Estimating Hemoglobin Levels using Drabkin’s or Cyanmethemoglobin Method

Introduction

Estimating hemoglobin levels using the Drabkin’s or cyanmethemoglobin method is a common laboratory procedure. Here is a practical protocol for estimating hemoglobin using the cyanmethemoglobin method:

Aim:

To estimate Hemoglobin by Drabkin’s or cyanmethemoglobin method.

Principle:

When blood is mixed with Drabkin’s reagent containing K cyanide & K ferricyanide hemoglobin reacts with ferricyanide to form methemoglobin which is converted to stable cyanmethemoglobin by cyanide. The intensity of
the color is proportional to hemoglobin concentration & is compared with a known cyanmethemoglobin standard at 540nm (green filter).

Requirements:

1. Hb-pipette or micro-pipette
2. Test tubes
3. Photometer or spectrophotometer.

Drabkin’s reagent-

K ferricyanide- 400mg
K dihydrogen phosphate 280mg
K cyanide 100mg
Distilled water- 1000m

4. Cyanmethemoglobin standard (Hb standard) commercially available. It is directly pipette in a cuvette & optical density measured at 540nm. The reading obtain corresponds to 15 gm% Hb

Specimen:

Capillary Blood or,
Whole with EDTA

Procedure:

3. Measure the absorbance of this solution at 540nm in a colorimeter after adjusting the OD at 0 by using Drabkin’s solution as blank.
4. Read absorbencies of standard (15 g/dl) by pipetting it directly in a cuvette.

Calculation:

Hemoglobin, g/dl = O.D. of TEST/O.D. of STD ×15

Normal values:

Men 14-18gm%
Women 12-16gm%

Clinical significance:

Hemoglobin is a critical component of blood with significant clinical significance. It plays several important roles in the body, and its measurement is a common part of clinical diagnostics. Here are some of the key clinical significances of hemoglobin:

1. Oxygen Transport: Hemoglobin binds to oxygen in the lungs and carries it to tissues throughout the body. This is essential for cells to produce energy through cellular respiration.
2. Carbon Dioxide Transport: Hemoglobin also helps transport carbon dioxide, a waste product of metabolism, from the tissues back to the lungs for exhalation.
3. Anemia Diagnosis: Hemoglobin levels are routinely measured in blood tests to diagnose anemia, a condition characterized by a lower than normal hemoglobin concentration. Anemia can be caused by various factors, including nutritional deficiencies, chronic diseases, or blood loss.
4. Oxygen Saturation Monitoring: Hemoglobin is used in conjunction with other parameters to measure oxygen saturation in the blood, which is important in assessing respiratory function and the effectiveness of oxygen therapy.
5. Monitoring Chronic Conditions: Hemoglobin levels can be used to monitor chronic medical conditions, such as kidney disease, chronic obstructive pulmonary disease (COPD), and heart disease, as changes in hemoglobin levels can indicate disease progression or treatment effectiveness.
6. Blood Transfusions: Hemoglobin levels are a key factor in determining the need for blood transfusions. When hemoglobin levels are critically low, transfusions may be necessary to restore oxygen-carrying capacity.
7. Surgical Planning: Hemoglobin levels are evaluated before surgery to assess the patient’s ability to tolerate the procedure and to predict the risk of excessive bleeding during surgery.
8. Monitoring Blood Disorders: Hemoglobin measurements are essential for diagnosing and monitoring various blood disorders, including sickle cell anemia and thalassemia.
9. Assessing Nutritional Status: Hemoglobin levels can be used to assess nutritional status, as deficiencies in iron, vitamin B12, or folate can lead to low hemoglobin levels.
10. Evaluating Blood Loss: Hemoglobin levels are used to estimate blood loss in cases of trauma, surgery, or gastrointestinal bleeding.
11. Monitoring Pregnancy: Hemoglobin levels are regularly checked during pregnancy to detect anemia, which can be common in expectant mothers.

Advantage:

Accuracy:
Wide Range:
Standardization:
Long-Term Stability:
Compatibility:
Minimal Interference:

In summary, the Drabkin’s and cyanmethemoglobin methods for estimating hemoglobin offer accurate and widely accepted techniques with several advantages, making them valuable tools in clinical diagnostics and research.

Disadvantage:

Cyanide Toxicity:
Chemical Waste:
Interference from Other Substances:
Time-Consuming:
Variability:
Limited Quantification:

Reference

“Hoffbrand’s Essential Haematology” by A. Victor Hoffbrand, Paul A.H. Moss, and John E. Pettit: This is a comprehensive and highly regarded textbook that covers the fundamental principles of hematology.
“Williams Hematology” by Kenneth Kaushansky, Marshall A. Lichtman, Josef T. Prchal, Marcel M. Levi, Oliver W. Press, and Linda J. Burns: This is a renowned textbook that provides in-depth coverage of both clinical and basic aspects of hematology.

“Wintrobe’s Clinical Hematology” by John P. Greer, Daniel A. Arber, Bertil E. Glader, Alan F. List, Robert T. Means Jr., Frixos Paraskevas, George M. Rodgers, and Michael H. Schmitt: This classic text has been a staple in the field of hematology for many years and offers comprehensive information on various hematological disorders.
“Hematology: Basic Principles and Practice” by Ronald Hoffman, Edward J. Benz Jr., Leslie E. Silberstein, Helen Heslop, John Anastasi, and Jeffrey Weitz: This textbook provides a detailed overview of the principles and practice of hematology, including the latest advancements in the field.

“Clinical Hematology and Fundamentals of Hemostasis” by Denise M. Harmening: This book is a good choice for those looking for a comprehensive guide to both clinical hematology and hemostasis.
“Rodak’s Hematology: Clinical Principles and Applications” by Elaine M. Keohane, Larry Smith, and Jeanine M. Walenga: This text focuses on the clinical application of hematology concepts and includes practical information for laboratory professionals.

“Hematology in Clinical Practice” by Robert S. Hillman, Kenneth A. Ault, and Henry M. Rinder: This book is designed for healthcare professionals and provides a practical approach to hematology in a clinical setting.
“Consultative Hematology: An Issue of Hematology/Oncology Clinics of North America” edited by Mikkael A. Sekeres and Christopher R. Cogle: This book focuses on the consultative aspects of hematology and is a valuable resource for clinicians.