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E Stimation of Hemoglobin

This document provides instructions for estimating hemoglobin levels using Sahli's method. It begins with an introduction on hemoglobin and its importance for oxygen carrying capacity. It then lists the apparatus required, including a Sahli's hemoglobinometer, hydrochloric acid, and a pricking apparatus. The principle involves hemolysis and formation of acid hematin when blood and hydrochloric acid are mixed. Steps of the procedure include collecting blood, mixing with acid, allowing time for conversion, diluting, and comparing color to a standard. Normal hemoglobin values at different ages and for males and females are provided. Precautions for accurate results and a reference are also included.

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460 views

E Stimation of Hemoglobin

This document provides instructions for estimating hemoglobin levels using Sahli's method. It begins with an introduction on hemoglobin and its importance for oxygen carrying capacity. It then lists the apparatus required, including a Sahli's hemoglobinometer, hydrochloric acid, and a pricking apparatus. The principle involves hemolysis and formation of acid hematin when blood and hydrochloric acid are mixed. Steps of the procedure include collecting blood, mixing with acid, allowing time for conversion, diluting, and comparing color to a standard. Normal hemoglobin values at different ages and for males and females are provided. Precautions for accurate results and a reference are also included.

Uploaded by

SANA
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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E​stimation of Hemoglobin

Ms Usra Hasan
Assistant Professor
Department of Physiology
School of Allied Health Sciences
Sharda University

Content
•
Aim
•
Apparatus Required
•
Principle
•
Procedure
•
Normal Values
•
Precautions
•
Reference
Hemoglobin (Hb) is a protein that carries oxygen in Red Blood Cells or Erythrocytes
Therefore,
Hb concentration in blood determines – oxygen-carrying capacity of blood

to estimate the concentration of hemoglobin (Hb) in the given blood sample, using
Sahli’s​ ​Method
AIM

Sahli’s Hemoglobinometer
Hydrochloric Acid (HCl) – N/10
Distilled Water
Pricking Apparatus – Lancet
Stirrer
Dropper
APPARATUS REQUIRED

Sahli’s Pipette
Hemoglobinometer

–
a single mark, hemoglobinometer
pipette –
measures 20 μL (0.02 mL) of blood
–
Hemoglobinometer Tube
round or square hemoglobinometer
tube –
graduated in %age (10 – 140) and gm/dL (2 to
22)
–
Stirrer
a glass rod – used to stir
–
Comparator
a rectangular box – fixed with two solid standard
coloured glass rods
opaque white background – provides necessary
contrast for easy comparison

Addition of HCl to blood causes:


i. ii.
Hemolysis Formation
of RBCs of Acid
Hematin

PRINCIPLE
Intensity of obtained is proportional to Hb content in blood

Hemoglobin + Hydrochloric Acid → Acid Hematin

PROCEDURE
–
with the help of a dropper, take N/10 HCl in graduated hemoglobinometer tube up to its lowest
mark, i.e., 10% or 2 gm/dL

–
Prick finger under aseptic precautions.
when a drop of reasonable size is collected, hold the pipette horizontally
apply its tip to the drop and draw exactly 20 μL of blood into pipette (without air
bubbles) take pipette away and wipe off any blood which adhere to the tip

–
immediately transfer blood from pipette into N/10 HCl in graduated
hemoglobinometer tube ​rinse pipette several times by drawing N/10 HCl used to mix
blood
–
mix contents thoroughly and leave solution to stand for about 10 minutes for max. conversion
of Hb in blood into acid hematin

–
dilute acid hematin by adding distilled water in drops
keep mixing with stirrer, thoroughly

–
continue dilution till its colour matches with that of standard
before comparing colour, stirrer should be lifted above solution only

–
note reading of meniscus from scale provided on hemoglobinometer tube
express Hb content as gm/100 mL of blood
At birth: 23 gm/dL
At end of 3 months: 10.5 gm/dL
After 3 months: 12.5 gm/dL
Adults
Males: 14 – 18 gm/dL
Females: 12 – 15.5 gm/dL

NORMAL VALUES
PRECAUTIONS

–
finger should not be squeezed to take out blood, otherwise fluid tissue might come out, thus
giving lower estimation of Hb

–
blood from pipette should be transferred quickly into tube containing N/10 HCl to prevent clotting
of blood in pipette
–
blood, after mixing with N/10 HCl, should be left for at-least 8 – 10 min. for complete formation
of acid hematin

–
distilled water should be added drop by drop to dilute acid hematin, otherwise over
dilution occurs –
matching of colours should be done only against natural source of light

https://youtu.be/5e5N3yEjFBQ
Manual for Practical Physiology by Dr A.K. Jain by Arya
Publications​REFERENCE

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