it is for the betterment of the goods

1. UNIT IV
BLOOD COLLECTION, EQUIPMENT, ADDITIVES, AND ORDER OF DRAW
Principles of Blood Collection
o Importance of Blood Collection
v Obtain blood for diagnostic purposes and to
monitor prescribed treatment.
v Remove blood for transfusions at a donor
center.
v Remove blood for therapeutic purposes, such
as treatment for polycythemia, a disorder
involving the overproduction of red blood cells.
o Methods in Blood Collection
v Venipuncture
• It is the process of collecting or “drawing”
blood from a vein and the most common way
to collect blood specimens for laboratory
testing.
v Arterial puncture
• The process of collecting blood preferably for
arterial blood gas or ABG.
• Physicians are medical professionals who
have the right to perform arterial puncture.
v Capillary Puncture
• Drops of blood for testing can be obtained by
puncturing or making an incision in the
capillary bed in the dermal layer of the skin
with a lancet, other sharp device, or laser.
Phlebotomy Materials
o Blood Drawing Station
v A dedicated area of a medical laboratory or
clinic equipped for performing phlebotomy
procedures on patients, primarily outpatients
sent by their physicians for laboratory testing. It
includes the following:
• A table for supplies.
• A special chair where the patient sits during
the blood collection procedure.
• A bed or reclining chair for patients with a
history of fainting, persons donating blood.
• Bed or padded table for infants
o Phlebotomy Chairs
v This should be comfortable for the patient and
have adjustable armrests to achieve proper
positioning of either arm.
o Equipment Carriers
v Equipment carriers make blood collection
equipment portable.
v This is especially important in a hospital setting
and other instances in which the patient cannot
come to the laboratory.
Two types are:
• Handheld Carrier - is a carrier or tray used to
contain enough equipment for numerous
blood draws. They are convenient for “stat”
or emergency situations or when relatively
few patients need blood work.
• Phlebotomy cart - is typically made of
stainless steel or strong synthetic material.
Normally, it has several shelves to carry
adequate supplies for specimen collection.
Commonly used for early-morning hospital
phlebotomy rounds. It is also a potential
source of nosocomial infection.
o Gloves
v A requirement in performing phlebotomy. A new
pair must be used for each patient and be
removed when the procedure is completed.
The acceptable gloves are:
• Nonsterile
• Disposable latex
• Nitrile
• Neoprene
• Polyethylene
• Vinyl examination
o Tourniquet
v It is a device that is applied or tied around a
patient’s arm prior to venipuncture to restrict
blood flow.
v The tourniquet is left in place for not more than
one minute.
Types of tourniquet are:
• Strap tourniquet - most common type which
is a flat strip of stretchable material, such as
latex, nitrile, or vinyl that is fairly inexpensive
and disposable.
• A Venoscope II transilluminator device - vein
appears as a dark line between the light-
emitting arms of the Venoscope II.
• AccuVein AV300
o Evacuated Tube
v Evacuated tubes are used to contain blood
specimen which come in various sizes, colors
and volumes depending on the type of test.
o Antiseptics
v Used to clean the puncture site prior to
collection.
o Cotton Balls and Gauze Pads
v Used to hold pressure over the site following
blood collection procedures.

2. UNIT IV
BLOOD COLLECTION, EQUIPMENT, ADDITIVES, AND ORDER OF DRAW
v Clean 2- by 2-inch gauze pads folded in fourths
is more preferred than cotton balls.
o Bandages
v Used to cover a blood collection site after the
bleeding has stopped.
v Latex-free bandages are available for those with
latex allergies.
o Different Blood Collection Devices
v Syringe Type Collection System
• It is an open system which only allows one
or single draw from the patient’s vein. It is
disposable type of blood collection device.
