1. A tourniquet prevents venous flow out of the arm. 8. SPS is sodium polyanetholesulfonate.

It is an additive that
prevents blood from clotting.
2. The gauge of a needle indicates the diameter of the
needle’s lumen.
9. Blood collected in a tube with an anticoagulant must be
mixed thoroughly after collection by gently and repeatedly
3. If a large-gauge needle is used in venipuncture, collection is inverting the tube.
slower and blood cells may be hemolyzed (destroyed) as
they pass through the narrower opening.
10. Thixotropic gel forms a barrier between blood cells and
serum or plasma, thus preventing contamination and
4. The rubber sleeve on the multisample needle keeps the allowing easy separation.
needle from becoming contaminated or injuring you or the
patient, and it keeps blood from leaking onto or into the
adapter or tube holder, especially when changing tubes. 11. Glycolysis is a cellular reaction used to harvest energy from
glucose.

5. An advantage of the syringe method is that blood appears in

the hub when the vein has been entered. A disadvantaged 12. Blood specimens used for analysis are whole blood, serum,
of the syringe method is that there is the potential for and plasma.
needle stick when depositing blood into the collection tube.

6. When blood tubes are evacuated, a vacuum is created Tan Lead analysis
within the tube so that a measured amount of blood will Red or pink Blood Bank
flow in easily. Light blue Coagulation
Lavender CBC
Gray Glucose tolerance test
7. Unused blood tubes must be discarded when they expire Black Sedimentation rate
because out-of-date tubes may have decreased vacuum, Gold BD Hemogard Chemistry testing
preventing a proper fill, or they may have additives that Green ABGs
degrade over time. Dark (royal) blue Trace metals
It is extremely important to correctly and positively identify the
patient in any phlebotomy procedure.
To properly identify a patient, match the information on the
requisition with the information on the patient’s identification band
(for inpatients) or with information provided by the patient (for
outpatients).
Information typically found on a requisition form includes the
patient’s name, date of birth, the patient’s hospital ID number (for
inpatients), the patient’s room number and bed (for inpatients), the
patient’s doctor’s name or code, the type of test requested, and the
test status.
When requisitions are received, you should examine them for the
necessary information, check for duplicates or errors, group them
together for the same patient, prioritize them, and gather all the
equipment you will need to perform the collections.
Hemoconcentration is an increase in the ratio of formed elements
to plasma caused by leaving the tourniquet on too long.
The three veins in the antecubital area suitable for venipuncture are
the median cubital, cephalic, and basilic.
The median cubital vein is the first choice for venipuncture because
it is large and well anchored and does not move when the needle is
inserted.
Veins feel spongy, bouncy, and firm on palpation; arteries pulsate;
and tendons feel rigid.
To help locate a vein, tap the arm, have the patient make a fist, or
warm the site with a warm towel or hot pack.
A hematoma is a reddened, swollen area in which blood collects
under the skin. It can form when the extra pressure from the
tourniquet forces blood out through the puncture.
The correct position for the arm after the venipuncture is straight or
slightly bend, but not bend back over the puncture site.
To correctly label blood tubes, label them at the bedside using a pen
or permanent marker. The label must have the patient’s name and
identification number, the date and time of collection, and the
collector’s initials or identification number. If labels are computer
generated, make sure all the information is present, and then add
the collector’s initials or identification number.
The requisition slip must contain the patient’s identification or chart
number, the patient’s last and first names, the patient’s date of
birth, the physician’s name, and the tests ordered.
Phlebotomists must prepare the patient and then wash their hands
so that when it is time to perform the venipuncture, the gloves and
everything they touch are as free from contamination as possible. If
phlebotomists wash their hands, put on gloves, and then prepare
the patient, they may inadvertently pick up contaminants from the
patient, which can then be transferred to the puncture site.
RED TOP
ADDITIVE: None
MODE OF ACTION: Blood clots, and the serum is separated by
centrifugation.
USES: Chemistries, Immunology and Serology, Blood Bank (Crossmatch)
GOLD TOP
ADDITIVE: None
MODE OF ACTION: Serum separator tube (SST) contains a gel at the
bottom to separate blood from serum on centrifugation.
USES: Chemistries, Immunology and Serology
LIGHT GREEN TOP
ADDITIVE: Plasma Separating Tube (PST) with Lithium heparin
MODE OF ACTION: Anticoagulates with lithium heparin; Plasma is
separated with PST gel at the bottom of the tube.
USES: Chemistries
PURPLE TOP
ADDITIVE: EDTA
MODE OF ACTION: Forms calcium salts to remove calcium.
USES: Hematology (CBC) and Blood Bank (Crossmatch); requires full draw –
invert 8 times to prevent clotting and platelet clumping.
MODE OF ACTION: Tube is designed to contain no contaminating metals.
USES: Trace element testing (zinc, copper, lead, mercury) and toxicology.
LIGHT GRAY TOP
ADDITIVE: Sodium fluoride and potassium oxalate
MODE OF ACTION: Antiglycolytic agent preserves glucose up to 5 days.
USES: Glucoses, requires full draw (may cause hemolysis if short draw).
YELLOW TOP
ADDITIVE: ACD (acid-citrate-dextrose)
MODE OF ACTION: Complement inactivation.
USES: HLA tissue typing, paternity testing, DNA studies.
YELLOW-BLACK TOP
ADDITIVE: Broth mixture
MODE OF ACTION: Preserves viability of microorganisms.
USES: Microbiology – aerobes, anaerobes, fungi.
BLACK TOP
ADDITIVE: Sodium citrate (buffered)
MODE OF ACTION: Forms calcium salts to remove calcium.
USES: Westergren Sedimentation Rate; requires full draw.

