Anti Coagulants & Plasma Proteins

• Presented By – • Prof.Dr.R.R.Deshpande • (M.D in Ayurvdic Medicine & M.D. in

Ayurvedic Physiology)• www.ayurvedicfriend.com• Mobile – 922 68 10 630• professordeshpande@gmail.com

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Contents of this PPT

• 1) What is Anti Coagulant ? • 2) Uses of Anti Coagulants

• 3) Examples of Anticoagulants –Coumerin & Heparin

• 4) Laboratory or in Vitro use of Anti Coagulants

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Contents of this PPT

• 5) Three Main Plasma Proteins as Albumin, Globulin & Fibrinogen

• 6) Properties of Plasma Proteins • 7) Functions of Plasma Proteins• 8) Pathology of Plasma Proteins • 9) Plasmapheresis or Therapeutic Plasma

Exchange

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What is Anticoagulant ?

• An anticoagulant is a substance that prevents coagulation or clotting of blood

• Such substances found naturally in leeches

• Anticoagulants can be used as a medication for thrombotic disorders

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Types of Anticoagulants

• 1) Which prevent blood clotting in vivo ( inside the body)

• 2) Which prevent blood clotting in vitro ( outside the body)

• 3) Which prevent blood clotting in vivo & in vitro both

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Heparin

• Naturally produced by mast cells in the body .• Mast cells are abundant in Liver & Lungs • Basophils also secrete Heparin

• Commercial heparin is prepared from the liver and other organs of animals. It is available in liquid form or dry form as sodium, calcium, ammonium or lithium salts.

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Heparin Action

• Prevents blood clotting by its anti thrombin activity. It suppresses the activity of thrombin

• Inactivates the active form of other clotting factors like IX, X, XI and XII

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Uses of Heparin

• 1) Anticoagulant –both in Vivo & in Vitro • 2) It prevents intravascular blood clotting during

surgery.• 3) To prevent clotting ,While passing the blood

through artificial kidney for dialysis.• 4) Prevent clotting ,during cardiac surgery, which

involves heart lung machine.• 5) Used to preserve the blood before transfusion

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Dose of Heparin

• 0.5 to 1 mg/kg body weight

• Postpones clotting for 3 to 4 hours

• Till Heparin is destroyed by the enzyme heparinase

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Coumarin Derivatives

• Warfarin and dicoumoral• Action -- Prevent blood clotting by inhibiting the

action of vitamin K. • Vitamin K is essential for the formation of various

clotting factors, namely II, VII, IX and X.• Commonly used as oral medicine• Warfarin --- used to prevent myocardial infarction

(heart attack), strokes and thrombosis.

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Coumarins (vitamin K antagonists)

• These oral anticoagulants are derived from coumarin, which is found in many plants

• A well known example from this group is warfarin (coumadin).

• It takes at least 48 to 72 hours for the anticoagulant effect to develop.

• Where an immediate effect is required, heparin Can be used

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EDTA

• EDTA = Ethylene diamine tetra acetic acid• This is strong anticoagulant.• Available in two forms:• i. Disodium salt (Na2 EDTA).• ii. Tripotassium salt (K3 EDTA).• Prevent blood clotting by removing calcium

from blood

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EDTA Uses

• 1) I.V – In lead poisoning

• 2) In Laboratory -- 0.5 to 2.0 mg of EDTA per mL of blood --- sufficient to preserve the blood for at least 6 hours

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Oxalate Compounds

• Oxalate combines with calcium and forms insoluble calcium oxalate

• So , oxalate removes calcium from blood and lack of calcium prevents coagulation

• Used only in Vitro –Because Oxalate is Poisonous ,so can not be used in Vivo

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Citrates

• Sodium, ammonium and potassium citrates

• Citrate removes calcium from blood and lack of calcium prevents coagulation

• It is used to store blood in the blood bank as: Acid citrate dextrose (ACD)

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Laboratory use

• Bulbs used for laboratory blood tests will have chemicals added as Anticoagulant

• Like Wintrobe bulb or Fluoride bulb

• Wintrobe Bulb = Double Oxalate bulb =Ammonium oxalate 6 mg + Potassium Oxalate 4 mg –Used for collecting blood for Haemogram + ESR PCV

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Laboratory use

• Fluoride Bulb = Sodium fluoride + Potassium Oxalate

• Used for collecting blood for Blood sugar Test

• EDTA Bulb = Ethylene Diamino Tetra Acetic Acid – Used for collecting blood for Platelet count

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Laboratory use

• But in Plain bulb there is no anticoagulant

• Plain bulb is used for collecting blood for serological Tests like Liver Function Test, Renal Function Test.

