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Experiment 4-5 Finals

QUALITATIVE TESTS FOR CARBOHYDRATES BASED ON FURFURAL
FORMATION AND OXIDATION-REDUCTION REACTION

A. Furfural Formation / Fragmentation with Strong Acids

When carbohydrates, particularly monosaccharides, are subjected to the action of strong acids
like hydrochloric acid (HCl) or sulfuric acid (H2SO4), dehydration of the sugars occur forming furfurals or
furfural derivatives. The furfural condenses with various aromatic compounds to give colored
derivatives. Ketones are more readily fragmented than aldoses.

1. Molisch α-naphthol reaction

This is a general test for carbohydrates in free or combined form. In
the presence of concentrated H2SO4, glycosidic bonds are hydrolyzed giving
monosaccharides which are then dehydrated. Theproducts react with α-
naphthol forming purple colored condensationproducts at the junction of
the two liquids.

2. Bial's orcinol test

This test is specific for pentoses which are decomposed by
concentrated HCl to a furfural which condenses with orcinol to formblue to
green colored compounds.

3. Seliwanoff test
This test depends upon the more rapid formation of furfural from
ketoses as compared to aldoses in the presence of heat and HCl.
The condensation product with ketoses is a dark red compound withor
without a brown-red precipitate.

B. Oxidation - Reduction / Reduction of Metallic Ions by Sugars

The reducing property of sugars depends on the presence of an actual or potential aldehyde
group in the molecule. When solutions of sugars are heated in the presence of certain metallic ions,
the aldehyde group is oxidized to the carboxyl group and the metallic ion is reduced to a lower
oxidation state.

1. Benedict's Test / Fehling's Test

Both reagents contain copper (II) complex ion. When reacted with
glucose, the reagents oxidize the sugar to an aldonic acid whilethe metal
ion is reduced to the brick-red copper (I) oxide, Cu2O.
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2. Tollen's Test
Another qualitative test for the presence of a reducing sugar uses
Tollen's reagent, which contains Ag+ ion in the form Ag(NH3)2+. In this
reaction, glucose is oxidized to gluconic acid and the silver complex ion is
reduced to free silver, which appears as a bright shinymirror on the inside
of the test tube.

3. Barfoed's Test
This is also a copper reduction test carried out in an acid medium;
the reagent being composed of cupric acetate, Cu(C2H3O2)2, in glatial
acetic acid. The test is based on the greater reactivity of monosaccharides
compared with the disaccharides andis thus used to distinguish the two
types of sugars.

The following activity will make use of some of these qualitative tests to identify an unknown
sugar sample. Known samples of sugar, monosaccharides and disacchardides, will be used as standards
to which the unknown sugar will be compared.

OBJECTIVES

1. To carry out qualitative tests for carbohydrates based on the reactivity of thehydroxyl
group or the potential aldehyde or keto group.
2. Identify an unknown sugar based on the reactions of known samples.
3. Differentiate reducing from non-reducing sugars.

MATERIALS

20 pcs test tubes 500 or 750 mL beaker pipette

test tube rack tripod aspirator
test tube holder wire gauze rubber bands
test tube brush bunsen burner masking tape
10 mL graduated cylinder medicine dropper pen marker

Reagents Test Solutions

Seliwanoff's reagent 0.5% glucose solution
Bial's Orcinol reagent 0.5% sucrose solution
Fehling's A and B solutions 0.5% fructose solution
Barfoed's reagent 0.5% xylose solution
Benedict's reagent 0.5% lactose solution Unknown
sugar solution (to be
provided by the instructor)
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PROCEDURES

Start by boiling 300 mL of water in a big beaker.

A. Furfural Formation

1. Seliwanoff Test

a. Place 1 mL of the test solutions and the unknown sugar solution

in 6 separate test tubes. Label each test tube usingthe masking
tape.
b. Into each test tube, add 5 mL of Seliwanoff's reagent.
c. Secure the test tubes together with a rubber band and place in
boiling water for 15 minutes.
d. Compare the color produced by the unknown sugar with thoseof
the known samples.
e. Record all data in the report sheet.

