1. Biochemistry of the blood 275 BCH
Tahani Al-Shehri
Building 8 – 3ed floor
Office 699
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2. Plasma protein , Blood detection . And Haematocrit determination
Experiment-1
Plasma protein , Blood detection .
And
Haematocrit determination

3. Heamatology (Hematology)
Blood
Color of blood ( Why ?)
Plasma
(function ?)
Serum
( Different between plasma & Serum )

4. Formed Elements and Plasma Introduction
Blood is suspension of cells in plasma
Blood composition :-
Formed Elements
and
Plasma

5. Formed Elements (45%)
The formed elements are cells and cell fragments suspended in the plasma.
The three classes of formed elements are:-
Erythrocytes (red blood cells),
Leukocytes (white blood cells),
Thrombocytes (platelets).

6. Plasma (55%)
The watery fluid portion of blood (90 % water) in which the corpuscular elements are suspended.
It transports nutrients as well as wastes throughout the body.
Various compounds, including proteins, electrolytes , carbohydrates, minerals, and fats, are dissolved in it

7. Blood Composition

8. Plasma protein The main plasma protein :- Albumin ( Mw 66.241)
Globulins (Mw of globulins Cover a wide range
Fibrinogen ( Mw )
Plasma protein can be separated from each other by :
Salting Out / Altracenterifuge/ Electrophoresis / chromatography

9. Objective To separate the principle proteins in serum and plasma
To detect the presence of blood
To detect haematocrit

10. Principle Principle of separation of plasma protein by salting out
The principle of the method is to use a salt which will cause dehydration and neutralization of protein molecule leading to precipitation .
The salt concentration needed for the protein to precipitate out of the solution differs from protein to protein.

11. Principle Principle of identification of plasma proteins :
Fibrinogen :
Biuret test
By appearance of clotting after adding equal volume of serum

13. Principle Globuloins : Biuret test By Heat Coagulation. Albumin :

14. Principle Biuret test Protein + Biuret reagent Blue Color
Heat coagulation
Protein + Dilute acid ( weak acid ) heating
protein ppt ( cloudiness )

15. II)The detection of blood
To detect the presence of small quantity of blood in urine , stomach fluid , etc
Principle of benzidine test
Blood in presence of peroxide (H2O2) catalyze oxidation of benzidine and guaiacum to give blue color .
Benzidine Minute blood Blue color

16. benzidine is toxic & carcinogenic
Caution :
benzidine is toxic & carcinogenic

17. Principle Disadvantage of benzidine test Not specific
Principle of Modified guaiacum test
Hb in blood alkali reacidification Hematin
extraction
Guaiauum hematin peroxide Blue color
Peroxidase present in milk , potatoes ,and pus as well as Fe+++, Cu++,K+ will give false positive results

18. Haematocrit (Hct) III) ( packed cell volume )
Simple screening test for anemia
Hct used in conjunction with MCV and MCHC
Principle of Haematocrit
Blood in heparinized capillary tube
centrifuge blood cell ( bottom )
Plasma ( top)

19. Methods
Using wintrobe’s tube
Using microhaematocrit capillaries

20. Calculation
Hct =length of red blood cell column total length of blood column Normal range : Men ( 0.4 – 0.54) : Women ( )

21. Interpretation Causes of reduced haematocrit – causes of anaemia
Causes of raised haematocrit – causes of polycythemia.
If pcv has been determined by wintrobe’ tube , one can obtain some more information .