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Red Blood Cells
Basco, Vince; Cruz, Ma. Lizette; Guilaran,
Diane; Masangkay, Anne and Perez, James Benedict
Department of Biology, College of Science, Polytechnic University of the Philippines
Abstract
A cells permeability to a solute is dependent to the size, polarity and
charge of the particular molecule. The hemolytic data on salt NaCl, urea
(CH4N2O) and fructose (C6H12O6) solutes in red blood cells (RBC) was
measured by light transmission of blood in a spectrophotometer at 510 nm
and 100% transmittance (T). Percent (%) T was used as an indirect measure
of RBC volume and changes in cell volume as used as an indicator of the
osmotic response of red blood cells to solutions with differing osmolarity. RBC
exposed to NaCl and fructose solutes at low concentrations are hemolyzed
due to their high %T, while RBC in urea exhibited outstanding %T regardless
of its concentration. NaCl has the lowest %T, with fructose having middling
%T and urea having high %T. Thus, RBCs membrane permeability is high in
urea, moderate in fructose and low in NaCl. The three solute solutions have
a negative or indirect relationship with the % Transmission. Each specific
molecule has its own way of entering the cell membrane; they differ on the
transport mechanism which enables them to enter the semi-permeable
membrane.
Keywords: red blood cell, cell membrane, spectrophotometer, hemolysis
INTRODUCTION
Red blood cell is the most
used model for membrane-solventsolute interactions (Scott, 1993)
where
the
cell
membrane
permeability is test in solute
transport understands how it
affects
the
cells
metabolic
processes,
and
physiological
systems (Langsdorf and Zydney,
1994). Haemolysis on the other
hand is a process where the red
Hemolysis is accompanied by
the changes of light absorbance of
cells suspension where the RBC
membrane burst and it settle to the
bottom causing the solution to be
clear (Hess et al., 2005). Light
absorbance can be measured
though
spectrophotometer
to
determine
the
percentage
of
transmitted
light
absorbance.
Spectrophotometer
indirectly
measures the amount of light in the
particular wavelength absorbed by
the substance (Abramoff and
Thomson, 1986), this is supported
by Beers Law, where the amount
of light which is absorbed or
transmitted is usually proportional
to the concentration of the
particular molecule in solution so
spectrophotometer is a proper tool
to measure the percentage of
transmitted light absorbance.
The
objective
of
this
experiment is to determine the
properties
of
the
cellular
membranes and the movement of
the water molecule across them;
understand what concentration of
solvent is viable to hemolysis or
crenation; and determine the rate
of osmosis of RBC in different
concentration of urea, fructose and
NaCl.
METHODOLOGY
Preparation of Stock Solution
Blood samples were collected
using a syringe, contained in
heparin coated tubes and stored in
of
Solute
in
highest
percent
transmission.
However, 0.32 M concentration of
NaCl showed the lowest percentage
100
transmission
among
the
80
concentrations. NaCl solution with
0.02 M concentration showed the
60
highest percent transmission and
% Transmission
as the time increases percentage
40
concentration decreases. This is
probably because the RBC in the
20
solution hemolysed due to low
concentration of NaCl. On the other
0
0.02 0.04 0.08 0.16 0.32 hand 0.04 to 0.32 M has lower
transmission but has
Concentrations of NaCl (M) percent
increasing trend.
120
0 mins
15 mins
30 mins
120
100
80
60
% Transmission
40
20
0
15 mins
30 mins
160
140
120
100
% Transmission
80
60
40
20
0
15 mins
30 mins
Pearson r - values
% Transmission against Solute Concentration
NaCl
-0.75
Urea
-0.16
Fructose
-0.87
% Transmission against Time
NaCl
0.43
Urea
0.40
Fructose
-0.82
Table 4 shows the correlation
values of % Transmission against
concentration and time. Specifically
the table has shown negative
correlation/relationship between %
transmission and concentration in
all of the three solutes. The table
also shows that NaCl and Urea`s %
Transmission
have
positive
correlation with time while in
contrary,
Fructose`s
%
Transmission have a negative
correlation with time.
The results have shown an
indirect relationship between %
Transmission
and
solute
concentration
which
may
be
explained by the shared idea above
because as the concentration of
the solute increases, the attraction
of water to that solution also
increases which eventually lead
into the movement of water from
the cell into the solution resulting
into a decrease in %transmission or
low hemolysis rate.
Transports in and out the cell
happens most of the times but it
can only be done by specific
molecules along with their specific
transports. There are different
kinds of transport mechanisms
which can be observed in a cell
materials
through
cell
membranes. Pages 109121,
in Laboratory outlines in
biology. W. H. Freeman, New
York, 529 pages
Cadwallader, D. E., & Phillips, J. R.
(1969).
Behavior
of
Erythrocytes
in
Various
Solvent Systems V: WaterLiquid Amides. Journal of
Pharmaceutical
Sciences,
58(10),
1220-1224.
doi:10.1002/jps.2600581012
Freeman, W. H. (2000). Diffusion of
Small
Molecules
across
Phospholipid
Bilayers.
Molecular Cell Biology. 4th
edition.
Retrieved
from
http://www.ncbi.nlm.nih.gov/
books/NBK21626/
Goodman, B.E. 2002. Transport of
small molecules across cell
membranes: water channels
and
urea
transporters.
Advan. Physiol. Educ. 26:
146-157.
Hess, J.R., Kagen, L.R., van der
Meer,
P.F.,
Simon,
T.,
Cardigan,
R.
(2005).
Interlaboratory comparison of
red-cell
ATP,
2,3diphosphoglycerate
and
haemolysis
measurements.
Vox Sang 89:4448
Inouye, C.Y. 2012. Water and solute
movement through red blood
cell membranes. Biology3151
Laboratory Manual.