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Bio2 Activity 3
Bio2 Activity 3
ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
Name: JENIFEL ARISTOSA Score: Course, Year & Section: Date performed:
Objectives
At the end of the experiment, student should be able to:
● Identify different the test for carbohydrates
● Differentiate the different classes of carbohydrates
Procedures:
A. Iodine Test for Polysaccharides
1. Perform this procedure using starch, glucose, sucrose, galactose, fructose and lactose .
Run a blank test using distilled water.
2. Place 3mL of starch solution in each of two test tubes. To one, add several drops of 40%
NaOH. Add a drop of very dilute iodine solution to each test tube. Note the color. Heat
the tube in which the color developed. Cool and note any change. Compare with the
blank tube.
3. Perform the iodine test on starch powder. Note the result. Compare the result with that of
the starch solution.
4. Using other polysaccharides, perform the test by adding one drop of iodine solution to
5mL of 1% solution of carbohydrate. Observe and record your results.
B. Molisch Test
1. To three separate test tubes, place 0.5mL of 1.0% solutions of glucose, sucrose, and
starch respectively.
2. Add 2mL water and 2 drops of Molish Reagent.
3. Mix each well. Incline each tube and slowly add 3mL concentrated sulfuric acid on the
side of the tube to form a layer below the sugar solution.
4. A purple ring at the junction is indicative of a carbohydrate.
1 |Biochemistry Manual
ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
6. Examine the crystals under low power microscope. The crystals may be washed several
times with distilled water before examination. Better crystals are obtained when allowed
to cool in a water bath.
7. Draw the crystals you obtained.
D. Benedict’s Test
1. Prepare 0.5% and 2.0% glucose, fructose, sucrose and starch.
2. To 1mL reagent, add exactly 2 drops of the sugar solution. Mix well.
3. Boil vigorously for two minutes (or place in boiling water bath for 3 minutes).
4. Allow the solution to cool spontaneously. Do not hasten cooling by immersion in cold
water.
E. Barfoed’s Test
1. Use 1.0% solution of the following sugars for this test: glucose, sucrose, and
fructose. To 1mL of the reagent, add two drops of the sugar solution.
2. Heat to boiling for 30 seconds. If no precipitate form, allow the tubes to stand for
about 15 minutes and examine.
3. To compare the reactions of monosaccharides and disaccharides, place the tubes at
the same time in a boiling water bath. Note the time when signs of reduction appear
in each tube.
4. Continue boiling for 15 minutes and remove the tubes from the bath. Note the
amounts of precipitate in each tube after standing for 15 minutes.
5. Record all observations.
F. Seliwanoff’s Test
1. Use 0.1% and 1.0% solutions of glucose, fructose and sucrose for this test.
2. To 5mL of the reagent, place 1mL of the sugar solution.
3. Place them in a boiling water bath for exactly 1 minute. Note the results.
4. Continue heating and observe the color change at one minute intervals for 4
minutes. Run a blank test.
G. Bial’s Test
1. Use 1.0% solutions of glucose, fructose and galactose for this test.
2. To 3mL of Bial’s reagent, add 2mL of the sugar solution.
3. Heat until it just begins to boil.
4. A blu egreen color forming at once or on cooling indicates a pentose. Ketohexoses and
methyl pentoses produce orange or olive-green colored solutions.
2 |Biochemistry Manual
ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
ACTIVITY:
A. Classification of Carbohydrates
1. Identify five samples of food found in your home.
2. List down the carbohydrate present in the ingredients.
3. Classify on what class of carbohydrate does it belong and identify its use.
3 |Biochemistry Manual
ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
Iodine Test This test is used to The iodine test is based on Violet to Black
differentiate the absorptive properties
polysaccharides from the possessed by large
rest of carbohydrates. It polysaccharide molecules.
is given positive by The glucose chains in most
starch and glycogen. It of polysaccharides are
can also be used to organized to form helices.
differentiate between The space between the
glycogen, starch, and turns of the helix can hold
cellulose. small iodine molecules. This
is seen with amylase chains
found in starch. Glycogen
and amylopectin can also
absorb these iodine
molecules on their surface.
The absorptive property of
4 |Biochemistry Manual
ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
polysaccharides decreases
upon heating.
Molisch Test It is a screening test for This test is based on the Purple Ring
confirming the presence reaction of the alpha-
or absence of naphthol with carbohydrate
carbohydrates in a given in the presence of sulfuric
solution. It is a highly acid. The sugars react with
sensitive test for alpha-naphthol in an acidic
carbohydrates. environment to form purple-
Monosaccharides, colored furfural or
oligosaccharides, and hydroxymethylfurfural
polysaccharides all give derivatives. The intensity of
positive Molisch’s test. the color is directly
proportional to the amount
of carbohydrate present in
the solution.
5 |Biochemistry Manual
ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
Benedict’s It is a test for reducing This test is based on the Bright Orange
Test sugars. Carbohydrates ability of reducing sugars to
having free functional undergo oxidation in alkaline
solutions. In the presence of
group, that is not
an alkali, reducing sugars
involved in a glycosidic undergo tautomerization to
bond, give this test form enediols. These
positive. All enediols reduce the cupric
monosaccharides and ions (Cu+2) to form cuprous
disaccharides except ions (Cu+). The cuprous ions
sucrose give positive from cuprous hydroxide.
Upon heating, it is converted
Benedict’s test. This test
to cuprous oxide that forms
is negative for precipitates.
polysaccharides. The citrate ions present in
the reagent release the
cuprous ions slowly for
reduction and prevent the
formation of Cu(OH)2 until
the oxidation-reduction
process is completed.
6 |Biochemistry Manual
ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
Seliwanoff’s This test is used to This test involves the Red Color
Test detect monosaccharides formation of furfural
with a ketonic functional derivatives by
group. It is widely used monosaccharides with
7 |Biochemistry Manual
ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
Bial’s Test It is a general test for Pentoses form furfural Blue Color
carbohydrates and is compounds in the presence
sensitive only for of concentrated acid. The
pentoses. Any furfural compounds formed
compound that contains by pentoses condense with
a pentose sugar will give orcinol to form blue colored
a positive Bial’s test. compounds.
9 |Biochemistry Manual
ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
metallic silver, as a
result of this reduction
leads to the formation of
a silver mirror on the
bottom and sides of the
test tube. However, an
α-hydroxy ketone gives
a positive Tollen’s test
as the Tollen’s reagent
oxidizes the α-hydroxy
ketone into an aldehyde.
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ST. ALEXIUS COLLEGE
Gen. San. Drive, City of Koronadal, South Cotabato, Philippines 09506, Tel.: (083) 228-2019, Fax: (083) 228-4015, Email: st.alexiuscollege@yahoo.com
11 |Biochemistry Manual