Estimation of hemoglobin (Hb) concentration


  

Sahli’s method

Principle

Hemoglobin in the red blood corpuscle is converted into a brownish compound called acid hematin by the action of hydrochloric acid. The higher the hemoglobin concentration, the more intense the hematin color will be. This brown color is matched against the standard brown-tinted glass in the comparator by direct vision. Reading is taken directly as g Hb/100 ml blood.

Apparatus and reagents


  • Blood sample
  • Hemoglobinometer
  • N/10 HCl (0.1 N)solution
  • Distilled water
  • Disposable lancet
  • cotton swab
  • Alcohol
  • Anticoagulant.

Procedure


  1. Study the hemoglobin tube and note the graduations.
  2. Fill a clean and dry graduate Sahli’s tube with N/10 HCl solution up to the mark 2 g (or 10 %)
  3. Lace the finger of the patient to obtain a drop of blood (or take the anticoagulant added blood sample).
  4. Place the tip of the Sahli’s pipette in the drop and draw a solid column of blood into the pipette up to the 20 cubic mm mark by gently sucking on the plastic mouthpiece fitted on the rubber tube.
  5. Wipe the tip of the pipette with tissue paper so that no blood is left sticking to its outside.
  6. Insert the tip of the pipette beneath the surface of HCl in the Sahli’s tube and gently blow out the blood.
  7. Rinse the pipette of any blood by thawing the solution in and out of the pipette ten times.
  8. Add blood by stirring with a rod.
  9. Place the tube in the comparator block and the tube stand for 10 minutes.
  10. Hold the comparator against a strong light and added distilled water drop by drop to the hematin solution until its color matches the color of the standard color on the comparator.
  11. Move out the stirrer and read the upper meniscus on the scale to obtain the percent of Hb or grams of Hb per 100 ml of blood.

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