NAME: GROUP NO.

: 6
BOLIVAR, Leonilyn SECTION: BSN1-A
LEGASPI, Ma. Lara Micaela INSTRUCTOR: Ms. Leslie Sampollo
SCORE:
OSUMO, Betiara Jazel
PELINGON, Robert Kris
PIO, Krea Joy
YSULAN, Cyky Mae DATE: December 11, 2020

Module 7 & 8: Carbohydrates

I. MATERIALS
PART A. Qualitative Analysis of Carbohydrates
Glucose
Test tube
Sucrose
Dropper
Fehling’s solution A
Beaker
Fehling’s solution B

Benedict’s reagent

Molisch’s reagent

Tollen’s reagent

Iodine solution

Distilled water

PART B. Carbohydrates
Test tubes Glucose

Fructose
Test tube rack
 Sucrose
Dropper
Maltose
Beaker
Starch

Seliwanoff’s regeant

Unknown solution

Barfoed’s regeant

Iodine solution

Water bath

PART C. Acid Hydrolysis of starch

Bunsen Burner Starch

Water
Test Tubes
Sulfuric Acid
Test tube

Test tube holder

PART D. Isolation of Glycogen from the liver of a replete and hungry animal
Graduated cylinder
Trichloroacetic acid
Lugol’s iodine
Top Loading balance
Distilled water
Spatula
Regeants
Test Tube
Balloons
Watch Glass

Beaker
Filter paper

Mortar and Pestle

Test Tube

Test Tube Rack

Spot plate

Beral Pipet

II. PROCEDURES
PART A. Qualitative Analysis of Carbohydrates
Solubility test
 Take a small amount of the given sample in a test tube and add 3 mL of distilled
water into it and shake the test tube well.

Molisch’s test
 Take about 2 mL of aqueous solution of the sample in a test tube and add few
drops of Molisch’s reagent into it. Pour 1 mL of conc. H2SO4 slowly along the side
of the test tube.
 Take 2 ml of aqueous solution of the sample in a test tube and add 1-2 mL each
of Fehling’s solution A and Fehling’s solution B into it. Keep the test tube in a
boiling water bath.

Benedict’s test
 Take 1-2 mL of aqueous solution of the sample in a test tube and add 1-2 mL of
Benedict’s reagent. Keep the test tube in a boiling water bath.

Tollen’s test
 Take 2-3 mL of aqueous solution of the sample in a test tube and add 2-3 mL of
Tollen’s reagent. Keep the test tube in a boiling water bath for 10 minutes.

Iodine test
  Take a small amount of aqueous solution of the sample in a test tube and add 1-
2 drops of iodine solution into it. 

PART B. Carbohydrates
 Known Sugar
 Add 1 mL of sugar solution to each of the 5 test tubes
 Label them 1(Glucose), 2(Fructose), 3(Sucrose), 4(Maltose), and
5(Starch).

Seliwanoff’s Test
 Add 1 mL of seliwanoff’s regeant using a dropper in each of the 5 test
tubes then heat it with hot water bath for 5 min then observe.

Barfoed’s Test
 Add 1 mL of Barfoed’s reagent using a dropper in each of the 5 test tubes
then heat it with hot water bath for 2-3 min then observe.

Iodine Test
 Add few drops of iodine solution using a dropper in each of the 5 test
tubes and observe.

 Unknown Sugar
 Add 1 mL of sugar solution to each of the 5 test tubes
 Label them A, B, C, D, and E

Iodine Test
 Add 1 mL of iodine solution using a dropper in each of the 5 test tubes
then observe.

PART C. Acid Hydrolysis of starch

 Put on protective gloves and safety goggles.
 Add water to a beaker. Heat the water on a Bunsen burner or other heat
source until it reaches the point of boiling. Once you've reached the boiling
point, reduce the heat slightly to keep the water boiling.
 Add a small amount of starch solution and sulfuric acid to a test tube
 Place the test tube into the beaker of boiling water. Allow the test tube to
boil for five minutes.
  Remove the test tubes from the beaker using test tube tongs. Place the
test tubes on a test tube cooling rack.
 Allow the test tubes to cool off until they can be handled easily.

PART D. Isolation of Glycogen from the liver of a replete and hungry animal
 Weigh liver replete sample and hungry liver sample have 1.085 g each
sample.
 Add 6mL of Trichloroacetic acid and pour the 6mL of Trichloroacetic acid
in the mortar.
 Add the liver sample in the mortar and pound for 10 minutes.
 Add distilled water to homogenate and label the two beakers.
 Get a two filter paper and fold each it into a cone shape and place each in
the beakers.
 Pour each sample in the beakers with filter paper and let it stand for
filtration.
 Label each three tubes the Control-Distilled Water, Replete Liver, and
Hungry Liver.
 Get 1mL per sample.
 Placed 1mL distilled water in the CTRL tube.
 Add 2 drops of Lugol’s Iodine and shake gently.
 Transfer 1mL each in a spot plate for better observation.

III. RESULTS AND DISCUSSION

PART A. Qualitative Analysis of Carbohydrates

Glucose Sucrose Starch

Solubility soluble soluble insoluble

Starch is a
Discussion Glucose is soluble in Sucrose is soluble in
homopolysaccharide the
(Solubility water because it is a water because it is a
reason why it is insoluble
Test) monosaccharide. polar molecule.
in water.

