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Hemoglobin Estimation Guide

This document outlines a lab procedure for estimating hemoglobin levels using the Cyanmethemoglobin method, which involves a spectrophotometer to measure light absorption. The process includes preparing Drabkin's reagent, mixing it with blood, and measuring absorbance at 540nm to calculate hemoglobin concentration. It notes that the current Drabkin reagent is old, potentially affecting the accuracy of results.

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25 views2 pages

Hemoglobin Estimation Guide

This document outlines a lab procedure for estimating hemoglobin levels using the Cyanmethemoglobin method, which involves a spectrophotometer to measure light absorption. The process includes preparing Drabkin's reagent, mixing it with blood, and measuring absorbance at 540nm to calculate hemoglobin concentration. It notes that the current Drabkin reagent is old, potentially affecting the accuracy of results.

Uploaded by

giboto5317
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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‫)‪Physiology labs for blood (3‬‬

‫(المعمل الثاني)‬ ‫قياس الهيموجلوبــــيـــــن‬


‫)‪Hemoglobin estimation ( lab 2‬‬

‫المجموعة السابعة‬

‫تلخيص ‪/‬‬
‫هيام الجوبي‬ ‫أنس العمراني‬

‫‪1‬‬
In this lab we will study about the new manual method for Hemoglobin(HB)
estimation , the Cyanmethemoglobin method .
In this method, a Spectrophotometer is used to measure the amount of light
absorbed by the colored compound Cyanmethemoglobin, because All forms of
Hb normally present in blood (i.e. oxyHb, reduced Hb, carboxyHb, and metHb)
are converted into the stable compound cyanmethemoglobin by the action of
Drabkin reagent
Characteristics of drabkin reagent:
1-contains potassium cyanide, potassium ferricyanide, and other substances
that (1) remove the lipid from the RBCs and destroy the RBCs causing the hemo
globin to expel out of them (2)convert the hemoglobin into cyanmethemoglobin
2-it must be diluted by distilled water by 1/90 ratio . The drabkin used in the
lab is already diluted
3- after dilation, it must be used within 6 hours and must not be exposed to
sunlight.
4- it must be clean ,pale yellow , not turbid and having PH of 7

Steps of this method:


1. Take 5ml of Drabkin’s solution in a test tube.
2. draw 20ul of blood into the pipette. Wipe the outer surface of the pipette
to remove excess blood.
3. Place the pipette into the tube containing Drabkin’s solution and slowly
expel the blood into the solution. Mix well and let it stand undisturbed for 3-5
minutes then put the solution in the special cuvette of the spectrophotometer.
4. after zeroing the spectrophotometer using Drabkin’s solution as blank.
put the cuvette containing the solution in the spectrophotometer to Measure
the absorbance of this solution at wave length 540nm
5. Calculate the hemoglobin concentration by multiplying the number that
appears in the screen by 37 . for example 0.31 x 37 = 11.47 g/dl
* currently the drabkin reagent in the lab is very old so the results are not very
accurate
2

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