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Hemoglobin

Sahli’s Method For The Estimation Of

Hemoglobin STAY CONNECTED

 November 18, 2022  Dhurba Giri  HEMATOLOGY 0

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Hemoglobin (Hb) is a conjugated protein present in Red Blood Cells (RBCs).  

It serves two important functions in the body- transportation of oxygen and
carbon dioxide in between tissues and lungs and acts as a bu!er in
maintaining blood pH. Normal Hgb is composed of four heme molecules
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nested in four globin molecules. Among four globin molecules, two chains
are alpha chains and two are non-alpha chains. Based on the type of non- Sahli’s Method For
alpha chain present, three di!erent types of hemoglobin are found in The Estimation Of
normal adults. Hemoglobin
 November 18, 2022 
1. HbA: 96-98%, Composed of two alpha(α) and two beta(β) globin 0

chains.
2. HbA2: 1.5-3.5%, Composed of two alpha(α) and two delta(δ) globin
Cyanmethemoglobin
chains. Method For The
3. HbF: < 1.0%, Composed of two alpha(α) and two gamma(γ) globin Estimation Of
chains. Hemoglobin
 November 17, 2022 
0
Hemoglobin can combine with other substances, some normally and some
:
 abnormally and can also occur as:
Colorimeter :
Principle,
1. Oxyhemoglobin: Oxygen combined with hemoglobin.
Instrumentation and
2. Carboxyhemoglobin: Carbon monoxide (CO) combined with Uses
hemoglobin.  June 24, 2022  0

3. Carbaminohemoglobin: Carbon dioxide (CO2) combined with

hemoglobin.
god-pod GOD-POD Method
4. Methemoglobin: Iron oxidized from its ferrous state to ferric state. For The Estimation
5. Sulfhemoglobin: Sulfur combined with the hemoglobin. Of Blood Glucose
 March 22, 2022  0
6. Cyanmethemoglobin: Methemoglobin bonded to cyanide ions.

Folin-Wu Method
Methods for Hemoglobin Estimation: For Estimation of
Blood Glucose
The di!erent methods used for the estimation of hemoglobin can be
 February 20, 2022 
divided as follows: 0

1. Visual methods:
Sahli’s method
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Dare method
Haden method Reverse Blood
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Wintrobe method
Procedure,
Tallqvist method Interpretation and
Limitations
2. Spectrophotometric methods:  December 21, 2017 
16
Oxyhemoglobin method
Cyanmethemoglobin method
Cross Matching :
3. Gasometric method Types, Principle,
4. Automated hemoglobinometry Procedure and
Interpretation
5. Other methods:
 July 2, 2019  7
Alkaline-hematin method
Specific gravity method
Lovibond Comparator method Reticulocyte Count :
Principle, Procedure,
Calculations and
Clinical Significances
Sahli’s method for hemoglobin estimation  November 6, 2018 
7
Sahli’s method, also called as acid hematin method is the visual comparator
method for the estimation of hemoglobin. As visual comparison may lead
to unacceptable imprecision and accuracy, this method is not
Urine Pregnancy Test
recommended nowadays and the use of spectrophotometric methods like (UPT): Principle,
Cyanmethemoglobin method is preferred to it. Procedure,
Interpretation and
Limitations
Principle:  December 25, 2017 
6
When the blood is added to dilute hydrochloric acid (HCl), hemoglobin
present in the RBCs is converted into brown-colored acid hematin. The acid
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 hematin solution is further diluted until it’s color matches exactly with the
permanent standard brown glass compared by direct vision. Differences Between
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histolytica and
Requirements: Entamoeba coli
 November 6, 2018 
6

Specimen:
Capillary or venous blood. Venous blood should be anticoagulated with 1.5-
1.8 mg EDTA per mL of blood and mixed immediately. CATEGORIES
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Instruments:
iVenus Open
BIOCHEMISTRY
1. Sahli’s hemoglobinometer
It is a set of devices that includes a comparator, hemoglobin tube, CYTOPATHOLOGY
hemoglobin pipette, and stirrer.
HEMATOLOGY

HISTOPATHOLOGY

IMMUNOLOGY

MICROBIOLOGY

PARASITOLOGY
Sahli'sHemoglobinometerSet
Uncategorized
Slotfor 100%=14.5%
Hbtube
160
•Hbtube

URINALYSIS
140

Golden-brown 120
glasssods
100 0.02ml
(20ul)

-Plasticbox

20

Comparator Hemoglobintube Hemoglobinpipette Stirrer

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Comparator: It is a rectangular plastic box with a slot in the

middle which accommodates a hemoglobin tube. Brown
standard glasses are provided on either side of the slot for
color matching. White opaque glass is present at the back to
provide uniform illumination.

