Constituent of Blood

DR. NOOR AZLINA ABU BAKAR

Learning outcomes:
The student will be able to:

1. List the components of blood and their function.

2. Describe the physical properties and chemical
components of blood.
3. Discuss ‘hemopoiesis’ and the important of
erythropoietin.
4. Outline the chemical components of normal
haemoglobin variants.
The importance of learning blood
physiology
• Many bodily dysfunction reflect the alteration in the blood –
haematological changes. i.e.: intensity and progress of the
diseases.
• Easily accessible and readily available tissue sample for
investigations.
Haematology
• Is the study of blood physiology and blood disorders.

• Study of blood physiology includes:

• Study the different components of blood.
• Their formation.
• Their role in different body functions and dysfunction.

• Study of blood disorders:

• Pathophysiology of disease of blood and their management
• Pathological alterations in blood including non-haematological diseases,
Blood examination fundamental aspects:
1. If the bone marrow is producing adequate number of mature cell
of haematopoietic lineages.
2. If development of each haematopoietic lineages is qualitatively
normal.
3. Investigation or haematological diagnosis:
• Different blood counts.
• Smear examination.
• Study of bone marrow.
 Composition of blood
Blood

Cellular component Fluid component

• Red blood cells (Erythrocytes). • Plasma (55% of total
• White blood cells (Leucocytes). blood volume).
• Platelet (Thrombocytes). • Made up of water and
solid particles.
 Composition of blood
• Upper plasma
layer is
separated from
lower red cell
mass by thin
buffy coat.

Separation of blood into cells and plasma by centrifugation. On

the left showing the appearance of whole blood consist of RBCs,
WBCs and platelets.
Buffy coat
• The buffy coat contains:
• Leucocytes and platelets.
• In the diseases, abnormal cells are found in the buffy coat (i.e:
systemic lupus erythromatosus, primitive red blood cells (atypical
mononuclear cells, promyelocytes, metamyelocytes, blast cells,
megakaryocytes etc.)
• Buffy coat preparation – detection of bacteria, fungi or parasites
within neutrophils, monocytes or circulating macrophages.
Functions of blood
1. Respiratory functions
2. Transport medium
3. Temperature regulation
4. Excretory function
5. Water homeostasis
6. Acid-base balance
7. Immunity
8. Storage
9. Body colour
10. Nutritive functions
11. Oncotic pressure
Functions of blood : Respiratory functions
• Oxygen transport from lungs to tissues and carbon dioxide from
tissues to lungs.
• Important function of respiration.
Functions of blood: Transport medium
• Various hormones from various endocrine tissues or glands.
• Chemical substances.
• Nutrients absorbed from GI tract to all tissues of the body.
• Vitamins.
• Thus, blood regulates growth and metabolism of the tissues.
Functions of blood: Temperature regulation
• Conducts heat from the interior body to the surface through blood
vessels.
• Vasoconstriction preserve body temperature.
• Vasodilation facilitates heat dissipation from the body.
Functions of blood: excretion function
• Excretion of waste materials by transporting them to the kidney.
Functions of blood: Water homeostasis
• 55% of blood contains plasma.
• 92% of plasma contains water.
• Loss of water can occur via diarrhoea, vomiting, excessive sweating
– decrease in blood volume (hypovolemia).
• Hypovolemia activates mechanism to increase water content in
blood thus maintaining blood volume.
Functions of blood: Acid-base balance
• Blood contains plasma protein and haemoglobin (Hb).
• Protein and Hb buffer play an important role in maintaining acid-
base balance of the body.
• Bicarbonate in blood buffers the acids (H+).
Functions of blood: Immunity
• Blood contains cells that play primary role in cellular immunity.
• Contains antibodies for humoral immunity.
• Contains cells and chemical substances that take part in non-
specific body defences.
Functions of blood: Storage
• Storage site for electrolytes, nutrients, chemicals,
hormones, etc.
Functions of blood: Body colour
• Provides natural colour to the body.
• In anaemia / hypovolemia – Body turns pale.
Functions of blood: Nutritive functions
• Deliver nutrients – glucose, lipids, proteins, vitamins, minerals etc.
Functions of blood: Oncotic pressure
• The albumin present in blood exert osmotic pressure (oncotic
pressure) – control capillary filtration and prevents oedema
formation.

