Josephine D.

Carandang
Zoo 120 ST-2L Group 1
DIFFUSION
Table 1.1 Summary of average diffusion rate based on different factors
MB
CR(C32H22N6Na2O
MO
(C16H18ClN3S)
(C14H14N3NaO3S)
6S2)
(cm/hr)
(cm/hr)
(cm/hr)
Agar
concentration
2%
5%

0.03096708
0.01856996

0.04989627
0.04788209

0.1
0.07916667

Temperature
29C, RT
0.03703704
0.03319502
0.11458333
9C, Refrigerator 0.0125
0.06458333
0.06458333
Discuss the effects of the ff factors on the movement of the dyes
a. Agar concentration
Two percent (2%) agar concentration resulted to a 0.0602877812 cm/hr
diffusion rate which is faster than that of the five percent (5%) agar
concentration which was only (0.0485395713 cm/hr). Physically, the particles
within the medium act as a barrier to diffusion. Collisions between diffusing
particles and the molecules of the medium lead to a reduction in the rate of
diffusion. This means that the greater the number of molecules or larger
particles within the medium, the lower the rate of diffusion.
b. Temperature
RT has higher rate of diffusion of 0.0616051304 cm/hr while ref/cooler
temperature only accounted for 0.0472222222 cm/hr diffusion rate. This is
because higher temperatures resulted in a faster rate of diffusion. Diffusion is
due to the random movement of molecules, and molecules move more quickly at
higher temperatures.
As temperatures decrease, molecular movement slows down, causing diffusion to
occur more slowly. At high temperatures, liquid water becomes a gas as the
molecules begin to move
rapidly and spread apart from each other. As temperatures cool, water molecules
slow their
movement and condense together to form liquid droplets.
c. Molecular weight
Methylene Blue
Molar mass: 319.85 g/mol
Formula: C16H18ClN3S
Methyl Orange
Molar mass: 327.33 g/mol
Formula: C14H14N3NaO3S
Congo red
Molar mass: 696.66 g/mol

Formula: C32H22N6Na2O6S2
Congo red has the heaviest MW, succeeded by methyl orange and the
lighest is the methylene blue. Based on the average diffusion rate, accounting
mainly the MW of the different dyes, methyl orange (0.0895833333 cm/hr) has
the highest diffusion followed by congo red (0.048889177 cm/hr) and lastly
methylene blue (0.0247685185 cm/hr). Theoretically, the larger the molecular
weight, the slower the rate of diffusion. This is because larger mass atoms have a
bigger physical appearance, thus decreasing the diffusion rate. Different result
obtained in the experiment may be due to possible sources of error, one of them
are bubbles in the agar gel and slanted formation of agar leading to possible
wrong reading of diffusion.
MOVEMENT ACROSS MEMBRANE
Table 1. 5. Relative number of stained yeast cells exposed to different
treatments.
Test Tube
Treatment
Stained cells*
A

+formalin

+heat

++

C
control
* relative# of stained cells

Figure 1.1 Yeast cell suspension with varying treatments

In this experimental set-up, it can be seen that relatively, the yeast
suspension with the least stained cell was from test tube A (treated with
formalin), followed by the test tube C (control) and the one with the most stained
cells was from test tube B (heated). The boiling treatment has the most stained
cell because as explained from the previous experiment, as temperature
increases the average kinetic energy of particles increases. Greater kinetic
energies lead to increased velocities. The increased velocity means that there is

a greater chance of collisions between particles, resulting in an increased rate of

diffusion. Generally, the rate of diffusion increases with temperature and thus
resulted to relatively high number of stained yeast cells. Formalin (30.031 g/mol)
treated test tube A, on the other hand, had the lowest relative number of stained
cells because formalin fixation is known to result to colour change or lost of
liveliness of color since is prone to polymerization, when it comes to fixation,
as a higher molecular weight and overall greater size mean a slower penetration
rate (molecular weight and rate of penetration are generally inversely
proportional to one another), plus the fact that the yeast cells are still alive. Test
tube C (control) also has a low count of stained yeast cells because they have
active yeasts that carry out active transport to remove stain from inside their
cells, so they will become colorless during the period of observation. When the
cell is injured by acid the dye penetrates more readily. Denatured yeast cells
absorb the stain, thus, the high stained cell count in test tube B.

