Estimation of Hemoglobin Content

ByDeepak Rajput

Aim: Estimation of Hemoglobin Content

References

  1. Wintrobe, M. M. (2012). Clinical Hematology. Lea & Febiger.
  2. Cheesbrough, M. (2006). District Laboratory Practice in Tropical Countries. Cambridge University Press.
  3. Articles from Google Scholar/PubMed on hemoglobin estimation methods.

Objective

To estimate the hemoglobin content in blood using the Sahli’s method (acid hematin method).

Principle

Hemoglobin estimation is crucial for diagnosing conditions like anemia and polycythemia. The Sahli’s method involves converting hemoglobin to acid hematin by adding dilute hydrochloric acid. The intensity of the resulting brownish-yellow solution is compared to a standard using a hemoglobinometer.

Materials Required

  1. Reagents: N/10 Hydrochloric acid (HCl)

Apparatus

Sahli’s hemoglobinometer (graduated tube, comparator), Hemoglobin pipette, Stirrer (glass rod), Cotton and antiseptic, Sterile lancet

Procedure

1. Sample Preparation

  • Prick the fingertip using a sterile lancet or collect capillary blood.
  • Wipe away the first drop; collect the second drop using a hemoglobin pipette.

2. Conversion to Acid Hematin

  • Fill the hemoglobinometer tube to the 2-gram mark with N/10 HCl.
  • Add 0.02 mL of blood (20 µL) to the HCl using the hemoglobin pipette.
  • Mix thoroughly with a stirrer to convert hemoglobin into acid hematin.

3. Dilution and Matching

  • Gradually add distilled water dropwise to the tube, mixing after each addition.
  • Continue until the color of the solution matches the standard in the comparator.

4. Reading Hemoglobin Content

  • Read the hemoglobin concentration directly from the graduated scale on the hemoglobinometer tube (g/dL).

Sample Data

Sample No.Hemoglobin Content (g/dL)
113.5
212.8
314.2

Interpretation

  • Normal Range:

Males: 13–18 g/dL

Females: 12–16 g/dL

Children: 11–14 g/dL

Newborns: 14–24 g/dL

  • Clinical Significance:

Low hemoglobin: Anemia, blood loss, malnutrition, chronic diseases.

High hemoglobin: Dehydration, polycythemia, or high-altitude adaptation

Precautions

  1. Ensure precise measurement of blood and reagents.
  2. Avoid bubbles while adding blood or water.
  3. Use a clean hemoglobinometer tube to prevent residue interference.
  4. Compare colors under natural light for accuracy.

Result: The hemoglobin content of the blood sample is 13.5 g/dL, which is within the normal range for males.

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