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Biological Macromolecules

Partnered Lab Report


Blaine Folsom and Kyle Goth
September 19, 2015

Introduction
The purpose of this experiment is to further our knowledge of macromolecules and the
four main classes of them; nucleic acids, carbohydrates, lipids and proteins. Nucleic acids
serves as a blueprint for proteins and the two types of them are DNA and RNA (Campbell et
al, 2012). Carbohydrates are a class of molecules which range from small sugar molecules to
large polysaccharides (Campbell et al, 2012). Lipids are compounds that do not mix well with
water (Campbell et al, 2012). Protein is a polymer of amino acids (Campbell et al, 2012).
Macromolecules are considered to be the most important molecules of all living things-from
bacteria to elephants (Campbell et al, 2012). Learning more about these macromolecules will be
achieved by testing a variety of substances to determine if they contain sugar, starch, lipids and
proteins.
Hypothesis

Exercise 1: If the water temperature is changing. Then the variable in the different test tubes will
react based on their glucose levels. If all samples are tested for sugar, then the glucose is most
likely to have a significant positive reaction to the Benedicts reagent. If all samples are tested
for starch. The variable is the different substance, and they will react to the Lugols iodine based
on their starch levels. Then the potato is most likely to test positive for starch/
Exercise 2: If all five samples are tested for proteins. Using Biurets reagent. The milk is most
likely to have a positive reaction. If all five samples are tested for lipids. Using the Sudan III.
The vegetable oil is most likely to have a positive reaction.

Materials and Procedure:


Exercise 1: First, put on safety glove and goggles. Use a graduated cylinder to measure 10 mL of
distilled water and transfer it in to the unknown dropper bottle, by removing the dropper lid.
Then replace the lid and cap before gently shaking until the unknown substance is fully
dissolved. Now, carefully crack open an egg and separate the white from the yolk. Collect the
yolk in a 9-oz plastic cup. Use a marker to label one of the jumbo pipets albumin, place it into
the egg white and whisk the egg white with a fork. Next use a marker to label six test tubes;
dH20, albumin, unknown, milk, glucose and egg white albumin stock. Hold the test
tubes by placing them in the 24-well plate. Using the albumin pipet, measure and transfer 0.5 mL
of egg whites from the cup into the tube labeled egg white albumin stock. Using the graduated
cylinder measure 4.5 mL of distilled water and pour it into the egg white albumin stock test tube.
Then, gently swirl the tube to mix the stock and using the albumin pipet transfer 1 mL of egg
white albumin stock into the tube labeled albumin. Next, label a jumbo pipet dH20 and use it
to transfer 1 mL of distilled water to the correct tube. Now, label a short pipet milk and transfer
1 mL of 2% milk to the correct tube. Next, add 24 drops of unknown in to the labeled test tube
and 24 drops of glucose in to the labeled tube. Record your observations about the color of each
of the samples on the corresponding table in the Lab Report Assistant.
Now, using the method of your choice boil water. Once boiling and using an oven mitt if
preferred, carefully fill a 250 mL glass beaker about half full with the water and set aside. Add
40 drops of Benedicts reagent to each of the five test tubes. Carefully place all of the tubes into
the beaker of boiling water and let them sit for ten minutes. After, ten minutes carefully remove
the tubes and return them to the well. In order to determine if a reducing sugar was present use
the provided Benedicts reagent color chart and record your observations about the color of each

