REPUBLIC OF THE PHILIPPINES

TARLAC STATE UNIVERSITY

COLLEGE OF SCIENCE
Lucinda Campus, Brgy. Ungot, Tarlac City Philippines 2300
Tel. No.: (045) 493-1865 Fax: (045) 982-0110 website: www.tsu.edu.ph

EXPERIMENT 3
CARBOHYDRATES

Performed by: Bernard Mark Mateo

Date Performed: March 02, 2020
BS Nursing 1-1

INTRODUCTION:
Carbohydrates are all about energy and are found in foods like fruits, vegetables,
breads, pasta, and dairy products.
Your body uses these foods to make glucose, which is your body's main energy
source. Glucose is a type of sugar that can be used right away for energy or
stored away to be used later.
"Carbohydrates provide the body with the energy it needs and are a good source
of many vitamins and minerals. However, not all carbohydrates are created
equal," says Donna Logan, RD, a registered dietitian at the University of Texas
Medical School in Houston.
Carbohydrate, class of naturally occurring compounds and derivatives formed
from them. In the early part of the 19th century, substances such
as wood, starch, and linen were found to be composed mainly of molecules
containing atoms of carbon (C), hydrogen (H), and oxygen (O) and to have the
general formula C6H12O6; other organic molecules with similar formulas were
found to have a similar ratio of hydrogen to oxygen. The general formula
Cx(H2O)y is commonly used to represent many carbohydrates, which means
“watered carbon.”
OBJECTIVE:
At the end of the experiment, one must be able to:
- Distinguish a carbohydrate from a non-carbohydrate
- Be familiar with the different procedures and the underlying principles
behind the comparative tests for the different simple sugar; and
- Be acquainted with the presence of simple sugars in clinical samples

RESULT AND OBSERVATIONS:

A. General Test For Carbohydrates
1. Molisch Test
Specific Objective: to identify carbohydrate from other biomolecules

Samples Observations
Glucose After mixing the sample and the reagent which is
 Molisch, there are some particles that seems oily
Carrageenan The result after placing 2 drops of the Molisch reagent,
it appeared a Jelly-like substance inside the tube
Chicken liver extract In the process of mixing, after a while the mixture
formed a purple ring at the top of the solution
Performed by: Bernard Mark Mateo & Roni Tamayo
Principle of Molisch’s test:
Molisch’s test is a general test for all carbohydrates. In this test, carbohydrates when
reacted with conc. H2SO4 get dehydrated to form furfural and its derivatives. When
monosaccharide are treated with conc H2SO4 or conc HCl, -OH group of sugar are
removed in the form of water and furfural is formed from pentose sugar and
hydroxymethyl furfural is formed from hexose sugar. These products reacts with
sulphonated α- naphthol to give a purple (violet red) colored complex.
Molisch test is a chemical test to detect carbohydrates. Carbohydrates undergo
dehydration when heated with concentrated H₂SO₄ to form furfural derivatives. Furfural
derivatives so obtained are condensed with alpha naphthol to give coloured compounds
and hence presence of carbohydrate is confirmed.
Result Interpretation of Molisch’s test:
Positive Molisch’s test: purple color complex
Negative Molisch’s test: no purple color
2. Iodine Test
Specific Objective: to identify the presence of starch in the given sample

Observations
Samples
Before Heating After Heating On Cooling
Potato starch Purple-blue in
Transparent and has It stays as it is
color purple blue particles
inside
Carrageenan Light yellow Became clearer Became clearer
Chicken Liver Extract Light yellow Became clearer Became clearer
Performed by: Bernard Mark Mateo and Amygail Vergara
Principle of Iodine test for carbohydrate:
Starch when reacted with I2 forms absorbed compound that gives blue color. On heating
or on addition of alkali like NaOH or KOH, color disappears. This reaction is only
physically association where I2 traps in the coiled structure of polysaccharide. On heating
or on addition of alkali; the coiled structure becomes linear and the I2 molecules are
released and the color disappears. The test will be answered by fructose, sucrose and
other keto containing carbohydrates. The iodine test is used to test for the presence of
starch. Starch turns into an intense "blue-black" colour upon addition of aqueous
solutions of the triiodide anion, due to the formation of an intermolecular charge-transfer
complex. In the absence of starch, the brown color of the aqueous solution remains.
Result interpretation of Iodine test:
Positive iodine test: dark blue color (starch)
Negative iodine test: glucose, fructose and sucrose.
B. Comparative Reactions of Carbohydrates
1. Fehling’s Test
 Specific Objective: to detect reducing sugar in a given solution
Color of Reagent: Sky Blue

