Date: October 27, 2022

Group Number: 6
Group Members: Guia Minerva Lim
Sarah Jane Julit
Aldriene Earl Horlador
Kim Emmanuel Jimenez
Marcellus Sta. Romana Jr.

Module Number and Title: 7 Carbohydrates

Results and Discussion

Part A. Qualitative Test for Carbohydrates

A.1 Solubility

Result Discussion
Glucose Glucose dissolves in water
because polar water
molecules bind to the
Based on the result glucose molecules. The
there is a grayish color numerous O-H (hydroxyl-)
in the solution which groups in glucose are pulled
means that Glucose is to the water molecules via
soluble in water dipole-dipole interactions.
These forces might be more
potent than interactions
between glucose.
Sucrose The sucrose molecules are
drawn together by the
positive and negative polar
Based on the result
regions. The sucrose
there is a grayish color
molecules oppositely
in solution which means
charged polar regions are
that sucrose is soluble
dragged to and pushed away
in water
by the polar water molecules,
causing the sucrose to
dissolve.
Starch Cold water will not dissolve
starch granules. When
heated in water, starch
granules absorb water and
Starch on the other
expand. Gelatinization is
hand is insoluble in
defined as the process by
water although the color
which crystalline portions
is graying white
inside granules absorb water,
disrupting the crystalline
structure and resulting in the
loss of birefringence.
 A.2 Molisch Test

Result Discussion
Glucose D-glucose interacts with
sulfuric acid and -naphthol
There is a form of a red- (Molisch reagent). D-glucose
violet (purple) coloured loses hydroxyl groups in the
product that appears as form of water when strong
a purple ring at the sulphuric acid is added to the
interface between the supplied solution. The loss of
acid and the test layers. three water molecules results
in the production of 5-
(hydroxymethyl) furfural.
Sucrose The dehydration of the
There is a form of a red-
carbohydrate by sulfuric acid
violet (purple) coloured
produces an aldehyde, which
product that appears as
condenses with two
a purple ring at the
molecules of -naphthol to
interface between the
form the purple ring seen at
acid and the test layers.
the interface.
Starch Based on the dehydration of
There is a form of a red-
the carbohydrate by sulfuric
violet (purple) coloured
acid, an aldehyde is formed,
product that appears as
which condenses with two
a purple ring at the
molecules of -naphthol to
interface between the
form the purple ring seen at
acid and the test layers.
the interface.

A.3 Fehling’s Test

Result Discussion
Glucose A Reddish precipitate is The reddish precipitate
formed indicates the presence of a
reducing sugar
Sucrose No formation of reddish There is non-reducing sugar
participate
Starch No formation of reddish There is non-reducing sugar
participate
 A.4 Benedict’s Test

Result Discussion
Glucose A reddish precipitate if The reddish precipitate
formed indicates the presence of a
reducing sugar.
Sucrose No formation of reddish There is non-reducing sugar
participate
Starch No formation of reddish There is non-reducing sugar
participate

A.5 Tollen’s Test

Result Discussion
Glucose A shinning silver mirror The silver mirror indicates the
is formed presence of a reducing sugar
Sucrose There is no formation of There is non-reducing sugar
silver mirror
Starch No formation of silver There is non-reducing sugar
mirror

A.6 Iodine Test

Result Discussion
Glucose No characteristic There is no starch present
change
Sucrose No characteristic There is no starch present
change
Starch A blue color is formed The blue color formed
indicates the presence of
starch
 Part B. Test for the Presence of Carbohydrates