2. Bial's Test

a. Place 10 drops of the test solutions and the unknown sugar

solution in another set of 6 separate test tubes. Label.
b. Add 4.5 mL of orcinol reagent into each test tube.
c. Secure the test tubes together with a rubber band and place in
boiling water for 15 minutes.
d. Compare the color produced by the unknown sugar with thoseof
the known samples.
e. Record all data in the report sheet.

B. Oxidation - Reduction Reaction

1. Fehling's Test

a. Mix 1 mL each of Fehling's A and B solutions in 6 separatetest

tubes. Add 4 mL of water.
b. Into each test tube, add 1.5 mL of each test solution and
unknown sugar solution. Label.
c. Secure the test tubes together with a rubber band and place in
boiling water for 1 minute.
d. Compare the color produced by the unknown sugar with thoseof
the known samples.
e. Record all data in the report sheet.
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2. Barfoed's Test

a. Prepare 6 test tubes each containing 3 mL of Barfoed's

reagent.
b. Into each test tube, separately place 15 drops of the test
solutions and unknown sugar solution. Label.
c. Secure the test tubes together with a rubber band and place in
boiling water for 10 minutes.
d. Compare the color produced by the unknown sugar with thoseof the
known samples.
e. Record all data in the report sheet.

2. Benedict's Test

a. Place 3 mL of Benedict's reagent in 6 separate test tubes.

b. Separately place 10 drops of the test solutions and unknownsugar
solution in each test tube. Label.
c. Secure the test tubes together with a rubber band and place in
boiling water for 2 minutes.
d. Compare the color produced by the unknown sugar with thoseof
the known samples.
e. Record all data in the report sheet.
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REPORT SHEET
Group No.: Group 1 Score:
Name: Sean Jodi M. Cosepe Date Performed: 12/5/23
C/Y/S: BSN 1A Date Submitted: 12/11/23

Experiment 4-5 Finals

QUALITATIVE TESTS FOR CARBOHYDRATES BASED ON FURFURAL

FORMATION AND OXIDATION-REDUCTION REACTION

DATA and RESULTS

Qualitative Parameter Carbohydrate

Test glucose sucrose fructose xylose lactose unknown
Seliwanoff's color Yellow Cherry Red Cherry Red Yellow Yellow Cherry Red
Test ketose / aldose Aldose Ketose Ketose Aldose Aldose Ketose
Bial'sOrcinol color Yellow Yellow Yellow Green Yellow Yellow
Test pentose? (+/-) - - - + - -
Fehling's color Red Blue Red Red Red Blue
Test reducing? (+/-) + - + + + -
Bardfoed's color Brick Red Dark Blue Brick Red Brick Red Brick Red Dark Blue
Test reducing? (+/-) + - + + + -
Benedict's color Reddish Light Blue Reddish Reddish Reddish Light Blue
Orange Orange Orange Orange
Test reducing? (+/-) + - + + + -

Identity of the unknown sugar: Sucrose

POST LABORATORY DISCUSSION

1. What is a reducing sugar?

A carbohydrate with a free aldehyde group (in the open-chain form) or a free ketone group is
referred to as a reducing sugar. The word "reducing" refers to the sugar's capacity to
contribute electrons and cause another molecule to be reduced. Benedict's test and Fehling's
test are two often used tests in which the presence of a reducing sugar leads in the
development of a colored precipitate.

2. What do the changes in the colors of the sugar solutions indicate when different test reagents
are used?

The color variations of sugar solutions when different test reagents are applied can reveal the
presence of reducing sugars. The reduction of metal ions (copper ions in Benedict's and
Fehling's tests, silver ions in Tollens' test) by the aldehyde or ketone groups of reducing sugars
causes these color changes. The colorful precipitates or mirrors that form as a result give
visual proof of the existence of these sugars.
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3. Is there a need to perform all the tests to completely identify the unknown
sugar? Explain.

