxy aldehyde or ketone unit. Disaccharides are carbohydrates composed of
two monosaccharide units linked together chemically. Oligosaccharides (less common
and of minor importance) contain from three to ten units. Polysaccharides consist of
very long chains of linked monosaccharide units (Harvey, 2011).

In this simulation, you will demonstrate and observe some qualitative test for
carbohydrates including the Solubility test, Molisch test, Fehling’s test, Benedict’s test,
Tollen’s test, and Iodine test.
BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
Far Eastern University – Manila 2nd semester

This material should not be duplicated nor distributed without permission.

Simulation Observation

General instruction: Click the “Simulator” tab on the link previously provided in the
module. From there, you can select which test you wish to perform. Fill out the table
below by providing the necessary information indicated per column.
BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
Far Eastern University – Manila 2nd semester

This material should not be duplicated nor distributed without permission.

 BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
Far Eastern University – Manila 2nd semester
Group #: N/A MTY1109 Laboratory
Date Submitted: February 16, 2022 EXP 1 Carbohydrate Chemistry
Name: Chrysandra S. Marin
GROUP DATA SHEET
Qualitative Analysis of Carbohydrates
Test Added reagent/s (12 pts) Samples Observation and Interpretation (24 pts)
Solubility test  Water Glucose The solubility test helps determine if a
compound will easily be dissolved in a
solvent. The final solution in tube A shows
that glucose is dissolved in water because it is
a polar molecule, making it water-soluble.
Thus, the result of the solubility test in tube A
is positive.
Lactose The solubility test helps determine if a
compound will easily be dissolved in a solvent.
The final solution in tube B shows that lactose
is dissolved in water because it contains C-O
bonds, -OH group, and permanent dipole
moment making it a polar molecule, which
means it is water-soluble. Thus, the result of
the solubility test in tube B is positive.
Sucrose The solubility test helps determine if a
compound will easily be dissolved in a
solvent. The final solution in tube C shows
that sucrose is dissolved in water because it is
a polar molecule, which means it is water-
soluble. Thus, the result of the solubility test in
tube C is positive.
Starch The solubility test helps determine if a
compound will easily be dissolved in a
solvent. The final solution in tube D shows
that starch did not dissolve in water because it
is a polymer, which means it has longer
chains. After all, longer chains are densely
entangled, making the molecule slowly
 BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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dissolve in a solvent. Thus, the result of the

solubility test in tube D is negative.

 Molisch’s reagent Glucose The usage of the Molisch test is for detecting
 Conc. H2SO4 any type of carbohydrate in a solution. In order
Molisch test to determine if a carbohydrate is present in a
solution, a reddish violet or purple-colored ring
appears in the middle of two solutions. Since
the final solution in tube A contains a purple-
colored ring, it means that glucose is a
carbohydrate by reacting with the Molisch's
reagent and Conc. H2SO4. Therefore, the
result of the Molisch test in tube A is positive.
Lactose The usage of the Molisch test is for detecting
any type of carbohydrate in a solution. In order
to determine if a carbohydrate is present in a
solution, a reddish violet or purple-colored ring
appears in the middle of two solutions. Since
the final solution in tube B contains a purple-
colored ring, it means that lactose is a
carbohydrate by reacting with the Molisch's
reagent and Conc. H2SO4. Therefore, the
result of the Molisch test in tube B is positive.
Sucrose The usage of the Molisch test is for detecting
any type of carbohydrate in a solution. In order
to determine if a carbohydrate is present in a
solution, a reddish violet or purple-colored ring
appears in the middle of two solutions. Since
the final solution in tube C contains a purple-
colored ring, it means that sucrose is a
carbohydrate by reacting with the Molisch's
reagent and Conc. H2SO4. Therefore, the
result of the Molisch test in tube C is positive.
Starch The usage of the Molisch test is for detecting
any type of carbohydrate in a solution. In order
 BIOCHEM LABORATORY: CARBOHYDRATES
Far Eastern University – Manila
Fehling’s test  Fehling’s solution (blue)
 Fehling’s solution (clear)
AY 2021-2022
2nd semester
to determine if a carbohydrate is present in a
solution, a reddish violet or purple-colored ring
appears in the middle of two solutions. Since
the final solution in tube D contains a purple-
colored ring, it means that starch is a
carbohydrate by reacting with the Molisch's
reagent and Conc. H2SO4. Therefore, the
result of the Molisch test in tube D is positive.
Glucose Fehling's test aims to determine the reducing
sugars in a solution by adding two Fehling's
solutions: the blue and clear solution. To
detect the reducing sugar in a solution, the
solution will turn red or contain an aldehyde.
On the other hand, the color of the solution will
not change if there is a presence of keto. For
instance, in tube A, the final solution turns into
color red, which denotes that there is a
presence of reducing sugar. Therefore, the
result in tube A using Fehling's test is positive
because the solute is an aldehyde.
Lactose Fehling's test aims to determine the reducing
sugars in a solution by adding two Fehling's
solutions: the blue and clear solution. To
detect the reducing sugar in a solution, the
solution will turn red or contain an aldehyde.
On the other hand, the color of the solution will
not change if there is a presence of keto. For
instance, in tube B, the final solution turns into
color red, which denotes that there is a
presence of reducing sugar. Therefore, the
result in tube B using Fehling's test is positive
because the solute is an aldehyde.
Sucrose Fehling's test aims to determine the reducing
sugars in a solution by adding two Fehling's
 BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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solutions: the blue and clear solution. To

