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Biochemistry of the blood 275 BCH

Experiment-1. Plasma protein , Blood detection .And Haematocrit determination . Heamatology (Hematology)BloodColor of blood ( Why ?)Plasma(function ?) Serum( Different between plasma

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Biochemistry of the blood 275 BCH

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    1. Biochemistry of the blood 275 BCH Tahani Al-Shehri Building 8 3ed floor Office 699 Tmalshehri@ksu.edu.sa Feel free to send any Question , Suggestion , or request related to Our Course . http://faculty.ksu.edu.sa/talshehri/default.aspx http://forums.ksu.edu.sa/forumdisplay.php?f=108

    2. 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. 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 340.000) 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 : By Heat Coagulation

    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. 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 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 wintrobes 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 ( 0.37 -0.47)

    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 .If pcv has been determined by wintrobe tube , one can obtain some more information .

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