Biochemistry Laboratory Experiment
Laboratory Experiment : Carbohydrates: Testing for Sugars
Learning Target(s):
(1) Draw the structures of carbohydrate molecules found in food;
(2) Determine the presence of sugars in food samples; and
(3) Perform laboratory experiment confirming presence of sugars in food
samples
Equivalent Period: 3 Laboratory Sessions (3 Lab Experiments)
Maximum Number of Points to be Acquired:
I. Introduction:
Sugars are an essential component of many foods and beverages, providing
sweetness, flavor, and texture. However, excessive consumption of added sugars has
been linked to various health problems, including obesity, type 2 diabetes, and
cardiovascular disease. As a result, there is a growing interest in accurately testing the
sugar content of food samples.
Testing the sugar content of food samples involves a range of analytical methods,
from simple visual tests to sophisticated laboratory techniques. These methods aim to
quantify the amount of various sugars, such as glucose, fructose, sucrose, lactose, and
maltose, present in the sample. Accurate and reliable sugar testing is essential for
ensuring food safety, compliance with regulations, and providing consumers with
accurate nutritional information.
Seliwanoff, Iodine and Benedict's tests are three common chemical tests used to
detect the presence of reducing sugars in food [Link]'s test involves adding
a few drops of Seliwanoff's reagent, which is a mixture of resorcinol and concentrated
hydrochloric acid, to the food sample. The mixture is then heated, and if the sample
contains ketoses, such as fructose, the solution will turn from yellow to cherry-red within
a few minutes. Benedict's test involves adding Benedict's reagent, which is a solution of
copper sulfate, sodium carbonate, and sodium citrate, to the food sample. The mixture is
then heated, and if the sample contains reducing sugars, such as glucose or maltose,
the copper ions in the reagent will be reduced to copper oxide, resulting in the formation
of a red, orange, or yellow precipitate.
Both Seliwanoff and Benedict's tests are widely used due to their simplicity, cost-
effectiveness, and ability to detect a broad range of reducing sugars. However, they may
give false-positive results in the presence of other reducing agents, and their sensitivity
and specificity may vary depending on the type and concentration of sugar being tested.
Therefore, it is important to use these tests in combination with other methods to ensure
accurate sugar testing. In the iodine test, a few drops of iodine solution, typically iodine-
potassium iodide solution, are added to the food sample. If the sample contains starch,
the iodine molecules will form a blue-black complex with the starch molecules, indicating
the presence of starch.
While the iodine test is useful for detecting the presence of starch, it is not specific to
sugars and will not detect the presence of other types of carbohydrates. Therefore, it is
important to use other tests, such as Seliwanoff's or Benedict's tests, to detect the
presence of reducing sugars in food samples.
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1 Seliwanoff’s Test
A. Laboratory Experiment No ____:
I. Materials:
10 mL Seliwanoff’s reagent 1 Test tube holder
Grape Extract 1 Test Tube Rack
Table Sugar Solution Water Bath
3 Test Tubes Bunsen burner
1 volumetric pipette Iron ring and stand
II. Procedure
Seliwanoff’s Test Precautions:
To perform the test you have got to follow some precautions. Here are some precautions
to successfully perform this test.
Before adding the reagent in the test tube, measure it carefully and make sure
that it is 5 mL.
Also, make sure that you are adding 1 ml of material to be tested because this is
for the exact compatibility of the both agents.
Make sure that what’s inside the test tube never makes a contact with any
human because it is highly corrosive and can bring serious damage to the skin.
1. In a clean, dry test tube, add 1mL of grape extract at 5% (ketose).
2. In the second test tube, add 1 mL of table sugar solution at 5% (aldose).
3. In the third test tube just add 1 mL of distilled water
4. For each tube, add 5 mL of Seliwanoff’s Reagent and mix thoroughly.
5. Heat the solution in the test tube. Boil the solution in boiling water for 5 minutes and
then wait on for the results.
6. Examine the appearance of the solution in the test tube Record your observations.
Positive Test: If the color changes to red, the Seliwanoff’s test is positive which means
ketosugar (Fructose along with Sucrose) exists in the solution.
Negative Test: If you find that no red color or even a slight pink tint is visible the result is
negative and indicates that Aldose sugar (Glucose) is present in the solution.
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III. Experiment Data and Observation
Sample Observation on the two Observation on the two samples
samples before the test after the test
Grape Extract
Table Sugar
Solution
IV. Post-Laboratory Questions. Make your answers clear and
sensible. Answers should be INDVIDUALLY constructed.
1. Which among the samples have was the ketose? Which one was aldose? What
was the basis for your evaluation?
2. Can the Seliwanoff’s Test be used to test all types of carbohydrates or does it have
limitation?
Reference:
3. Discuss the mechanism of the Seliwanoff’s Test as to how it can detect the
presence of sugar in a sample.
Reference:
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4. What is the role of heat in performing the Seliwanoff’s Test?
Reference:
V. Documentation. Take photos of the conduct of the experiment. Paste at
least 6 photos and write a detailed caption for each. Note: You may use one
phone camera but each member should independently write their caption.
You may also use different photos.
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