detect the reducing sugar in a solution, the
solution will turn red or contain an aldehyde.
On the other hand, the color of the solution will
not change if there is a presence of keto. For
instance, in tube C, the final solution turns into
color blue, which denotes that there is no
presence of reducing sugar. Therefore, the
result in tube C using Fehling's test is negative
because the solute is a ketone.
Starch Fehling's test aims to determine the reducing
sugars in a solution by adding two Fehling's
solutions: the blue and clear solution. To
detect the reducing sugar in a solution, the
solution will turn red or contain an aldehyde.
On the other hand, the color of the solution will
not change if there is a presence of keto. For
instance, in tube D, the final solution turns into
color blue, which denotes that there is no
presence of reducing sugar. Therefore, the
result in tube D using Fehling's test is negative
because the solute is a ketone.

Benedict’s test  Benedict reagent Glucose Benedict's test is just the same as Fehling's
test, which determines or detects the presence
of reducing sugar in a solution. The only
difference between the two tests is the solvent
used. In tube A, the final solution turns into
color red, which denotes that there is a
presence of reducing sugar. Therefore, the
result in tube A using Benedict's test is positive
because the solute is an aldehyde.
Lactose Benedict's test is just the same as Fehling's
test, which determines or detects the presence
of reducing sugar in a solution. The only
 BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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difference between the two tests is the solvent

used. In tube B, the final solution turns into
color red, which denotes that there is a
presence of reducing sugar. Therefore, the
result in tube B using Benedict's test is positive
because the solute is an aldehyde.
Sucrose Benedict's test is just the same as Fehling's
test, which determines or detects the presence
of reducing sugar in a solution. The only
difference between the two tests is the solvent
used. In tube C, the final solution turns into a
color blue, which denotes that there is no
presence of reducing sugar. Therefore, the
result in tube C using Benedict's test is
negative because the solute is a ketone.
Starch Benedict's test is just the same as Fehling's
test, which determines or detects the presence
of reducing sugar in a solution. The only
difference between the two tests is the solvent
used. In tube D, the final solution turns into a
color blue, which denotes that there is no
presence of reducing sugar. Therefore, the
result in tube D using Benedict's test is
negative because the solute is a ketone.
 Tollen’s reagent Glucose The Tollen's test, known as the silver-minor
test, is also a type of test in determining the
Tollen’s test presence of an aldehyde or a ketone in a
solution. The test is called a silver-minor
because it gives a metallic appearance when
the Tollen's reagent is added to a solution that
contains an aldehyde. Aldehyde is readily
oxidized; that is why it reacts with Tollen's
reagent because the said reagent is a solution
of silver nitrate and Ammonia. In tube A,
glucose reacts with the reagent forming a
metallic appearance. Thus, it is an aldehyde,
BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
Far Eastern University – Manila 2nd semester

and the result using Tollen's test is positive.

