Nursing Biochemistry

Laboratory Report

Biochemical Processes
Activity No. 2

Amiruddin, Algenne Marhee P. (Principal Author)

Daud, Ayesha Munira A.
Del Rio, Gianella B.
Locker No.31; NurBio Lab C
1st Semester, SY 2019-2020
Nursing Biochemistry
Laboratory Report

RATIONALE
The main focus of activity 2 is to define and determine the different biochemical processes through the
performed experiment. There are four biochemical processes conducted in this laboratory activity namely
Dialysis, Osmosis, Diffusion, and Surface Tension.

Dialysis is a process used to separate colloidal particles from dissolved ions or molecule through a
semipermeable membrane (Lau, 2019). Semipermeable membrane is defined as a membrane that allows
some molecules to pass without letting the other unnecessary things to pass. An example of it is the
cellophane which we used in the experiment. It prevents the loss of proteins from body fluids while
excreting liquids, salts, and small waste molecules. Dialysis can also help impaired renal function
(Dialysis, n.d.).

Osmosis is one of the most important processes in chemistry and biology. It is the process where solvent
molecules move through a semipermeable membrane from a dilute solution into a more concentrated
solution (Helmenstine, 2019). It equalizes concentration on both sides of a membrane. An example for
this is when red blood cells are placed into fresh water.

Osmosis and diffusion go hand in hand. Osmosis can be referred to as the diffusion of water while
diffusion is the motion of matter from a region of high concentration to low concentration. An example
for diffusion would be the spreading of a drop of food coloring evenly through a liquid.

Surface Tension is defined as the capability of the property of the surface of a liquid to resist an external
force which is because of the cohesive nature of water molecules (Georgia State University, n.d.).

The group came up with three objectives for this laboratory activity. First is to elucidate biochemical
processes and its definition. Second, to perform the biochemical processes and list the essentials of the
laboratory activity. Last, to understand the importance of these biochemical processes in our lives. It is
important to conduct this activity because we get new knowledge and learnings about the topic.
Biochemical processes is said to give rise to the complexity of life. It gives knowledge about the
processes that occur within living cells, which we can relate to the study and further understanding of
tissues, organs, and its functions.

DATA INTERPRETATION

There were four biochemical processes performed in the laboratory: (1) dialysis, (2) diffusion, (3)
osmosis, and (4) surface tension.

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Nursing Biochemistry
Laboratory Report

A. Dialysis

SAMPLE REAGENT ADDED OBSERVATION

Dialysate + 25% Trichloroacetic No appearance of white

Test for Proteins
Acid precipitate

Appearance of a precipitate on
Dialysis + Fehling’s reagents A
Test for Sugars the surface and brick red near
&B
fire

In table A, the group presented the data and results of dialysis. Dialysis is the separation and purification
of particles in a liquid on the basis of differences in their ability, the proteins, to pass through a semi-
permeable membrane. In this case, the water cellophane served as the semi-permeable membrane where
small molecules of milk pass through the pores of the water cellophane. The goal of the experiment was
to test and to know if there were any sugars or any proteins present when the reagent is added to the
dialysate. For the dialysate that was added a 25% Trichloroacetic Acid, the result was no appearance of
white precipitate. In the test for proteins, there was no appearance of white precipitate this is because of a
human error. It may be because of careless stirring of the milk dialysate; it had lapses because the interval
of stirs and stops were not the same. Also, another human error is that the doer of the experiment runs out
of time that is why the sensitivity of the test was not checked; thus having no basis for comparison. But
through the test that challenged the sensitivity of the test, there were precipitates.. On the other hand, for
the test for sugars, there was an appearance of a yellowish brown precipitate on the surface and a brick
red precipitate on the side where most amount of the fire was directed. This simply means that reducing
sugar is present due to the formation of yellowish brown and brick red precipitates.

