Blood Bottles Guide

geekymedics.com/blood-bottles-guide/

Laura Jayne Watson February 20, 2014

Knowing which blood bottles to use for different tests may not be that important for passing your
finals, but will be absolutely essential when you start work on the wards afterwards. It is one of those
practical things which tends to be poorly taught at medical school, if at all. Instead, you are
seemingly expected to just pick it up on your first day as a junior doctor, when taking blood tests
correctly will suddenly become a massive cornerstone of your workload. Even though most hospitals
now have printable stickers for bottles which tell you which ones to use, it is still vital to have a basic
understanding of which bottles are used and why to prevent embarrassing mistakes and/or awkward
phone calls from the lab.

The colours of the vacutainer bottles are standardised, but depending which hospital you end up
working in there may be some variation in preferences between individual laboratories, especially
when it comes to tests which are performed less commonly. I have used as many sources as possible
to compile the information below, so hopefully, it is as accurate as possible, but obviously I cannot
guarantee it will be applicable to all hospitals. Always check local guidelines if you’re not sure.

You may also be interested in ourblood test reference ranges guide. For more information on how to
perform venepuncture, see our guide.

Purple blood bottle (a.k.a. “lavender”)

Overview
Purple blood bottles are generally used for haematology tests where whole blood is required for
analysis.

Additive
Purple blood bottles contain EDTA (ethylenediaminetetraacetic acid), which acts as a
potent anticoagulant by binding to calcium in the blood. EDTA also binds metal ions in the blood
and is used in chelation therapy to treat iron, lead or mercury poisoning. Its blood-binding capacity
also means it can be labelled with radioisotopes and used as an EDTA scan to assess renal
glomerular filtration rate.

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Tests

Common tests
Common tests performed using the purple blood bottle include:

Full blood count (FBC)

Erythrocyte sedimentation rate (ESR)
Blood film for abnormal cells or malaria parasites
Reticulocytes
Red cell folate
Monospot test for EBV
HbA1C for diabetic control
Parathyroid hormone (PTH)*

Less common tests

Less common tests performed using the purple blood bottle include:

Ciclosporin/tacrolimus levels
Some viral PCR tests
G6PD
ACTH level*
Porphyria screen*
Plasma metanephrines*
Fasting gut hormone screen*

Tips for use

The purple tube needs inverting about 8 times to mix the sample with the EDTA. About1ml of
blood is sufficient to perform a full blood count , however, to get an ESR you will need a full
purple bottle.

Pink blood bottle

Overview
The pink bottles work in the same way as the purple ones but are specifically used only forwhole
blood samples being sent to the transfusion lab.

Additive
Pink blood bottles also contain the anticoagulant EDTA.

Tests

Common tests
Common tests performed using the pink blood bottle include:

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 Group and save (G&S): this simply means the patient’s blood is typed and tested for
antibodies, then saved in the lab in case it is required; it DOES NOT get you blood products
for transfusion. If you need blood products you have to request a crossmatch.
Crossmatch (XM): this means that the patient’s blood is typed and tested as above, then
matched to specific units of blood, platelets or other products for transfusion. You need to
specify on the form how many units you need, why you need them and when they are
required. A full crossmatch takes about 45-60 minutes in the lab – if you have an unstable
bleeding patient and think you’ll need blood products sooner than this, you still need to send a
crossmatch sample, but you can ask the lab for units of type-specific blood (which take 10-
20 minutes), or in a genuine emergency you can use their stocks of O negative blood from
the fridge.
Direct Coomb’s test (a.k.a. direct antiglobulin test) for autoimmune haemolytic anaemia.

Less common tests

Less common tests performed using the pink blood bottle include:

Testing for specific red cell antibodies (usually 3 bottles are required).
Haematology tests such as FBC (if the ward runs out of purple bottles).

Tips for use

The pink tube needs inverting about 8 times to mix the sample with the EDTA. It should contain
at least 1ml of blood, but more is preferred by the labs if at all possible. It has a special label which
needs to be carefully filled in by hand at the bedside to ensure the correct patient details are
used, preventing potentially catastrophic mismatched blood transfusions. If you need blood for a
patient urgently or have any unusual or complicated requests, you must ring the transfusion lab and
let them know, or you risk invoking their wrath.