sample on the corresponding table in the Lab Report Assistant. After making observations, pour
contents of all five test tubes (NOT the egg white albumin stock), down the drain with running
water and wash and dry the tubes.
Next, re-label the clean tests tubes as dH20, albumin, unknown, milk and
starch and place them in the well-plate. Now use the labeled pipets to add 1 mL of distilled
water to its correct tube and 1 mL of milk to its correct tube. Add 24 drops of Starch from the
dropper bottle to the labeled tube and 24 drops of unknown to the labeled tube. Use a plastic
spoon to collect a small amount of raw potato, leaving the potato in the center of the spoon.
Record your observations about the color of each of the samples in the tubes and the potato on
the corresponding table in the Lab Report Assistant. Label a clean graduated pipet with IKI
and then use the pipet to transfer one drop of IKI from the glass bottle into each of the five tubes
and the raw potato. Record your observations about the color of each of the samples on the
corresponding table in the Lab Report Assistant. If the sample turns black, this indicates the
presence of starch. After making observations, pour contents of all five test tubes (NOT the egg
white albumin stock), down the drain with running water and wash and dry the tubes.
Exercise 2: Use the marker to label four clean test tubes, dH20, albumin, unknown and
milk, then place them in the well-plate. Using the albumin pipet measure 1 mL of egg white
albumin stock solution into the test tube labeled albumin. Now use the labeled pipets to add 1
mL of distilled water to its correct tube and 1 mL of milk to its correct tube. Add 24 drops of
unknown from the dropper into the corresponding tube. Record your observations about the color
of each of the samples on the corresponding table in the Lab Report Assistant. Add 5 drops of
Biurets reagent to each of the four samples. Then record your observations about the color of
each of the samples on the corresponding table in the Lab Report Assistant. Then allow the

samples to incubate for five minutes and record your observations about the color of each of the
samples on the corresponding table in the Lab Report Assistant. The presence of a purple color
indicates a positive for protein. Record your results on the corresponding table.
Now, place a piece of aluminum foil on a flat surface and place one piece of round filter
paper on top of the foil. Using a jumbo graduated pipet, carefully (it can stain clothing) draw up
Sudan III and place one drop in five places, about an inch apart on the filter paper. Allow each of
the drops to dry completely and then using a pencil, not a pen, outline each of the five circles.
Then label the circles as follows, unknown, albumin, distilled water, milk and vegetable
oil. Now using the labeled pipet add one drop of distilled water to the corresponding circle. Use
the milk pipet to add one drop of milk to the correct circle. Use the albumin pipet to add on drop
of albumin to the correct circle. Add one drop of unknown to the correct circle. And add one drop
of vegetable/canola oil to the corresponding circle. Allow the reactions to occur for about 30
minutes. A positive lipid test is indicated if the red color carries outside of the pencil line. After
30 minutes, make your observations about the five reaction drops and record your observations
of each of the samples on the corresponding table in the Lab Report Assistant. From these
observations make notes about the presence of lipids in the appropriate section. Wash all of your
equipment and return to the kit.

Results
In exercise 1A, we where testing to see if the reducing of sugar was present throughout
the various items compounds that we used. The reducing of sugar reducing sugar is a sugar that
acts as a reducing agent and can donate electrons to another molecule. Specifically, a reducing
sugar is a type of carbohydrate or natural sugar that contains a free aldehyde or ketone group.
Reducing sugars can react with other parts of the food, like amino acids, to change the color or
taste of the food.(Rizzo, "The Definition of Reducing Sugars" September 18, 2015). In this
experiment there was a positive and a negative control in each experiment. In the first one
positive control is glucose as we are testing for the reducing of sugar this was to be expected in
the negative control. After adding Benedict's Reagent and setting the test tubes into the boiling
water. After 5 minutes the results were in. In the first round of testing we found that the dH O,
2

unknown and the albumin didnt contain any of the sugar reducing properties they remained the
clear color blue from start to finish. The milk and glucose did show sugar reducing properties as
they changed color after adding the Benedicts Reagent and placing in the boiling water. The
milk turned a yellowish-peach color, and the glucose turned a orangish-red color. This shows me
that milk and glucose has sugar reducing properties.

Sample

Initial Color

Final Color

Reducing Sugar Present in


Sample? Y or N

dH O

Clear

Blue- Clear

No

Albumin

Clear with
white chunks.

Dark Blue with


white chunks.

No

Unknown

Clear

Blue- Clear

No

Milk

White

Peach/Yellow

Yes

Glucose

Clear

Orangish-Red

Yes

In exercise 1B, were testing for the presence of starch in several different items. In this
experiment that there was also a positive and negative control. The positive control was the
starch and the negative control was once again was dH O. After adding the IKI solution we
2

watched for the items to turn black. Black indicates the presence of starch. The water, albumin,
unknown, and milk where are negative for starches. They all remained the brown color from
when the IKI solution was added. The starch and the potato both turned black showing the
presence of starch.

