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Questions and Answers
Procedure if a great excess has been drawn in, inaccuracy will result, in this case the pipette must be ______, ______ and refilled.
clean, dried
Wipe off with gauze the blood on the outside of the ______ Pipette.
pipette
Hb is converted into acid haematin by ______.
Hcl
By keeping the point of the pipette to the bottom of the tube and gently blowing off the blood without causing ______.
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Mix the solution in the tube by ______ rod.
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Leave the tube for ______ min.
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Now dilute the solution by adding ______ water, drop by drop, stirring and mixing all the time with a glass rod until the color matches with standard.
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Hemoglobin estimation by Automated method - Principle - Adding lyse in the blood, the red blood cell will be broken and release hemoglobin. Hemoglobin and lyse form a new mixture, which can absorb the wavelength ______ nm.
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The bottom of the meniscus is read on ______ scale.
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Through the comparison of the absorbency between the pure diluent and the sample - the concentration of the sample hemoglobin is ______.
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Study Notes
Hemoglobin Overview
- Hemoglobin (Hb) is a protein molecule in red blood cells that carries oxygen from the lungs to the body's tissues and returns carbon dioxide from the tissues back to the lungs.
- Hemoglobin concentration is reported in grams per deciliter of blood.
Structure of Hemoglobin
- Hemoglobin is a chromoprotein consisting of a colorless globin molecule attached to four red-colored haem molecules.
- The globin molecule consists of two alpha and two beta polypeptide chains.
Types of Hemoglobin
- HbF (fetal Hb): normally synthesized only during fetal development, consisting of 2 alpha and 2 gamma chains.
- HbA or HbA1: the normal Hb in adults, representing about 97% of total Hb, consisting of 2 α and 2 β chains.
- HbA2: a minor component of normal adult Hb, consisting of 2 alpha and 2 delta chains, found in low levels in normal human blood.
- Myoglobin: found in muscles, consisting of one heme and one globin molecule.
- HBA1C: has glucose residues attached to β-globin chains, increased in diabetes mellitus (DM).
Normal Hemoglobin Values
- Women: 12-16 g/dL
- Men: 14-17.4 g/dL
- Children:
- 0-2 weeks: 14.5-24.5 g/dL
- 2-8 weeks: 12.5-20.5 g/dL
- 2-6 months: 10.7-17.3 g/dL
- 6 months-1 year: 9.9-14.5 g/dL
- 1-6 years: 9.5-14.1 g/dL
- 6-16 years: 10.3-14.9 g/dL
- 16-18 years: 11.1-15.7 g/dL
Hemoglobin Interpretation
- Increased values:
- Physiological: high altitude, young age
- Pathological: dehydration
- Decreased values:
- Physiological: fluid therapy
- Pathological: anemia, tumors, hemorrhage, blood parasites, malignant
Methods for Hemoglobin Estimation
- Sahli's method
- Drabkin's method
- Automated method
Sahli's Method
- Principle: Hemoglobin + (0.1 N) HCl → Acid hematin (brown color)
- Apparatus and reagents: Sahli-type Haemoglobinometer, Hb tube, Hb pipette, whole blood with EDTA, lancet, cotton, distilled water, HCl solution (0.1 N)
- Procedure:
- Fill Hb tube to 20 mark with HCl.
- Fill Hb pipette exactly to 20 microliter.
- Wipe off blood on outside of pipette.
- Empty pipette into tube.
- Rinse pipette at least three times.
- Mix solution in tube by glass rod.
- Leave tube for 10 minutes.
- Dilute solution by adding distilled water, drop by drop, stirring and mixing until color matches standard.
Automated Method
- Principle: adding lyse to blood breaks red blood cells, releasing hemoglobin, and forming a new mixture that absorbs wavelength 540 nm.
- Measure absorbency and compare with pure diluent to calculate hemoglobin concentration.
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Description
Test your knowledge on the estimation of hemoglobin concentration (Hb) and the function of hemoglobin in the body. Learn about the significance of hemoglobin in transporting oxygen and carbon dioxide between the lungs and tissues.