Lactose The Tollen's test, known as the silver-minor
test, is also a type of test in determining the
presence of an aldehyde or a ketone in a
solution. The test is called a silver-minor
because it gives a metallic appearance when
the Tollen's reagent is added to a solution that
contains an aldehyde. Aldehyde is readily
oxidized; that is why it reacts with Tollen's
reagent because the said reagent is a solution
of silver nitrate and Ammonia. In tube B,
lactose reacts with the reagent forming a
metallic appearance. Thus, it is an aldehyde,
and the result using Tollen's test is positive.
Sucrose The Tollen's test, known as the silver-minor
test, is also a type of test in determining the
presence of an aldehyde or a ketone in a
solution. The test is called a silver-minor
because it gives a metallic appearance when
the Tollen's reagent is added to a solution that
contains an aldehyde. Aldehyde is readily
oxidized; that is why it reacts with Tollen's
reagent because the said reagent is a solution
of silver nitrate and Ammonia. In tube C,
sucrose did not react with the reagent, forming
no changes in appearance. Thus, it is a
ketone, and the result using Tollen's test is
negative.
Starch The Tollen's test, known as the silver-minor
test, is also a type of test in determining the
presence of an aldehyde or a ketone in a
solution. The test is called a silver-minor
because it gives a metallic appearance when
the Tollen's reagent is added to a solution that
contains an aldehyde. Aldehyde is readily
oxidized; that is why it reacts with Tollen's
 BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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reagent because the said reagent is a solution

of silver nitrate and Ammonia. In tube D,
starch did not react with the reagent, forming
no changes in appearance. Thus, it is a
ketone, and the result using Tollen's test is
negative.
 Iodine solution Glucose The purpose of the Iodine test is to determine
if there is a presence of starch in a solution. In
Iodine test tube A, the reaction between glucose and
iodine solution results in having a yellowish
solution, which means that there is no
presence of starch. Hence, the Iodine test for
carbohydrate detection is negative.
Lactose The purpose of the Iodine test is to determine
if there is a presence of starch in a solution. In
tube B, the reaction between lactose and
iodine solution results in having a yellowish,
which means that there is no presence of
starch. Hence, the Iodine test for carbohydrate
detection is negative.
Sucrose The purpose of the Iodine test is to determine
if there is a presence of starch in a solution. In
tube C, the reaction between glucose and
iodine solution results in having a yellowish,
which means that there is no presence of
starch. Hence, the Iodine test for carbohydrate
detection is negative.
Starch The purpose of the Iodine test is to determine
if there is a presence of starch in a solution. In
tube D, the reaction between starch and iodine
solution results in having a blue color solution,
which means that there is a presence of
starch. Hence, the Iodine test for carbohydrate
detection is positive.
BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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This material should not be duplicated nor distributed without permission.

BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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GUIDE QUESTIONS:

1. Some of the test(s) you tried from this virtual lab are important basis for
determining glucose levels of diabetic patients from blood and/or urine. Can you
identify which are these tests and how is modern method used glucose
monitoring different? (Show the reactions) (7 pts)

The qualitative tests useful for determining a person's glucose level are Fehling's
test, Benedict's test, and Tollen's test. These three qualitative tests help detect
the reducing sugar, wherein glucose is considered a reducing sugar since its
structure contains an aldehyde group with an open-chain form, making the
molecule react with the chemical reagents of the three qualitative tests. We all
know that when a solution contains an aldehyde mixed with the chemical
reagents of the three qualitative tests, it will have a red color solution, which
means there is a presence of reducing sugar.

However, many high technologies have been developed to help determine the
glucose level more conveniently and effectively in the modern world, such as
Continuous glucose monitoring, Noninvasive sensor, Glucose sensing via the
eye, and many more. Using these technologies produces a more accurate
glucose level of a person because the percentage is indicated right away after
using the device. For example, blood glucose monitoring is a device that detects
the glucose level by pricking a finger with a lancet and then putting the blood in a
test strip. Afterward, the test strip will be inserted into the device to determine the
glucose level. Overall, the difference between the method of the three qualitative
tests and the modern method of detecting glucose levels is that the modern
method is easier to use and less hassle since there is no need for using chemical
reagents, unlike the qualitative tests.

