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Lab Report
Lab Report
AP Biology
Schulte
Experiment # 1
The effects of acids and bases, inorganic salts, organic solvents, and temperature on the physical
properties of proteins
Abstract
In part A of this lab, the experimenters were testing the effect of strong acids and bases.
First, three test tubes were prepared which contained a different protein solution in each one:
albumin, casein, and gelatin. It was then tested the effect of the acid’s, hydrochloric acid,
addition to these three solutions. Out of the three solutions, casein was the most reactive to the
hydrochloric acid and gelatine had been the least reactive due to the lack of change in the
gelatine as it had only became slightly cloudy while the casein obtained an oily texture and much
precipitate. Albumin became slightly cloudy but not as much as casein had. NaOH was also
added to these three solutions, after being rid of the hydrochloric acid, in increasing amounts.
Most of the reactions remained the same but albumin had become slightly more reactive. This is
because the enzymes and substrates are more reactive between albumin and sodium hydroxide.
Copper sulfate, silver nitrate and isopropyl alcohol was added to three different test tubes which
contained 2% albumin. They all had different reactions but silver nitrate seemed to have the
most reaction which caused albumin denaturation.
In part B, 10-mL of 2% albumin was added to a 50-mL beaker and made a mixture with
25-mL of saturated ammonium sulfate. The mixture was then filtered and the filtrate was
collected. Three test tubes were created: albumin solution, the filtrate, and a small portion of the
precipitate from the funnel with distilled water. The experiment called for the addition of
ammonium sulfate to each of the test tubes. The results varied greatly as the colors and textures
changed for each. Test tube 2 was the most reactive as it had changed color to a periwinkle blue
and became thicker and cloudier. This is because the substances were ‘salted out’ or purified
and the precipitate was removed.
In part C, a test tube was placed in a water bath and increased the temperature while
recording observations of the albumin solution. Data was recorded at 38, 48 and 88 degrees
Fahrenheit during times when precipitate first appeared, the solution appeared milky white, and
at the very end. There was more of a reaction when the water both had reached 88 degrees
which might be due to enzyme denaturation.
The purpose of this lab is to observe the reactions of the enzymes when affected by
different factors such as acids, bases, and extreme temperature.
Results
Test Tube 1 2 3
Test Tube 1 2 3
Results Blue, fizzy, little bit White, bubbly, foamy A little cloudy, there is
of precipitant a yellow tiny
Part A is shown in the graph above. The graph shows the results of both parts and how the
solutions reacted to the additives. The first chart reveals the effects of strong acids and bases and
the second chart shows the effect of inorganic and organic additives.
Part B
Observations
The chart above represents the data which was recorded in part B of this experiment. It is to
show how the solution had filtered or ‘salted out’ the contents in the different test tubes.
Part C
Temperature Additional Observations
The chart above represents part C of the lab. It describes what was observed as the temperature
in the hot water bath increased.
Analysis/Conclusion
In part A, the purpose of finding the effects of strong acids and bases was fulfilled as
noted that the more solution that was added would result in increased reactions such as a cloudy
color, precipitate being formed, and coagulation, and results of the test tube contents. My
hypothesis was if there was an increased addition of the acids and bases in the solution, then the
reaction will be greater; this was proven correct because there was more of a reaction in each of
the test tubes as HCL, the acid, and NaOH, the base, was added in increasing amounts. The
experiment was also to find the effect of inorganic and organic additives. My hypothesis was
fulfilled because I said if an inorganic additive is added to an organic solution then there would
be more of a reaction rather than if an organic additive was added to an organic solution. This
was showing in the results because the two inorganic additives, copper sulfate and silver nitrate,
had more of a reaction then the organic additive, Isopropyl alcohol. I believe there was some
error in this part of the lab. When AgNO3, an inorganic substance, mixes with an organic
substance, which in this case is albumin, it should turn into a darker color rather than the cloudy
white it had turned into. I believe this is because the experimenters put in an insufficient amount
of the AgNO3 or had not cleaned out the test tubes enough.
In part B, our purpose was to find whether or not the proteins were precipitated and
purified. Our purpose of this part of the lab seemed to be fulfilled as there was some purifying
done. Our solutions were salted out as solutions which had precipitate no longer did after the
sodium hydroxide and copper sulfate were added and the solutions were more clear than before
they were added. The public often see salting out in aqueous solutions that have high ionic
strength which are able to make the molecule unable to precipitate proteins. My hypothesis was
proven to be incorrect as I had predicted that if the ammonium sulfate was added, the solutions
would become cloudier and more precipitate would be added; however, I was proven wrong as
the solutions had become clearer.
In part C, the effect of heat on the various solutions was found. My hypothesis was
validated by the data as my hypothesis was that if there was an increase of temperature, then
there would be an increased reaction. It was validated because when the temperature increased,
these reactions listed were heightened. Extreme temperatures are able to denature an enzyme,
whether that be high or low. In the case of this experiment, an increasing temperature is seen.
Although it was not high enough to completely denature the enzyme, a reaction had still taken
place as the temperature increased. The experiment has achieved the slight denaturation of the
enzymes in this protein solution.
To continue this investigation, the method of how extreme cold temperatures affect the
solutions just as was done in part C will be tested. When enzymes denature, the cell is unable to
function due to the enzyme not being able to react with its substrate. This can cause the cell to
shut down, resulting in organs to shut down, harming a human and damaging their system.
Background research: The Nuffield Foundation has tested the reaction of enzymes in various
conditions. It was tested in hydrochloric acid with the catalase along with the effect of pH.
Background Research and Bibliography
Factors affecting enzyme activity. (n.d.). Retrieved December 07, 2017, from
http://www.nuffieldfoundation.org/practical-biology/factors-affecting-enzyme-activity
L. (2016, July 29). Salting Out. Retrieved December 04, 2017, from
https://chem.libretexts.org/Core/Physical_and_Theoretical_Chemistry/Thermodynamics/
Non-Ideal_Systems/Salting_Out