Water
Withdraw/ exert hydrostatic H2O dissipate
conserve H2O pressure out of capillary -
oedema

Plasma protein – Albumin Plasma protein – Albumin

Exert oncotic pressure Exert oncotic pressure
Blood volume
• Total blood volume : 70 – 80 ml/kg body weight.
• In adult male : 5 – 6 L.
• In adult female : 4.5 – 5.5 L.
• In children : significantly less volume than the adults.
Determination of blood volume
• Blood volume is determine by the ratio of plasma volume to the cell
volume separately.
• Normal ratio plasma to cell volume is 55:45.

Composition of plasma.
Water : 92%
Solids or solutes (8%) – Plasma protein make 90% of solids,

a. Inorganic
i. Anions : Chloride, bicarbonate, phosphates, sulphates, etc.
ii. Cations : Na+, K+, Ca2+, Mg2+, etc.

b. Organic
i. Colloids : Plasma proteins
ii. Crystalloids : Glucose, lipid, urea, uric acid, etc.
Measurement of cell volume
• Cell volume is measure by measuring the volume of red cells.
• The volume of leucocytes and platelet is ignored as its
constitute the minor fraction of the total cell volume.
• Method : Radioactive isotope study.

• Red cell volume – volume occupied by circulating red cells.

• Calculated by subtracting plasma volume from total blood volume.
• Directly calculated by injecting tagged red cell using radioisotope of
chromium (51Cr), isotope of iron (59Fe), Phosphorus (32P).
Determination of plasma volume
• Is measure by using dyes (Evans blue T-1824).
• Also measure using injection of serum albumin labelled with
radioactive iodine.
• Average plasma volume : 3.5 L (5% of the body weight of 70kg man).
• Total blood volume:

100
3.5 L x = 5.8 L
(100 – Haematocrit)

• Haematocrit = 40.
Effective blood volume
• Effective blood volume = Total blood volume – Volume that is
sequestrated.

• Means that the volume which present actually in the circulation.

Difficult to estimate the sequestrated volume of blood in the visceral
organs.
Specific gravity and viscosity of blood
• Specific gravity or density of whole blood is approximately 1.050.
• Depends on the number of cells and composition of plasma.
• Measured by the extend of light bending in a refractometer or part
of a test strip dipped into blood.
• Loss of water from blood (dehydration) increases the specific
gravity.
• Viscosity: is a measure of resistance to flow.
• Blood viscosity is 4 - 5 time that of water. (Due to the presence of
cells and macromolecules like protein in it).
Plasma 55%
(-200C)

• Is the fluid component of blood which constitutes about 55% of

total blood volume.
45%
• If whole plasma is to be used for investigation, blood is
collected with anticoagulants.
• Can be stored for years for future investigations.
• Fresh frozen plasma (FFP) stored below – 20oC.
• To preserve clotting factors, plasma should be frozen within 6
hours after collection.
• FFP used for plasmapheresis (therapeutic plasma exchange).
Composition of plasma
Protein (7%) Albumin : 54%
Globulin : 38%
Fibrinogen : 7%
Plasma Others : 1%
55%

Water (92%)

Electrolytes
Cell Nutrients
45%
Other Waste products
solute (1%) Regulatory substances
 Serum
• Serum = Plasma – clotting factors
• Plasma is separated by
centrifuging
anticoagulated blood.
• Serum is separated from
blood by allowing the
(Blood plasma Serum (no fibrinogen) blood to coagulate.
(contains fibrinogen)

Buffy coat
(WBC and platelets) Clotted blood

Red blood cells

Plasma protein : 3 types
Plasma protein are formed mainly in the liver.

