Estimation Of Hemoglobin By Sahli’s Methods

Estimation of hemoglobin

estimation of hemoglobin by sahli’s method

1. Introduction of haemoglobin
2. Function of hemoglobin
3. Types of hemoglobin
4. Principle of hemoglobin
5. Requirement of hemoglobin
6. Testing procedure
7. Normal range
8. Clinical significance
9. Precaution

1. Introduction of hemoglobin ( estimation of Hemoglobin)
   Hemoglobin is the red respiratory pigment of the RBC. it is composed of haem (iron) 4% and globulin (protein) 96%.
   Hemoglobin carries O2 from the lungs to the tissue cell and CO2 from tissue to the lungs. when the hemoglobin reacts with the oxygen to form oxyhemoglobin 97% of O2 is transported in this form similarly when hemoglobin reacts with CO2 to form carboxyhemoglobin 30% of CO2 is transported in this form.
3. Function On haemoglobin: –
   Hemoglobin acts as a good buffer so it maintains acid-base balance.so Estimation of Hemoglobin is important.
   it is globular in shape the molecular weight of HB is 68000 dalton.
4. Types of hemoglobin:-
   Normally two types
   a. HbF ( fetal hemoglobin)
   b. HbA ( adult hemoglobin)

a. HbF:-(fetal Hemoglobin)
It is present in newborns and children.
it consists of Two Alpha chains and two Gamma chains normally.
it is replaced by HbA (adult hemoglobin ) by the end of the first year after birth range of hbF less than 1%.

b. HbA:-(adult hemoglobin)
This is present in adults. it consists of 2 alpha Chains and 2 Beta chains this type of hemoglobin consists throughout life.

Estimation of hemoglobin by Sahli’s method
The different methods used in the laboratory for the estimation of hemoglobin are as follows.

1. Sahli’s method or acid hematin method
2. Cynameth hemoglobin or Drawkin’s method
3. Oxy hemoglobin method
4. Alkaline hemoglobin method

Note:- in all four methods Cynameth hemoglobin method is the more reliable method and Sahli’s method or acid hematin method is the most common method in a rural area.

4. Principle of Hemoglobin
   Hemoglobin is converted into acid hematin by the action of N/10 HCL and then waits 5 to 10 minutes for the change of color and matches with the comparator box.
5. Requirement of hemoglobin: –
   Hemo meter or comparator box
   N/10 HCL
   Hb tube or Sahli’s tube
   Hb pipette
   EDTA vial
   Cotton
   Cotton swab
   Tourniquet
   Syringe
   Distilled water
   Dropper
   Timer
   Applicator
   -Spirit
   Specimen:- Capillary blood or venous blood 0.02ml.
   Preparation of N/10 hcl:- 8.6 ml concentration of HCL solution is mixed with 100 ml of distilled water
6. Testing procedure of Estimation of Hemoglobin :-
   . First of all Sahli’s tube and Hb tube are filled by N/10 HCL on the 2 mark.
   . Then 20 microlitres of blood are mixed with N/10 HCL.
   . Wait 10 minutes for the reaction of acid hematin.
   . After 10 minutes colour is brown of the solution.
   . Brown colors match with the compared box the color does not match then add distilled water drop by drop and mix with the applicator until matching with the comparator box.
   . The color is matched with comparator wax then Distilled Water adding stop.
   . Then I saw the Hb tube and the reports were mentioned properly.
   .Range of hemoglobin seen on the Hb tubes.
7. Normal range.
   In adult males:- 11.5 to 15.5 gm/dl
   In adult females:- 11.5 to 13.5 gm/dl
   During pregnancy: – 9.5 to 12.5 gm/do
   Newborn or children: – 12.5 to.19.5 gm/dl
8. Clinical significance: –
   Increase Hemoglobin then
   Polycythemia
   Vomiting
   Diarrhea
   Decrease Hemoglobin then
   Anemia( due to decrease the concentration of hemoglobin)
   Chronic diseases
   Pregnancy
9. Precaution: –
   The HB tube should be dry and clean.
   Blood must be taken exactly up to 20 micro L in the HB pipette.
   N/10 HCL must be taken exactly up to 2 Mark
   Blood and N/10 HCL mixed property
   Distilled water must be mixed after 10 minutes.
   Give the result carefully.

Note:- when he wants a percentage result you must multiply 6.8 .eg 14 x 6.8 = 95.2%