• Syringe volume is selected according to the
size and condition of the patient’s vein and
the amount of blood to be collected.
v Evacuated Tube System (ETS)
• It is a closed system which allows numerous
tubes to be collected with a single
venipuncture.
v Winged Infusion Set
• It is an indispensable tool for collecting blood
from small or difficult veins. It allows much
more flexibility and precision than a needle
and syringe.
o Needles and Sharp Disposal Containers
v Marked with a biohazard symbol, rigid,
puncture-resistant, leakproof, disposable, and
have locking lids to seal the biohazardous
wastes.
v Used needles, lancets, and other sharp objects
must be disposed of immediately.
Other Phlebotomy Materials
o Disinfectants
v Disinfectants are not for skin use; only for
surfaces and instruments.
• 5.25% NaOCl with a ratio of 1:100 for
nonporous surfaces and 1:10 for large
biospills with 10 minutes of contact time.
• 70% isopropyl alcohol or diluted spray.
o Hand sanitizers
v To be used as a substitute only if the hands are
not visibly soiled.
o Microscopic slides
v Precleaned 25- by 75-mm (1- by 3-in.) glass
microscope slides are used to make blood films
for hematology determinations.
• Plain Slide
• Frosted Slide - for labelling
o Labeling materials
v Indelible pen or fine point non-smear markers
for labeling purposes.
v Labeling tape or masking tape
o Timer
v It is needed to accurately determine specimen
collection times and time certain tests.
o Post it memo pads or memo pads
v It is used for organizing and remembering the
list of patients who are scheduled to be drawn.
Parts of the Needle
o Parts of the Needle
v Bevel
• The end that pierces the vein
• Allows the needle to easily slip into the skin
and vein without coring
v Shaft
• Long cylindrical portion
v Hub
• The end attaching to the blood collection
device
v Lumen
• Internal space of the needle.
o Parts of Different Blood Collection Devices
v Syringe Type Collection System
Together with the needle, the other parts of the
syringe system are:
• Graduated Barrel - is a cylinder with
graduated markings in either milliliters (mL)
or cubic centimeters (cc).
• Plunger - a rod-like device that fits tightly into
the barrel.
v Evacuated Tube System (ETS)
The three basic components of ETS are:
• A Multisample Needle has additional parts
which are the threaded hub and rubber
sleeve over needle.
• The Tube Holder is a clear, plastic,
disposable cylinder with a small threaded
opening at one end where the needle is
screwed into it and a large opening at the
other end where the collection tube is
placed.

3. UNIT IV
BLOOD COLLECTION, EQUIPMENT, ADDITIVES, AND ORDER OF DRAW
• Evacuated Tubes are used with both the
ETS and the syringe method of obtaining
blood specimens.
v Winged Infusion Set or Butterfly
• Plastic extension or wings are attached to
the needle. It holds the needle in its place.
• Tubing- 5- to 12-in. length where the blood
flows while drawing.
• Luer Attachment - connects to the syringe or
ETS. Multisample Luer adapters are
available separately.
Different Types of Needles
o Based on Gauge/Color
v The needle gauge is indicated by a number
related to the diameter of the lumen.
v It has an inverse relationship; wherein a higher
gauge indicates a smaller diameter of the needle.
Needle gauge must be selected according to the:
• Size and condition of the patient’s vein,
• The type of procedure, and
• Equipment being used.
v Appropriate needles for a typical blood collection
for most blood specimens range from G20
through 23; however, G21 is considered the
standard for most routine adult antecubital
venipuncture.
v Shown on the table are the common venipuncture
needle gauges with the needle type and use.
Gauge Needle Type Typical Use
15-17 Special
needle
attached to a
collection
bag
For blood donation and
therapeutic phlebotomy.
18 Hypodermic Used as a transfer needle
rather than for blood
collection, but safety issued
diminished its use.
20 Multisample
Hypodermic
Sometimes used when
large-volume tubes are
collected or large-volume
syringes are used on
patients with normal-sized
veins.
21 Multisample
Hypodermic
Considered the standard
routine venipuncture needle
for patients with normal
veins or for syringe blood
culture collection.
