Anticoagulants are substances that prevent blood from clotting in vivo or in vitro. They work through chelating or non-chelating actions. Heparin is produced naturally in the liver and mast cells, while hirudin is produced by leeches. Anticoagulants are used for studying blood constituents and coagulation, in blood banks, and to treat conditions like heart attacks and mini-strokes. An ideal anticoagulant must keep blood fluid, minimize platelet aggregation and cell destruction, and not cause hemolysis or change red blood cell size. They are classified based on their nature, form, or use, and can chelate calcium ions or have non-chel

1. Anticoagulants

2. Anticoagulants are chemical substance that perevent
coagulation of blood either invivo or invitro. Each di er in
mode of action ie;
1) Chelating action
2) Non chelating action
Heparin is mostly produced by liver and mast cell inHB,
&Hirudine is produced by leech

3. Uses:
✩ Study of various constituents of blood
✩ Study of coagulation of blood
✩ Used in blood banks
✩ Treatment of transinet ischemic attack( mini -strokes),
heart attacks.

4. Porperties of ideal anticoagulant:
✩ It must be:
✩ Soluble in blood
✩ Keep blood in fluid condition
✩ Minimize destruction of leukocytes
✩ Minimize aggregration of platelets
✩ It must not:
✩ Bring hemolysis
✩ Change size of rbc

5. Classification
✩ Depending on nature
✩ 1) natural : eg; heparin, hirudine
✩ 2)artificial: eg; EDTA, oxalate, citrate
✩ Depending on forms
✩ 1)dry:eg; EDTA, oxalates, fluorides
✩ 2)wet:eg; ACD, sodium citate
✩ Depending on use
✩ 1)routinely : eg; EDTA, oxalate, sodium citrate

6. ✩ Depending on action
✩ 1)ca chelators:eg; EDTA, trisodium citrate, double oxalate
✩ 2)non ca chelating: eg; heparin, warfarin

7. EDTA:ETHYLENE DIAMINE TETRA
ACETIC ACID

9. Action
➢ Powerful ca chealiting agent. blood cannot clot without
ca ie; ca is converted in to insoluble salt.
➢ there are 3 types of edta: Na, K, Li
➢ K salt are more soluble hence mainly pre ered.

10. Concentration :
➢ K salts are used in 1.5-2mg/ml of blood
➢ 10% solution is made and dried.

11. Uses
✩ Tc-total count
✩ Dc-di erential count
✩ Hb estimation
✩ Pcv
✩ indentifying parasites
✩ blood bank

13. Disadvantages
✩ Expensive
✩ Excess cause shrinkage
✩ causes degenrate changes
✩ excess amount cause increase in pcv decrease in MCHC
✩ unsuitable for coagulation studies

14. Trisodium citrate

15. ➢ Wet anticoagulant
➢ action: chelation, less powerful compared to edta
➢ They can be:
➢ a) tri sodium citrate
➢ b) ACD -acid citrate dextrose: used in blood bank
➢ c)CPD: 21 days
➢ d)CPD-A: 35 days

16. TRI SODIUM CITRATE
• Commonly used as 3.8% in estimation and 3.2% in
coagulation studies.
• For ESR 3.8% is used in 1:4 ratio(anticoagulant:blood)
• For coagulation studies 3.2% is used in 1:9 ratio.

17. prepration of Na3C6H5O7: 3.8%
➢ Distilled water-100ml
➢ Trisodium citate-3.8gm
➢ Dissolve both and mix well
➢ . 4ml for 2ml of blood
➢ 1:4 ratio ie;. 4ml anticoagulant+1.6ml blood

18. prepration of Na3C6H5O7: 3.2%
• Distilled water-100ml
• Tri sodium citrate-3.2gm
• Combine both &mix well
• 1:9 ratio ie; .1ml anticoagulant +. 9ml blood

19. Advantages and uses
▪ used for ESR and coagulation studies
▪ blood banks

20. Disadvanyages
▪ Cannot be used for estimation of cell counts, hb etc

21. Oxalates

22. Types: Na, K, NH3

23. • Ammonium oxalate:1.2gm
• Potassium oxalte: 0.8gm
• Distilled water: 100ml
• . 5ml for 5ml blood

25. Uses & advantages
▪ PCV
▪ ESR
▪ TLC
▪ specific gravity
▪ prevent swelling and shrinkage e ect

26. Disadvantage
▪ Morphology of wbc is not preserved
▪ Ca oxalate ppt is formed which is toxic hence not used in
blood banks

27. Sodium fluoride

28. • Choice of estimation of sugars
• Ca chelators :action
• They prevent glycolysis ie; break down of glucose, by
inhibiting the enolase enzyme
• Conc: 30mg /5ml blood

29. Prepration
➢ Distilled water-100ml
➢ Sodium flouride-3gm
➢ Dissolve and mix well
➢ 1ml of solution is dried & su cinet for 5ml of blood
➢ Flouride oxalate mixture: Potassium oxalate &sodium flouride
in ratio 3:1 is used for blood sugar determination
➢ conc :2mg/ml

30. Advantages and uses
• Blood sugar estimation, alcohol and other biochemical test

31. Disadvantages
• It can inactivate cardiac and liver enzymes. hence not used
for these studies

32. Heparin

33. ➢ Natural anticoagulant
➢ Cannot be made artificially ;hence expensive
➢ Non chelating agent
➢ Action : inhibits thromboplastin formation and also destroys
thrombin , &distrubs clotting mechanism
➢ Conc : .2mg/ml of blood

34. Uses and advantages
▪ Used for blood gas analysis
▪ Determine osmotic fragility
▪ ESR, PCV, chemical estimation can also be done by this
▪ Does not a ect cell size or composition of the blood

35. Disadvantages
• Expensive
• Blood film prep cannot be done;leukocytes clamps & give a
bluish background
• Prevent coagulation only for a partocular period of time

36. Hirudin
• Hirudin is a naturally occurring peptide in the salivary glands
of blood-sucking leeches (such as Hirudo medicinalis) that
has a blood anticoagulant property.
• This is fundamental for the leeches' habit of feeding on
blood, since it keeps a host's blood flowing after the worm's
initial puncture of the skin.
• The present study investigated platelet function tests using
hirudin
• It is used to prevent blood clots following surgery

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