Samples Observations
Glucose At first, it was blue in color but when it was heated with
glucose, it turned into dark orange appearance
Sucrose When the sucrose and Fehling's solution was mixed it was blue
but it backwards orange in color when it was heated
Fructose In mixing the fructose and Fehling's reagent and when was
heated it appeared in a dirty orange color
Lactose After heating the two sample and reagent the mixture bacame
color orange also
Performed by: Angelo San Juan & Aidene Salas
Principle of Fehling’s test:
Fehling’s test is one of the sensitive test for detection of reducing sugars. Fehling’s
reagents comprises of two solution Fehling’s solution A and solution B. Fehling’s
solution A is aqueous copper sulphate and Fehling’s solution B is alkaline sodium
potassium tartarate ( Rochelle salt). Rochelle salts (sodium potassium tartarate) present in
the reagent acts as the chelating agent in this reaction.These two solution are mixed in
equal amount before test. On heating an aldehyde or reducing sugar with Fehling’s
solution give reddish brown prepitate. Formation of red precipitate of cuprous oxide
denotes the presence of reducing sugar
Fehling's solution is used as a chemical test used to differentiate between water-soluble
aldehyde and ketone functional groups, and as a test for monosaccharides. ... Fehling's
can be used to determine whether a carbonyl-containing compound is an aldehyde or a
ketone.
Result interpretation:
Positive Fehling’s test: reddish brown ppt ( glucose, fructose, lactose)
Negative Fehling’s test: No red ppt (sucrose, starch)
2. Benedict’s Test for Test Sugars
Specific Objective: to detect reducing sugar
Color of Reagent: Blue

Samples Observations
Glucose The mixture of Benedict's reagent and glucose turned into
color yellow
Sucrose After combining the sucrose and the reagent, the color became
green
Fructose After mixing the reagent and the fructose it appeared in
orange color
Lactose The colour of the mixture resulted with a yellow green color

Performed by: Aidene Salas & Allysa Macalino

Principle:
The Reducing sugar under alkaline condition form enediols. Benedict’s solution contains
milder alkali Na2CO3. Enediols are powerful reducing agents. They can reduce cupric
ions to cuprous ions which is the basis for Benedict’s reaction. The cuprous hydroxide
during the process of heating is converted to red cuprous oxide.
Benedict's Test is used to test for simple carbohydrates. The Benedict's test identifies
reducing sugars (monosaccharide's and some disaccharides), which have free ketone or
aldehyde functional groups. Benedict's solution can be used to test for the presence of
glucose in urine.
Positive benedict’s test: color change from blue to brick red ppt ( glucose) Negative
Benedict’s test: no change in color( sucrose)
3. Benedict’s Test for Urine Samples
Specific Objective: to detect reducing sugar
Color of Reagent: Aqua Blue

Samples
Fasting Urine Sample
Random Urine Sample
Observations
At first there's no changes occur, but when it was heated
under 60°C water, it turned out to be in a dark blue green
color.
After being heated the color that appeared was blue green

Diabetic Urine Sample When it has been heated, the mixture became the color of
yellow green
Performed by: Angelo San Juan & Amygail Vergara
C. Specific Test
1. Barfoed’s Test
Specific Objective: to detect reducing sugar and to distinguish
monosaccharide from reducing disaccharide
Color of Reagent: Sky Blue

Samples Observations
Glucose After mixing tge glucose and Barfoed's reagent, there was no
changes happened at all
Sucrose When dropping the Benedict's reagent into the sucrose, it
remains as it is
Fructose There's no reaction happened after placing the reagent into the
fructose sample
Lactose The result after adding the Barfoed's reagent into the sample,
it remains the same.
Performed by: Allysa Macalino & Roni Tamayo
Principle of Barfoed’s test:
Barfoed’s test is used for distinguishing monosaccharides from reducing disaccharides.
Monosaccharides usually react in about 1-2 minute while the reducing disaccharides take
much longer time between 7-12 minutes to react with the reagent. Brick red color is
obtained in this test which is due to formation of cuprous oxide.
Barfoed's test is used to detect the presence of monosaccharide (reducing) sugars in
solution. Barfoed's reagent, a mixture of ethanoic (acetic) acid and copper(II) acetate, is
combined with the test solution and boiled. A red copper(II) oxide precipitate is formed
.will indicates the presence of reducing sugar
2. Seliwanoff’s Test
Specific Objective: to distinguish between aldose and ketose sugar Color of
Reagent: Transparent

Samples Observations
Glucose There are no changes when mixing the Glucose and
 Seliwanoff's reagent, but when it was heated it turned into
light red in color
Sucrose At first there's no reaction at all and when it was heated it
bacame color red
Fructose After heating the mixture turned into tge color of dark red

Performed by: Angelo San Juan & Aidene Salas

Principle of Seliwanoff’s test:
Seliwanoff’s test is used to distinguish aldoses from ketoses. On treatment with conc.
Acid, ketoses are dehydrated more rapidly to give furfural derivatives and on
condensation with resorcinol give cherry red complex.
The test will be answered by fructose, sucrose and other keto containing carbohydrates.
Seliwanoff's test is a chemical test which distinguishes between aldose and ketose sugars.
... If a sugar contains an aldehyde group, it is an aldose. This test relies on the principle
that, when heated, ketoses are more rapidly dehydrated than aldoses.
Result Interpretation of Seliwanoff’s test:
Positive seliwanoff’s test: Fructose and sucrose
Negative seliwanoff’s test: glucose, distilled water *If the reaction is allowed for longer
time, aldoses also produce positive results
3. Bial’s Orcinol Test
Specific Objective: to distinguish pentoses from hexoses; this distinction is
based on the color that develops in the presence of orcinol and iron (III)
chloride
Color of Reagent: Green

