The document discusses various anticoagulants used in blood collection, highlighting their actions, uses, advantages, and disadvantages. It details substances like EDTA, double oxalate, tri sodium citrate, heparin, acid citrate dextrose, fluoride, and plain bulbs, explaining their specific applications in laboratory testing. Key concepts of haemostasis and the difference between serum and plasma are also described.

1. ANTICOAGULANT USED IN
BLOOD COLLECTION
Ms. Ankita Bhatiya
Assistant Professor
Shree.P.M.Patel college of
Parmedical science & Technology

2. • ANTICOAGULANT:
When blood is collected it clots after some time. Anticoagulants are the chemicals, which stope
clotting, of blood
When mixed with blood in proper proportion.
• CLOT FORMATION:
Haemostasis: Prevention of blood loss. OR when injury occur bleeding stop by the action is known as
Haemostasis
The Thromboplastin released by damaged tissue, or platelets convert inactive prothrombin into active
thrombin in the presence of calcium ions. Thrombin converts soluble fibrinogen into insoluble fibrin clot in
the presence of calcium ions.
Prothromnbin ------------------------------ Thrombin
(Inactive) (active)
Fibrinogen ------------------------------------ Fibrin
(Insoluble) (soluble)
Fibrin + blood cells --> Clot (fine threads)

3. The more commonly used anticoagulated bulbs in the pathological laboratory are as follows:
1) EDTA 2) Double oxalate 3) Tri sodium citrate 4) Heparin 5) ACD or CPD 6) Fluoride
1. EDTA: (Ethylene diamine tetra acetic acid, disodium salt)
5-10% of EDTA solution is prepared & two drops of it is added in each bulb.
Many of such bulb are prepared place all this bulb in the oven at 80-degree C
for 1 hour. After 24 hr. take out all the bulbs this is for full drying of bulbs.
Action: -It acts as a strong calcium chelating agent. The calcium in blood is bound in a unionized and soluble
complex with EDTA.
Uses: - Tests, which are performed by using EDTA bulbs, are Hb, WBC, RBC, PCV, ESR, Platelets, and DC etc.

4. • Advantages:
1.It preserve of cellular morphology. Good morphology of the cells is observed even after 2 to 3 hrs
of blood collection.
2. Since platelet clumping is inhibited, for platelet counts this anticoagulant is preferred.
• Disadvantages:
1. Excess of EDTA causing shrinkage of WBC & RBC.
2. Excess of EDTA causing decrease in PCV & increase in MCHC.
3.Platelets swell disintegrate due to excess of EDTA & artificially high platelet count may be
obtained due to disintegrated platelets.

5. 2. Double Oxalate:
3 part of ammonium oxalate and 2 parts of potassium oxalate are combined together to balance
swelling effect of ammonium oxalate shrinking effect of potassium oxalate on the red blood cells. The
solution of double oxalate is prepared as follows.
a) Ammonium Oxalate - 2.4 g or 3g
b) Potassium Oxalate - 1.6 g or 2g
c) Distilled water - 100 ml
0.2 ml of this solution contains 8 mg or chemicals, which prevent clotting of about 3 to 4 ml blood.0.2
ml of this anticoagulated solution is added in each one of bottles. The bottles are heated in an
incubator at 60-degree C to 80-degree C for one hour. The bottles are taken out of incubator when
layer appears at the bottom.
Uses: The tests performed by using double oxalates are Hb, WBC, RBC, ESR, and PCV etc.

6. • Action: -Oxalates combine with Ca +2 in blood to form insoluble precipitate of calcium oxalate.
• Disadvantages:
1. WBC morphology is not preserved well hence it is not used for blood smear.
2.It is toxic & since calcium oxalate precipitate may cause harm, it is never used for blood banking
& blood transfusion.

7. 3. Tri Sodium Citrate:
This anticoagulant is used in the liquid form.
Action: -It converts ionized calcium into unionized soluble complex.
Use:
1. It is use in ESR determination by Westergren's method (0.4 ml of 3.8 g/dl sodium citrate and blood is added up to
the mark 1.6 ml).
2. In the PT, APTT, PTT determination (0.2 ml of 3.8 g/dl sodium citrate solution taken & blood is added 1.8 ml).
Disadvantages:
Since it is used as a liquid, due to dilution of cellular elements, it is unsuitable for haemoglobin determination & for
blood cell counts.

8. 4. Heparin:
This anticoagulant is used when plasma is required urgently for certain emergency determination such as blood sugar,
urea & electrolytes. It is used in a concentration of 1 to 2 drops for 3 to 5 ml of blood.
Action:
It acts as inhibitor (to stop) of thrombin formation
Disadvantages:
1. It is expensive.
2. It is unsuitable roc count & smears.
3. It prevents coagulation for only a limited period of time.

9. 5. Acid Citrate Dextrose (ACD)) or Citrate Phospho Dextrose (CPD):
It is used in the blood bank for collecting blood for transfusion. The most commonly used
solution is a mixture of citric acid, sodium citrate glucose. The glucose provides some
'Nourishment' for the cells.
Disadvantages:
Since it is used as a liquid, due to dilution of cellular elements, it is unsuitable for
haemoglobin determination the blood cell counts.

10. 6. Fluoride:
5% sodium fluoride & 5% EDTA solution. Mix well & it will give a 10% fluoride solution is delivered
two drops in each bulbs. It inhibits enolase enzyme and ceases glycolysis. (Glycolysis by RBCs
decreases the plasma glucose level) be prevented by collecting blood in the fluoride bulbs. It inhibits
enolase enzyme, which helps the glucose to get converted to Glucose - 6 -PO4.
Glucose ---------------------Glucose - 6 - Phosphate
Use:
This bulb is especially used in blood Glucose (sugar) test.

11. 7. Plain bulb:
Use:
It is used for serological & biochemical tests. Take a sterile bulb do not add any anticoagulant
chemical in it. So that the blood will clot & the yellow coloured fluid known as serum will appear.
Definitions:
1.Serum: It is a pale yellow colour separation from clotted blood.
Blood is collected in a plain bulb for serum.
Serum do not contain Fibrinogen in its composition, because fibrinogen is used in clotting of blood.
2.Plasma: It is a pale yellow colour separation from anticoagulated blood. Generally, blood is collected
in any anticoagulated bulb for plasma. Plasma contain fibrinogen in its composition because blood is
not clotted over here.