ANTICOAGULANT
Definition
Anticoagulant is an agent that is used to prevent the formation of blood clots.
Note: Some of them occur naturally in blood-eating animals such as leeches and mosquitoes.
They are commonly called blood thinner. Some are used for the prevention or treatment of
disorders characterized by abnormal blood clots and emboli. There are several different types
of anticoagulants. Each type works at a different level on the blood coagulation pathway.
Laboratory uses and types:
Sometimes are used in medical equipment, such as test tubes, blood transfusion bags and
dialysis equipment
Characteristics of anticoagulants
An anticoagulant selected for use in hematological examination must have the following
qualities:
[Link] must not alter the size of the cell
[Link] must not cause hemolysis
[Link] must minimize platelet aggregation
[Link] must minimize disruption of staining and morphology of leukocytes
[Link] must be readily soluble in water
[Link] should be soluble in blood
[Link] must be keeping the blood in fluid condition
Common Anticoagulant Materials
1. Oxalates
2. Double Oxalates
3. Potassium Oxalate
4. EDTA (Ethtylene diamine tetra-acetic acid)
5. Heparin
6. Sodium Citrate
7. Sodium Fluoride
8. ACD (Acid Dextrose Citrate)
9. CPD (Citrate Phosphate Dextrose)
10. CPDA-1 (Citrate Phosphate Dextrose Adenosine)
1. Ethylenediamine tetra-acetic acid
EDTA is a standard hematology anticoagulant because of its
- very effective and complete anticoagulation
- lack of effect on the size (morphology) or number of blood cells in the specimen.
Mode of Action:
This anticoagulant removes free calcium ions, which is essential for coagulation by chelating
them
Concentration:
0.5-2.0 mg EDTA per/ml of blood will preserve blood excellently for at least 6hrs.
Types/ Forms of EDTA:
o K3-EDTA
o K2-EDTA
o Na2-EDTA
o Na3-EDTA
o Li2-EDTA, etc.
Advantages/Uses
o Making a blood smear for cell morphology studies
o used for Tests for CBC, microfilaria, coomb’s test.
o EDTA preserves the staining and morphology of Leukocytes
Disadvantages
o Not used for coagulation studies because it destroys factor V and VIII
o Occasionally occurs platelets satellitism with neutrophils
o Occasionally occurs platelet aggregation
o Excessive conc% of EDTA will cause shrinkage of RBC’s and erroneous PCV, MCV,
and MCHC results.
2. Sodium Citrate
The anticoagulant of choice in coagulation studies (PT and APTT) and ESR.
Mode of Action:
The anticoagulant removes the free Calcium ions by binding to them to form calcium citrate
complex.
Advantages/Uses
o Sodium Citrate is the anticoagulant for choice for studies of platelets function and
morphology.
o Acts as both a diluent and an anticoagulant
1. Coagulation studies {concentration of 3.2% and 1part and 9parts (1:9)}
2. ESR {concentration of 3.8% and 1part and 4parts (1:4)}
Disadvantages
o It interferes with many chemical tests
o Used alone it preserves blood for only few min.
o It has a tendency to shrink cells. Sodium citrate is generally not used for CBC.
3. Heparin
o It is a natural anticoagulant in the body, found in the liver, and may also be with in
basophils and mast cells Heparin is also called anti thromboplastin or antithrombin.
o It is available in a liquid or dry form a s sodium, calcium, ammonium and lithium salt
Each of these will interfere with determination of their respective ions in the plasma
Mode of Action
It interferes with the formation and/or activity of thrombin and the activity of clotting factors
IX, X, XI, XII.
Concentration:
The optimum con% is 0.1-0.2 mg/ml of blood.
Advantages/Uses
o Heparin is the choice of Anticoagulant for blood pH and blood gas Analysis and Acid
base balance.
o It may be used for special trace elements studies and some cytology.
o Excessive heparin does not alter the RBC volume.
Disadvantages
o It causes clumping of leukocytes
o It interferes with staining of leukocytes.
o It is the most expensive anticoagulant
o Blood clot in 8-12 hrs because clotting is only delayed and not prevented.
o It is not suitable for agglutination tests, and coagulation studies
o It may interfere with some automated biochemical analysis of plasma.
4. Oxalates
Mode of Action:
o These acts by chelating calcium. Calcium oxalate is formed as insoluble precipitate.
o These are used for blood chemistry and hematocrit.
Types:
o Potassium oxalate
o Ammonium oxalate
o Double oxalate
Concentration
This is used at conc.% of 2 mg/ml of blood.
Uses
This anticoagulant is most often used for chemical analysis
Disadvantages
Potassium oxalate shrinks the RBC, about an 8% shrinkage in the PCV and therefore it is not
recommended for use with blood for PCV and ESR.
5. Double Oxalates (Potassium oxalate & ammonium oxalate)
Double oxalates is used for ESR and HCT
Concentration
Potassium oxalate and ammonium oxalate are used together in a ratio 2:3, this is done to
counter the swelling effect of ammonium oxalate and shrinkage effect of potassium oxalate
on the RBC
Advantages/Uses
Double oxalates can be used for Hb%, TLC, RBC count and ESR by Wintrobes method.
Disadvantages:
o Leukocyte morphology is not well preserved.
o The calcium chelated is precipitated in calcium oxalate which is a toxic substance.
6. Sodium fluoride
Mode of Action
Sodium fluoride inhibits the glycolytic enzymes responsible for the breakdown of glucose in
the blood. (At room temperature about 10% glucose is lost per hour from an untreated
sample)
Concentration
The optimum concentration: 1 mg of mixture per 1 ml of blood.
Use
Glucose determination
Disadvantages
o It is poisonous.
o Its inhibition of unease, and glycolytic enzymes may interfere with urea and glucose
determinations that employ enzyme activity.
o Alkaline phosphatase, amylase and uric acid cannot be determined in blood
containing sodium fluoride.
7. ACD (Acid Dextrose Citrate)
CPD (Citrate Phosphate Dextrose)
CPDA-1 (Citrate Phosphate Dextrose Adenosine)
Uses
o in blood transfusion
Color code tube selection of anticoagulants