Samples Observations
Glucose After mixing Glucose and Bial's Orcinol it turned into
transparent green and after being heated, it became light
yellow in color
Fructose When Bial's Orcinol added to fructose it became also
transparent greenvand when it was heated it appeared in color
dark yellow
Xylose

Performed by: Angelo San Juan & Roni Tamayo

Principle of Bial’s test:

Bial’s test is useful in distinguishing pentoses sugar from hexoses sugars. Pentosses
( such as ribose sugar) form furfural in acidic medium which condense with orcinol in
presence of ferric ion to give blue green colored complex which is soluble in butyl
alcohol.
Bial's test is used to distinguish pentoses from hexoses; this distinction is based on the
color that develops in the presence of orcinol and iron (III) chloride. Furfural from
pentoses gives a blue or green color
4. Mucic Acid Test
 Specific Objective: to identify the presence of the sugar galactoseColor of
Reagent: Transparent
Performed by: Bernard Mark Mateo and Allysa Mac

The mucic acid test is used to identify the presence of the sugar galactose in food or in
synthetics manufacture. When concentrated nitric acid is heated together with galactose,
a dicarboxylic acid called mucic acid forms as a white precipitate, which counts as a
positive result. The mucic acid test gives a positive result with several galactose-
containing substances such as lactose and dulcite. With these other sugars, there is no
formation of a white precipitate when heated with nitric acid, but an acid that is soluble in
water. To test the presence of other sugars, the Benedict's test, Barfoed's test and other
tests in sugar analysis are usually conducted.
5. Osazone Formation
Specific Objective: to identify and differentiate sugars
Color of Reagent:
Performed by:

Conclusion:
After the experiment, I therefore conclude that we are able to identify carbohydrate from
other biomolecules using different sample and of course the Molisch Reagent. Also with
the color of the mixture or solution it self we identify whether a specific sample has
presence of starch. On the other hand other test such as Fehling’s Test and Benedict’s
Test and Barfoed’s Test helped us to detect reducing sugar and to distinguish
monosaccharide from reducing disaccharides in a given solution with a given color
produced after the experiment.

Reference:
BIOCHEMISTRY Laboratory Manual
EVERYDAY HELATH https://www.everydayhealth.com/weight/why-carbohydrates-
are-important-for-your-diet.aspx
BRITANNICA https://www.britannica.com/science/carbohydrate

Critique Paper:

Benedict's Reagent: A Test for Reducing Sugar

(https://www.nku.edu/~whitsonma/Bio150LSite/Lab%203%20Organic/Bio150LRevMolec.html)
Carbohydrates are divided into two groups based on the complexity of their structure.
Simple carbohydrates can form either a single ring structure (monosaccharides) or a
double ring structure (disaccharides -- formed when a pair of monosaccharides bond).
Simple carbohydrates include familiar sugars such the monosaccharides glucose (the
basic fuel of cells) and fructose (found in fruits). Common disaccharides include sucrose
(table sugar) and lactose (the sugar in milk).
Complex carbohydrates (polysaccharides) are chains of many bonded simple
carbohydrates, and are often used for energy storage. These include starch, cellulose, and
glycogen.
One test for the presence of many simple carbohydrates is to use Benedict's reagent. It
turns from turquoise to yellow or orange when it reacts with reducing sugars. These are
simple carbohydrates with unbound aldehyde or ketone groups. In lab, we used
Benedict's reagent to test for one particular reducing sugar: glucose.
Interpreting Benedict's Reagent Results
Benedict's reagent starts out aqua-blue. As it is heated in the presence of reducing sugars,
it turns yellow to orange. The "hotter" the final color of the reagent, the higher the
concentration of reducing sugar. In general, blue to blue-green or yellow-green is
negative, yellowish to bright yellow is a moderate positive, and bright orange is a very
strong positive. (See below).
Critique:
As the same to this article, we also conducted a test regarding Benedict’s Test and this
experiment help us to detect reducing sugar from different carbohydrate samples. This
Benedict’s Test is used to test for simple carbohydrates. The Benedict’s test identifies
reducing sugars (monosaccharide’s and some disaccharides), which have free ketone or
aldehyde functional groups. Benedict’s solution can be used to test for the presence of
glucose in urine.
Some sugars such as glucose are called reducing sugars because they are capable of
transferring hydrogens (electrons) to other compounds, a process called reduction. When
reducing sugars are mixed with Benedicts reagent and heated, a reduction reaction causes
the Benedicts reagent to change color. The color varies from green to dark red (brick) or
rusty-brown, depending on the amount of and type of sugar.