1. Albumin : 4 – 5.5 g%
2. Globulin : 1.5 – 3 g%
3. Fibrinogen : 0.3 g%
Plasma protein : Albumin (4 – 5.5 g%)
• Major constituent of plasma protein.
• Formed in the liver.
• Half life : 20 days
• Molecular weight : 66000
• Smallest in diameter among plasma protein, in kidney disease or
glomerular injury it appears early in the urine (albuminuria).
• In kidney disease, albuminuria decrease the albumin plasma protein thus
causes hypoalbuminemia.
• Hypoalbuminemia can also occur in liver disease due to low production of
albumin. Which this can result in decrease colloidal osmotic pressure
leading to oedema.
Plasma protein : Globulin (1.5 – 3 g%)
• Formed in the liver.
• Divided into 3 categories:
1. ⍺ (⍺1 , ⍺2)
2. β (β1, β2)
3. "
• Molecular weight is 90000 – 156000.
• Its includes different transport proteins : transferrin, ceruloplasmin, hemopexin, etc.
• Form different lipoprotein in combination with lipids in plasma.
• Antibodies (immunoglobulin) are "-globulin.
• Normal albumin-globulin ratio (AGR) is 1.5 – 2.5:1.
• AGR is altered in many disease i.e. immunological and inflammatory disease.
Plasma protein : Fibrinogen (0.3 g%)
• Produced in the liver.
• Molecular weight : 340000
• Important role in blood coagulation.
• Contributes to the viscosity of plasma and determination of ESR.
Function of plasma proteins
1. Osmotic pressure
2. Viscosity
3. Immunity
4. Coagulation
5. Transport
6. Buffering
7. Protein store
8. Synthetic function
9. Determination of ESR.
Function of plasma proteins: Osmotic
pressure
• Osmotic pressure due to plasma protein is called oncotic pressure.
• Normal oncotic pressure is 25 mmHg (helps in maintaining volume of
vascular compartment – prevent loss of fluid from capillaries into the
interstitial space.
• Therefore, when oncotic pressure decrease due to
hypoproteinaemia (liver and kidney disease) oedema manifests
due to escape of fluid into the interstitial space.
Function of plasma proteins: Viscosity
• Plasma protein contributes to 50% of blood viscosity (depends on
the molecular shape of the plasma protein).
• Reason why fibrinogen molecules (elongated and fibrillar in shape)
contributes more to blood viscosity than albumin molecules
(ellipsoid in structure).
Function of plasma proteins: Immunity
• Antibodies are plasma proteins (!-globulin).
• Provides humoral immunity and prevent from infections
(extracellular pathogens or effect of toxic substances).
Function of plasma proteins: Coagulation
• Depends on concentration of fibrinogen that form fibrin thread.
• Other plasma protein for clotting factors is prothrombin.
Function of plasma proteins: Transport
• Serve as carrier molecule for various substance such as hormones,
drugs, metal, etc.
Function of plasma proteins: Buffering
• Forms an important buffering system of the body known as protein
buffers.
• Aids in acid-base balance.
Function of plasma proteins: Protein store
• Plasma protein serve as mobile protein reserve which can be utilized
for tissue growth especially during protein depletion.
Function of plasma proteins: Synthetic
function
• Provide substrate for synthesis of protein hormones like
erythropoietin, etc. and various enzyme.
Function of plasma proteins:
Determination of ESR
• Depends on the concentration of fibrinogen in the plasma.
• Fibrinogen facilitates rouleaux formation – increase rate of
sedimentation of red cell.
Separation of plasma protein
• Salt separation
• Paper electrophoresis
• Cohn’s fractionation
• Ultracentrifugation
• Immunohistochemical analysis, etc.

• Determination of concentration of various fraction is useful for

estimation of albumin-globulin ratio (AGR) and for diagnosis and
prognosis of different diseases.
Hemopoiesis
• Foetal life : occur in spleen and liver and subsequently in
red bone marrow (medullary cavity of all bones).
• From childhood red bone marrow is progressively
replaced by fat tissue (yellow marrow).
• In adult : hemopoiesis take place in vertebrae, sternum,
ribs, clavicles, pelvic bones, skull and end of humerus and
femurs.
Bone marrow
• Site of normal hemopoiesis / haematopoiesis (formation of
blood cells).
• Is an active process to maintain the normal number of
blood cells (red cell, white cell, platelets) in the peripheral
blood.
• Should be able to response to increase demands i.e.
haemorrhage, infection, etc.
Erythropoietin
• Is a hormones that regulate the production of erythrocytes.
• Is a glycoprotein secreted mainly by the kidney (85%) and liver (15%).
• In kidney: the cell found in the interstitium of the inner cortex and
outer medulla, lying outside the tubular basement membrane.
• Increases erythropoiesis by acting on bone marrow and foetal yolk
sac, liver, spleen.
• Its promotes:
• Differentiation of erythropoietic stem cells to proerythroblast
• Proliferation of committed stem cells
• Haemoglobin synthesis.
Factors promote erythropoietin secretion
1. Hypoxia
2. Alkalosis
3. Hormones (growth hormone, prolactin, thyroid hormones,
cathecholamines, corticosteroids, androgens.
4. Hemolysate (product released following red cell destruction)
Thank you