All tests in the experiment that rely on furfural production and oxidation-reduction processes
may only be required on rare occasions for a comprehensive identification of the unknown
sugar. The selection of tests is determined by the initial observations and the
required information. Further testing may be avoided if a specific test clearly shows or
identifies the unknown sugar. However, if things are unclear or there may be numerous sugar
kinds, performing repeated tests is critical for a precise identification.

4. Differentiate furfural formation reaction from oxidation-reduction reaction.

Furfural formation is a dehydration process, whereas oxidation-reduction reactions include

an electron transfer. Furfural creation entails the elimination of water molecules in order to
generate a new chemical (furfural). Electrons are exchanged between reactants in oxidation-
reduction processes. Furfural is formed by reacting particular pentose sugars with furfural as
the result. Oxidation-reduction processes can involve a broad variety of chemicals, and the
results are determined by the individual reactants used. Furfural formation is used in the
manufacture of furfural for a variety of industrial applications. Many chemical and biological
processes rely on oxidation-reduction reactions. Furfural creation is a chemical process that
produces furfural from pentose sugars, whereas oxidation-reduction reactions are a large
family of reactions that include electron transfer between reactants.

5. Name other sugars which may be positive to Bial's test.

Bial's test is a chemical test used to determine the presence of pentoses, which are five-
carbon monosaccharides. Bial's test is exclusive to pentoses and may not be positive for
hexoses, which are six-carbon-atom monosaccharides. Arabinose is another pentose sugar
that might result in a Bial's test result. It's common in plant polysaccharides like
hemicellulose. Lyxose is an uncommon pentose sugar found in natural products such as
bacterial polysaccharides.

CONCLUSION

The presence of reducing sugars, such as glucose or fructose, is indicated by a color shift in
Benedict's or Fehling's tests from blue to green, yellow, orange, or brick-red. In these tests, no
color change may indicate the lack of reducing sugars. The presence of ketoses, particularly
fructose, is shown by the red color shift in Seliwanoff's Test, whereas the presence of carbs is
indicated by the violet ring in Molisch's Test. The findings of these tests when combined
provide for a thorough conclusion on the types of carbohydrates present.
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CLINICAL APPLICATIONS

1. Cite examples of sugars which may be administered intravenously topatients.

Sugars are often administered intravenously (IV) in the form of intravenous fluids containing
different electrolytes and carbs. Dextrose (glucose), fructose, and galactose are sugars that
can be given to patients intravenously. Dextrose is the most often utilized sugar in
intravenous solutions; it is given as a direct and rapid source of energy. It is frequently used to
treat dehydration, hypoglycemia, and as part of parenteral nutrition. Fructose is utilized in
some medical setting, however it is not as often used in intravenous solutions as glucose.
Galactose is less frequent in intravenous solutions. Its intravenous administration is
uncommon, however it may be explored in certain medical settings.

2. What sugar is found in the bloodstream? Which sugar is stored as energy?

Glucose is the sugar present in the bloodstream. Glucose is a simple sugar, or

monosaccharide, that is essential for delivering energy to the cells of the body. When it
comes to storing sugar as energy, glucose is transformed and stored as glycogen. Glycogen is
a polysaccharide, or complex carbohydrate, that acts as a short-term energy storage form in
the liver and muscles. When energy is required, glycogen may be broken down into glucose
via a process known as glycogenolysis, and this glucose is then released into the circulation to
be utilized for energy by cells.

3. Which tests are useful for the diagnosis of elevated blood sugar level?

The Fasting Blood Glucose Test examines blood glucose levels after a fast of at least 8 hours.
Oral Glucose Tolerance Test (OGTT), in which the patient fasts overnight before measuring
blood glucose levels before and after drinking a glucose-rich drink. Hemoglobin A1c (HbA1c)
Test, which examines the average blood glucose levels during the previous 2-3 months. These
are tests that can be used to diagnose high blood sugar levels.