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General indications – To Conclude

• Therapeutic uses of anticoagulants include --• Atrial fibrillation • Pulmonary embolism• Deep vein thrombosis• Venous thrombo embolism• Congestive heart failure• Stroke, myocardial infarction• Genetic or acquired hypercoagulability

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Food & herbal supplements

• Foods & food supplements with blood-thinning effects include ---

• Alfalfa, avocado, beer, celery, cranberries, fish oil

• Garlic, ginger, ginseng, grapefruit, green tea,, licorice, niacin, onion, papaya, pomegranate, soybean, turmeric, wheatgrass

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Other Anti Coagulants

• Proteins from venom of viper snake -- Peptone, C-type lectin

• Hirudin --- from the leach -- Hirudinaria manillensis

• Reducing the temperature to about 5°C postpones the coagulation of blood.

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Blood Clotting Tests

• To diagnose blood disorders.

• To monitor the patients treated with anticoagulant drugs such as heparin and warfarin.

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Bleeding Time( BT)

• Time interval from oozing of blood after a prick till arrest of bleeding.

• Determined by Duke method using blotting paper

• Normal = 3 to 6 minutes.• It is prolonged in Purpura.

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Clotting Time ( CT)

• Time interval from oozing of blood after a prick till the formation of clot.

• Determined by capillary tube method.

• Normal = 3 to 8 minutes

• It is prolonged in Hemophilia.10/10/2017 Prof.Dr.R.R.Deshpande 24

Prothrombin Time ( PT)

• Time taken by blood to clot after adding tissue thromboplastin to it

• Normal = 10 to 12 seconds. • It is prolonged in deficiency of prothrombin

and other factors like factors I, V, VII and X. • However, it is normal in hemophilia.• Prothrombin time indicates the total quantity

of prothrombin present in the blood

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General indications for Anti coagulants

• Therapeutic uses of anticoagulants include --• Atrial fibrillation • Pulmonary embolism• Deep vein thrombosis• Venous thrombo embolism• Congestive heart failure• Stroke, myocardial infarction• Genetic or acquired hypercoagulability

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Market Products Sr.No Generic Name Brand Name Availability

1 Heparin Nuparin 1000 & 5000 Units /ml in 5 ml vials for Injection

2 Low molecular Weight Heparins

Enoxaparin = Clexane 20 mg (0.2 ml) & 40 mg (0.4 ml) Prefilled syringes –20 to 40 mg OD –S/C

3 Oral Anti coagulants Warfarin =Uniwarfin 1,2,5 mg tab

4 Oral Anti coagulants Bishydroxycoumarin ( Dicumarol) = Dicoumarol

50 mg Tab

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3 Main Plasma Proteins

• 1. Serum albumin

• 2. Serum globulin

• 3. Fibrinogen• Note –Fibrinogen is not present in serum

,because it is converted into fibrin during blood clotting

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Normal Values

• Total proteins = 7.3 g/ 100 ml

• Serum albumin : 4.7 g/ 100 ml• Serum globulin : 2.3 g/ 100 ml• Normal A/G ratio is 2 : 1.

• Fibrinogen : 0.3 g/ 100 ml

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Separation of Plasma Proteins

• 1. Precipitaion method• 2. Salting out method• 3. Electrophoretic method• 4. Cohn’s fractional precipitation method

• 5. Ultracentrifugation method• 6. Gel Filtration Chromatography• 7. Immuno electrophoretic method

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Where Plasma Proteins are Synthesized ?

• 1) In Embryo –By mesenchyme cells. First albumin is synthesized and then other proteins are synthesized later.

