1A post lab questions

1. Explain the process of homogenization and the purposes of each solvent added.
a. Homogenization is the process wherein two substances that are insoluble to
each other become a uniform substance all throughout. In the experiment, these
two substances were the sucrose solution and the minced liver. The sucrose
solution was used as the solvent because it allows the tiny blended pellets of the
liver to be suspended in the sucrose to create a uniform solution. Furthermore,
sucrose prevents the organelles from disrupting because its molecules are too
big to enter the organelle or the cell unlike using water for homogenization, it
might enter the organelle which could lead to a lysis. We homogenized these
two substances by using a blender.
2. What is the significance of the different speed being used in each level of
centrifugation?
a. Using varying speeds of centrifugation allows us to separate certain substances
of varying densities. Substances with heavier densities tend to be sedimented
earlier on than less dense substances found in the solution.
3. What organelles are separated in each centrifugation?
a. Centrifugation at 2rpm separates the nuclei and cytoskeleton of the cell. Next at
5rpm, sediment may contain mitochondria, lysosome, and/or peroxisome.
Finally, at 8 rpm the sediment may contain microsomes or small vesicles.
4. Conclusions
a. We were able to separate/ isolate different components of the cell using
differential centrifugation. Furthermore, by identifying which organelles were
separated, we were able to recognize which biomolecules are the main
components of each organelle. For example the cytoskeleton is composed
mainly of protein biomolecules while mitochondria have a fat and protein
biomolecules that makes up its membrane.
1B

TESTS OBSERVATIONS RESULTS

DISCHE THE SAMPLES SHOWED NO CHANGE OF COLOR AND Absence of
REMAINED AS IS. either RNA or
DNA
MOLISCH SEDIMENT 1: STRONG PURPLE COLOR Sediment 1-3:
SEDIMENT 2: VERY FAINT PURPLE presence of
SEDIMENT 3: LIGHT PURPLE COLOR carbohydrates
SUPERNATAN 3: A DARK PURPLE COLOR WITH A HINT
OF RED. MIGHT BE MAROON
BIURET SEDIMENT 1: FAINT PURPLE Presence of
SEDIMENT 2: FAINT PURPLE protein
 SEDIMENT 3: STRONG PURPLE COLOR
SUPERNATANT 3: FAINT PURPLE ALMOST PINK-ISH
SUDAN SEDIMENT 1: FAINT RED Presence of
SEDIMENT 2: OILY SAMPLE. STRONG RED STAIN SETTLED fat
ABOVE SAMPLE
SEDIMENT 3: OILY SAMPLE. RED STAIN REACHED THE
BOTTOM OF THE SAMPLE
SUPERNATANT 3: FAINT RED. RED STAIN REACHED THE
BOTTOM OF THE SAMPLE ALMOST MIXING WITH THE
SOLUTION.

1. What are the substances tested in each test? What are its theoretical results?
a. Dische Test
i. Dische Diphenylamine test is used to detect the presence of DNA. This is
achieved through the interaction of diphenylamine and deoxyribose creating a
blue color. The intensity of the color is equals to the concentration of DNA
found in the solution. However, is presence of a greenish color is observed, that
might indicate the presence of RNA.
b. Molisch Test
i. This test allows for the detection of carbohydrates in the solution. This occurs
when the sugars found in the solution is dehydrated by the concentrated
sulfuric acid. This forms furfural (from pentoses) or hydroxymethyl furfural
(from hexoses). The furfurals then react with the Molisch reagent creating a
purple or violet color.
c. Biuret Test
i. This test detects the presence of peptide bonds in a solution thereby detection
of proteins. The copper sulfate reacts with the peptide bonds which forms a
chelate complex (bonding of ions and molecules to metal ions). This chelate
complex produces a violet color in alkaline conditions which is achieved through
the addition of sodium hydroxide.
d. Sudan III Test
i. This test is used to detect the presence of fat in the solution. This is achieved by
first extracting or isolating the lipid in the solution. Next, the lipid reacts with
the Sudan III reagent to develop a red color.
2. What biomolecules are present in the samples analyzed based on the results of your
experiment?
a. Based on our experiment the biomolecules present in our samples were carbohydrates,
protein, and fat. The presence of nucleic acids was not observed.
3. Explain the results gathered from each test in comparison to the theoretical or expected results.
a. The results gathered in Molisch, Biuret, and Sudan III showed the expected results of
what biomolecule we might find. However, in the Dische test for nucleic acid, we were
not able to find or observe the presence of DNA in our samples. This could mean that
some errors might have been present during the experimentation that could lead to this
result.
4. Conclusion
a. We were able to perform the different tests that could help us identify is there is
presence of biomolecules in our homogenized solution. In doing so, we observed the
presence of carbohydrates, fats, and proteins in our homogenized solution with the
exception of nucleic acids. With this experiment, we have become more familiar with
the different test that we might encounter in the future.