Activity 3 Biochem

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
Name: Score: Course, Year & Section: Date performed:
Activity 3: Test for Carbohydrates
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.
C. Osazone/ Phenylhydrazine Test

1. In seven separate test tubes, add 1 mL of 1.0% solutions of glucose, fructose, galactose,
lactose, and sucrose and label properly.
2. Add 1mL of phenylhydrazine reagent. Mix the contents well and stopper with cotton loosely.
3. Put all the tubes at the same time in a large beaker of boiling water. Note the time of immersion
and the time of formation of each osazone.
4. Shake the tube occasionally to avoid supersaturation.
5. Remove the tubes after 30 minutes and cool. Note the sugars that form osazones only after
cooling.
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.
1 |Biochemistry Manual
ST. ALEXIUS COLLEGE
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 bluegreen color forming at once or on cooling indicates a pentose. Ketohexoses and methyl
pentoses produce orange or olive-green colored solutions.
H. Tollen’s Pholoroglucinol Test

1. Use 1.0% solutions of glucose, fructose and galactose for this test.
2. To 1 ml of each of the sugar solution, add 1 mL of concentrated hydrochloric acid and 2-3 drops
of Tollen’s Phloroglucinol reagent.
3. Heat it in a boiling water bath. A positive test is indicated by a red color.
I. Mucic Acid Test

1. Use 1.0% solutions of galactose, glucose and lactose for this test.
2. Place 10mL of sugar solution in a small beaker.
3. Add 5mL concentrated nitric acid and evaporate to approximately 5mL on the steam bath. Do not
evaporate to dryness.
4. Allow the solution to cool, or preferably to stand in your locker until the next laboratory period.
5. Note and draw the appearance of any crystal formed under the microscope.
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.
ST. ALEXIUS COLLEGE
Sample Carbohydrate Classification of Use

Present Carbohydrate
Yakult Glucose Monosaccharide Used to treat

hyperkalemia
ST. ALEXIUS COLLEGE
B. Test for Carbohydrates

1. Write down the significance or purpose for each given test.
2. Identify the principles involved for each test.
3. Indicate the positive result for each test.
Test Significance/Purpose Principle Involved Positive

Result
Iodine Test
Molisch Test
ST. ALEXIUS COLLEGE
Osazone Test
Benedict’s
Test
Barfoed’s
Test
Seliwanoff’s
Test
ST. ALEXIUS COLLEGE
Bial’s Test
Tollen’s
Phloroglucino
l Test
Mucic Acid
Test

Activity 3 Biochem

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Name: Score: Course, Year & Section: Date performed:

Activity 3: Test for Carbohydrates

C. Osazone/ Phenylhydrazine Test

H. Tollen’s Pholoroglucinol Test

I. Mucic Acid Test

Sample Carbohydrate Classification of Use

Yakult Glucose Monosaccharide Used to treat

B. Test for Carbohydrates

Test Significance/Purpose Principle Involved Positive

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