• 2) In Adults – Mainly from reticuloendothelial cells of liver. Also from spleen, bone marrow, disintegrating blood cells and general tissue cells. Gamma globulin is synthesized from B lymphocytes.

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Plasma Proteins –Properties

• 1) Molecular Weight

• 2) Oncotic pressure

• 3) Specific Gravity = 1.026

• 4) Buffer Action

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Molecular Weights of plasma Proteins

• Albumin = 69,000

• Globulin = 1,56,000

• Fibrinogen = 4,00,000

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Oncotic pressure of plasma Proteins

• Osmotic pressure exerted by proteins in the plasma is called colloidal osmotic (oncotic) pressure

• In this -- Albumin plays a major role

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Buffer Action of plasma Proteins

• Buffer action means -- acceptance of hydrogen ions

• 1/6 of total buffering action of the blood is contributed by Plasma Proteins

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Functions of Plasma Proteins

• 1) Coagulation of Blood – Fibrinogen has a major role

• 2) Immunity or Defense mechanism – Gamma Globulins or Immuno-globulins act as anti bodies to fight with microorganisms ,which cause diseases like Typhoid, Mumps, Influenza, Measles, Hepatitis, Polio

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Functions of Plasma Proteins

• 3) Transport -- Albumin, alpha globulin and beta globulin do the transport of the hormones, enzymes, etc.

• The alpha and beta globulins transport of metals in the blood

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Functions of Plasma Proteins

• 4) Maintain Osmotic pressure in the blood --• At the capillary level, substances are exchanged between the

blood and the tissues. But the plasma proteins cannot pass through the capillary membrane easily because of their large size and remain in the blood. In the blood, these proteins exert the colloidal osmotic or oncotic pressure.

• Osmotic pressure exerted by the plasma proteins is about 25 mm Hg.

• Albumin ,Globulin ,Fibrinogen exert pressure in descending order

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Functions of Plasma Proteins

• 5) Regulation of Acid Base Balance – Albumin is important in regulating the acid base balance in the blood due to it’s buffering action

• 6) Maintain Viscosity of blood -- Albumin• provides maximum viscosity than the other

plasma proteins, which helps in maintaining Blood Pressure

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Functions of Plasma Proteins

• 7) Useful For ESR Test – ESR is Good Prognostic Test in the disease like TB ,RA . Rouleaux formation is responsible for ESR .

• Rouleaux formation is accelerated by Globulin and fibrinogen

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Functions of Plasma Proteins

• 8) Help in Suspension Stability of RBCs –

• Globulin and fibrinogen help RBCs to remain during circulationsuspended uniformly in the blood

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Functions of Plasma Proteins

• 9) Last source of Energy -- The plasma proteins are used by the body as the last source of energy during fasting or inadequate food intake or inadequate protein intake. Tissue Macrophages split Plasma proteins into amino acids ,which are taken up by blood .They are distributed in the body to form cellular proteins. So Plasma proteins are called as Reserve Proteins

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Plasma Proteins –Pathology

Sr.No

Plasma Proteins

When Increased ? When Decreased ?

1 Total Proteins Dehydration ,Acute infections like acute hepatitis and acute nephritis ,Excess of glucocorticoids Leukemia Rheumatoid arthritis

DiarrheaHemorrhageBurnsPregnancyMalnutrition

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Plasma Proteins –Pathology Sr.No Plasma Proteins When Increased ? When Decreased ?

2 Albumin Dehydration ,CCF Cirrhosis,Malnutrition

3 Globulin Cirrhosis, RA GlomerulonephritisEmphysema

4 Fibrinogen MI ,Stroke Liver dysfunction

5 Albumin –Globulin Ratio

Hypothyroidism Liver Dysfunction

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What is Plasmapheresis ?

• Experimental procedure done in animals to demonstrate the importance of plasma proteins.

• Clinical significance of Plasmapheresis = – Therapeutic Plasma Exchange

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Plasmapheresis

• Used as a blood purification procedure for an

• Effective temporary treatment of many auto immune diseases like ---

• Myasthenia gravis , Thrombocytopenic purpura , GuillainBarré syndrome

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Prof.Dr.R.R.Deshpande

• Sharing of Knowledge

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• Propagating Ayurved

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