BIOLOGICAL EXPERIMENT REPORT

Setting up Model for digestive organ

Chiranth

INVESTIGATION QUERY:

How does the presence of the enzyme amylase within a visking tube containing starch
surrounded by distilled water effect the iodine test for starch and benedict’s solution test
for maltose in the contents within and outside the tube?

BACKGROUND INFORMATION:

1. Dialysis tubing:

A Dialysis tubing, also known as a dialysis tubing or semipermeable membrane tubing, is a

laboratory apparatus made of a special type of cellulose tubing that is selectively
permeable. It is used in various biological experiments and has significant relevance in the
field of biology and biochemistry due to its ability to mimic semi-permeable membranes
found in living organisms.
2. Amylase catalyst:

Amylase is an enzyme that plays a key role in the digestion of carbohydrates, particularly
starch, in various living organisms, including humans.
There are different forms of amylase, including salivary amylase found in saliva and
pancreatic amylase produced by the pancreas. These enzymes are responsible for breaking
down starch into simpler sugars.

3. Starch's Nature:

Starch is a complex carbohydrate made up of long chains of glucose molecules linked

together. It is the primary storage form of energy in plants.
Starch is too large and complex to be absorbed and utilized directly by cells. Therefore, it
needs to be broken down into smaller, more easily absorbable molecules.

4. Turning into Maltose:

Amylase catalyzes the hydrolysis (breakdown) of the starch molecules into maltose, which
is a disaccharide composed of two glucose molecules linked together.
This process involves the enzymatic cleavage of the glycosidic bonds between glucose units
in the starch, resulting in the release of maltose as a product.

5. Starch Detection with Iodine:

Iodine solution is used to test for the presence of starch, a complex carbohydrate composed
of glucose molecules linked togeth
er. Iodine solution, typically iodine dissolved in potassium iodide, forms a complex with
starch molecules. This complex, often appearing as a dark blue or purple color, is a visual
indicator of the presence of starch.

6. Benedict's Solution Testing for Maltose:

Benedict's solution is used to test for the presence of reducing sugars, including maltose,
which is a disaccharide composed of two glucose molecules linked together. Benedict's
solution contains copper ions, which can be reduced by certain reducing sugars. In the
presence of reducing sugars, the blue solution turns from blue to green, yellow, orange, or
red, depending on the amount of reducing sugar present.

HYPOTHESIS:
The presence of amylase inside the visking tube will result in the starch breaking down to
maltose and thus the solution with amylase will test negative for starch and positive for
maltose inside the tube. The solution outside the tube for this set-up will test negative for
starch as it is too large to diffuse outwards and will test positive for maltose due to diffusion
through the visking tube. The control set-up with no amylase will test negative for maltose
inside and outside the tube due to no hydrolysis of starch occurring and will test positive
for starch test inside the tube only.

MATERIALS REQUIRED:

Benedict’s solution
Iodine solution
Water bath at 90° C
Test tubes
Pipettes
5% amylase solution
1% starch solution
Water
2 visking tubes
Elastic bands
Beakers
Safety goggles

PROCEDURE:

Both visking tubes are taken and a knot is tied at one end using the elastic bands followed
by twisting to secure the knot.
5 mL of the starch solution is added to each tube and 3 mL of the amylase solution is added
to only one visking tube which will be referred to as tube A (with the control being tube B).
Both visking tubes are then added to their separate test tubes that are labelled A and B and
are tied at the other end and twisted to ensure that they are sealed appropriately.
The test
tubes are then filled with distilled water and allowed to rest to allow diffusion and
hydrolysis to occur.
Using pipettes take the solution from outside the visking tube and place them in separate
test tubes for iodine solution testing and benedict’s solution testing (Note that benedicts
solution testing requires placing the reagent solution mixture in a 90° C water bath for 5
minutes to observable changes, if any)
The contents from inside the visking tubes are carefully emptied into separate beakers by
untying the knots and are then transferred into test-tubes for iodine solution testing and
Benedict’s solution testing.
Only 2 drops of iodine solution and Benedict’s solution are to be added to each test-tube.
Observations are made and are Analyzed.

VARIABLES:

Independent variable  Presence of 5% amylase solution inside the visking

tube

Dependent variable  Change in color of solutions after iodine/ Benedict’s

solution testing

Controlled variables  Temperature of experimental set-up

 Volume of starch solution

 Volume of distilled water

 Visking tube (material used)

 Test-tubes

 Volume of Benedict’s solution and iodine solution

added for testing

RISK ASSESSMENT:

RISK SAFETY PRECAUTION

Use of 90° C water  Ensure that your hand is well above the water level in the
bath water bath while placing the test-tubes to avoid any
potential burns

General Precautions  Safety goggles to be always worn.

  Lab coat to be always worn.

DATA COLLECTION:

Data collected:

Type of testing TUBE A TUBE B

Inside Visking Outside Visking Inside Visking Outside Visking

tube tube tube tube

Iodine solution There is no There is no The solution There is no

change in the change in the turns blue-black change in the
Testing for color of the color of the color of the
starch solution solution solution
(remains (remains (remains
orangish- orangish- orangish-
brown) brown) brown)

Benedict’s The solution The solution There is no There is no

solution testing turns orange shows a slight change of color. change of color.
for maltose color change to
green (remains blue) (remains blue)

INTERPRETATION AND CONCLUSION:

Thus, it can be inferred that amylase has an enzymatic effect on starch whereby it catalyzes
its break down into a disaccharide called maltose. The solution within the visking tube A
tested negative for starch with the presence of amylase as it had been completely broken
down to maltose and the solution in the visking tube B tested positive for starch as it had no
enzyme to break it down.

Furthermore, the disaccharide maltose, unlike starch, is small enough to diffuse out the
visking tube. Thus, the solution inside and outside the visking tube A tests positive for
maltose. The liquid outside and inside visking tube B tests negative for maltose due to a lack
of amylase to break the starch down.

This model accurately represents the digestion of macromolecules and the absorption of
their monomers in the small intestine with the following similarities:
Visking Tube (Semi-Permeable Membrane): The Dialysis tubing serves as a model for the
semi-permeable membrane of the small intestine. The small intestine's walls have a semi-
permeable membrane that allows certain substances to pass through while restricting
others.

Liquid Inside the Visking Tube (Starch and Amylase): This liquid mixture represents the
contents of the small intestine after the ingestion of food. In this case, it contains starch (a
complex carbohydrate) and amylase (an enzyme that breaks down starch into simpler
sugars, like glucose). The amylase in the tube models the digestive enzymes produced by
the pancreas and released into the small intestine.

Liquid Outside the Tube: The liquid outside the Dialysis tubing represents the environment
surrounding the small intestine, including the bloodstream. It serves as a model for the
extracellular fluid that surrounds the small intestine and into which absorbed nutrients are
released.

EVALUATION:

Suggest in your opinion whether the result is satisfying or not. Why?

Once the experiment has been analyzed, it is clear that the conclusion is in accordance with
theoretical knowledge and thus the result is satisfying. However, multiple improvements
can be made to the experimental procedure. The result is purely qualitative, and the
numerical values of starch and maltose is not analyzed. For example – we measure the
relative amounts of maltose by observing a range of colors from green to orange rather than
obtaining a numerical reading.

Random Errors:
Pipettes were used for more than one solution that may have yielded a random error due to
partial maxing of solutions.
The time for which the Benedict’s solution - experimental solution was placed in the water
bath was not timed accurately using stop watch.
Systematic Errors:

The volume of solution extracted by the pipettes was not highly accurate thus yielding a
minor systematic error.