Sample

Initial Color

Final Color

Starch Present in Sample? Y


or N

dH O

Clear

Brown-Clear

No

Albumin

Clear and white


chunks.

Brown-Clear

No

Unknown

Clear

Clear with a trace of


brown

No

Milk

White

White

No

Starch

Cloudy White

Black

Yes

Raw
Potato

White

Black

Yes

In exercise 2A, we are looking to see if any of the items that we have contain protein. We
once again use dH2O as the negative control and the positive control is milk. After adding the
Biurets Reagent to the different things the changes began. The different compounds will turn
purple if there is protein. The dH O remained unchanged. The albumin, milk, and unknown all
2

contained protein in some way. These all turned a purple or purple mixed in with them.

Sample
dH O
2

Initial
Color
Clear

Intermediate Color

Final Color

Protein Present in
Sample? Y or N

Clear

Clear

Albumin

White and
Chunks

Purple- Clear

Purple- Clear

Unknown

Clear

Purple- Clear

Purple- Clear

Milk

White

White with a layer of


purple on top.

Purple and
White mixed.

In exercise 2B, we are looking to see if any of the various compounds contain lipids. We
first took and put the compounds on filter paper that contained Sudan III dots. After taking and
putting the compounds on the filter paper we let them set for 30 minutes to observe if they
contain lipids. If they spread outside of the pencil marks that we placed then it contains lipid.

After observing we found that the dH O, albumin, and the unknown didnt contain any lipids.
2

The milk and vegetable oil contained lipids.

Sample

Sample Observations

Lipid Present in
Sample? Y or N

Distilled
water

Stayed the same from beginning to end. The circle


didnt go outside of the pencil mark.

Albumin

It stayed about the same red and didnt spread outside


of the circle

Unknown

There was no change that I could observe.

Milk

Spread outside of the circle and become a lighter red.

Vegetable
oil

Light red and went slightly outside of the circle.

Conclusion
Throughout this experiment we have learned the many different types of macromolecules
that are found in various things and how they react with different compounds of life.
So if all samples are tested for sugar then the glucose is most likely to have a significant
positive reaction to the Benedicts reagent. Through exercise 1A we have proved this hypothesis
to be a true hypothesis. Because we know that glucose is a form of sugar and is most likely going
to have a sugar reducing property of some type. So if all samples are tested for starch then the
potato is most likely to have a significant positive reaction to the Lugols iodine. In this
experiment this for the most part is true. There was one other thing besides the potato that you
would expect to show starch and that was the starch itself. So the hypothesis is still correct for
this exercise. So If all samples are tested for proteins using Biurets reagent then the milk is most
likely to have a positive reaction. There were two other compounds that contained proteins in
this exercise. So this hypothesis is not really one that will stand. So If all samples are tested for
lipids using the Sudan III then the vegetable oil is most likely to have a positive reaction. This is
another one that does have one other compound that contains lipids but out of the two oil is most
likely going to contain lipids. The hypothesis can stand and be proven throughout the exercise.
The only thing that could have caused some issues was the fact that I was missing one of
my test tubes. I just substituted with the large test tube in order to complete the experiment. All
other things with the experiment went smoothly.

Works Cited
Campbell, N.A., Dickey, J.L, Reece, J.B., Simon, E.J. & Taylor, M.R. (2012) Biology concepts
& connections (7th ed.) San Francisco, CA: Pearson Benjamin Cummings.

Hands-On-Labs. (2015). HOL Courseware and Resources. Denver, CO. Web


Rizzo, Natalie. "The Definition of Reducing Sugars." LIVESTRONG.COM. Demand Media, Inc,
6 Mar. 2015. Web. 18 Sept. 2015.
"LabArchives, Your Electronic Lab Notebook." LabArchives, Your Electronic Lab Notebook.
LabPaq. Web. 19 Sept. 2015.

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