ANTICOAGULANTS

NEEDLE DISPOSAL
 Assoon as needle safety device is
activated dispose of entire assembly
in a biohazard sharps container.
VACUUM COLLECTION TUBES
 Vacuum collection tubes are glass or plastic tubes
sealed with a partial vacuum inside by rubber
stoppers.
 The air pressure inside the tube is negative, less
than the normal environment.
 After inserting the longer needle into the vein,
the phlebotomist pushes the tube into the holder
so that the shorter needle pierces the stopper.
 The difference in pressure between the inside of
the tube and the vein causes blood to fill the tube.
 The tubes are available in various sizes for adult
and pediatric phlebotomies
ADDITIVES

Different blood tests requires

different types of blood
specimens.
Most tubes have additives called
anticoagulants which prevent
clotting/coagulation of the blood.
Plastic tubes may have an
additive to enhance clotting of
the blood
ANTICOAGULANTS
 Anticoagulants are already in the tubes in the
precise amount needed to mix with the amount of
blood that will fill the tube.
 The color of the stopper on each tube indicates
what, if any, anticoagulant the tube contains.
 It is important to completely fill each tube so that
the proportion of blood to chemical additive is
correct, otherwise, the test results may not be
accurate or the specimen will be rejected and will
need to be recollected.
 It is also important to thoroughly mix the blood
with the additive by gentle inversion
OXALATE
Combines with calcium to
form an insoluble salt
Interfere with Na, K, and
most BUN (Urease)
measurements.
Concentration: 1-2mg/mL of
blood.
CITRATE
It combines with calcium in a
non-ionized form
Concentration:3.2-
3.8g/dL(0.105M or 0.129 M) in
a ratio of 1 part to 9parts of
blood
 The blue bottle is used for
haematology tests involving the
clotting system, which require
inactivated whole blood for analysis.
 This contains buffered sodium
citrate, which acts as a reversible
anticoagulant by binding to calcium
ions in the blood and subsequently
disrupting the clotting cascade.
COMMON TESTS:
 Coagulation screen including bleeding time
for platelet function, prothrombin time
(PT)for extrinsic pathway, activated partial
thromboplastin time (APTT) for intrinsic
pathway, and thrombin time (TT) or
fibrinogen assay for the final common
pathway .
 D-dimer for thrombosis e.g. due to DVT or
PE
 INR for monitoring patients on warfarin
(this is calculated from the prothrombin
time
 Activated partial thromboplastin ratio
(APTR) for monitoring patients on IV
heparin infusions (this is calculated from
the APTT)
 Anti-Xa assay for monitoring patients on
high-dose low molecular weight heparins
like tinzaparin
ETHYLENEDIAMINE TETRAACETIC
ACID (EDTA)
 It combines with calcium in a process
called chelation
 2 forms: versene and sequestrene

 CONCENTRATION: 1-2mg/mL of blood

 Uses: carcinoembryonic antigen (CEA),

TDM and lead testing
 Blood specimens for nucleic acid
testing are generally collected into EDTA
anticoagulant to inhibit enzymes that
might break them down.
 These bottles are generally used for
haematology tests where whole blood is
required for analysis.
 COMMON TESTS:
 full blood count(FBC)
 erythrocyte sedimentation rate (ESR)
 blood film for abnormal cells or malaria parasites
 reticulocytes
 red cell folate
 Monospottest for EBV
 HbA1C for diabetic control
 parathyroid hormone (PTH)
 The pink bottles work in the same way as the
purple ones, but are specifically used only for
whole blood samples being sent to the
transfusion lab
 This tube also contains the anticoagulant EDTA

 COMMON TESTS:

 group and save (G&S)

 crossmatch (XM)

 direct Coomb’s test (aka direct antiglobulin test)

for autoimmune haemolytic anamea
FLOURIDE
Itforms weekly dissociated
calcium components.
Interfere with Na, K, and Most
BUN (urease) testing.
Concentration:10mg/mL of
blood
HEPARIN (MUCOITIN POLYSULFURIC
ACID)
 It acts as antithrombin and
antithromboplasmin, anti-factor X, Ideal
universal anticoagulant
 It accelerates the action of antithrombin III,
Neutralizing thrombin and preventing the
formation of fibrin.
 It is available as sodium, lithium, potassium and
ammonium salts
 Concentration: 0.2mg/mL OF BLOOD

 Lithium heparin may be used for most

chemistry test except for lithium and folate
levels.
HEPARIN (MUCOITIN POLYSULFURIC
ACID)
 Sodium heparin is the injectable form
used for anticoagulant therapy.
 Uses: NH3, Carboxy/methhemoglobin,
plasma hemoglobin, Ph and blood gas,
cytogenetic studies (sterile tubes)
 Lithium heparin: glucose, BUN,
Ionized calcium, electrolyte studies and
creatinine.
 This less commonly used bottle is
for biochemistry tests which require
heparinised plasma or whole blood
for analysis.

 This
contains sodium heparin,
which acts as an anticoagulant
COMMON TESTS:

ammonia
Insulin
renin and aldosterone
 This rare species of bottle is used for
biochemistry tests requiring separated
heparinised plasma.
 I have never actually used one but have
seen them on the dermatology ward.
This bottle is known as the plasma
separator tube (PST).
It contains lithium heparin, which
acts as an anticoagulant, and a
plasma separator gel similar to that
used in the yellow bottle, which acts
to separate out the plasma layer.
COMMON TESTS:

Itcan be used for routine

biochemistry, but most hospitals
seem to use the yellow bottle for
this.