VIRTUAL LAB
BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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MODERN TECHNOLOGIES
Blood glucose monitoring
BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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Continuous glucose monitoring

2. What are reducing sugars? Why are monosaccharides considered as reducing

sugars? Which qualitative tests can help you determine the presence of these
reducing sugars and the principle behind it? (7 pts)

Reducing sugar are carbohydrates that function as a reducing agent because

they can reduce the oxidizing agent by donating electrons to it. According to
Rizzo (n.d.), a carbohydrate that contains either a free aldehyde or ketone group
in a chemical structure is considered a reducing sugar. However, the aldehyde is
easy to oxidize compared to ketones. It is because the chemical structure of an
aldehyde contains hydrogen, which makes aldehyde to be easily oxidized. On the
other hand, a ketone does not have any hydrogen in its structure, leading to
resistance from oxidizing agents. For example, all monosaccharides contain
hemiacetals, which it has an open chain that includes either an aldehyde or
ketone group. Thus, making monosaccharides to be a reducing sugar.

Moreover, there are three qualitative tests that can detect the presence of
reducing sugar in a solution, namely, Fehling's test, Benedict's test, and Tollen's
test. As stated by Supriya (n.d.), in Fehling's test, the reduction occurs when two
Fehling's solutions are added into a solution that contains either an aldehyde or a
ketone group. The color blue Fehling's solution comprises copper (II) sulphate,
while the other solution contains sodium tartrate. If these two chemical reagents
are added to a solution with an aldehyde group and heated afterward, the color of
the solution will turn into the color red, which means that reducing sugar is
present in the solution. On the contrary, if a solution contains a ketone group, the
color of the solution will not cause any changes, meaning a reducing sugar is not
present.
BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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Another test that detects the presence of reducing sugar is Benedict's solution.
This test is similar to Fehling's test because if a solution has an aldehyde and is
mixed with Benedict's reagent, which is composed of blue copper (II) ions (Cu2+,
cupric ions), the solution will turn red when heated. Meaning there is reducing
sugar in the solution. On the other hand, if a ketone group is in the solution mixed
with Benedict's reagent and heated afterward, the mixture of the two chemicals
will not cause any changes because reducing sugar is not present in the solution.

And lastly, Tollen's test is also for detecting the reducing sugar by using Tollen's
reagent, which is composed of ammoniacal silver nitrate. This test is the same
with the two solutions; however, the only difference is that the final solution turns
into a silver-mirror appearance when Tollen's reagent is mixed with an aldehyde,
which means a reducing sugar is present. And when a solution with ketone
groups mixed with Tollen's reagent causes no changes, it means that there is no
reducing sugar in the final solution.

3. Explain why all mono- and disaccharides are soluble in water? (5 pts)

Monosaccharides and disaccharides easily dissolve in water because of their

molecular structure, which contains many free -OH groups (hydroxyl and
carbonyl). The -OH group is the reason why both saccharides are water-soluble
because it has the capability of forming hydrogen bonds, which makes the
molecule to be polar. Polar molecules are molecules that include both partially
negative and positive charges on each end of the molecule. When the partial
negative charge of a molecule interacts with the partial positive charge of another
molecule, this will make both polar substances dissolve one another. For this
reason, polar molecules are considered to be hydrophilic or water-loving.

4. What are some examples of artificial sweeteners? How are they helpful for
diabetics? (5 pts)

There are lots of artificial sweeteners that have been made, such as Aspartame,
Sucralose, Acesulfame K, Saccharin, Xylitol, and many more According to
Bramlet (n.d.), artificial sweeteners are low-calorie sugars that are considered to
be a substitute for natural sugars, like honey, molasses, coconut sugar, and
such. Cotis & Harris (2015) stated that artificial sugar is safe and helpful to
people with diabetes because it can reduce calorie and carbohydrate intake,
which helps maintain a person's glucose level. As stated by Castro (2021), the
following are the recommended artificial sweeteners for people with diabetes,
and these are the:
 Saccharin (Sweet'N Low)
 Aspartame (NutraSweet)
BIOCHEM LABORATORY: CARBOHYDRATES AY 2021-2022
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 Acesulfame potassium (Sunett)

 Neotame (Newtame)
 Advantame
 Sucralose (Splenda)
 Stevia (Pure Via, Truvia)

These artificial sweeteners do not cause any effects on the glucose level of a
person since it has low calories. Hence, it is safe to use.

