Anticoagulants

 Anticoagulants are chemical substances which can prevent the coagulation of blood
either in vivo or in vitro
 Anticoagulants are used to prevent coagulation of blood samples
 Whole blood and plasma are obtained using anticoagulants
 Tube containing adequate (pre-weighed) amount of anticoagulants are known as
vacutainers
 Each anticoagulant has specified for particular test /testing
 One anticoagulant can interfere with tests carried out by another anticoagulant

Anticoagulants

Natural Artificial

(chemical)
Substances

Heparin EDTA, Oxalates

Hirudin Citrates

 Depending up on form

Dry Wet (liquid)

(Solid)

EDTA, oxalate ACD

Citric acid
Fluorides
Dextrose
Sodium
citrate

 Based on use

Routinely used Special propose

ACD – Blood Banking
EDTA , Oxalate Fluoride Oxalate – Blood
Sodium Citrate sugar estimation
 Each anticoagulant differ in its modes of action
 Most of the anticoagulants prevent clotting by calcium chelating
Binding calcium as an insoluble salt which
precipitates or converting it to a soluble or non – ionized form

 EDTA (Ethylene Diamine Tetra Acetate)

 Routinely used anticoagulant in hematology lab

 Commercially it is known as versene/sequestrene
 Most powerful calcium chelate-It prevents coagulation by binding calcium and
converting it to a non-ionized soluble complex
 It is used at a concentration of 1-2 mg/100 ml of blood( 1.5-2.2 mg/ml of Blood)

Advantages :Preseves cell morphology and preferred for all cell counts

 It gives best preservation of cell morphology and prevents the clumping of

platelets.
 Smear can be made up to 3-4 hours after collecting blood in EDTA
 Disadvantage :- Only disadvantage is that it is slightly more expensive than other
commonly used anticoagulants
 Oxalates :- They combine with calcium in the blood to form insoluble calcium oxalate which is
precipitated thus preventing the coagulation of blood

Potassium Oxalate Ammonium Oxalate Double Oxalate

Used to a concentration
of 2mg/ml of blood
Used for chemical analysis
Not recommended for
PVC, ESR and for study of
cell morphology , cause
shrinkage of RBC’s
Used as a concentration
of 2mg/ml of blood
Used for chemical analysis
Not recommended for
PCV, ESR. They cause
swelling of RBC
Balanced Oxalate
(Wintrobes mixture)
Ammonium Oxalate and
potassium oxalate in the
ratio 3:2
Conc 2mg/ml*. used for
Hb, PCV, ESR
morphological studies
 *From VHSE text
books
 In PG text books
0.4mg/ml

 Citrate : - Prevent coagulation by binding calcium in to a non-ionized soluble complex

 Used only as a wet anticoagulant
 Used in Blood culture
 Cannot be used for doing blood cell counts, Hb estimation us it causes dilution of blood

Trisodium Citrate ACD

(Anticoagulant choice of ESR estimation) Special anticoagulant used in blood
banking
ESR estimation by Westergren method Citric acid,Trisodiumcitrate, Dextrose
3.8% sodium citrate mixed with blood in
the ratio 1:4
For coagulation studies 3.2 % is mixed
with 9 parts of whole blood (1:9)

 Heparin :- Natural biological anticoagulant which is normally present in the body in

small amounts
 It is antithrombin III in action. Inhibits the formation of thrombin from
prothrombin and thereby inhibiting action of thrombin on fibrinogen
 Required only in very small amount like 0.1 – 0.2 mg/ml of blood
 It is not used routinely and is very much expensive
 Advantage : It does not affect cell size or the composition of the blood and it is
the best anticoagulant for investigations like osmotic fragility test blood gas
analysis
 Electrolyte estimation
 Disadvantage:- not satisfactory for doing cell counts as it causes clumping of cells
and for smear preparation as it gives a colored background
 Disadvbantage: Anticoagulation only in limited period , Blue colouration, Not stable
in hot climate, Expensive.

Q) All are examples of anticoagulants which act as chelators except

a) Heparin
b) Oxalates
c) Citrate
d) EDTA
 Sodium fluoride
 Required in large quantities
 A mixture of potassium oxalate and sodium fluoride in the ratio 3 : 1 used for collecting
blood for blood glucose estimation
 MOA :- Sodium fluoride being an enzyme poison prevents glycolysis

MUST KNOW

 ESR – Must be done with in 2hrs after collection in anticoagulants

 PT, Plasma coagulation studies platelet count with in 2hrs
 Hb, RBC, WBC, Reticulocyte – done within 24 hours
 Smear must be made within 3 – 4 hours using EDTA blood