OSMOSIS IB BIOLOGICAL MEMBRANE

Table 1.6. Weight (g) of eggs placed in different solutions for 24 hours.
Solutions
Day 0
Day 1
Day 2
Water (W)
180.94 g
188.28 g
189.79 g
Saline (Sa)
179.73 g
186.02 g
186.45 g
Syrup (Sy)
175.96 g
180.73 g
187.33 g
As shown in the table, the weight of the eggs in all the solutions increased
as days passed. Inside the egg membrane is a concentrated solution of proteins
and water. When the egg is soaked in distilled water, osmosis causes water to
diffuse into the egg to equalize the concentration of water on both sides of the
membrane, and the egg increases in volume, hence the increase from 180.94 g to
188.28 g after 24 hours. The egg soaked in saline solution increased too (179.73
g to 186.02 g) after 24 hours. However, the saline solutions increase was slightly
lower (6.29 g) than that of waters (7.34 g). Consequently, the egg in syrups
solution also increased after 24 hours (from 175.96 g to 180.73 g). This is
theoretically incorrect since, osmosis in a hypertonic (e.g.syrup)solution would
cause the water to diffuse back out of the egg, and the egg should have
decreased in volume or shrink because the concentration of water inside the egg
was higher than the concentration of water in the corn syrup, therefore the
water moved from the egg to the beaker. Given the fact that the shell-less eggs
were boiled and solid (meaning water content of egg decreased) may account for
this varying result. On the part of the third set-up, where in water was drained
from all the set-ups, and refilled with 150 ml distilled water, all the eggs still
increased in size. This is because the egg increases in size the longer you leave it
in the water until the water concentration inside and outside the egg is equal.
One possible source of error in this experiment may be mishandling of the egg
that could lead to breakage and would lead to either smaller or bigger weight
and size of the egg.

OSMOTIC PRESSURE IN RBC

Criterion to establish hemolysis? degree of transparency of water-blood solution
Table 1.7. Hemolyzing concentrations and isotonic coefficient i of different
solutions
Solutions Molecular Weight
[Hemolyzing]
i*
KCl
74.5513 g/mol
0.05 M
4
CaCl2
110.98 g/mol
0.05 M
4
Na2SO4
142.04 g/mol
0.01 M
2
*Hemolyzing concentration of sucrose (342.2965 g/mol)= 0.2 M
The non-electrolyte is the sucrose while the three reagents (KCl, CaCl2,
Na2SO4) are all electrolytes. Na2SO4 has the lowest isotonic coefficient (2) and
hemolyzing concentration(0.01M) which may be attributed to its comparatively
higher molecular weight (142.04 g/mol). On the other hand, KCl and CaCl2 has
the same isotonic coefficient (4) and hemolyzing concentration [0.05M] with
74.5513 g/mol and 110.98 g/mol molecular weight respectively. Generally,
molecular weight of a substance gives some idea of the approximate diameter of
its molecules -- the larger the molecule the greater the molecular weight. Steric
configuration and symmetry, or its lack, also influence the effective diameter of a
molecule. Very large molecules (e.g. proteins) have great difficulty entering a
cell, while small molecules (e.g. amino acids) may enter cells more freely.
The molar concentrations of non-electrolytes that prevent hemolysis are
approximately equal.
Osmotic pressure depends on the number of particles in solution. Since
electrolytes dissociate in aqueous solution, hemolysis will occur at lower molar
concentrations for electrolytes than for non-electrolytes.
Solutions of nonpenetrating nonelectrolytes (e.g., sucrose) cause hemolysis (due
to influx of water by osmosis) at approximately the same molar concentrations.
This is because these solutions have the same number of molecules per liter.
Thus nonelectrolyte solutions of the same molar concentrations demonstrate the
same osmotic pressure. On the other hand, solutions of nonpenetrating
electrolytes (e.g., NaCl) cause hemolysis (due to influx of water by osmosis) at
lower molar concentrations than the nonpenetrating nonelectrolyte. This is
because an electrolyte can dissociate into two ions (e.g., NaCl into Na+ and Cl-),
and every ion in the dissociated solution exerts the same osmotic pressure as is
produced by the entire NaCl molecule. Therefore, at the same molar
concentrations, there would be more molecules per liter in the electrolyte
solution than in the nonelectrolyte solution and the solutions would demonstrate
different osmotic pressures. Further, different electrolyte solutions can vary in
the osmotic pressure they exert, depending on the degree of dissociation of the
particular electrolyte in the solvent.

APPENDIX