For more details on performing blood transfusions safely see ourOSCE guide.

Blue blood bottle

Overview
The blue bottle is used for haematology tests involving the clotting system, which require
inactivated whole blood for analysis.

Additive
Blue blood bottles contain buffered sodium citrate, which acts as a reversible anticoagulant by
binding to calcium ions in the blood and subsequently disrupting the clotting cascade. Sodium
citrate is also added to blood products for transfusion and acts as a preservative by stopping them
from clotting in the bag.

Tests

Common tests
Common tests performed using the blue blood bottle include:

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 Coagulation screen: including bleeding time for platelet function, prothrombin time
(PT) for assessment of the extrinsic pathway, activated partial thromboplastin time
(APTT) for assessment of the intrinsic pathway, and athrombin time (TT) or fibrinogen
assay for assessment of the final common pathway.
D-dimer to assist with ruling out thrombosis (e.g. deep vein thrombosis).
INR for monitoring patients on warfarin (this is calculated from the prothrombin time).
Activated partial thromboplastin ratio (APTR) for monitoring patients on intravenous
heparin infusions (this is calculated from the APTT).
Anti-Xa assay for monitoring patients on high-dose low molecular weight heparins like
tinzaparin.

Less common tests

Less common tests performed using the blue blood bottle include:

Specific clotting factors (e.g. factor VIII, factor IX, von Willebrand factor)
Thrombophilia screen
Lupus anticoagulant

Tips for use

The blue bottle needs to be inverted 3-4 times to mix the sample with the anticoagulant. The
sodium citrate liquid in the bottle dilutes the blood sample, and the machines in the lab are
specifically calibrated to interpret results based on a set ratio of blood to anticoagulant. It is,
therefore, essential that the bottle is filled to the line marked around its edge to ensure the
tests are interpreted accurately – otherwise, the samples may be over-anticoagulated.

If your first bottle is light blue and you are using a butterfly needle you will likely need to use two of
these blood bottles to obtain a full sample. This is because when you attach the first bottle it will
draw in air from the tubing, resulting in a sample that is unable to fill with enough blood to reach the
required level. To avoid this, you can initially attach a light blue blood bottle to remove the air (i.e.
fill the tubing with blood), then detach this and fill a second blue blood bottle to the required level.

Some clotting tests need to be taken at specific times ; INRs should ideally be done in the
morning, and anti-Xa assays must be taken 3-4 hours after tinzaparin is given. APTR timings are
often indicated on the prescription algorithm.

Yellow blood bottle (a.k.a. gold)

Overview
These bottles are used for a huge variety of tests requiring separated serum for analysis,
including biochemistry, endocrinology, oncology, toxicology, microbiology and immunology.

Additive
This tube is known in the lab as theSST (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. The serum separator consists of an inert polymer gel which floats
as a layer between the blood cells and plasma to form a physical barrier between them. This
means that the sample can be centrifuged (spun) in the lab and the separated serum easily removed.
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Tests

Biochemistry tests
Biochemistry tests are the ones you will encounter most commonly:

Urea and electrolytes (U&Es): 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 and sometimes free T3.
Vitamins: including 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 tests
Endocrinology tests that can be performed using a yellow blood bottle include:

Beta-hCG
Calcitonin*
Cortisol
EPO
Sex hormones
Growth hormone
IGF-1

Tumour marker tests

Tumour marker tests that can be performed using a yellow blood bottle include:

PSA
CEA
CA-125
CA19-9
AFP
Lactate dehydrogenase (LDH)

Toxicology tests
Toxicology tests that can be performed using a yellow blood bottle include:

Ethanol
Cannabis
Opiates
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 Benzodiazepines
Other drugs (e.g. cocaine, amphetamines)

Drug level tests

Drug level tests that can be performed using a yellow blood bottle include:

Paracetamol
Salicylates (aspirin)
Digoxin
Lithium
Gentamicin
Carbamazepine

Microbiology/virology tests
The yellow blood bottle can be used to test for a wide variety of bacterial, viral, fungal and parasitic
infections including HIV and viral hepatitis.

