Laboratory Activity 6 : HEMOGLOBIN DETERMINATION

Question 1

The (?) is the gold standard technique for hemoglobin determination.

Answer 1

Cyanmethemoglobin method

Question 2

This is a type of colorimetric method wherein blood is mixed with a solution containing (?).

Answer 2

potassium ferricyanide and potassium cyanide

Question 3

The (?) oxidizes iron to form methemoglobin.

Answer 3

potassium ferricyanide

Question 4

The (?) then combines with methemoglobin to form cyanmethemoglobin, a stable color pigment measured photometrically at 540nm.

Answer 4

potassium cyanide

Question 5

Manual hemoglobin determination can also be done using the (?) which gives an estimation of hemoglobin based on its color.

Answer 5

Sahlis method

Question 6

(?) is based on converting hemoglobin to acid hematin (brown in color) and then visually matching its color against a solid glass standard.

Answer 6

Sahlis method

Question 7

A. Cyanmethemoglobin Method
1. Using a serological pipet, transfer (?) into a clean test tube.
2. Make a (?) and discard the first 2 drops.
3. Using a Sahli pipette, draw blood up to the (?) mark. Wipe off any blood which adheres to the outside of the pipet.
4. Dispense the blood into the tube of (?) and carefully wash blood into the reagent by repeated rinsing of the pipette.
5. Mix well and allow to stand at room temperature for (?).
6. Read the absorbance of the resulting solution in a spectrophotometer at (?) against reagent blank.
7. Convert the absorbance into (?) by using a constant determined from the absorbance of a series of standards of known concentrations (calibration curve), or compute for Hemoglobin concentration appling the Beers Law
8. Express results (concentrations) in (?)

Answer 7

5 ml. of Drabkins solution

finger puncture

20 cumm

Drabkins reagent

3 minutes

540 nm

concentration of hemoglobin

g/dL and in g/L

Question 8

Hb Concentration =

Question 9

Acid Hematin Method (by Sahli-Helige)
1. Place into a Sahli round (or square) tube 0.1 N HCl up to (?) mark.
2. Make a finger puncture and discard the first (?).
3. Using a Sahli pipette, draw blood up to the (?) mark. Wipe off any blood which adheres to the outside of the pipet.
4. Dispense the blood into round (or square) tube and carefully wash blood into the reagent by (?) of the pipette.
5. Mix well and allow to stand at room temperature for (?)
6. (?) with distilled water until the brown color matches with the comparator block of the hemoglobinometer.
7. (?) from round (or square) tube in gms%

Answer 9

2

2 drops

20 cumm

repeated rinsing

5 minutes

Dilute colored solution

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