Hemoglobin tube: Sahli’s graduated hemoglobin tube is

graduated in one side in gram percentage (g%) from 2 to 24,
and on the other side in percentage (%) from 20 to 140. The
tube is also called Sahli-Adams tube.

Sahli’s pipette or hemoglobin pipette: It contains only one

mark at 20μl or 0.02ml. Unlike WBC and RBC diluting pipettes, it
contains no bulb.

Stirrer: It is a thin glass rod used for stirring the mixture inside
the hemoglobin tube.
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 Reagents:
1. N/10 Hydrochloric acid (HCl): Mixing 36 grams HCl in distilled water
to 1 liter gives 1 N HCl. Diluting it 10 times will give N/10 HCl.
2. Distilled water

Procedure:
1. Ensure that the hemoglobinometer tubes and pipette are clean and
dry.
2. Fill the hemoglobinometer tube with N/10 HCl up to its lowest mark
i.e. 2 g% or 10% mark with the help of a dropper.
3. Take blood up to mark in the Sahli’s pipette (20 μl). Wipe the extra
blood outside the pipette and deliver it to N/10 HCl in the hemoglobin
tube.
4. Mix and leave it for 10 minutes in order for a complete conversion of
hemoglobin to hematin.
5. Add distilled water drop by drop and stir till color matches with the
standard glass of the comparator.
6. Take the reading at lower meniscus, which directly gives the
hemoglobin concentration in 100 ml of blood.

Advantages and Disadvantages of Sahli’s

method

Advantages:
1. Easy to perform and convenient.
2. Not very time consuming. Can be performed within maximum 15
minutes.
3. Reagents and apparatus are cheap and easily available. Reagents are
less harmful.
4. Can be used in mass surveys. Doesn’t require electricity.

Disadvantages:
1. Acid hematin is a suspension, not a true solution. So, some turbidity
may result.
2. This method can’t measure all hemoglobins. It estimates only
oxyhemoglobin and reduced hemoglobins. But carboxyhemoglobin,
methemoglobin and sulfhemoglobin (which all constitute about 2-
12% of total hemoglobin) are not converted to acid hematin.
3. HbF is not converted to acid hematin. Therefore, Shali’s method is not
suitable for measuring hemoglobin in infants upto 3 months.
4. WBC counts >100,000/cumm will produce turbid solution of acid
hematin, which will increase the hemoglobin report by 5-10%.
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 5. Chances of visual error are high. Color of acid hematin also fades
gradually.
6. The color of glass standard may fade over time.

References:
1. Pal, G.K., 2006. Textbook Of Practical Physiology-2Nd Edn. Orient
Blackswan.
2. Ghai, C.L., 2012. A textbook of practical physiology. JP Medical Ltd.
3. A. V. Naigaonkar. A Manual Of Medical Laboratory Technology.
4. Cheesebrough, M., 1998. District laboratory practice in tropical
countries, part II. Cambridgeshire Tropical Health Technology,
Cambridge, UK, 231.
5. Laboratory Procedures in Clinical Hematology By United States.
Department of the Army
6. Mohan, H. and Mohan, S., 2011. Practical Pathology for Dental
Students. JP Medical Ltd.

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 ACID HEMATIN METHOD ADVANTAGES OF SAHLI'S METHOD

ALL METHODS OF HEMOGLOBIN ESTIMATION

DISADVANTAGES OF SAHLI'S METHOD PRINCIPLE OF SAHLI'S METHOD

PROCEDURE OF SAHLI'S METHOD SAHLI'S METHOD

About Dhurba Giri  33 Articles

Dhurba Giri is the founder and content creator of LaboratoryTests.org. He
is a Medical Laboratory Technologist and Scientific Blogger from Pokhara,
Nepal. Connect with him:

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