B.1 Seliwanoff’s Test

Result Discussion

Glucose There was barely any Adding one milliliter of

color change in the Seliwanoffs solution and 5
known sugar solution. minutes boiling in the
In the unknown solution, water, In the known
glucose sugar's color solution the sugar slightly
changed to dark red changes the Color, while
after 5 minutes in boilingin unknown sugar the
water. Color turns into dark red
which means it is positive
in ketoses.
Fructose In a known solution, After adding one ml of
after 5 minutes, the Seliwanoff's solution and
color of fructose sugar allowing the mixture to boil
becomes dark red. for five minutes, the known
There was a noticeable sugar's color changed to
change in fructose sugar dark red, indicating that it
after 5 minutes in the was positive for ketones,
unkown solution whilst the unknown sugar's
color clearly changed.
Sucrose A dark red color The procedure of the test
appeared in the solution. results in a dark red color,
indicating the presence of
ketosis in sucrose, which
indicates the test is
positive.
Maltose The color of the known After adding one ml of
sugar solution has not Seliwanoff's solution and
changed after five allowing the mixture to boil
minutes. The color of for five minutes, the known
the maltose sugar in the sugar's color changed to
unknown solution dark red, indicating that it
remained the same after was positive for ketones,
5 minutes in boiling whilst the unknown sugar's
water. color clearly changed.
Starch The color of known The known sugar's color
solution hasn't was barely noticeable after
significantly changed in adding one milliliter of
the water after 5 Seliwanoff solution and
minutes of boiling. The heating the water for five
starch's color changes minutes, but the color of
to red after 5 minutes in the unknown sugar
boiling water in the solution changed to red,
unknown solution. indicating that this sugar is
positive for ketosis.
 B.2 Barfoed Test

Result Discussion

Glucose The appearance of a Produces a red precipitate

brick red hue within 3-5 indicating the presence of
minutes, glucose is monosaccharides in the
positive by Barfoed's solution.
Test.
Fructose The appearance of a Produces a red precipitate
brick red hue within 3-5 indicating the presence of
minutes, fructose is monosaccharides in the
positive by Barfoed's solution.
Test.
Sucrose The lack of red color, Since monosaccharides
sucrose is negative are absent, the solution
for Barfoed's test. has not changed.
Maltose The lack of red color, Since monosaccharides
maltose is negative are absent, the solution
for Barfoed's test. has not changed.
Starch The lack of red color, Since monosaccharides
starch is negative are absent, the solution
for Barfoed's test. has not changed.

B.3 Iodine Test

Result Discussion

Glucose By adding a few drops Lack of polysaccharide

of iodine solution the indicates that the solution
glucose color turns into has not changed.
yellow which means this
sugar is negative for
polysaccharides
Fructose Fructose's color Lack of polysaccharide
changes to yellow in the indicates that the solution
unknown solution, has not changed.
indicating that, like
glucose sugar, it is
negative in
polysaccharides.
Fructose becomes
purple in known
solution, indicating the
presence of sucrose
sugar in
polysaccharides.
Sucrose When iodine solution is Lack of polysaccharide
added to the unknown indicates that the solution
sucrose solution, the has not changed.
 color changes to yellow,
indicating that the
polysaccharide content
of the sucrose is
likewise negative. When
we added iodine to the
known solution of
sucrose, the color
became yellow,
indicating that the
solution was deficient in
polysaccharides.
Maltose Maltose is the same in Lack of polysaccharide
Glucose, fructose, and indicates that the solution
sucrose, the Color of has not changed.
this sugar turns yellow
which means this is
negative for
polysaccharides.
Starch When a few drops of The presence of
iodine solution are polysaccharides is shown
added, the color of the by the solution's change in
starch changes to color.
bluish-purple.

Observation Conclusion
(Iodine Test)
Test Tube A There is no reaction. Starch may be
distinguished from
Test Tube B There is no reaction. monosaccharides,
disaccharides, and other
Test Tube C There is no reaction. polysaccharides using
iodine staining. Because
Test Tube D There is no reaction. starch is a coiled polymer
of glucose, the iodine used
Test Tube E After adding a few drops in this test reacts with the
of iodine, the starch coiled molecules to
solution immediately provide bluish-black
reacts and turns into results. Iodine does not
purple. Which means react with other non-coiled
that polysaccharides are carbohydrates and instead
present. stays yellowish brown. In
order to test for starch, a
bluish black hue is a good
result, but a yellowish
brown tint is a negative
result.
 Part C. Acid Hydrolysis of Starch

Observation After Observation after

Adding H2SO4 heating Discussion

A cloudy solution After heating, a The hydrogen

is formed. cloudy solution of bonds between
Starch starch becomes starch molecules
transparent. were destroyed by
acid hydrolysis.
When the amount of
acid was increased,
the number of free
hydroxyl groups first
increased, then
decreased.
Individual starch
granules absorb the
liquid and swell
when combined with
water or another
liquid and heated.

