Anticoagulants

Anticoagulants
• Anticoagulants are chemical substances that are added to blood to prevent coagulation.
Blood must remain fluid within the vasculature and yet clot quickly when exposed
to subendothelial surfaces at sites of vascular injury. When intravascular thrombi
form, rapid activation of the fibrinolytic system restores fluidity. Under normal
circumstances, a delicate balance between coagulation and fibrinolysis prevents
both thrombosis and hemorrhages.

• The substances responsible for this removal or inactivation are called anticoagulants. It is
used in hematological laboratory where complete blood or plasma is required.
Types of anticoagulants:-

1. Ethylenediamine tetra acetic acid (EDTA)

• (EDTA) has become the standard hematology anticoagulant
because of its very efficient and complete anticoagulation and its
lack of effect on the size (morphology) or number of blood cells
in the specimen. Therefore, it can be used for blood cell count,
ESR, etc. It exerts its effect by tightly binding (chelating)
calcium ion thus effectively blocking coagulation. It has
recommended a concentration of 1.5±0.25mg of Na2 or K3 EDTA
per 1ml of blood (e.g. 0.02ml of 10% (W/V) solution of K3EDTA
is used for 1ml of blood). This concentration does not appear to
adversely affect any of the erythrocyte or leukocyte parameters.
2. Sodium Citrate (Tri-sodium citrate, Di- sodium
citrate)

Sodium citrate combines with calcium, thereby

preventing the conversion of prothrombin to thrombin, and
coagulation does not occur. This anticoagulant is prepared by
dissolving 3.8gm of trisodium citrate in 100 ml of distilled water.
0.5 ml of anticoagulant is required for 2 ml of blood. Addition of
dextrose to anticoagulant provides nutrition to the red cells and
helps in longer storage. Such anticoagulant is known as acid
citrate dextrose (ACD). This anticoagulant is used in the solution
form, as it is in blood bank. The storage of blood with this
anticoagulant is maximum up to 21 days.
3. CPD (citrate phosphate dextrose)

• In this anticoagulant, the citrate is dissolved in phosphate

buffer, which maintains pH more accurately than ACD acid
citrate dextrose solution. The dextrose present in this
provides nutrition to other cells. The storage period of blood
with CPD is 21 days. The disadvantage is that, it is costly and
difficult to prepare and adjust pH.
4. Oxalate
• Salts of oxalic acid by virtue of their ability to bind and
precipitate calcium as calcium oxalate serve as suitable
anticoagulants for many hematologic investigations. Three
parts of ammonium oxalate is balanced with two parts of
potassium oxalate (ammonium oxalate causes cellular
swelling and potassium oxalate causes erythrocyte
shrinkage). It is used in the proportion of 1-2mg/ml of blood. It
is prepared by dissolving 1.2 gm of ammonium Oxalate and 0.8
ml of potassium oxalate to 100 ml of distilled water. 0.2 ml of
anticoagulant is required for 2 ml of blood.
Biological Anticoagulant:-
1-Heparin
• Heparin is an excellent natural anticoagulant extracted
from mammalian liver or pancreas. It is more expensive
than the artificial ones. Heparin prevents clotting by
inactivating thrombin, thus preventing conversion of
fibrinogen to fibrin. It is the best anticoagulant when
absolute minimal hemolysis is required (e.g., osmotic
fragility test and hematocrit determination). It is used in
concentration of 10-15 units/ml blood.
2- Coumarin, Dicoumarol and warfarin:-

• The coumarin derivatives are vitamin k

antagonists and thus inhibit the action of this
vitamin that is essential as a cofactor for the
synthesis of six glutamic acid containing
proteins namely, factor ll (prothrombin),Vll, lX,
and X, protein C, and protein S. The action of
this anticoagulant is, however, slower that of
heparin.
• Vitamin K is essential for the production of active clotting

factors II, VII, IX, and X. In humans, vitamin K is obtained

primarily from the diet in the form of phylloquinones, which are

found in greatest concentration in green leafy vegetables.