5. Why is dietary fiber good for you? What are some of its sources? What’s the
difference between soluble and insoluble fiber and some of their health benefits?
(5 pts)

Dietary fibers are known as roughage or bulk because it helps in controlling our
weight by making us feel full faster. These dietary fibers are essential for people
to consume because it helps in maintaining our health by regularizing bowel
movements, lowering cholesterol level, helps in controlling sugar level, and
maintaining body weight. According to Staff (2021), dietary fibers are usually
found in fruits, vegetables, whole grains, and legumes. Moreover, there are two
types of dietary fibers: soluble and insoluble fibers. The soluble fibers are soluble
in water, or they can dissolve in water. This kind of fiber can be found in plant
pectin and gums. It benefits people's health, especially with diabetes, because it
can help control the blood glucose level. It also helps in the reduction of blood
sugar and cholesterol levels.

On the other hand, insoluble water is hydrophobic, meaning it does not react or
dissolve in water, and can be found in plant cellulose and hemicellulose. The
benefits that people can get from this kind of fiber are that it also helps in
lessening the possibility of diabetes because it can maintain or control insulin
sensitivity. In addition, this fiber permits water to get into someone's void, which
makes the void discharge easily. For this reason, it helps to maintain and control
bowel movement.
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This material should not be duplicated nor distributed without permission.

Bibliography
Castro, R. M. (2021, April 7). Artificial Sweeteners: Any effect on blood sugar? Retrieved from MAYO
CLINIC: https://www.mayoclinic.org/diseases-conditions/diabetes/expert-answers/artificial-
sweeteners/faq-20058038

Mark, A. (2014, April 3). Chemistry. Retrieved from Socratic Q&A: https://socratic.org/questions/how-
do-polar-covalent-molecules-dissolve-in-water

Rizzo, N. (n.d.). The Definition of Reducing Sugars. Retrieved from LIVESTRONG:

https://www.livestrong.com/article/386795-the-definition-of-reducing-sugars/

BIOLOGY ONLINE EDITORS. (2021, September 6). Reducing Sugar. Retrieved from Biology Online:
https://www.biologyonline.com/dictionary/reducing-sugar

Bramlet, K. (n.d.). ARTIFICIAL SWEETENERS AND NATURAL SWEETENERS: WHAT TO KNOW. Retrieved
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BYJU's. (n.d.). Is Glucose A Reducing Sugar. Retrieved from BYJU'S: https://byjus.com/jee-questions/is-

glucose-a-reducing-sugar/

Castro, R. M. (2021, April 7). Artificial Sweeteners: Any effect on blood sugar? Retrieved from MAYO
CLINIC: https://www.mayoclinic.org/diseases-conditions/diabetes/expert-answers/artificial-
sweeteners/faq-20058038

Clark, J. (2020, September 13). Oxidation of Aldehydes and Ketones. Retrieved from LibreTexts:
https://chem.libretexts.org/Bookshelves/Organic_Chemistry/Supplemental_Modules_(Organic_
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Oxidation_of_Aldehydes_and_Ketones

Dansinger, M. (2021, June 23). Home Blood Sugar Testing. Retrieved from WebMD:
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Lakna. (2019, October 15). What is the Difference Between Reducing Sugar and Starch. Retrieved from
PEDIAA: https://pediaa.com/what-is-the-difference-between-reducing-sugar-and-starch/

Mark, A. (2014, April 3). Chemistry. Retrieved from Socratic Q&A: https://socratic.org/questions/how-
do-polar-covalent-molecules-dissolve-in-water

Rizzo, N. (n.d.). The Definition of Reducing Sugars. Retrieved from LIVESTRONG:

https://www.livestrong.com/article/386795-the-definition-of-reducing-sugars/

Sapkota, A. (2020, November 18). Fehling's Test - Definition, Principle, Procedure, Result, Uses. Retrieved
from Microbe Notes: https://microbenotes.com/fehlings-test/

VARSITY TUTORS. (n.d.). Help with Organic Carbohydrates. Retrieved from Varsity Tutors:
https://www.varsitytutors.com/organic_chemistry-help/help-with-organic-carbohydrates?
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