BIOCHEMISTRY 285 PHL Introduction Blood Glucose

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Presentation transcript:

BIOCHEMISTRY 285 PHL Introduction Blood Glucose

Blood Blood is a connective tissue that circulates in the closed system of blood vessels Functions: Transportation Regulation of acid-base balance Regulation of body temperature Immunity Coagulation

Blood: Composition Blood Plasma Cellular Elements Water Solids : Diffusible - Anabolic - Catabolic Non- diffusible RBCs WBCs Platelets

Types of Samples Whole blood Plasma Serum Water (plasma) + cells (RBCs, WBCs, platelets) Plasma Water + solids (e.g. glucose, urea, albumin, fibrinogen) No cells Serum Serum = plasma – clotting factors

Centrifuging tubes Centrifugator

Preparation of sample: Plasma anticoagulant Transfer the clear supernatant to specimen tube Add venous blood Mix then centrifuge cells Cent. tube

Preparation of sample: Serum Venous Blood Transfer the clear supernatant to specimen tube Allow blood to clot (20min.) Remove the clot and centrifuge cells Cent. tube

Comparison Plasma Serum Anticoagulant No anticoagulant plasma serum Contains clotting factors Advantage: time saving No anticoagulant Contains no clotting factors Advantage: less interference plasma serum

Anticoagulants Definition: Types : 1- Heparin Anticoagulants are chemicals which prevent blood clotting Types : 1- Heparin MOA: Prothrombin thrombin Advantage: less interference with chemical tests Disadvantage: high cost

Anticoagulants: cont. 2-EDTA (ethylene diamine tetraacetic acid): MOA: Binds to calcium Advantage: prevents platelets clumping 3-Oxalates: (Na, K, Li, or NH4 salts) Form insoluble complex with calcium Disadvantage: interfere with lactate dehydrogenase N.B Na, K salts should not be used in electrolytes determination

Anticoagulants: cont. 4-Citrate: 5-Na fluoride: (enzyme poison) e.g. trisodium citrate Used in ESR 5-Na fluoride: (enzyme poison) Used in blood sugar determination b/c it inhibits glycolysis N.B: it inhibits urease enzyme., therefore it should not be used in urea determination

Deproteinization Purposes: 1- Proteins have UV absorption 2- Proteins make the solution turbid & difficult to read 3- Determination of non-protein nitrogen & glucose 4- Use the precipitate in plasma protein determination e.g. albumin

Deproteinization Agents Acids 1- Trichloroacetic acid 2- Tungestic acid MOA: ↓ pH proteins become cations ppt as insoluble salts of acids Bases 1- Zinc hydroxide 2- Cu, Ba, Cd hydroxide -↑ pH, proteins become anions ppt as insoluble salts of heavy metals

Deproteinization Agents Organic substances e.g. ethanol or ether MOA: remove water from protein mol.