TRANSPOSRT ACROSS THE MEMBRANE

Abiog M.L., Arcellana J.E., Pangilinan C.J.

INTRODUCTION
The membrane of the cell is semi-permeable. A semi-permeable membrane is a
membrane that serves as a barrier between the cell and the extracellular environment. It
allows certain molecules and ions to pass through it and prevent others from doing so
(Nelson D.L, Cox M. 2013). Its permeability properties ensure that essential molecules
such as glucose, amino acids, and lipids readily enter the cell, metabolic intermediates
remain in the cell, and waste compounds leave the cell. It regulates the flow of materials
through a two-way traffic (Singleton P, 1999). The two-way traffic allows the nutrients to
enter the cell and the wastes to leave the cell in order to sustain its metabolism and
provide electrical currency formed by the entry and exit of ions. The membrane also
gives mechanical strength, shape, and protection to the cell. In short, the selective
permeability of the plasma membrane allows the cell to maintain a constant internal
environment (Zipursky Ls et al. 2004).
Particles may enter the cell through active transport and passive transport. In
active transport, there is the need of energy since the substance will be transported in a
concentration gradient from an area of low solute concentration to an area of high solute
concentration. There are cases where the cell itself takes substances inside through
endocytosis. Endocytosis has 2 types, the phagocytosis and the pinocytosis.
Phagocytosis is the process by which large particles or solid matter is engulfed by the
cell. On the other hand, pinocytosis is the process in which liquid matter or dissolved
substances are ingested by the cell (MellmanI 1996). In passive transport, the
movement of biochemical and other particles across the cell membranes does not need
the input of energy from the cell (Alcamo, I. Edward 1997). Diffusion, osmosis and
dialysis are some common ways by which molecules move across the cell membrane.
Diffusion is the net movement of particles or ions from an area of high solute
concentration to an area of low solute concentration. The difference of concentration
between the two areas is often referred to as the concentration gradient, and diffusion
will continue until this gradient has been eliminated or when homeostasis is reached.
Since diffusion moves materials from an area of higher concentration to an area of
lower concentration, it is described as moving solutes "down the concentration gradient"
or the movement of materials and particles against the concentration gradient. A
distinguishing feature of diffusion is that it results in mixing or mass transport. There are
different factors affecting the rate of diffusion. First is the magnitude of concentration
difference across the membrane. Second is the size and shape of the molecules of the
solute. Third is the temperature. The lower the temperature, the rate of diffusion is
slower due to the decrease of kinetic energy of molecules (J. Philibert 2005). There are
two types of diffusion, the Simple diffusion which is the kinetic movement of ions and
particles through the spaces of membrane without the binding to a carrier protein along

the membrane and the facilitated diffusion which requires ions and particles to bind to a
carrier protein that will transport them to the cell (H. Mehrer; N.A. Stolwijk 2009).
Osmosis is a special type of diffusion of solvent particles across a membrane. It is the
spontaneous net movement of solvent through a semi permeable membrane in the
direction that will equalize the solute concentration on both sides. In this process, the
water molecules move from the solution that is more diluted to the solution which has
higher concentration. In osmosis, there is what we call the osmotic pressure which is
higher in a solution with higher solute concentration. Therefore the movement of solvent
will stop when the 2 solution reaches the equilibrium of when the osmotic pressure
became equal from the both side of the selective permeable membrane (Oxford
University Press 2005.). Dialysis is the process of separating molecules on the basis of
size and ability to diffuse through a semi-permeable membrane. In dialysis, big particles
or molecules like proteins will not be allowed to pass through the membrane. It makes
use of diffusion across a selective permeable membrane. In dialysis, parchment and
cellophane are used. They have small pores that allow the ion or crystalloid, such as
sugar and salt which forms true solutions with water, to pass through the membrane but
not colloidal particles because they have larger size. In line with this, The mixture of
crystalloid and colloids that is placed in a dialyzing bag and immersed in distilled water
dialyzed the crystalloids out through the cellophane and left the colloids inside the bag
(Reed, R 2007).
This scientific paper aims to let the learners know some type of biological
transport processes within the cell. The learners should also be able to understand and
relate the processes of osmosis, diffusion and dialysis to the selective permeability
property of the cell membrane. And to be knowledgeable on the tests that will confirm
the presence of sugar and protein in the dialysate of the experiment performed that will
allow the learners perform those kinds of tests.