Itcan also be used for blood ethanol

provided the sample is not for legal
purposes.
 These bottles are used for a huge variety of tests
requiring separated serum for analysis,
including biochemistry, endocrinology, oncology,
toxicology, microbiology and immunology.
 This tube is known in the lab as the SST (serum
separating tube).
 It contains two agents; silica particles and a
serum separating gel.
 The silica particles work to activate clotting and
cause the blood cells to clump together
COMMON TESTS:
 Biochemistry tests are the ones you will
encounter most commonly:
 Urea and electrolytes (U+E) – this includes urea,
creatinine, sodium and potassium
 C-reactive protein (CRP)

 Liver function tests (LFTs) – this includes

bilirubin, ALP, AST/ALT, GGT, total protein and
albumin
 Amylase assay

 Bone profile – this includes calcium, phosphate,

ALP and albumin
 magnesium assay
 iron studies – this includes serum iron, ferritin,
transferrin saturation and total iron binding
capacity
 lipid profile – this includes cholesterol, LDL,
HDL and triglycerides
 thyroid function tests (TFTs) – this includes
TSH, free T4 +/- free T3
 vitamins e.g. vitamin B12
 troponins – this requires 2 samples to be taken at
different times to assess the acute trend
 creatine kinase (CK)

 urate

 serum osmolality – this requires a urine sample

to be taken at the same time
 endocrinology: beta-hCG, calcitonin*, cortisol,
EPO, sex hormones, growth hormone, IGF-1
 tumour markers: PSA, CEA, CA-125, CA19-9,
AFP, lactate dehydrogenase (LDH)
 toxicology: ethanol, cannabis, opiates,
benzodiazepines, other drugs e.g. cocaine,
amphetamines
 drug levels: paracetamol, salicylates (aspirin),
digoxin, lithium, gentamicin, carbamazepine
 microbiology/virology: serology for a wide variety
of bacterial, viral, fungal and parasitic infections
including HIV and viral hepatitis
 immunology: immunoglobulins, complement,
autoantibody screen, rheumatoid factor, thyroid
antibodies, α1AT, ACE
 The grey bottle is only used for two tests, so
compared to the yellow one it’s fairly easy to
remember!
 It is used for biochemistry tests requiring whole
blood for analysis.
 This contains two main agents.

 Sodium fluoride acts as an antiglycolytic

agent to ensure that no further glucose
breakdown occurs within the sample after it is
taken.
 Potassium oxalate acts as an anticoagulant.
COMMON TESTS:

glucose – this can be fasting or

non-fasting, or part of a glucose
tolerance test (GTT)
lactate
The red bottle is less common –
it is used for biochemistry tests
requiring serum which might be
adversely affected by the separator
gel used in the yellow bottle.
This contains silica particles
which act as clot activators
COMMON TESTS:
 The use of this bottle varies greatly –
some hospitals use it for many sensitive
tests, including hormones, toxicology,
drug levels, bacterial and viral serology
and antibodies
 Whereas others seem to only use it for a
few very specific purposes and use the
yellow bottle for most things.
 Some hospital definitely uses it for ionised
calcium, but not much else
IMPORTANT NOTES
 Trop I- SERUM
 Trop T- Heparinized plasma

 Under filling blood collection tubes can

affect RBC morphology and lipids in
EDTA tubes, and binding of electrolytes
and troponin to heparin in some plasma
tubes.
 Excess citrate in plasma from
insufficient blood volume leads to falsely
elevated clotting times.
 Heparin is a poor choice for nucleic acid testing
because it can be coextracted with DNA and
inhibits DNA polymerase chain reaction (PCR’s)
 Gray top can also be used for lactate and alcohol
determinations.
 Respinning gel tubes can cause increase
potassium.
 For microcollection tubes, lithium and
ammonium salts of heparin are the preferred
anticoagulants.
 Polypropylene is used in several tube designs,
including specimen tubes and test tubes.
 Polystyrene or other high-impact plastic
type containers are commonly used for
specimen collection (urine)
 Hospitalized patient are likely to be
receiving heparin (especially under
critical care), which can delay clotting in
blood collection tubes even with
activators and lead to fibrin strands that
can clog up aspiration probes on
instrumentation.
TUBE INVERSION
NOTE: ALWAYS FOLLOW YOUR FACILITY’S
PROTOCOL FOR ORDER OF DRAW

 Handle all biologic samples and blood collection

“sharps” (lancets, needles, luer adapters and
blood collection sets) according to the policies
and procedures of your facility.
 Obtain appropriate medical attention in the
event of any exposure to biologic samples (for
example, through a puncture injury) since they
may transmit viral hepatitis, HIV (AIDS), or
other infectious diseases.
 Utilize any built-in used needle protector if
the blood collection device provides one.
 BD does not recommend reshielding used
needles, but the policies and procedures of
your facility may differ and must always be
followed.
 Discard any blood collection “sharps” in
biohazard containers approved for their
disposal.