B. Diffusion

SAMPLE REAGENT ADDED TIME TO PRODUCE UNIFORM COLOR

10% Methylene Blue 9 minutes and 30 seconds

Distilled Water
10% D and C Red #40 More than 30 minutes

Figure 1. (Left) Test for Protein, (right) Test for Sugar

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry
Laboratory Report

In table B, the group presented the data and results of diffusion. Diffusion is the movement of a liquid
from higher concentration to lower concentration. In this experiment, instead of using 10% Copper (II)
Sulfate and 10% Prussian blue, the group utilized 10% Methylene Blue and 10% D and C Red #40
respectively. As per the table below, it took 9 minutes and 30 seconds before the 10% Methylene Blue
and the distilled water produced a uniform color. On the other hand, it took more than 30 minutes to
produce a uniform color of the 10% D and C Red #40 and the distilled water. The concept behind this is
that the smaller the molecule, the faster it diffuses. As for this case, the molar mass of methylene blue is
319.85 g/mol while on the other hand, the molar mass of D and C Red #40 is 496.42 g/mol. Going back to
the concept, 10% Methylene Blue diffused faster than 10% D and C Red #40 because of its molar mass –
which is lighter than the latter.

C. Osmosis

SAMPLE REAGENT ADDED OBSERVATION

High power lens: Clear and white colored with

0.9 % NaCl solution
dots and black little particles; glittery

High power lens: The texture looks rough with

0.1M NaCl solution some scratches on it.
Figure 2. (Left) D and C Red #40, (Right) Methylene Blue
Greenish in color
Defribinated Blood

1.0M NaCl solution Lens 40x: The texture is patterned in circular form

High power lens: Plain and demonstrates a static

“Control”
picture

In table C, the group presented the data and results of osmosis. Osmosis is the movement of water or
other solvent through a plasma membrane from a region of low solute concentration to a region of high
solute concentration. In this experiment, the group prepared 4 test tubes. Each test tube contains 5 drops
of defibrinated blood. For test tube 1, it contains 5 drops of defibrinated blood and 0.9% NaCl solution.
For test tube 2, it contains 5 drops of defibrinated blood and 0.1M NaCl solution. For test tube 3, it
contains 5 drops of defibrinated blood and 1.0M NaCl solution. For the 4th test tube, labeled as “control”,
it serves as the basis of comparison for the other samples. Each sample was viewed under a microscope.
The observation for the sample with 0.9% NaCl Solution is that under a high power lens, the little objects
are clear and white with dots and little black particles; it also looks like glitters. A 0.9% NaCl solution is
said to be an isotonic because when blood cells exist in such location, the extracellular and intracellular

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry
Laboratory Report

fluids are in osmosis equilibrium. Next is that, the observation for the sample with 0.1M NaCl solution is
that in high power lens, it has a rough texture and greenish in color. The 0.1M NaCl solution is said to be
a hypotonic solution in which the cells bulged. It is to be considered that in a hypotonic solution, the
solute molecules inside the cell are greater than the solute molecules outside the cell. Next is that, the
observation for the sample with 1.0M NaCl solution is that under Lens 40x, the texture is patterned in
circular form. With the 1.0M NaCl solution, the cell experienced shrinking; thus making it a hypertonic
solution. Finally, for the “control” test tube, it is left alone so that the other samples may be compared to
it.

Figure 3. Test Tube 1 Test Tube 2 Test Tube 3 Control

0.9% NaCl 0.1M NaCl 1.0M NaCl

D. Surface Tension

OBSERVATION (TIME TO
TEST TUBE REAGENT ADDED COALESCE AFTER
SHAKING)

1 mL Distilled Water + 1mL Did not mix/immiscible almost

No. 1
Chloroform immediately after shaking

1 mL Distilled Water + 1 mL
The solution coalesce after 5
No. 2 chloroform + 1 mL Soap
minutes
Solution

In table D, the group presented the data and results of surface tension. Surface tension is a phenomenon in
which the surface of a liquid, where the liquid is in contact with a gas, acts as a thin elastic sheet. For the
first activity under surface tension, test tube no. 1 contains 1 mL distilled water and 1 mL Chloroform
while test tube no. 2 contains 1 mL distilled water, 1 mL chloroform, and 1mL soap solution. The only
difference is that test tube no.2 has a 1mL soap solution which is not present in test tube no. 2. The
observation for test tube no. 1 is that the particles were immiscible almost immediately after shaking.
While for the test tube no. 2, it coalesces after 5 minutes. Water is a polar solvent while chloroform is a
non-polar solvent. Like dissolves like that is why water and chloroform does not go together; chloroform

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry
Laboratory Report

is insoluble in water and will just form a layer when mixed. However, by the presence of soap solution, it
can be mixed together. Although it shows different layers (by having different colors of layer; blue on
top, apple green below), the soap served as an emulsifying agent that reduced the surface tension.

