This document discusses hemoglobin estimation and provides information about hemoglobin. It notes that hemoglobin has a molecular weight of 68,000 and weighs about 33% of a red blood cell. The hemoglobin molecule has 4 subunits, with each subunit containing a heme group bound to a polypeptide chain. Hemoglobin is able to transport 4 oxygen molecules and has a higher affinity for carbon monoxide than oxygen. Normal hemoglobin levels are provided for males, females, children, and pregnant women. Common techniques for hemoglobin estimation include the acid hematin, photometric cyanmethemoglobin, and automated methods.

1. Hemoglobin Estimation
Third Year
Dr. Ahmed EL Tahir
2008

2. Biochemical features of
Hemoglobin
Molecular weight is 68,000
Weighs about 33% of cell weight (RBC)
Hemoglobin molecule has 4 subunits (a
multimetric protein)
A subunit = Heme + a polypeptide chain
– Heme has Fe2+ that binds O2 for
transport
Globin (polypeptide chains) identify
various hemoglobins
One Hb molecule transports 4 oxygen
molecules (molecule to molecule)

3. –Heme is a cyclic tetrapyrrole
consisting of four molecules of pyrrole
linked by alpha –methyl bridges.
Hemoglobin uses the same binding
site to transport O2 and CO;
the binding site has a higher affinity
for CO
Membrane proteins (antigens) are
used to distinguish blood groups.

4. Notes:

5. Notes:

6. The Formed Elements: Red Blood
Cells
Function:
Notes:
Males: 4.5-6.3 million RBC/mm3
Females: 4.2-5.5 million RBC/mm3
Life span 120 days

7. Molecular Structure, and Function
of Red Blood Cells - Hemoglobin
95% of protein in RBC’s
14-18 g/dl for males
12-16 g/dl for females
Approximately 280 million
molecules/cell
Functions:

8. Hemoglobin Structure
Notes:

9. Abnormal Hemoglobin

10. Red Blood Cell Turnover

11. Hb Estimation
Hemoglobin concentration measurement is
among the most commonly performed bood
tests ,usually as part of a complete blood
count
It is typically tested before blood donation
Results are reported in g/L ,g/dl or mol/L.
1 g/dL equals about 0.6206 mmol/L, and 1
g/L equals about 0.06206 mmol/L.
Normal levels are:
Women: 12.1 to 15.1 g/dl
Men: 13.5 to 16.5 g/dl
Children: 11 to 16 g/dl
Pregnant women: 11 to 12 g/dl

12. King Fahad Medical City
Riyadh
Department of Haematology and
Flow Cytometry Section
Location: G-1331, 1st Floor, Central
Laboratories, Main Hospital

13. Examining the components of
Blood

14. From Haemtology Dept. -KFMC
Request form:
Request without the minimum information
listed below may be delayed or cancelled.
– 1. Name of Hospital / Ward
– 2. Patient Name
– 3. Patient ID Number
– 4. Date of Birth / Age
– 5. Sex
– 6. Time / Date of Collection
– 7. Required Tests
– 8. Indicate specimen source, if applicable.
– 9. Indicate volume if applicable.
– 10. The clinical diagnosis.

15. SPECIMENS
Policy and Procedure
– All specimen containers must be properly closed
to avoid leakage and clearly labelled with:
a. Name
b. Patient Hospital Number
c. Sex
d. Age / Date of Birth
e. Date / Time of Specimen Collection
2. Each specimen must be enclosed in a
separate biohazard transport bag.
a. Each bag must be securely sealed.
b. The transport bag has two separate sections; one
for the specimen and one for the request form.
c. Do not enclose the request form in contact with the
specimen container or staple it to the bag. An
exception to this rule is the 24-hour urine where the
form is placed in a biohazard transport bag and
attached to specimen container.

16. Criteria for Rejection of Specimens
Specimens not received in the appropriate
collection tube.
Specimens received without proper labelling
on the collection tube.
Specimens received without correct details
on the laboratory requisition form (Name
and Hospital Number on the collection tube
and requisition form not matching).
Quantity of sample not sufficient (incorrect
blood to anticoagulant ratio).
Specimens collected on the previous day.
Haemolysed and / or clotted samples.
Specimens for special tests, e.g.
cytochemical stains, special coagulation
tests without prior appointment.

17. Hemoglobin refrence values:
HGB
Men 13.0-18.0 g/dL
Women 11.5-16.5 g/dL
Infants (full-term, cord blood) 13.5-
19.5 g/dL
Infants, 3 months 9.5-13.5 g/dL
Infants, 1 yr. 10.5-13.5 g/dL
Children, 3-6 yr. 12.0-14.0 g/dL
Children, 10-12 yr. 11.5-14.5 g/dL

18. Men 40-54 %
Women 37-47 %
Infants (full-term, cord blood) 44-64
%
Infants, 3 months 32-44 %
Children, 3-6 yr. 36-44 %
Children, 10-12 yr. 37-45 %
HCT Hematocrit (PCV):

19. Mean Cell Hemoglobin
This represents the absolute amount of
hemoglobin in the average red cell in a sample.
Its units are picograms (pg) per cell.
The MCH is calculated from the [Hb] and the RBC
using the following equation :
MCH (pg) = (Hb x 10) ÷ RBC
This value is generally not useful because the
combination of MCV and MCHC provides more
specific information. For example, a low MCH
could be due to smaller than normal cells with
normal Hb concentration, or normal sized cells
with lower than normal Hb concentration. It is
better to know about cell volume and hemoglobin
concentration directly .

20. Techniques:
Different methods
Acid haematin,
Photometric cyanmethemoglobin
estimation
and automated estimation with the
help of counters.
Sahli's and Drabkin's method

21. Cyanmethemoglobin method
Principle:
Procedure
Calculation
Result: Interpretation:
It is advised in every admitted case and in
all the females undergoing antenatal
check up.