UNIVERSITI TEKNOLOGI MARA

FACULTY OF APPLIED SCIENCES

BACHELOR OF SCIENCE (HONOURS) BIOLOGY

LAB 4 REPORT

BLOOD PHYSIOLOGY I: ERYTHROCYTE FUNCTIONS

LECTURER: DR. SITI NUR HUSNA BINTI MUHAMAD

GROUP: RAS2013B
DATE: 7/12/2022
GROUP MEMBERS: 1. SITI NUR SHAZWANI BINTI SHUKOR (2021617436)
2. AIDA QASRINA BINTI NOOR HISHAM (2021453958)
3. EIDA ADLIENA BINTI QAMARUL AZMAN (2021476542)
4. AININ SOFIYA BINTI ABD RAHMAN (2021823294)
5. NUR FARAH LIANA BINTI MOHD ISA (2021834796)
6. NUR SYAWANIE BINTI ABDUL AZIZ (2021871398)

INSTRUCTIONS
1. Completed report sheets will provide the basis for your course work (lab report –
10%) assessment mark in BIO560.
2. Make sure your name is clearly written in the space provided on the front of this
report sheet.
3. In completing the report sheets:
a) Do not expand any written answer beyond the space provided.
b) Sketches and diagrams should fill the space provided and should be made
with a pencil.
c) All written work should be in ink.
4. Report sheets should be handed in for marking, a week from the date of this
practical.
5. Reports handed in late WILL NOT be marked.
1. INTRODUCTION

Plasma and cells compose up the blood, which circulates throughout the body. It
removes waste from cells and provides vital elements such as glucose, oxygen, and
hormones to cells and organs. The main components of blood are plasma, red blood cells,
white blood cells and platelets. The plasma contents in blood fluid are 55% which is plasma
made up from 92% water and the remaining 8% contains glucose, hormones, proteins,
mineral salts, fats and vitamins. Meanwhile the remaining 45% of blood is composed of red
blood, white blood, and platelets. The main function of the red blood cell is delivered and
eliminates oxygen from the lungs. Haemoglobin was transported through the body via
oxygen. The red blood cell has a four-month lifespan and the body will replace it regularly.
There are several functions of blood such as providing oxygen for tissues and cells,
supplying essential nutrients such as glucose, fatty acids and amino acids, eliminating waste
products including urea, lactic acid and carbon dioxide and controlling body temperature.
There are several diseases related to red blood cells which are known as anaemia. Anaemia
happens if there is a decrease in the amount or size of red blood cells or the amount of
haemoglobin in the blood, and is defined by a decreased capacity to carry oxygen. The
whole status of the erythrocytes must be examined to identify the causes of anaemia. There
are many ways to examine the haemoglobin such as haematocrit, blood haemoglobin
concentration, RBC count, RBC size and percent haemoglobin per cell. Hematocrit test (Hct)
determines the percentage of red blood cells in your blood. Due to their role in transporting
oxygen throughout the body, red blood cells are essential. Results of tests that show low or
high hematocrit levels could reveal blood diseases or other diseases. Second, haemoglobin
determination is known as a colorimetric technique like the cyanmethemoglobin method
which is typically used to measure blood haemoglobin (Hb). The results obtain a rough
estimate of the Hb value, other, simpler tests are frequently carried out in the lab. Lastly,
blood test is a complete blood count (CBC). It helps medical professionals in diagnosing a
variety of illnesses and conditions. It also looks for signs of pharmaceutical side effects in
your blood. This test is used by healthcare professionals to detect diseases and modify
treatment.
2. FLOW OF PROCEDURE ACTIVITY 1

1. Finger was cleaned and soaked in 70% alcohol with cotton wool. Finger was
pricked using a sterile lancet and a drop of blood was obtained. The drop that forms has
been wiped off and a second drop was allowed to accumulate.

2. The red-circled end of a heparinized capillary tube was touched to the drop.
The tube was holded in a horizontal position and the blood was allowed to enter until the
tube was one-half to three-fourths full.

