LABORATORY EXERCISE

CARBOHYDRATES

Introduction
Carbohydrates are polyhydroxy alcohol with a carbonyl group (an aldehyde or
ketone group). These biomolecules can be classified based on the carbon atom
present in the sugar and the number of monosaccharide units (monosaccharides,
disaccharides, oligosaccharides, and polysaccharides). Monosaccharide is the
simplest sugar and cannot be hydrolyzed further. On hydrolysis, disaccharides
yield two monosaccharide units. Polysaccharides are divided into two types:
homopolysaccharides and heteropolysaccharides.

LEARNING OUTCOMES
At the end of the experiment, the learners should be able to:
1. Perform qualitative tests for carbohydrates
2. Discuss the chemical reaction involved in each qualitative test
3. Analyze the observed result in each chemical test
4. Predict the acidity or alkalinity from the given concentration

Materials and Equipment

1. Test tubes
2. Test tube brush
3. Test tube rack
4. Water bath
5. Beaker 250 ml and 500 ml
6. Wire gauze
7. Test tube holder
8. Pipette
9. Aspirator
10. Glass slide
11. Distilled water
12. 2% glucose solution
13. 2% fructose solution
14. 2% lactose solution
15. 2% galactose solution
16. 2% sucrose solution
17. 2% maltose solution
18. 2% xylose solution
19. 2% starch solution
20. Sucrose, solid
21.Seliwanoff's reagent
22. Molisch reagent
23. Bial's orcinol test
24. Sulfuric acid
25. Fehling's A and B solution
26. Tollen's reagent
27. 10% NaOH solution
28. Lugol's solution
29. 10% ferric chloride solution
30. Phenylhydrazine HCI
31. Glacial acetic acid
32. Sodium acetate
33. Polarimeter
34.Electric stove
35. Electric water bath
36. Microscope
Safety Precautions
In this activity, the student will be handling different chemicals, so it is of utmost
importance that safety precautions be practiced.

1. The laboratory gown must be worn at all times. NO LABORATORY GOWN, NO

EXPERIMENT.
2. Wear a face mask and disposable nitrile gloves DURING EXPERIMENT.
3. Read the procedure ahead of the designated laboratory period.
4. Most of the qualitative tests in the experiment are expressed in approximate
amounts; consult your instructor about ANY ALTERATION IN THE VOLUME of the
reagents added to the test sample if the theoretical result is not observed.
5. DO NOT leave the pipette in reagent bottles to avoid cross-contamination
6. Always apply a control test tube (Distilled water instead of the test solution
that will be examined) to validate any doubt in the observed result
7. Prepare a hot water in a large beaker containing a boiling chip

Procedures

General Test for Carbohydrates

A. Seliwanoff's Test
1. Label 9 clean test tubes and add 1 ml of the sugar solution in each tube: 2%
glucose, fructose, lactose, galactose, sucrose, maltose, xylose, and starch.
2. Prepare a control tube containing distilled water instead of a sugar solution.
3. Add 2 ml of Seliwanoffs solution to each test tube and mix the contents by
gently agitating the samples.
4. Place the test tubes in a boiling water bath and stand for 10 minutes at the
same time.
5. Record the change in color after the boiling is complete.
B. Molisch's Test
1. Place 1 ml of each sugar solution, add 2 ml of Molisch's reagent, and mix the
samples by gentle agitation of the test tube.
2. Prepare a control tube
3. Tilt the tube and carefully add 2 ml of conc. Sulfuric acid, allows the acid to
run down the side of the test tube.
4. Observe the color of the ring formed at the junction of the two liquids and
record the visible results.

C. Bial's Orcinol Test

1. Add 2 ml of Bial's reagent to 1 ml of sugar solution to be tested, and gently
mix the
samples.
2. Carefully heat the test solution over an alcohol lamp until the mixture starts to
boil.
3. Add 10 drops of 10% ferric chloride solution.
4. Record any changes in color in the laboratory sheet.

Reducing Property Test of Sugars

A. Benedict's test
1. Label 9 clean test tubes and add 1 ml of the sugar solution in each tube.
(Prepare a control tube containing distilled water instead of sugar solution).
2. Add 2 ml of Benedict's solution to each test tube and mix the contents by
gently agitating the samples.
3. Place the test tubes in a boiling water bath and stand for 3 minutes at the
same time.
4. Record the change in color after the boiling is complete.
B. Barfoed’s test
1. Label 9 clean test tubes and add 1 ml of the sugar solution in each tube.
(Prepare a control tube containing distilled water instead of sugar solution)
2. Add 2 ml of Barfoed's solution to each test tube and mix the contents by
gently agitating the samples.
3. Place the test tubes in a boiling water bath and stand for 10 minutes at the
same time.
4. Record the change in color after the boiling is complete.