Immunology tests
Immunology tests that can be performed using a yellow blood bottle include:

Immunoglobulin levels
Complement levels
Autoantibody screen
Rheumatoid factor
Thyroid antibodies
α1AT
ACE

Tips for use

The yellow bottle needs to be inverted about 5 times to mix the sample with the silica and
separator. Don’t panic if the blood starts to clot or separate in the bottle, it’s supposed to! The
amount of blood required will depend on how many tests you’re doing, but at least 1ml is ideal. You
can usually get about 12 tests from one full yellow bottle. Remember that different labs may be
located in different areas and technicians don’t like sharing – this means you’ll need to put your
biochemistry and microbiology samples in separate yellow bottles to go to their respective
laboratories.

Grey blood bottle

Overview
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.

Additive
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
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anticoagulant. Some variants of the grey bottle use EDTA as the anticoagulant instead.

Tests

Common tests
Common tests performed using the grey blood bottle include:

Glucose: this can be fasting or non-fasting, or part of a glucose tolerance test (GTT)
Lactate

Less common tests

Less common tests performed using the grey blood bottle include:

Blood ethanol (if not for legal purposes)

Tips for use

The grey bottle needs to be inverted about 8 times to mix the sample with the fluoride and
oxalate. Only a tiny amount of blood is required for a glucose test, but to assess lactate levels the
bottle should ideally contain at least 1ml of blood . Venous glucose results are generally more
accurate than finger-prick capillary blood glucose tests, especially in hyperglycaemic patients, but
can take a while to come back from the lab. If you require a blood glucose test urgently then kindly
ask one of the nurses to perform a capillary blood glucose for you on the ward. Samples for venous
lactate need to be sent to the lab immediately. Again, the results tend to take a while to come
back, so if you’re desperate to quickly assess a patient’s lactate level see if you can get access to an
ABG machine that does arterial lactates – these are often available in A&E or ITU and take about 2
minutes to process.

Red blood bottle

Overview
The red bottle is less common – it is used forbiochemistry tests requiring serum which might be
adversely affected by the separator gel used in the yellow bottle.

Additive
The additive in red blood bottles contains silica particles which act as clot activators.

Tests

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.

My hospital definitely uses it for ionised calcium , but not much else.

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Less common tests
Less common tests performed using the red blood bottle include:

Fluoride
Cryoglobulins
Cold agglutinins

Tips for use

The red bottle needs inverting 5 times to mix the sample with the clot activator. There is also
another version of the red bottle made out of glass, which contains no additives whatsoever.

Dark green blood bottle

Overview
This less commonly used bottle is for biochemistry tests which require heparinized plasma or
whole blood for analysis.

Additive
The additive of dark green blood bottles contains sodium heparin, which acts as an
anticoagulant.

Tests

Common tests
Common tests performed using the dark green blood bottle include:

Ammonia*
Insulin*
Renin and aldosterone

Less common tests

Less common tests performed using the dark green blood bottle include:

Aluminium
Gut hormones
Amino acids
Homocysteine
Chromosomal tests

Tips for use

The green bottle needs to be inverted about 8 times to mix the sample with the heparin. This
bottle cannot be reliably used to assess sodium levels.

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Light green blood bottle

Overview
This rare species of blood bottle is used for biochemistry tests requiringseparated
heparinized plasma. I have never actually used one but have seen them on a dermatology ward
once.

Additive
This bottle is known as the plasma separator tube (PST). It containslithium 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.

Tests
It can be used for routine biochemistry, but most hospitals seem to use the yellow bottle for this.
It can also be used for blood ethanol provided the sample is not for legal purposes.

Tips for use

The light green bottle needs inverting about 8 times to mix the sample with the heparin and
separator. This bottle cannot be reliably used to assess lithium levels.

Other blood bottles

Despite 18 months of adventures on the wards, there are still some blood bottles I have not yet
encountered. I can only assume this means they are hardly ever used and therefore not ones you’ll
need to know in detail.

These other blood bottle types include:

Dark blue: used for toxicology and trace elements such as zinc, selenium and
copper (however, the ever-versatile yellow bottle can also be used for these).
Tan: used to test for lead.
Orange: contains a thrombin-based clot activator which allows stat serum testing.
Light yellow: used for HLA phenotyping, tissue typing, DNA analysis and paternity testing.
White: used for molecular diagnostics such as PCR and DNA amplification studies.
Black: for paediatric ESR.
Clear lid: used as a discard tube, for example when taking blood from a central line.

