Republic of the Philippines

SOUTHERN LEYTE STATE UNIVERSITY-MAIN CAMPUS

College of Technology
Department of Food Science and Technology
Sogod, Southern Leyte

Name: John Snipner P. Pinto Date Performed: April 4, 2023

Lab Schedule: MTh 1:00 PM - 5:00 PM Date Submitted: April 11, 2023

FT 304- Food Chemistry 2

Laboratory Exercise No. 2
Microscopic Examination and Effect of Different Factors on the Structure of Starches

Introduction

Starch is a type of complex carbohydrate that can be found in a variety of foods,

including grains, vegetables, and fruits. It is a crucial source of energy for both people and
animals. Amylose and amylopectin are the two different types of molecules that make up starch.
To create a granule structure, these molecules are organized in a particular way.

Microscopic examination is an essential technique for understanding the structure of

starch. It entails using a microscope to examine the starch granules' dimensions, composition,
and shape. Depending on the starch source, the granules' size and shape can vary. For instance,
the granules of corn starch are more rounded and smaller than those of potato starch.

The structure of starch can also be impacted by many variables. The temperature is one
such element. Heat causes the granules of starch to absorb water and expand, changing the
structure of the substance. For this reason, starch is frequently employed in cooking as a
thickening agent. The pH scale, which has the greatest stability for starch between 5.0 and 7.0, is
another component that can have an impact on its structure. The structure of the starch can be
changed and lose its ability to thicken at pH levels below 5.0 or over 7.0.

Furthermore, enzymes can modify the structure of starch. Starch molecules can be broken
down by enzymes like amylase, which alter the structure. Because of this, amylase is used to
make sweeteners like high fructose corn syrup.

In conclusion, microscopic examination is a useful instrument for learning about the

composition of starch. In a variety of industries, including food, agriculture, and biotechnology,
it's critical to comprehend the variables that can impact the structure of starch.
Objectives:

1. To determine the structure of different scratches under the microscope.

2. To investigate the effect of pH and heat on starch structure.

Materials:

Microscope Hydrocloric Acid (HCl)

Glass Slide and coverslips Sodium hydroxide (NaOH)
Wire Loop Test tubes
Iodine Solution Test tube rack
All Purpose Flour, Cornstarch, and Rice Starch Alcohol lamp
Dropper or pipette Flying Tiger (alcohol)
Distilled water

Methodology

A. Microscopic Examination of Starch Solution

A method for observing and evaluating a substance's characteristics under a microscope

is called microscopic examination. The size, shape, and structure of the starch granules in
a starch solution can be determined by microscopic examination. In this experiment, it is
important to prepare an all-starch solution (all-purpose flour, cornstarch, and rice starch)
with 50 mL of water and 2.5 grams of each starch before doing a microscopic
examination. To make a starch solution, we perform the following steps: 1.) Add a little
amount of starch powder to a test tube. 2.) Add a few drops of water to the test tube, then
stir the mixture until it forms a paste-like consistency. 3.) Fill the test tube with extra
water and stir the ingredients until a milky, thin solution forms. Once all the starch
solution is prepared, it can be examined under a microscope. To conduct a microscopic
examination, we also perform the following steps: 1.) Drop some of the starch solution
onto a glass slide. 2. Add the iodine solution. 3. As we didn't cover the glass slide in our
experiment, assume that we placed a cover slip over the drop of starch solution. 4. Let the
solution dry. 5.) Set the glass slide on the microscope's stage and adjust the focus until the
starch granules are clearly visible. As we can see, starch granules are tiny, oval-shaped
objects that can be seen under a microscope. The source of the starch might affect the size
and form of the starch granules.

B. Effect of Heat on Starches

We gather the necessary materials before beginning this experiment. This leaves us with
25 mL of starch solution in a beaker. Also, we heat them on a hot plate until the starch
 starts to gelatinize. The beakers are then taken out of the alcohol lamp and placed aside.
Next, we prepare and clean a glass slide for each sample, then we add a small amount of
each heated sample onto the glass slide. Then, using a stirring rod, added a drop of iodine
solution to each sample. We next dried all of the samples, and at 10x or 40x
magnification, we looked at the starch granules' shape and size.

C. Effect of Iodine Solution To The Color of Starches

We first gather the supplies we'll need for this experiment. For each variety of starch, we
additionally prepare three test tubes with the labels A, B, and C. Afterwards, we fill each
test tube with 5 ml of the starch solution. After that, test tube A received 5 ml of distilled
water, test tube B received HCl, and test tube C received NaOH. We now add 1-2 drops
of iodine solution to each test tube once they have been thoroughly combined, while also
checking the color of each solution. Follow the steps again for the remaining two
starches.

