Questions and Answers
What are the primary components of hemoglobin?
What is the main function of hemoglobin in the body?
Where does the formation of hemoglobin occur?
What does each gram of hemoglobin carry in terms of oxygen volume?
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What role does ferricyanide play in the hemoglobin determination process?
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What is formed when methemoglobin reacts with potassium cyanide?
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What is the visual indicator of cyanmethemoglobin in the determination process?
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What method is used to measure the absorption of light in this hemoglobin determination experiment?
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What happens to hemoglobin (Hgb) levels at high altitudes?
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How does pregnancy affect RBC count and Hgb levels?
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Which condition is associated with decreased hemoglobin levels?
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What results in an increase in hemoglobin levels?
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Which factor does NOT contribute to mechanical sources of error in hemoglobin testing?
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What can cause a normal RBC count but a low hemoglobin level?
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Which condition is characterized by increased hemoglobin levels?
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What occurs at low altitudes regarding RBC count and Hgb levels?
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What is the first step in the hemoglobin determination procedure using Drabkin's reagent?
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Which of the following materials is NOT listed as needed for hemoglobin determination?
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What is the purpose of determining hemoglobin concentration in a patient?
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What range represents normal hemoglobin values for adult females?
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Which substance is produced during the oxidation of hemoglobin?
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What is the effect of altitude on hemoglobin values?
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Which component is NOT part of Drabkin's solution?
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How does sex affect hemoglobin values in adults?
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Study Notes
Hemoglobin Determination
- Hemoglobin (Hb) consists of heme (4% - iron-containing pigment) and globin (96% - colorless protein).
- Primary function of hemoglobin is to transport oxygen from lungs to tissues and carbon dioxide from tissues to lungs.
- Hemoglobin formation occurs in developing red blood cells (RBCs) in the bone marrow.
Principle of Cyanmethemoglobin Method
- Blood is mixed with Drabkin’s solution (contains ferricyanide and cyanide).
- Ferricyanide oxidizes ferrous iron (Fe2+) in hemoglobin to ferric (Fe3+), creating methemoglobin.
- Methemoglobin reacts with cyanide ions to produce cyanmethemoglobin (HiCN), which generates a measurable brown color.
- Spectrophotometry measures light absorption of cyanmethemoglobin at 540 nm.
Normal Hemoglobin Values
- Adult males: 14 – 18 gm/dl
- Adult females: 12 – 16 gm/dl
- Newborns: 17 – 19 gm/dl
Required Materials
- Alcohol wipes, Hb pipettes or micropipettes, cotton, tape
- Colorimeter or spectrophotometer, test tubes (cuvettes)
- Drabkin’s solution (includes potassium ferricyanide, potassium cyanide, etc.)
- Hemoglobin standard curve for concentration determination
Procedure Overview
- Combine 5 ml of Drabkin’s reagent with 20 µl of blood.
- Wait 5 to 10 minutes for the reaction.
- Measure absorbance against a blank at 540 nm and compare with the standard Hb curve for concentration.
Clinical Significance of Hemoglobin Testing
- Used for screening and assessing severity of anemia, as well as evaluating treatment response.
- Normal values can be affected by age, sex, pregnancy, and altitude:
- Newborns and men typically have higher values than women.
- Increased altitude results in elevated RBC and Hb levels due to lower oxygen availability.
- Pregnancy leads to lower Hb concentration because of fluid gain and reduced RBC concentration.
Pathological Variations
- Decreased Hb levels may result from blood loss, bone marrow suppression, or abnormal hemoglobin conditions (hemoglobinopathies).
- Conditions like iron deficiency anemia can maintain normal RBC count but exhibit low Hb levels.
- Increased Hb levels indicate elevated circulating RBC counts due to conditions such as polycythemia vera or dehydration.
Common Mechanical Errors
- Potential errors can arise from pipetting mistakes, using dirty or scratched cuvettes, or utilizing deteriorated reagents.
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Description
Test your knowledge on hemoglobin and the cyanmethemoglobin method. This quiz covers the structure of hemoglobin, its components like heme and globin, and the methodology behind hemoglobin determination. Perfect for students in biology or medical fields.
