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ANTICOAGULANTS

Potassium EDTA
EDTA chelated divalent cations. Potassium EDTA is the standard anticoagulant
used for hematology, because it preserves the cellular components of the blood (but
is not a fixative). This anticoagulant can be used for some biochemistry but is not
suitable for:
Alkaline phosphatase - due to chelation of metallic cofactors which inhibits the
enzyme activity.
Potassium and sodium - due to the addition of potassium to the sample.
Calcium and magnesium - these are chelated by the EDTA.
EDTA is also an unsuitable additive for samples requiring bacterial culture,
since the chelation of the divalent cations inhibits the growth of bacteria. EDTA
is sometimes used to prevent cells clumping in fluid samples requiring cell
counts to accompany a cytology evaluation but it does not actually 'fix' the
cells. A sample fixed in formalin or alcohol/ethanol is required for accurate
cytology examination. EDTA is not suitable for samples requiring virus
isolation in cell/tissue culture because it forms a gel when added to the cell
culture medium and this disrupts the cell monolayer.
Plain (no anticoagulant)
Plain (or clotted) samples are used to provide serum for serology and most
biochemistry or endocrine assays. Serum is plasma without fibrinogen since
the fibrinogen has been used' in the formation of the clot.
Lithium Heparin
Lithium heparin accelerates the action of antithrombin III which neutralizes
thrombin and thus prevents the formation of fibrin from fibrinogen (clot
formation). This effect makes heparin samples unsuitable for determination of
fibrinogen or clotting factors.
Lithium heparin is a standard anticoagulant used to obtain plasma for
biochemistry analysis. Lithium heparin is the most suitable anticoagulant for
the isolation of viruses in cell/tissue culture. This anticoagulant is not suitable
for hematology as the heparin alters the cell morphology. Whilst measurement
of hemoglobin, and blood cell counts can be obtained using this anticoagulant
an accurate white cell differential and morphology comments are not possible.
Fluoride/Oxalate
Fluoride/oxalate samples are used for glucose (and lactate) determination
only. Sodium fluoride functions by stabilizing the blood cell membrane and
inhibiting the enzyme systems involved in glycolysis, which prevents red blood
cells metabolising any glucose present in the sample. For this reason it is the
only suitable sample for accurate glucose analysis. Fluoride is a potent
inhibitor of many enzymes and the inhibition of glycolysis tends to cause fluid
shifts. Fluoride is a weak anticoagulant on its own, so potassium oxalate

(another powerful enzyme inhibitor) is usually added to supplement its action.


Other plasma or serum samples may be used for glucose analysis ONLY if the
plasma/serum is separated from the cells within 1 hour of sample collection.
Without an antiglycolytic agent, the blood glucose concentration decreases by
approximately 0.56 mmol/l per hour at 25C.
Sodium citrate
Sodium citrate is the standard anticoagulant for coagulation assays. Citrate
functions by chelating calcium. The effect is easily reversed by the addition of
calcium to the sample. Other anticoagulants are not suitable for coagulation
assays as they interfere with coagulation factors. Citrate also inhibits ALP, ALT
and interferes with the measurement of phosphate.