ANTICOAGULANTS

Introduction
 Is a Substance that prevents clotting of blood
 Most of the anticoagulants used in the lab act by
binding calcium as an insoluble salt or soluble but
unionized salt
Purpose

 For studying morphology of blood components

 For studying coagulation profile
 Preservation of blood in blood bank
Properties
 Must be soluble in blood
 Must be able to keep blood in fluid condition
 Must not bring hemolysis in blood cells
 Must preserve the morphology of cells
 Must minimize aggregation of platelets
1. oxalate
A. potassium oxalate

 Used at a concentration of 2mg/ml of blood

 Shrinks RBC

 Not recommended for PCV, ESR and smear

preparation
B. Ammonium oxalate

 2mg/ml

 Causes swelling of RBC

 Not recommended for PCV, ESR and smear

preparation
 C. Balanced oxalate

 Mixture of 3 parts of ammonium oxalate and 2 parts of

potassium oxalate

 2mg/ml

 Used for Hb, TLC,RBC count, ESR(wintrobe’s method)

 Not good for blood smears since WBC morphology is often

distorted in blood anticoagulated with oxalate
Mode of action
 Calcium in the blood combines with oxalate to
form an insoluble calcium oxalate compound
which precipitates
Disadvantage
 Doesn’t preserve the morphology of the cells too
well

 Can’t be used as an anticoagulant to be injected

into the body as it is toxic and calcium oxalate
precipitate may cause harm

 Can’t be used for coagulation profile as the labile

factors(V and VIII are unstable in oxalate)
2. EDTA
 Commercial name : versene and sequestrene

 Used as the disodium salt or dipotassium salt of the acid

 Used at a concentration of 1-2mg/ml

 Gives the best preservation of cell morphology

 Preferred for all cell counts and smears

 Smears can be made upto 3 hours after collecting blood and good
morphology is still preserved
 EDTA reduces platelet activation and clumping

 Studies have demonstrated that the WBC count

remained stable for at least 3 days when EDTA
anticoagulated blood was stored at room temperature

 Anticoagulant recommended by ICSH is the

dipotassium salt at a concentration of 1.50±0.25
mg/ml
Action
 Calcium is bound in an unionized form, but still
soluble complex with the EDTA
Advantages
 Cellular morphology is preserved better, even 2-3
hours after blood collection

 Platelet clumping is prevented so EDTA is a better

anticoaglant for platelet count
Disadvantages
 Slightly expensive than oxalate

 Unsuitable for use in cagulation profile as it directly inhibit the

coagulation process

 Responsible for the activity of antiplatelet autoantibody which

may sometimes cause platelet adherence to neutrophils in blood
films

 EDTA is not suitable forchemistry test as it inhibits the alkaline

phosphatase, creatine kinase, and leucine aminopeptidase
activities.
3. Citrate
A. Trisodium citrate

 Commonly used as 3.8% solution for ESR by

westergren method and for PT

 For ESR: 1 part of 3.8% TSC is mixed with 4 parts of

blood

 For coagulation profile: 1 part of 3.2% TSC is mixed

with 9 parts of blood
ACTION
 Prevents coagulation by binding calcium in an
unionized form
Disadvantage
 Is used as a liquid so that it dilutes the blood with
which it is mixed
 Therefore, not acceptable for any blood cell counts
or for Hb estimation
B. ACD
 Commonly used anticoagulant in blood banking

 Blood can be stored upto 21 days at 2-6°c

 Composition:
Trisodium citrate - 22gm
Citric acid - 8 gm
Dextrose -25gm
Water -1 litre
B. CPD
 Blood can be stored upto 28 days

 Better than ACD due to addition of phosphate

buffer
C. CPDA

 Blood can be stored upto 35 days

 Composition
Trisodium citrate - 26-30gm
Citric acid - 3.27 gm
Na dihydrogen phosphate - 3.22gm
Dextrose -31.8gm
Adenine - 0.275gm
Water -1 litre
D. CPD-SAGM

 Blood can be stored upto 42 days

 It has integrally connected plastic bag system

 One bag contains CPD solution and another bag contains SAGM
additive solution

 SAGM(Saline Adenine Glucose Mannitol solution)

Nacl
Adenine
Dextrose
Mannitol
D/w
Heparin
 Highly acid substance which is normally present in the
body in small amounts

 Is a natural biological anticoagulants

 Used for some haematology special tests ,Also for

testing electrolytes or blood gases

 Earlier heparin blood was used in transfusion (open

heart surgery) but now it is not in much use
 It is not a calcium chelator
 Used at a concentration of 0.1-0.2 mg/ml
 Sometimes expressed in international
units(1000IU=10mg)
 Causes minimum hemolysis
Action
 It binds to the anti-thrombin III that results in its
activation
 Thus, inhibits the action of thrombin on fibrinogen
by complexing with anti-thrombin III and prevents
coagulation
 Also acts as an inhibitor of thromboplastin
formation
Disadvantage
 Is expensive
 Unsuitable for blood films as it gives a faint
discoloration to a background
 Can’t be used for leukocyte count as it tends to
cause the leukocyte clump
 Prevents antioagulation only for limited time and
so heparinised blood should be processed promptly
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