MATERIALS AND METHODS

A. Diffusion
100 ml of distilled water and warm water were placed into 2 separate 250 ml
beaker. 2 to 3 crystals of Potassium permanganate were added to each beaker. The
test was observed and recorded.

B. Osmosis
A thistle tube, secured with cellophane on the large end and the other end caped
with a finger, were filled until about 10cm with 30% sugar solution. The thistle tube was

dangled in a 500ml beaker filled with distilled water with the level that coincides with the
sugar solution inside the thistle tube. Afterwards, the level of sugar solution inside the
thistle tube was measured with ruler for 2 hours in a 30-minute interval. The test was
observed and recorded.

C. Dialysis
1 square foot of cellophane was soaked in water, afterwards folded like a filter
paper to fit into a glass funnel. 50 ml of milk were poured into the cellophane and tied at
the edges with string leaving some airspace above the milk. The bag is suspended into
a 500 ml beaker that it almost touches the bottom. Distilled water was added into the
beaker equal to the surface of the milk.
The milk was dialyzed by stirring the water outside for an hour. Presence of
protein and sugar were tested using the mended dialysate.
1. Test for proteins:
1ml of dialysate was placed to the test tube by using pipette and 1 ml of
25% trichloroacetic acid solution was added. White precipitate or turbidity
confirms the presence of proteins in the solution.
2. Test for sugar:
1 ml of fehlings A and fehlings B were combined and heated in a boiling
water bath. 1 ml of dialysate were added then boiled for 1-2 minutes. Yellow or
red precipitate confirms the existence of reducing sugar in the solution.

RESULTS AND DISCUSSION

A. Diffusion

1. Distilled water + Potassium permanganate

Potassium permanganate merges slower. A decrease in
temperature will cause molecules to slow down. This will slow
diffusion and make the spread of molecules less even.

2. Warm water + Potassium permanganate

Potassium permanganate blends faster because the solution
used the energy from the heat to move around quickly

B. Osmosis
The sugar solution is unable to pass through the membrane
because of the higher percentage of solute, but the distilled water pass
through. There is a net movement of water toward the inside of thistle
tube. Due to incoming water molecules, the level of solution rises in
245 cm in the first 30 minutes.

C. Dialysis
The cellophane sheet has very fine pores. These pores
permit ions (or small molecules) to pass through but not the
large colloidal particles. When a solution containing dissolved
ions (electrolyte) or molecules is placed in a bag of permeable
membrane dipping in pure water, the ions diffuse through the
membrane

1. Test for Proteins

The solution was clear upon adding the 25% trichloroacetic acid solution.
This indicates that the proteins are not present in the solution. Big molecules like
protein are not allowed to pass through since the cellophane sheet has very fine
pores.

2. Test for Sugars

Yellow precipitate appeared in the solution confirming the presence
of reducing sugar.

Post-Lab Questions
1. Explain the effect of temperature on the rate of diffusion of solute particles.
Temperature affects the rate of diffusion. The warmer the water the faster the
diffusion rate, the colder the water the slower the diffusion rate. The reason is that
molecules are constantly moving. When they are heated, they use the energy from the
heat to move around and bounce off to each other.