Iodine test and Trommer’s Test on Starch

Observation Discussion Conclusion

Iodine Test When the A blue-black The iodine reagent
absence of blue colour results is made by
hue occurs, from the dissolving iodine in
starch hydrolysis presence of water using
has finished. starch. If starch potassium iodide
amylose isn't since iodine is not
present, the color very soluble in
will stay orange water. This
or yellow. produces a strongly
Cellulose, starch blue-black hued
amylopectin, or substance by
disaccharides like forming a soluble
sucrose in sugar linear triiodide ion
do not contribute complex that glides
to the color. into the starch coil.
Trommer’s Test The color does Only the Trommer's test and
(NaOH + CuSO4) not become red, intermediate Benedict's test for
signifying that the stages of reducing sugars are
Trommer's test hydrolysis have quite similar in that
was been performed. both processes
unsuccessful. As a result, the cause copper
 solution still lacks oxides to precipitate
the final out of solution and
hydrolysis change color. The
product, glucose. Trommer's test
involves subjecting
a sample to sulfuric
acid, a strong acid
that hydrolyzes
disaccharides into
monosaccharides.
After the solution
has been
neutralized (typically
with potash or soda,
but hydroxide may
also be employed,
as in the Benedict's
test), a copper
sulfate solution is
then added.

Part D. Test for the Presence of Glycogen

Observation Discussion Conclusion

Control The solution is a Glycogen is not Thus, there is a
yellowish lighter present in this deficiency of
brown color. solution. glycogen when
Liver of replete The solution is a The solution's fasting. The liver's
dark brown color brown yellowish lighter
and contains little appearance brown hue denotes
liver particles. suggests that the absence of
glycogen is glycogen, whereas
present. the liver's brown
Hungry animal The solution is a There is no color shows the
yellowish lighter glycogen in this presence of
brown color. solution since it glycogen. When the
practically has liver is hungry, there
the same color as is a scarcity of
purified water. glycogen, and the
liver is filled with
glycogen.
 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 Results Interpretation

Solubility When a clear solution In order to identify and

without any cloudiness distinguish these proteins,
or precipitation protein solubility tests help
develops, the solubility to understand the solubility
test yields a positive pattern of proteins. The
result. The solubility of proteins
unsatisfactory result of provides information on
the solubility test is the solubility of proteins
demonstrated by the during the development
appearance of a cloudy and testing of novel
solution with protein compositions.
precipitation. Because there is a chance
that the findings will be
untruthfully negative,
solubility tests need to be
carefully monitored. The
test tubes should be kept
standing for a bit to
provide solubility time.
Molisch Test The purple-colored ring The Molisch test is used to
forms when the test identify the presence of
solution and sulfuric acid carbohydrates in certain
collide. Sulfuric acid samples. It may be used to
stays above the test distinguish between
solution because it has different biomolecules and
a higher density than the the carbohydrates that are
solution. Lack of color formed as a consequence
indicates a bad result. of various processes.
Because they lack the five
carbon atoms necessary
for the reaction to be
effective, trioses and
tetroses are unable to
make furfural. The Molisch
test does not precisely
detect carbohydrates.
Some organic acids,
including citric, lactic,
oxalic, and formic acids,
among others, have the
ability to create furfurals