LIGHT BLUE TOP ORANGE TOP

ADDITIVE: Sodium citrate ADDITIVE: Thrombin
MODE OF ACTION: Forms calcium salts to remove calcium. MODE OF ACTION: Quickly clots blood.
USES: Coagulation tests (protime and prothrombin time), full draw USES: STAT serum chemistries.
required.
LIGHT BROWN TOP
GREEN TOP ADDITIVE: Sodium heparin
ADDITIVE: Sodium heparin or lithium heparin MODE OF ACTION: Inactivates thrombin and thromboplastin, contains
MODE OF ACTION: Inactivates thrombin and thromboplastin virtually no lead.
USES: For lithium level, uses sodium heparin. For ammonia level, use USES: Serum lead determination.
sodium or lithium heparin.
PINK TOP
DARK BLUE TOP ADDITIVE: Potassium EDTA
ADDITIVE: EDTA MODE OF ACTION: Forms calcium salts.
USES: Immunohematology

WHITE TOP
ADDITIVE: Potassium EDTA
MODE OF ACTION: Forms calcium salts.
USES: Molecular / PCR and bDNA testing.
ORDER OF DRAW: Order of Draw Tube Stopper Color Rationale for Collection
Order
1. Blood culture tubes which are sterile are drawn first. This Blood cultures (sterile Yellow SPS Minimizes chance of
collection) Sterile media bottles microbial contamination
prevents the transfer of unsterilized materials from other tubes Coagulation tubes Light Blue The first additive tube in the
into the sterile tube. order because all other
2. Light blue-topped tubes (for coagulation tests) are next. These additive tubes affect
coagulation tests
tubes are always drawn before tubes containing other kinds of Glass nonadditive tubes Red Prevents contamination by
anticoagulants or clot activators, because other additives could additives in other tubes
contaminate this tube and interfere with coagulation testing. If Plastic clot activator tubes Red Filled after coagulation tests
because silica particles
coagulation tests are the only tests ordered, you only need a light Serum separator tubes Red & Gray rubber activate clotting and affect
blue-topped tube. Your institution may have you draw a plain red- (SSTs) Gold plastic coagulation tests (carryover
topped tube first and discard it (this may help prevent of silica particles into
subsequent tubes can be
contamination by tissue fluids). The discard tube may be a plain overridden by anticoagulant
red glass tube without any additive. Also, if you are drawing with a in them)
butterfly, you must draw a discard tube in order to draw out the Plasma separator tubes Green & Gray rubber Heparin affects coagulation
(PSTs) Light-green plastic tests and interferes in
air in the tubing, and prevent a short draw. collection of serum
3. Glass red-topped tubes or plastic red-top tubes may be drawn Green specimens; causes the least
now. Heparin tubes interference in tests other
than coagulation tests
4. Red-gray (gold BD Hemogard) tubes and plastic red-topped tubes EDTA tubes Lavender Responsible for more
are next. These contain clot activators, which would interfere with Pink carryover problems than
many other samples if passed into other tubes. any other additive: elevates
Plasma preparation tubes Pearl top Na and K levels, chelates
5. Green tubes are drawn next. The heparin from the green tube is (PPTs) and decreases calcium and
less likely to interfere with EDTA-containing tubes than vice versa. iron levels, elevates PT and
6. Lavender tubes are next. EDTA binds many metals, in addition to PTT results
Oxalate/fluoride tubes Gray Sodium fluoride and
calcium, so it can cause problems with many test results, including potassium oxalate affect
giving falsely low calcium and falsely high potassium readings. For sodium and potassium
this reason, lavender tubes are drawn near the end. levels, respectively. After
hematology tubes because
7. The gray-topped tube is last. This tube contains potassium oxalate damages cell
oxalate. The potassium would elevate the potassium levels membranes and causes
measured in electrolyte analysis, and oxalate can damage cell abnormal RBC morphology.
Oxalate interferes in
membranes. Also, another additive, sodium fluoride, elevates enzyme reactions.
sodium levels and inhibits many enzymes.
Stopper Color Additive Department(s) Royal blue None (red label) Chemistry
Light Blue Sodium citrate Coagulation EDTA (lavender
label)
Red (glass) None Chemistry, Blood bank, Sodium heparin
Serology/Immunology (green label)

Red (plastic) Clot activator Chemistry Tan (glass tube) Sodium heparin Chemistry
Tan (plastic) EDTA
Red/light-gray Nonadditive NA (Discard tube only)
(plastic) Yellow Sodium Microbiology
polyanethol
Red/black (tiger) Clot activator Chemistry sulfonate (SPS)
Gold and gel
Red/gold separator Yellow Acid citrate Blood
dextrose (ACD) bank/Immunohematology
Green/gray Lithium heparin Chemistry
Light green and gel
separator

Green Lithium heparin Chemistry

Sodium heparin

Lavender EDTA Hematology

Pink Blood bank

Gray Sodium fluoride Chemistry

and potassium
oxalate
Sodium fluoride
and EDTA
Sodium fluoride

Orange Thrombin Chemistry

Gray/yellow