3. One end of the tube was sealed by pushing it into a tablet of sealing
compound and it was rotated to form a plug.

4. The capillary tube was placed in a microhaematocrit centrifuge with the plug
end to the outside, and it was centrifuged at 9.000 rpm for 4 minutes.

5. The haematocrit value was read directly using the scale provided at the end
of 4 minutes.

6. The haematocrit value was measured the following way: The height of the red
cell column and the height of the cells plus plasma was measured in millimetres. The
haematocrite was calculated using the following formula.

Height of red cells (mm)

Hct (%) = x 100

Height of red cells and plasma (mm)
 ACTIVITY 2

1. Tallquist Method

This test uses a book of special Tallquist blotting papers and a colour
comparison chart having different intensities of red. These intensities
correspond to different concentrations of Hb found in human blood.

a. A drop of blood was obtained and place it on a piece of blotting paper.

b. Before the blood becomes dry or coagulated, its colour was matched with
the closest colour on the comparison chart. The number by each colour represents the
percent of Hb in the blood. The number is multiplied by the Tallquist standard of 16.5 g to
give you grams of Hb per 100 ml of blood.

3. RESULTS

Blood physiology I: Erythrocyte Function

NAME HAEMATOCRIT (%) Hb (g/100 ml)

Tallquist Method

SHAZWANI 67.7 231

Calculation of Haematocrit Percentage

Hct (%) = Height of red cells (mm) x 100

Height of red cells and plasma (mm)

= 21 mm x100

31 mm

= 67.7 %

Calculation of Tallquist Method

Tallquist standard: 16.5g

Shazwani : 14 x 16.5g

= 231 g/mL
 POST LAB QUESTION

a. What is anaemia? Which blood measurements provide information on a possible

anaemic condition?

Anaemia is a disorder in which there are insufficient healthy red blood cells to
transport oxygen to your body's tissues. Being anaemic, or having low
haemoglobin, can make you feel exhausted and frail. Complete blood count
(CBC) measurements is a blood measurement that can provide information on a
possible anaemic condition. This test is used to determine how many blood cells
are present in a sample of your blood.

b. Briefly explain the function of the following in erythrogenesis:

Vitamin B12 :Helps in formation of red blood cells and anaemia

prevention. It is also used in proliferation during
differentiation. In addition, Vitamin B12 deficiency prevents
DNA synthesis, inhibits purine and thymidylate synthesis and
leads to erythroblast death which results in anaemia from
inefficient erythroblast.

Erythropoietin : Vital hormone that stimulates the bone marrow to create red
blood cells in the body. It helps to maintain the balance of the
blood.

Iron : Essential for the production of haemoglobin that allows red

blood cells to transport oxygen throughout the body.

Its function is to absorb vitamin B , which is required for the

Intrinsic factor : 12

production of red blood cells.

c. Polycythemia (excess number of red cells) occurs in patients with chronic

emphysema. Explain the mechanism responsible for this response.

Polycythemia is the body's reaction to an increased demand for oxygen. It

occurs when haemoglobin is unable to absorb large amounts of oxygen
from the lungs. This could be due to decreased atmospheric pressure, as
at high altitudes, or to impaired pulmonary ventilation. The sustained
increase in red cells in people who live permanently at high altitudes is a
direct result of the environment's reduced oxygen pressure. Emphysema is
a chronic pulmonary disease that causes abnormal distension of the lungs
 with air, resulting in chronic hypoxemia which is low oxygen tension in the
blood and absolute polycythemia. Extreme obesity can also severely
impair pulmonary ventilation, resulting in absolute polycythemia, also
known as Pickwickian syndrome.

d. How does haemoglobin carry both oxygen and carbon dioxide in the blood?