Fehling’s Test A positive outcome and
the presence of
reducing sugars are
indicated by the
precipitate's color, which
is reddish-brown. The
absence of the reddish
precipitate or the
presence of deep blue
hue point to a poor
result and a lack of
reducing sugars.
Benedict’s Test Within three minutes, a
red precipitate forms.
reducing any existing
sugars.
Tollen’s Test A dark gray precipitate
or silver mirror forming
on the bottom and
edges of the test tube
indicates a positive
result, indicating that the
provided sample
contains reducing
sugars or aldoses. The
absence of reducing
sugars, aldoses, or -
hydroxy ketoses in the
test sample is indicated
themselves or compounds
that do so.
You can distinguish
between the presence of
aldehydes and ketones in
carbohydrates using
Fehling's test, which does
not react to any other
ketone sugars save alpha-
hydroxy-ketone. Medical
institutions utilize Fehling's
test to determine whether
there is glucose present in
urine. This helps in figuring
out whether the patient
has diabetes. Aromatic
aldehydes cannot be
detected by this assay.
This process only takes
place in an alkaline
environment.
If it becomes yellow, the
mixture contains between
0.5% and 1% sugar. If it
turns orange, there is 1%
to 1%.5% sugar in the
mixture. There is 1.5 to 2.0
percent sugar in the
solution if the color
becomes crimson. There
would be 0.1 to 0.5
percent sugar in solution if
the color turned to green
after boiling. Additionally, if
the color turns brick red,
the solution contains more
than 2% sugar.
In chemical laboratories,
the Tollen's test is widely
used to distinguish
between aldehydes and
ketones during qualitative
organic analysis. This test
also distinguishes between
reducing sugars and non-
reducing sugars. Some
carbohydrates that do not
contain an aldehyde group
may test positive for
Tollen's test because they
 by a negative result. areomerize in an alkaline
This sudden absence environment.
serves as proof.
Iodine Test When a blue-black or Starch may be detected in
purple tint occurs, the a number of materials with
test is considered this assay. Similar to that,
successful since starch this test is conducted to
is present. The result is gauge how effectively
undesirable and photosynthesis occurs in
indicates that there is noplants. This test cannot be
starch present if the performed in an acidic
color does not change. environment due to starch
hydrolysis. The test is
qualitative in nature and
does not measure starch
concentration.
Seliwanoff’s Test Success is shown by the Seliwanoff's color reaction
growth of the cherry-red is used in the method for
colored complex, which the colorimetric
demonstrates that the measurement of fructose
given sample contains in fermentation medium. A
ketoses. The test modified version of this
sample doesn't contain method can be used to
ketosis if the result is determine the
negative, or if the color concentration of ketoses in
doesn't develop at all or a specific sample.
only after a significant Seliwanoff's reagent may
amount of time has create molecules with
passed. identical colors in
conditions of high glucose
or other sugar
concentrations, which
might result in
interference. This test is
broad and cannot
discriminate between
various forms of ketoses,
hence a separate test is
required to detect the
precise ketose sugar.
Barfoed Test Searching for red This test allows for the
precipitate will help you identification of reducing
find any reducing monosaccharides and the
monosaccharides in separation of reducing
your sample. If the color disaccharides from
appears within the first reducing
few minutes, the sample monosaccharides. This
contains reducing test cannot be used to
monosaccharides. identify sugar in urine due
However, if the color to the presence of Cl- ions,
 doesn't appear during which may disrupt the
the first three minutes, reaction. If a sample has a
the sample is of higher concentration of
diminishing disaccharides, it can
disaccharides. produce a positive
outcome.
Trommer’s Test Trommer's test and The Trommer's test
Benedict's test for involves subjecting a
reducing sugars are sample to sulfuric acid, a
quite similar in that both strong acid that hydrolyzes
processes cause copper disaccharides into
oxides to precipitate out monosaccharides. After
of solution and change the solution has been
color. neutralized, a copper
sulfate solution is then
added.

2) What is the difference between Fehling's test and Benedict's test?

Which is more sensitive, why?

In Fehling's solution, which is sodium hydroxide solution, tartrate ions form a

combination with copper(II) ions. When copper(II) and tartrate ions are
complexed, copper(II) hydroxide does not precipitate. Benedict's solution,
which is sodium carbonate solution, is a combination of copper(II) ions with
citrate ions. Since Benedict's changes color over time when exposed to heat,
it is more sensitive (from blue to green to yellow, and finally orange).

3) What is the product of the acid hydrolysis of starch? Prove your

answer.

Since starch is a polymer of glucose, when it is hydrolyzed, glucose is the end

result. Starches are carbohydrates that have a lot of bonded glucose
molecules in them. A strong acid like hydrochloric or sulphuric acid can be
used to separate these simple glucose sugars from one another. When
glucose is heated in neutral solution, it slowly degrades; sulphuric acid speeds
up the process, while hydrochloric acid accelerates it considerably more.
Therefore, sulphuric acid has been selected as an appropriate starch-
hydrolysing agent, and conditions of hydrolysis have been created that allow
for highly accurate estimation of starches. It is expected that the hydrolysis
will proceed in phases and produce a number of intermediate intermediates.
These are also known as maltose, soluble starch, and other dextrins. When
cellulose polymers' -1, 4-glycosidic connections are broken by acids, the
sugar molecules glucose or oligosaccharides are produced.

4) What are the reagents needed to conduct the test for the presence of
glycogen? List the steps involve.

The reagents needed to conduct the test for the presence of glycogen is the
Lugol's iodine, 5% elemental iodine is combined with 10% potassium iodide.
 Put 1 ml of the sample that is provided in a dry, clean test tube.

 In another tube, take control of 1 cc of distilled water.

 To both tubes, add a few drops of Lugol's solution, then swirl them
together.

 Watch how the test tubes' look of color changes.

 The test tubes should be heated in the water bath until the color is gone.

 Remove the test tubes so they can cool.

 Make a note of the color appearance in the test tubes.