Anticoagulants

Faculty, Department of Pathology, SMIMS.

This presentation is meant for circulation amongst DMLT students of
SMU only.
What are anticoagulants

• Anticoagulants are the chemical agents which prevent the coagulation

of blood
Different anticoagulants used in
hematology investigations
I. EDTA (Ethylene Di-Amine Tetra Acetic Acid)
ii. Double oxalate
iii. Sodium citrate
iv. Heparin
v. Sodium fluoride
EDTA
•  What is EDTA and what are its different forms and what is its mechanism
of action?
• There are three different forms which are
i. Tripotassium EDTA
ii. Disodium EDTA
iii. Di Lithium EDTA
Tripotassium EDTA is more soluble and it is more preferred than the other
forms.
Mechanism of action: EDTA acts by Chelation of calcium ions/ removing the
free calcium ions. We know that calcium is very much needed for clotting of
blood. Hence, when the calcium ions are removed, the blood does not clot.
What are the advantages and
disadvantages of EDTA as anticoagulant in
Hematology
• The advantages are
i. The morphology of the blood cells are well preserved up to 4 hours and
hence it is the anticoagulant of choice to peripheral smear preparations and
cell counts.
• Disadvantages
i. If there is excess EDTA/ the volume of blood filled is less so that the relative
volume of EDTA is excess, then it results in shrinkage of RBC’s. This shrinkage
can result in erroneously high MCHC and reduced hematocrit with decrease in
MCV
ii. Platelets can also disintegrate and can result in erroneously high values/
Pseudothrombocytopenia.
iii. EDTA is not suitable for coagulation studies
Double oxalate
• It is a combination of ammonium and potassium oxalate in the ratio
of 3:2 ( 1.2mg and 0.8 mg respectively in 100 ml of distilled water).
0.5 ml of this solution is sufficient for anticoagulating 5 ml of blood.

• What is the mechanism of action of double oxalate

The oxalates combine with calcium in the blood to form insoluble
precipitate of calcium oxalate. Thus there is depletion of calcium
which is necessary for coagulation of blood.
Double oxalate contd…
• What are the advantages of double oxalate combination
Ammonium oxalate causes swelling of the cells, whereas potassium
oxalate results in shrinking. So, the combined effect is that the cells
retain their original shape and size.

•  What are the uses of Double oxalate

This anticoagulant is used in estimation of ESR by Wintrobes’s method
and PCV.
Sodium citrate
•  What is the mechanism of action of sodium citrate
The sodium citrate binds with the free calcium resulting in sodium
citrate complex. Thus depleting calcium and preventing coagulation.

•  What are the uses of Sodium citrate

i. Used for estimation of ESR by Westergren’s method. The ratio of
citrate and blood has to be maintained at 1:4
ii. Used in coagulation studies. The ratio of citrate and blood has to be
maintained at 1:9
iii. As a component of Acid Citrate Dextrose solution in blood banking
Sodium Citrate
Heparin
• Heparin an acid mucopolysaccharide, has an affinity for blood
proteins and act as antithrombin and antithromboplastin. It prevents
conversion of prothrombin to thrombin, thus resulting in
anticoagulation.
• What are the uses of heparin as anticoagulant in hematology
Heparin is used as anticoagulant in the following hematology
investigations
i. Osmotic fragility test
ii. For L E Cell preparation
iii. In Microhematocrit capillary tubes.
Heparin contd…
Thankyou