Haemoglobin has an oxygen binding sites because haemoglobin is made up of

four subunits, which are two alpha and two beta. Each subunit contains an iron-
containing central heme group that binds one oxygen molecule, allowing each
haemoglobin molecule to bind four oxygen molecules. Because oxygen binds to
haemoglobin, it changes its shape. More H+ is produced and the pH decreases as
the level of carbon dioxide in the blood rises (at the tissue level). The increase in
carbon dioxide and subsequent decrease in pH reduces haemoglobin's affinity for
oxygen. The oxygen molecule separates from the haemoglobin molecule. Next,
carbon dioxide is transported in the blood from body tissues to the lungs by
binding to haemoglobin, with approximately 10% of the carbon dioxide binding to
haemoglobin. Carbaminohemoglobin is formed when carbon dioxide binds to
haemoglobin. The binding of carbon dioxide to haemoglobin is reversible. When
carbon dioxide reaches the lungs, it can easily dissociate from haemoglobin and
be expelled from the body.

e. Why is the inhalation of car exhaust fumes life threatening? Explain the physiology
involved.
Car exhaust fumes contain high levels of carbon monoxide (CO) gas. Carbon
monoxide is a gas that is both colourless and odourless. Carbon monoxide can
increase to dangerous levels when combustion fumes become trapped in a poorly
ventilated or enclosed space. The inhalation of this carbon monoxide will build up
in the bloodstream which can cause serious tissue damage. Poisoning involving
carbon monoxide is exceedingly risky and perhaps fatal. If you breathe in
excessive amounts of carbon monoxide, your body will start to replace the oxygen
in your blood with carbon monoxide. You might pass unconscious when this
happens, and in these circumstances, death is possible.
f. Why are the haematocrits, haemoglobin concentrations, and erythrocyte counts
generally lower for females than males?

Males have higher haemoglobin concentrations (hematocrit) than females. Due to

their greater capacity to carry oxygen. However, females have more muscle mass
in their bodies. Typically , erythrocyte counts in women are lower compared to
men because women lose iron during menstruation. Males have higher red cell
mass due to the higher tissue oxygenation efficiency per unit red cell count.

4. DISCUSSION Activity 1.

According to the result of activity 1, the calculation haematocrit percentage of the

subjects is 67.7%. The haematocrit percentage of subjects is quite high because the
range of normal level haematocrit percentage is from 36% to 45% and the average is
41% for the adult females. While for the adult males range of haematocrit percentage
starts from 43% to 49% and its average is 46%. The activity is done by doing
haematocrit test, the small samples of blood are used to get hematocrit and it was
evaluated by using microhematocrit centrifuge. Hematocrit is the percentage of red
blood cells in the total blood volume while hemoglobin is part of our red blood cell.

The result of a haematocrit test showing low or high hematocrit levels can be an
early indicator of certain conditions, help diagnose certain diseases and may be a
reflected response for certain treatment. A high hematocrit level can be an indicator
of several conditions such as polycythemia, heart disease, dehydration, and
smoking. Polycythemia is an uncommon disorder in which humans have too many
red blood cells and the human blood becomes too thick. The symptoms of
polycythemia may include headache, bleeding gums and heavy bleeding from small
cuts. The studies show that high hematocrit levels increase the risk of developing
cardiovascular disease which is heart disease. Dehydration can lower all the water in
the body , including the plasma in the blood which is lower plasma levels can
increase the ratio of the red blood cells to blood volume. Hematocrit levels can be
affected by smoking in two ways. Body may produce more erythropoietin by
smoking, the kidney hormone that stimulates red blood cell production and it will
make more red blood cells which mean higher hematocrit levels.

A hematocrit level that is lower than normal may be a sign that the human body
did not have enough red blood cells and there are many different medical conditions
such as anemia, leukemia, hemolytic anemia, hyponatremia and blood loss due to
injury or illness. There are factors that may cause hematocrit levels to drop which is
losing the blood from injury, the human’s bone marrow not making enough red blood
cells and the human red blood cells dying earlier than usual. Anemia is a condition in
which red blood cells are not enough. Leukemia is caused by lower red blood cells
than normal red blood cells and it is because leukemia cells in the bone marrow are
displacing healthy red blood cells. Hemolytic anaemia is the condition in which the
red blood cells are breaking down or dying faster than the body can replace them.
There are conditions for blood to be watered down which is hyponatremia because
there is too much water in the body. Blood loss can affect the number of red blood
cells and the lower red blood cells mean lower hematocrit levels.