EVAPORATING DISH REAGENT ADDED OBSERVATION

After 2 minutes, tiny particles of

1 mL Dilute Bile Solution + 1
No. 1 sulfur formed below the diluted
Pinch Sulfur
bile solution

1 mL Distilled Water + 1 Pinch The sulfur powder floated on the

No. 2
Sulfur distilled water

In this part of the surface tension activity, it has been observed that with 1mL Dilute Bile Solution and 1
Pinch Sulfur is that it just took a few minutes for the tiny particles of sulfur is formed below the dilute
bile solution. On the other hand, in 1 mL distilled water and 1 Pinch Sulfur is that the sulfur powder

floated on it.

ANSWERS TO QUESTIONS
1. What are the types of transport processes by which metabolites through biomembrane?

ANSWER: The movement of materials across a biological membrane may occur in two ways
and these are actively and passively. The active transport involves the movement of materials

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Figure
Natural Sciences 4. Dilute
Department, Bile
College Solution
of Science andand Distilled
Information Water. Sulfur
Technology, Ateneo de Zamboanga University
powder is sprinkled on each sample.
Nursing Biochemistry
Laboratory Report

against a concentration gradient. In order for molecules to move across a membrane, it

requires that a cell expends energy such as ATP hydrolysis. There are two main types of
active transport and these are 1) primary (direct) active transport which includes the direct
use of metabolic energy to mediate transport, and 2) secondary (indirect) active transport
which includes coupling the molecule with another moving along an electrochemical
gradient. In passive transport, it involves movement of materials along a concentration
gradient and does not require the expenditure of energy. There are three main types of passive
transport; 1) Simple diffusion, 2) Osmosis, and 3) Facilitated diffusion. Simple diffusion
happens when small molecules pass through the lipid bilayer of a cell membrane (Darnell,
Lodish, & Baltimore, 1986). Molecules diffuse down a gradient, from an area of high
concentration to one with a lower concentration of the molecule (Bank, n.d.). According to
Stillwell (2016), Osmosis is a special type of diffusion, that is to say the diffusion of water
across a semipermeable membrane. Water readily crosses a membrane down its potential
gradient from high to low potential. Facilitated diffusion is defined as the movement of large
or charged molecules via membrane protein.

2. What is the effect of the following on red blood cells?

a) Hypotonic salt solution
ANSWER: If red blood cells are in hypotonic (lower concentration outside the cell)
solution, it will result in more water flowing into the cell than out which results in
swelling of the cell and potential hemolysis (bursting) of the cell (Soult, 2019).

b) Hypertonic salt solution

ANSWER: Red blood cells carry oxygen throughout the body and removes carbon
dioxide, transporting it to the lungs for a person to exhale (Freeborn, Cunningham, &
LoCicero, n.d.). Red blood cells that are placed in hypertonic (higher concentration)
solution, water flows out of the cell faster than it comes in which results in crenation
(shriveling) of the blood cell (Soult, 2019).

c) Isotonic salt solution

ANSWER: In an isotonic solution, the flow of water in and out of the cell is
happening at the same rate.

3. How does the kidney maintain the body’s internal environment?

ANSWER: The kidney’s role in our body is to maintain homeostasis, which to maintain the
internal environment of our body while constantly changing. There are three main
components of the kidney’s function in our body, (1) maintain the volume of body fluids, (2)

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry
Laboratory Report

maintain the balance of salt ions in body fluids, and (3) to excrete unhealthy molecules that
contain nitrogen such as urea, ammonia, and uric acid (CK12, 2019).