Molisch's
Red violet ring Red violet ring Red violet ring
test

Discussion Carbohydrates Carbohydrates Carbohydrates undergo

 undergo dehydration undergo dehydration
by conc. H2SO4 to by conc. H2SO4 to
dehydration by conc.
form furfural form furfural
H2SO4 to form furfural
(furfuraldehyde) or its 9furfuraldehyde0 or
9furfuraldehyde0 or its
derivative. The its derivative. The
derivative. The furfurals
furfurals further react furfurals further react
(Molisch's further react with α-
with α-naphthol with α-naphthol
test) naphthol present in the
present in the present in the
Molisch’s reagent to form
Molisch’s reagent to Molisch’s reagent to
a purple ring at the
form a purple ring at form a purple ring at
interface between the
the interface between the interface between
acid and the test layers.
the acid and the test the acid and the test
layers. layers.

A reddish precipitate No formation of No formation of reddish

Fehling's test
is formed. reddish precipitate. precipitate.

Glucose is a reducing
sugar that reduce the There is no such There is no such
Discussion
Copper (II) ions in the precipitate is formed precipitate is formed by
(Fehling's
test reagent to form by the non-reducing the non-reducing sugar
test)
red precipitate of sugar sucrose. starch.
cuprous oxide.

Benedict's A reddish precipitate No formation of No formation of reddish

test is formed. reddish precipitate. precipitate.

Glucose is a reducing
sugar that reduce the There is no such There is no such
Discussion
Copper (II) ions in the precipitate is formed precipitate is formed by
(Benedict's
test reagent to form by the non-reducing the non-reducing sugar
test)
red precipitate of sugar sucrose. starch.
cuprous oxide.

A shining silver mirror No formation of silver No formation of silver

Tollen's Test
is formed. mirror. mirror.

Discussion The reducing sugar There is no such There is no such silver

(Tollen's glucose reduce silver silver mirror is mirror is produce by a
test) ions in the test produce by a non- non-reducing sugars.
reagent to elemental reducing sugars.
 silver, appears as
silver mirror on the
inner surface of
vessels.

No characteristic No characteristic
Iodine's test A blue color is formed.
change. change.

Iodine reacts with starch

No blue coloration No blue coloration
discussion to form a starch/iodine
appeared because of appeared because of
(iodine's complex, which gives a
the absence of the the absence of the
test) characteristic blue color
starch. starch.
to the reaction mixture.

PART B. Carbohydrates
 Known Sugar
Seliwanoff’ Discussion Barfoed’ Discussion Iodine Discussion
s Test (Seliwanoff’ s Test (Barfoed’s Test (Iodine Test)
s Test) Test)
Glucose No color No reaction/ Blue color Dark red No No reaction/
change Negative turns to coloration is color Negative result
result dark red positive for the chang
presence of e
Monosaccharid
e
Fructos Dark red Dark red Blue color Dark red No No reaction/
e color coloration is turns to coloration is color Negative result
positive for dark red positive for the chang
presence of presence of e
ketoses Monosaccharid
e
Sucrose Dark red Dark red No color No reaction No No reaction/
color coloration is change color Negative result
positive for chang
presence of e
ketoses
Maltose No color No reaction/ No color No reaction No No reaction/
change Negative change color Negative result
result chang
 e
Starch No color No reaction No color No reaction Color Dark blue
change change turns coloration is
into positive for the
dark presence of
blue Polysaccharide
s

 Unknown Sugar
Unknown Solution Observation (Iodine Test) Conclusion

A No color change No reaction/ Negative result

B Dark blue coloration Dark blue coloration is

positive for the presence of
Polysaccharides
C No color change No reaction/ Negative result

D No color change No reaction/ Negative result

E No color change No reaction/ Negative result

PART C. Acid Hydrolysis of starch

Observation Observation
After Adding Discussion
H2SO4 After heating

The starch present in the

Cloudy solution of starch solution has been broken
Cloudy solution is
Starch becomes transparent after down by the acid,
formed
heating resulting in a transparent
liquid

 
 Observation
Blue does not appear and it seems
Iodine test like starch hydrolysis has passed.
Yellow orange coloration
Trommer's test
Blue coloration
(NaOH + CuSO4)
PART D. Isolation of Glycogen from the liver of a replete and hungry animal
Observation
Control The color turns into a light
yellow
Liver of Replete The color turns light green
with little brown particles
on the center
Hungry animal The color turns into yellow
with little light orange
particles on the center
POST LAB QUESTIONS:
1. In a tabular form, summarize all the color tests used in the experiments. Indicate
their expected results and their corresponding interpretation
Color Test Expected Result
Seliwanoff’s Test Dark red coloration
Barfoed’s Test Blue to dark red coloration
Iodine Test Dark blue coloration
Molisch’s Test Red violet/purple ring
Fehling’s Test Reddish precipitate
Discussion
Starch amylose is not present
Negative results due to absence of
glucose
Discussion
No particles left-
completely isolated
solution
brown particles-is not
completely isolated, still
has glycogen
Light orange particles,
semi isolated, few
glycogen
Interpretation
Positive result for
monosaccharide
Positive result for ketoses
Positive result for
polysaccharide
Positive result for all
carbohydrates
Positive result for
Aldehyde
Benedict’s Test Reddish precipitate Positive result for 2.0% of
sugar
Tollen’s Test Silver / Dark grey Positive result for
precipitate Aldehydes
Trommer’s Test Red brown precipitate Positive result for
Aldehydes

2. What is the difference between Fehling's test and Benedict's test? Which is more
sensitive?
The main difference between Benedict's solution and Fehling's solution is that
Benedict's solution contains copper(II) citrate, while Fehling's solution contains
copper(II) tartrate. Moreover, Benedict's test is more sensitive because Benedict's
reagent was developed after Fehling's. The citrate complex in Benedict's reagent is
much more stable that Cu(OH)2, so Benedict's reagent doesn't precipitate in standing
for long unlike Fehling's.