2. How does passive transport differ from active transport?

Active and passive transport are biological processes that move oxygen, water
and nutrients into cells and remove waste products. Passive transport moves molecules
down the concentration gradient. It goes from high to low concentration and it does not
require energy. Passive transport also includes diffusion, facilitated diffusion and
osmosis. While Active transport, involves the passage of ions through a concentration
gradient with the use of ATP and contains endocytosis, sodium-potassium pump and
exocytosis. (Bettelheim F.A. etal. 2007)
3. Explain how renal insufficiency may be corrected by the use of dialysis.
The dialysis is a way for a person with renal failure to be alive. It gives their
kidney a natural rest that may result to the returning of the function of their kidney. But,
Dialysis is not intended to correct renal problems but to help support the body and
replace the kidneys function until the person either finds a donor, get better or die.
(Pendse S. 2008)
4. What will happen to a red blood cell if you put it in a hypertonic, hypotonic, and
isotonic solution?
When blood cells are bathed in a solution that is isotonic to them, they remained
unchanged. If a hypotonic solution is introduced, the cells experience an inflow of water
and usually burst or usually called hemolysis that is caused by osmosis. If hypertonic
solution is introduced, the cells lose water and shrivel. Shriveling that result from
osmotic loss of water is called crenation. (Patton. 2013)

SUMMARY

There are different types of biological processes within the cell. The experiments have
shown the types of biological processes named diffusion, osmosis and dialysis. In diffusion,
temperature has a significant impact on the rate of diffusion the warmer the water, the faster the
diffusion rate. So when warm water was added to potassium permanganate, the blending or
rate of diffusion is faster because of the energy of the heat that makes the particles to move
quickly unlike when distilled water were added to the potassium permanganate the movement of
the particles is not that fast. Next is osmosis. Osmosis is the movement of water from low
concentration to high concentration. That is why the sugar solution has risen because the
molecules of water passed through the membrane to the inside environment or to the high
concentration solution water molecules move from the solution that is more diluted to the
solution which has higher concentration because of the osmotic pressure that is present in
higher solute concentration. The movement will stop when equilibrium or the osmotic pressure
is already equal. And lastly the dialysis test. The cellophane has very small pores that large
colloidal particles like the milks protein or the casein cant make it through it. So when the test
for proteins were conducted to the water outside the membrane, the results has revealed that
the proteins in the solution were absent so there is no protein molecules that went through the
membrane to the outside, And when the test for sugar were done to the solution inside the

membrane, it appeared that the sugar particles were reduced meaning there is the addition of
water.

REFERENCES

Arcizet, et al. "Temporal Analysis of Active and Passive Transport in Living Cells".
Pp. 101-105
Alcamo, I. -Edward (1997). "Chapter 25: Passive transport". Biology coloring
workbook. Illustrations by John Bergdahl. New York: Random House. pp. 2425.
J. Philibert (2005). One and a half century of diffusion: Fick, Einstein, before and
beyond. Diffusion Fundamentals, 2, 1.11.10.
H. Mehrer; N.A. Stolwijk (2009). "Heroes and Highlights in the History of Diffusion"
(PDF). Diffusion Fundamentals. 11 (1): 132.
"Osmosis" Oxford English Dictionary (3rd ed.). Oxford University Press September
2005.
Bettelheim, F. A., Brown, W.H., Campbell, M. K. & Farrell, S. O. (2007).
Introduction to general, organic, and biochemistry (10th ed.). Cenage Learning.
Patton. (2013) Laboratory Manual Essentials of Seeleys Anatomy and physiology (8 th
ed) . McGraw- hill international edition pp. 18
Pendse S, Singh A, Zawada E. Initiation of Dialysis. In: Handbook of Dialysis. 4th ed. New York,
NY; 2008:1421

Reed, R (2007). Practical Skills in Biomolecular Sciences, 3rd ed. Essex: Pearson
Education Limited.
Singleton P (1999). Bacteria in Biology, Biotechnology and Medicine (5th ed.). New
York: Wiley.
Lodish H, Berk A, Zipursky LS, et al. (2004). Molecular Cell Biology (4th ed.). New York:
Scientific American Books.