Experiment 5 otbnabogilo

To Determine the Haemoglobin Content and

Oxygen Carrying Capacity of
Own Blood Sample
Aim:
To determine the Haemoglobin content and Oxygen carrying capacity of own blood
sample.
Requirements:
Sahli's Haemoglobinometer containing comparator, haemoglobin tube,
haemoglobinpipette, dropper and stirreraheees
Pricking needle
Absorbent cotton
0.1 N Hydrochloric acid (HCI)
Distilled water
70% Alcohol ubo

H160
140
120
100
80
W10 HCI 60
40
20

Haemoglobinometer
tube

Haemoglobinometer
(comparator)

20 cu.mm

Haemoglobinometer plpette
Fig. 5.1: Sahli haemoglobinometer with accessories
(5.1)
 To Determine the Haemoglobin content ...
Human Anatomy and Physiology (Practical)
5.2

Theory:
erineiple next pa )
Hemoglobin (Hb) is a conjugated protein present in the red blood cells.
The haemoglobin molecule is made up of heme and globin.
constitutes about 4% of the
Heme is a pigment that contains one iron atom which
haemoglobin molecule while, globin is acolourless protein constituting about 96%
of total haemoglobin molecule.
It consists of two alpha (c) and two beta (B) polypeptide chains.
from iron, amino
The synthesis of hemoglobin takes place in the red bone marrow
acids, ribonucleic acid and enzymes.
the lungs to tissues
The function of hemoglobin is transportation of oxygen from
and carbon dioxide from the tissues to the lungs.
oxygen molecules. it is
When the one hemoglobin molecule combines with four
called as oxyhemoglobin.
One gram of hemoglobin molecule carries 13.4 ml of oxygen.
called as reduced
Hemoglobin molecule returning carbon dioxide from the tissues is
hemoglobin.
Abnormal Hb.
Hemoglobin ismainly divided into two types; Normal Hb and
o Normal Hb: Adult Hb, Fetal Hb and Embryonic Hb.
Hb.
o Abnormal Hb: Hb S, Hb C, Hb D, Hb Eand Unstable
such as;
Different methods are available for determination of hemoglobin
Visual methods: Sahli's method
Gasometric method
Spectrophotometric method
Automated hemoglobinometery
o Alkaline-hematin method
Specific gravity method
o Comparator method
B-polypeptide
chains CH,
8 um
CH

HO-C
Ho

Surface vlew
C-CH2
cH, HO-C

a-polypeptide H,Q
chains
Side view
o0c
(a) RBC shape (b)Hemoglobin molecule (c) Iron-containing heme

Fig. 5.2:Structure of haemoglobin

 Human Anatomy and Physiology(Practical) 5.3 To Determine the Haemoglobin content
Normal Values:
Women: 12 to 16 gm/100 mlof blood (83 to 1 1 0 % ) . s d a e t
Men: 14 to 18 gm/100 mlof blood (97 to 124%).
Infants: 14 to 20 gm/100 ml of blood (97 to 138%).
Clinical Significance:
Determination of hemoglobin is the most frequently performed laboratory test in
the clinical practice.
" It is mandatory to evaluate the level of hemoglobin prior to the patient going for
any surgery.
Estimation of hemoglobin is done to detect the anaemic condition of patient.
Haemoglobin estimation below normal level indicates that the patient is anaemic.
In this condition, not enough haemoglobin is available to carry sufficient
from the lungs to tissue.
oxygen
The symptoms of anemia are non-specific such as weakness or fatigue.
The physical examination of person shows palor, changes in the nail beds, glossitis
(red tongue), or hepatosplenomegaly (enlarged liver or spleen) may give a clue
about etiology of the anemia.
Conditions that Decreases Hb Concentration:
Different types of anemia
Cancer
Gastritis (Inflammation of the stomach lining)
Hypothyroidism (Underactive thyroid)
Iron deficiency anemia
Chronic kidney disease
Cystitis (Urinary bladder inflammation)
Pregnancy due to hemodilution
Conditions that Increases Hb Concentration:
Newborns and infants have high count
High altitude due to lack of oxygen
Excessive sweating
Severe diarrhoea and vomiting
Principle:
When the blood get mixes with 0.1 N HCI for 5 to 7
minutes, the red blood cells are
haemolysed and hemoglobin is converted into acid hematin (reddish
colour). brown in
The acid hematin solution is further diluted with drop by
drop addition of distilled
water until its colour matches with standard tubes of
Haemoglobinometer.
The percentage of hemoglobin directly reads from the graduated tube.
On the graduated tube 100% of the scale corresponds to 14.5 gm %
of hemoglobin.
 (Practical) 5.4 To Determine the Haemoglobin content ...
Human Anatomy and Physiology
Procedure: (Sahli's method)
Clean and dry the hemoglobinometer pipette properly.
the graduated tube with 0.1 N hydrochloric acid upto the mark 10 (10%) with the
"Fill
help of dropper.
aseptically to get free flow of
Then sterilize the finger with 70% alcohol and prick it
blood.
drop of blood is discarded and then the blood is sucked into
The first
hemoglobinometer pipette upto mark 20 mm².
graduated tube
Immediately transfer the blood of hemoglobinometer pipette into
containing0.1 NHCI.
of stirrer and wait for 10-15 minutes for
The blood and acid is mixed with the help
complete conversion of Hb to acid hematin.
mixture is then diluted with drop by drop addition of distilled water and stirs it
The
properly.
of graduated tube matches with the
The distilled water is added until the colour
standard tube in hemoglobinometer.
the percentages of hemoglobin by
When the matching of tubes is completed, read level under source light.
holding the hemoglobinometer and tubs at eye
Calculation:
Percentage of Hemoglobin ():
Hemoglobin (Y)
Gms of Hemoglobin (X) = Percentage of
14.5 g of Hg = 100 %
X(Gms of Hb) = Y
100
Y = Xx 14.5
():
Oxygen carrying capacity of Hemoglobin
100 % Hb = 18.5 ccof oxygen
Y% = Z
18.5
Z = YX 100
Result:
The own blood sample contains;
() Gms of Hemoglobin (gms)=
(i) Percentage of Hemoglobin (%) =
(ii) Oxygen carrying capacity (cc) =
Report:
Patient name:
Sample number: A8
Human Anatomy and Physiology (Practical) Haemoglobin content..
5.5 To Determine the
Observed count:
Gms of Hemoglobin(gms):
Percentage of Hemoglobin
o Oxygen carrying
capacity (%):odiesdut belhi
(cc):
Normal Hb count:
Women:
Men:
Infants:
Method used:ioig stmon doiporod lo boole eds ieavi
Date:

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