C. Fehling's Test
1. In a clean test tube, transfer 1 ml of each sugar solution in separate test
tubes.
2. Add 1 ml of Fehling's A and B and place the test tubes in a boiling water bath
for 5 minutes
3. Note any formation of precipitate and record your observation

D. Tollen's test
1. Wash the test tubes with a 10% sodium hydroxide solution
2. Transfer 1 ml of each sugar solution in separate test tubes and add 2 ml 5%
silver nitrate solution followed by 1 ml of 10% sodium hydroxide solution
3. Shake constantly the mixture to mix the reagents
4. Observe for the formation of silver mirrors in the inner surface of the test
tubes.
D. Ozasone Test
1. Transfer 0.1 g of sodium acetate, 0.5 g of phenylhydrazine HCl in each test,
and 10 drops of glacial acetic acid
2. Add 1 ml of the sugar solutions (glucose, fructose, lactose, and maltose) in a
separate tube and heat for 30 minutes in a boiling water bath.
3. After 30 minutes, remove the tubes from the water bath and cool down the
mixture to room temperature.
4. Collect the precipitate by aspiration using a dropper and transfer the crystals
to a clean glass slide
5. Examine under the microscope the structure of the crystal forms in each sugar
sample. Record your observations

After Activity Instruction

1. Answer the questions in the Report Form.
2. Detach the Report Form NEATLY from the manual by cutting along the broken
lines on the left side of the page.
3. Submit the Report Form to your instructor.

Proper Waste Disposal

Excess sugar solution can be discarded directly into the sink. Test solutions
containing the reagents must be discarded in designated acid and base waste
containers.
 TOTAL SCORE:
LABORATORY REPORT

Carbohydrates
Name: _____________________________ Date: ________________
Course, Year & Section: ______________ Group No.: _____________

General Test for Carbohydrates

Seliwanoff’s Test
Sugar Solution Visible Result
Glucose
Fructose
Lactose
Galactose
Sucrose
Maltose
Xylose
Starch

Molisch’s Test
Sugar Solution Visible Result
Glucose
Fructose
Lactose
Galactose
Sucrose
Maltose
Xylose
Starch
Bial’s Orcinol Test
Sugar Solution Visible Result
Glucose
Fructose
Lactose
Galactose
Sucrose
Maltose
Xylose
Starch

Reducing Property Test of Sugar

Benedict’s Test
Sugar Solution Visible Result
Glucose
Fructose
Lactose
Galactose
Sucrose
Maltose
Xylose
Starch
Barfoed’s Test
Sugar Solution Visible Result
Glucose
Fructose
Lactose
Galactose
Sucrose
Maltose
Xylose
Starch

Fehling’s Test
Sugar Solution Visible Result
Glucose
Fructose
Lactose
Galactose
Sucrose
Maltose
Xylose
Starch
Ozasone Test

Draw the crystal structure as seen under the microscope.

Glucose Fructose

Observations:

Maltose Lactose

Observations:
Observations:
Questions:
1. Enumerate the sugar that gives a negative result with Seliwanoff's test.
Aldoses, such as glucose and galactose, give a negative result, showing no color change.
Therefore, glucose and galactose are examples of sugars that give a negative result with
Seliwanoff's test.

2. Discuss the difference between the reaction in Benedict's and Barfoed's tests.

3. Explain why starch gives false negative results with Benedict's test
Benedict's test is primarily designed to detect reducing sugars, which are sugars that
have a free aldehyde or ketone group capable of reducing copper ions. Starch is a
polysaccharide composed of glucose units linked together through glycosidic bonds,
and it does not contain any free aldehyde or ketone groups in its structure. Therefore,
starch does not have the reducing properties required to react with Benedict's reagent
and produce a positive result. As a result, Benedict's test will typically yield a false-
negative result when testing for starch. To detect starch, other tests like the iodine test
(which forms a blue-black complex with starch) are more appropriate.
 TOTAL SCORE:

Grading Rubrics
Activity: Carbohydrates
Name: ______________________ Section: _________ Group No.: _______
Instructor: ___________________ Date Submitted: _________________

Rate the Student's performance by checking the appropriate box using the
following criteria:
Rate Description
4 – Excellent The student carries our procedures efficiently,
systematically, and independently.
Appropriate behavior is observed at all times.
The student exhibits strong knowledge about the
corresponding core competency.
3 – Satisfactory The student carries out procedures efficiently and
systematically but requires minimal
guidance and supervision.
Appropriate behavior is observed most of the time.
The student exhibits moderate knowledge about the
corresponding core competency.
2 – Fair The student carries out procedures efficiently and
systematically but requires close
guidance and supervision.
Appropriate behavior is seldom observed.
The student exhibits minimal knowledge about the
corresponding core competency.
1 – Poor The student carries out procedures inefficiently,
unsystematically even under close
guidance and supervision.
Appropriate behavior is never observed.
The student does not exhibit any knowledge about the
corresponding core competency.
 RATE
CRITERIA
4 3 2 1
Planning and Assessment
1. The materials needed for the activity were
prepared.
Skill Assessment
1. The chemical reactions involved in each chemical
test were explained.
2. The actual results were interpreted correctly
3. The polarimeter was properly used during
measurement of specific rotation.
4. Diligence and independence were observed
during the activity.
Post Laboratory Assessment
1. The student was able to interpret the results
obtained.
2. The working area was kept clean and orderly.
Total Score

Comments and Suggestions:

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