To balance hematocrit levels, someone who has lower hematocrit levels can
consume iron-rich foods to improve the red blood cells such as red meat, liver,
spinach, peas, whole wheat bread and so on. The healthcare provider may also
suggest using over-the-counter iron supplements. They also may suggest changes
to our lifestyle such as quitting smoking or adopting a heart-healthy diet. To lower the
hematocrit, we must stay hydrated, avoid drinking alcohol , start moderate exercise
and so on.

Activity 2:

Based on the results obtained from our experiment for Activity 2, the subject had
achieved 231g/ml of the blood by using Tallquist Method calculation. The amount of
haemoglobin found in a person's red blood cell was determined by their haemoglobin
levels to carry oxygen. The complex protein known as haemoglobin, or simply
haemoglobin, is contained in red blood cells and aids in carrying carbon dioxide from
tissues to the lungs and circulating oxygen throughout the body.

The haemoglobin levels of a patient can be quickly evaluated using the Tallquist
haemoglobin scale. A small piece of Tallquist paper that has been soaked with a
sample of blood is used to compare with a Tallquist colour chart using the Tallquist
method of quantifying haemoglobin The number by each colour represents the
 percent of haemoglobin in the blood. In order to compare the blood's colour to the
colour scale and determine the haemoglobin concentration in blood under normal
and anaemic conditions, allow blood to be absorbed into one of the test papers.

A quick point-of-care diagnostic assay, the haemoglobin colour scale, determines

haemoglobin concentrations between 4 and 14 g/dL. According to the haemoglobin
colour scale, it was observed that the blood stains of our subject matched the
number shades of red which was 14 g/dl. This shows that the haemoglobin levels of
this subject are normal by the age of adult women. This is because sex, age, and
general health are often factors that influence normal haemoglobin levels in the
blood. Haemoglobin levels that are excessively low or excessively high in
comparison to normal haemoglobin levels can have several negative effects on one's
health.

High haemoglobin levels are very uncommon, whereas low haemoglobin levels,
or anaemia, are relatively prevalent and can affect persons of all ages. Nevertheless,
pregnant women and those dealing with a few other diseases are more likely to
experience them. Treatment for the underlying reason is typically necessary to treat
haemoglobin levels, whether they are high or low.

Conclusion :

To sum up, the result obtained from the haematocrit test was 67.7% which is
quite high from standard range of normal haematocrit. Range for normal haematocrit
for females is from 36% to 45% while for male from 43% to 49%. This test required
small blood samples. The results from the hematocrit test can be used to measure
certain conditions such as help to diagnose certain diseases. There are several
conditions that will occur if the level of haematocrit is high such as polycythemia,
heart disease, and dehydration. The research found that, a person with a high level
of haematocrit has a high risk of developing cardiovascular disease such as heart
disease. Lower level of haematocrit is the sign that the body does not have an
insufficient amount of red blood cells. There are several diseases caused by lower
haematocrit level such as anaemia, leukaemia, hemolytic anaemia and
hyponatremia. The normal level of haematocrit can be obtained if they consume iron-
rich food especially for those with low hematocrit while to decrease the level of
hematocrit, they should start healthy exercise and stay hydrated.
Result obtained from experiment 2 was 231g/ml by using Tallquist Method
Calculation. Haemoglobin is a protein complex which contains red blood cells and serves
to carry oxygen throughout the entire body. The Tallquist haemoglobin scale is
the fastest way to measure the haemoglobin . The Tallquist colour chart has been
used to measure the blood sample on the Tallquist paper. The percent of
haemoglobin can be determined by the number that represents each colour. The
scale of haemoglobin colour concentration between 4 and 14g/dL. The results from
haemoglobin colour matched the shades of number 14g/dl which is normal
haemoglobin level for adult women. There are several factors that influence the
haemoglobin level such as sex, age and general health.