Blood culture bottles

Blood culture bottles contain a culture medium to encourage the growth of any bacteria in the
blood sample. There are different bottles available with culture media for aerobic (blue lid) and
anaerobic (purple lid) organisms, alongside a variety of others, including one with ablack lid for
mycobacterial cultures. Until recently, many hospitals require both aerobic and anaerobic culture
samples from a patient, whilst others were happy with just aerobic samples. However, there is now a
move towards using the purple top (a.k.a. “burgundy”) anaerobic bottles as the standard
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receptacle for all blood cultures, as studies have shown that they are more effective. Check local

Blood culture bottles

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guidelines if you’re not sure.

Tips for use

Blood cultures must be obtained using an aseptic non-touch technique (ANTT) to prevent
contamination of the samples with the patient’s skin flora or any bugs that might be lurking on your
hands. They should also be taken before any antibiotics are started. The blood culture bottle
should always be the first one you fill, and ideally needs 8-10ml of blood to ensure a good chance
of catching any organisms (if using an anaerobic and aerobic bottle, the aerobic bottle should be
filled first). In a perfect world, you would also take another culture from a different site to maximise
the diagnostic yield. The results take about 5 days to come back, so if your patient is septic you need
to ring microbiology and start them on some empirical antibiotic treatment in the meantime.

Check out our guide to performing blood cultures for more information.

Arterial blood gas syringes

ABG syringe

Overview
An ABG is a very useful test when you find yourself confronted with a critically unwell patient, as it
tells you about their oxygenation, their acid-base balance, and in some cases their potassium
and lactate as well, and the results are available within minutes.

Tips for use

ABG syringes contain heparin to prevent the sample from clotting – you need to expel this through
the needle before taking your sample. Some fill automatically as the plunger is pushed back by the
pressure of the blood coming through the needle, whilst others require you to pull on the plunger
yourself to fill the syringe. Try to get the self-filling ones – like those shown above – if you can, as
they are infinitely better. Compared to taking venous blood, ABGs are technically more challenging,
riskier and much more uncomfortable for the patient. For more details and tips on how to take an
ABG, see our guide.

Once you have obtained your sample it needs to betaken straight to the lab, as it will become
denatured and useless within 10 minutes. To learn how to interpret an ABG, see our guide.

Tips for easy venepuncture

Invert doesn’t mean shake: be gentle with your samples or they’ll haemolyse.

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Tests above with a star (*) next to them need to be sent to the labimmediately on ice – to do this
you either need to get a special ice bag directly from the lab or if your ward has an ice freezer you
can make one yourself by filling a normal sample bag with crushed ice. If you’re not sure you’re
doing it right, ring the lab and check before you take the blood sample.

The bottles can be used for other non-blood fluids too, for example, pleural aspirates, ascitic taps
and CSF samples obtained by lumbar puncture. The tests each bottle is used for are the same: the
purple one is for cell count, the yellow one is for electrolytes, albumin and LDH, the grey one is for
glucose, and blood culture bottles can be used for fluid cultures. Don’t forget to specify on your
request form what kind of fluid is in the bottle and which part of the patient you got it from.

If you ever find yourself having to get blood from a child, there are specialpaediatric blood
bottles which are much smaller than the adult ones. The colours can vary compared to adult
bottles, so always check with the local lab.

If you are unsure about what you need to do for a particular blood test, ask yourfriendly local lab
technician. They are usually more than happy to give you advice as it means they can prevent you
from making mistakes that create lots of hassle and annoying paperwork. It’s also better for you if
you check, as it saves you having to do your blood tests all over again if you do it wrong, and saves
your patient from the unpleasantness of unnecessary additional stabbings.

Blood bottle summary

We have summarised the different types of blood bottles below organised by the order of draw,
including the relevant tests and the number of required inversions.

I hope you found this guide helpful. Good luck!

Blood bottle types

References
1. BD Vacutainer Product FAQs. Available from: [LINK].
2. BD Vacutainer Venous Blood Collection Tube Guide. Available from [LINK].

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3. UCI Pathology Services Manual. Specimen Tube Containers. Available from: [LINK].

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