Results and Discussion

A. MICROSCOPIC EXAMINATION OF STARCH SOLUTION

SAMPLE OBSERVATION MICROSCOPIC IMAGE

All Purpose Flour Dark blue spots

Cornstarch Dirty white with scattered

black spots
Rice Starch Off white with black lines in
its surface

The structure and characteristics of starch molecules can be determined by microscopic

examination of a starch solution. As the starch solution is being evaluated, all samples are
examined under a microscope. The first sample was the all-purpose flour, and when it had been
dried and placed on a glass slide with an iodine solution, I had noticed under a microscope that
the sample had numerous dark blue dots all over it. It looks like a crater to me. On the other
hand, I noticed under a microscope that the cornstarch that had been mixed with iodine solution
and allowed to dry had a filthy white appearance with black dots similar to what we would see if
we tested sperm cells. The rice starch finally showed up under a microscope as an off-white
powder with black lines on its surface after drying and the addition of iodine solution. Its
appearance is comparable to that of a full moon. Therefore, the size, shape, and surface
characteristics of the starch granules, as well as their behavior under specific circumstances, can
be determined through microscopic examination of a starch solution. Understanding the
characteristics of starch and how they affect how starch-containing foods behave during
processing and cooking can be done with the use of this information.

B. EFFECT OF HEAT ON STARCHES

SAMPLE OBSERVATION MICROSCOPIC IMAGE

All Purpose Flour After adding iodine solution,

the hue of the bubbles changed
to a blue with black spots and a
yellow outline. The bubbles
are rising.
Cornstarch There are bubbles forming,
which later turn into
translucent gel. It becomes
purple when an iodine solution
is added. It has a mixture of
blue and purple tint after
drying up.

Rice Starch The sample forms top bubbles

and is rising. Iodine solution is
added, and the color changes to
blue-green. It becomes pale
blue with fading orange spots
after drying.

A sample of all-purpose flour was used; when heated, it produced bubbles, and when an iodine
solution was added, the sample colored the flour blue. After drying the samples, I noticed black
spots and a yellow outline under the microscope. On the other hand, bubbles and a translucent
gel are also created when cornstarch is heated. The material appears blue and purple under the
microscope after drying and becomes purple when iodine solution is added. Last but not least,
the rice starch sample forms bubbles, changes color when iodine solution is applied and takes on
a blue-green hue, and when dried, the sample appears in the microscope as light blue with faded
orange spots. In conclusion, the final result can be significantly impacted by the complicated
effects of heat on starches. To get the correct texture and flavor while cooking and baking, it's
crucial to understand the characteristics of starch and how heat affects them.
 C. EFFECT OF IODINE SOLUTION TO THE COLOR OF STARCHES

SAMPLE COLOR OBSERVATION MICROSCOPIC IMAGE

All Purpose A Light Purple It has fewer spots and a

Flour purple color

B Dark Purple It has numerous spots

that are gathered
together in a dark purple
color

C Off-white Unclear, off-white color

surrounded by many dots
Cornstarch A Dark brown Brown spots with
translucent colour and
scattered

B Dark purple Numerous purple spots

are conjugate together

C Dirty white Small, almost tiny, black

spots are scattered
Rice starch A Light red Black with fewer spots
scattered

B Dark red Black with lesser spots

scattered

C Translucent Dark black with many

spots that are forming
bonds

Where: A = distilled water, B = HCl, and C = NaOH

▪️ Starch in a sample can frequently be detected using an iodine solution. The samples are diluted
with iodine solution for the test, and the color change is then observed. The formation of a
complex between iodine and starch molecules can explain how an iodine solution affects the
color of starches. In order to start, we added distilled water to each sample of starch. To observe
how adding iodine solution to distilled water affected the color of the starches in each of the
three samples. The color will change similarly to how light purple or bright red did in the past.
They claimed that the amylase enzyme works more quickly the sooner the blue hue of starch is
gone. It demonstrates that iodine was able to pass through the membrane and interact with the
solution's starch. The starch solution changed from its initial milky white appearance to a dark
purple / black hue.

▪️ The iodine molecules can fit into the starch molecule's helical structure and create a complex
that absorbs light in the visible spectrum, which causes the colour shift. The complex has a
blue-black appearance because most of the light that is absorbed is in the blue and violet
spectrum. Moreover, when mixed with HCl, iodine solution significantly affects the color of
starches. Acid treatment caused damage to the starch's microcrystals by dissolving the glycosidic
bond, lowering the starch's molecular weight, reducing the amount of amylopectin, and
increasing the amount of amylose with true hydroxyl groups. When the concentration of HCl
reached 23%, the binding strength was at its highest point.

▪️ The amount of starch in the sample directly correlates with how intense the blue-black hue is.
The hue becomes more intense as the starch content rises. On the other hand, there won't be any
color change when iodine solution is added if the sample doesn't include any starch. Similar to
when NaOH was added to the samples, there is no difference. It is merely a process of
gelatinization. This suggests that the starch gelatinized as a result of the greater NaOH
concentration, that the starch's EH-type crystal structure was broken by the NaOH, and that this
led to amylose leaching and the formation of an amorphous structure.

In conclusion, a complex between the molecules of iodine and starch is what causes an iodine
solution to affect the color of starches. The ensuing blue-black hue serves as a helpful clue if
starch is present in a sample.

Conclusion

Since roughly 400 years ago, microscopy has been a fundamental instrument of science, and it
continues to be one of the most popular techniques in the material and life sciences. The
development of microscopic techniques has made it possible for us to get comprehensive data on
the specimen being studied. The current experiment has contributed to our understanding of the
microscopic examination and impact of many factors on starch structure. It became clear that the
concentration of distilled water, HCl, and NaOH had a significant impact on how all starches
changed color in response to the addition of iodine solution. Moreover, heating the starches has
the potential to change their consistency and cause them to gelatinize.
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