4. Explain how soap solution lowers the surface tension of fats.

ANSWER: According to Scientific American (2019), most liquid dish soaps are formulated
as surfactants. Liquid dish soaps can lower the surface tension of fats because naturally
surfactants have a hydrophilic end which means they love water and they also have a
hydrophobic end which does not like water but wants to link with fats (Hessong, 2018).
Soaps has both non-polar and polar ionic bonds which gives them the ability to lower the
surface tension of water and fats.

5. What is the role of bile salts in fat digestion?

ANSWER: According to Wilson, bile salts came from the hepatocyte cells in the liver and are
from cholesterol (2018). The bile salts help with the digestion of fats by breaking down large
molecules so that digestion occurs easily. Bile salts also helps our body absorb vitamins A, D,
E, and K.

CONCLUSION

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ullamcorper suscipit lobortis nisl ut aliquip ex ea commodo consequat.

REFERENCES
Bank, E. (n.d.). What Kinds of Molecules Can Pass through the Plasma Membrane through

Simple Diffusion. Retrieved from https://education.seattlepi.com/kinds-molecules-can-pass-

through-plasma-membrane-through-simple-diffusion-4587.html

Chemistry of buffers and buffers in our blood. (n.d.). Retrieved from

https://www.khanacademy.org/test-prep/mcat/chemical-processes/acid-base-
equilibria/a/chemistry-of-buffers-and-buffers-in-blood

CK-12. (n,d). Kidneys. Retrieved from

https://www.ck12.org/biology/kidneys/lesson/Kidneys-MS-LS/
Dialysis. (n.d.). Retrieved from https://www.davita.com/education/kidney-vocabulary/dialysis

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry
Laboratory Report

Darnell, J., H. Lodish, and D. Baltimore. 1986. Molecular Cell Biology. Scientific American

Books, Inc., New York

Freeborn, D., Cunningham, L., & LoCicero, R. (n.d.). What Are Red Blood Cells. Retrieved from

https://www.urmc.rochester.edu/encyclopedia/content.aspx?ContentID=34&ContentTypeID=160

Georgia State University. (n.d.). Retrieved from

https://usgs.gov/special-topic/water-science-school/science/surface-tension-and-water?qt-
science_center_objects=0#qt-science_center_objects

Helmenstine, A. M. (2019, May 04). Chemistry Basics: What Is a Buffer? Retrieved from
https://www.thoughtco.com/definition-of-buffer-604393

Helmenstine, A. (2019). Osmosis Definition in Chemistry. Retrieved from

https://www.thoughtco.com/definition-of-osmosis-605890

Helmenstine, A. M. (2019, May 07). The pH Level in Chemistry Distinguishes Acids From
Alkalines. Retrieved from https://www.thoughtco.com/definition-of-ph-in-chemistry-604605

Hessong, A. (2018, April 28). How does detergent break surface tension?. Retrieved from

https://sciencing.com/detergent-break-surface-tension-5452223.html

Lau, K. (2019). Dialysis. Retrieved from

https://chem.libretexts.org/Bookshelves/Physical_and_Theoretical_Chemistry_Textbook_Maps/S
upplemental_Modules_(Physical_and_Theoretical_Chemistry)/Physical_Properties_of_Matter/So
lutions_and_Mixtures/Case_Studies/Dialysis

Scientific American. (2014, March 13). Surfactant Science: Make a Milk Rainbow.
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rainbow/

Soult, A. (2019). Osmosis and Diffusion. Retrieved from

https://chem.libretexts.org/Courses/University_of_Kentucky/UK%3A_CHE_103_-
_Chemistry_for_Allied_Health_(Soult)/Chapters/Chapter_8%3A_Properties_of_Solutions/8.4%3
A_Osmosis_and_Diffusion

Stillwell, W. (2016). An Introduction to Biological Membranes (Second Edition). Retrieved from

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Nursing Biochemistry
Laboratory Report

https://www.sciencedirect.com/topics/medicine-and-dentistry/osmosis

Wilson, D. R. (2018, June 15). Everything you want to know about bile salts. Retrieved
from https://www.healthline.com/health/bile-salts

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University