22 Multisample
Hypodermic
Used on older children and
adult patients with small
veins for syringe draws on
difficult veins.
23 Butterfly Used on infants and
children and on difficult or
hand veins of adults.
o Based on Length
v Most multisample needles come in 1- or 1.5 in.
lengths. Syringe needles have a wider range.
Butterfly needles are usually ½ to ¾ in. long.
v Some of the new safety needles come in slightly
longer lengths to accommodate resheathing
features.
v 1- to 1.5 in. are the ones most commonly used for
venipuncture.
v Length selection depends on user preference and
the depth of the vein.
• 1-in. needles for routine venipuncture
because it is less intimidating to the patient.
• 1.5-in. needles for those with larger hands to
achieve the proper angle for vein entry.
o Based on Procedure
v Syringe Type Collection Needles
• RATIONALE: This type is used for patients
with small or difficult veins.
• The syringe type of needle is used on
venipuncture procedures that only utilize one
tube.
• As previously discussed, it has various
gauges that is color coded for easier
identification.
v Evacuated Tube System (ETS) Multisample
Needles
• RATIONALE: This type allows multiple tubes
of blood to be collected during a single
venipuncture.
• They are threaded in the middle and have a
beveled point on each end.
• The threaded portion screws into a tube
holder.
• The end of the needle that pierces the vein
is longer and has a longer bevel.
• The shorted end penetrates the tube stopper
during specimen collection.
• It is covered by a sleeve that retracts as the
needle goes through the tube stopper so that
blood can flow into the tube.
• When the tube is removed, the sleeve slides
back to cover the needle to prevent leakage.
• They are available with or without safety
features.
• These needles are also color coded.

4. UNIT IV
BLOOD COLLECTION, EQUIPMENT, ADDITIVES, AND ORDER OF DRAW
v Winged Infusion (Butterfly) Needles
• RATIONALE: It is used for collecting blood
from small or difficult veins such as hand
veins and veins of elderly and pediatric
patients
• It allows more flexibility and precision than a
needle and syringe.
• It has a ½- to ¾-in. stainless steel needle
permanently connected to a 5- to 12-in.
length of tubing with either an attachment for
syringe or to a multisample adapter for the
ETS system.
• Plastic extensions that resemble wings are
attached to the needle for easier handling
and needle insertion.
• During use, the needle may be held from
above by gripping the “wings” together
between the thumb and index finger.
• Similar to the previous two types, it is also
available in various gauges. Although a G23
needle is most commonly used for
phlebotomy.
• In rare occasions, a G25 needle is used by
special personnel to collect blood from the
scalp or from other tiny veins of premature
infants and other neonates.
Different Types of Antiseptics Used in Venipuncture
o 70% isopropyl alcohol (isopropanol)
v Cotton balls are dipped in 70% isopropyl alcohol
and drained.
v Individually wrapped alcohol pads are soaked in
70% isopropyl alcohol.
v Isopropanol or ethanol-based prep pads should
not be used when samples are obtained for
legal blood alcohol testing. A non-alcohol
antiseptic should be used, such as povidone-
iodine.
o Povidone-iodine
v Povidone-iodine prep pads or swab sticks are
used for blood samples to be collected for
culture.
Blood Collection Additives
o Anticoagulants
• The most common use of anticoagulants is
for the prevention of clotting.
• The mechanism is either by chelating
(binding) or precipitating calcium to make it
unavailable for the coagulation process or
by inhibiting thrombin formation that is
required for the conversion of fibrinogen to
fibrin in coagulation.
• A test requiring whole blood or plasma
requires a tube with an anticoagulant that
must be gently mixed immediately to
prevent microclot formation and hemolysis
which affect test results negatively.
v Ethylenediaminetetraacetic acid (EDTA)
• EDTA is commonly available as a
powdered di-potassium (K2) or liquid tri-
potassium (K3) salt.
• Mechanism: Binding or chelating calcium
• It is widely used in Hematology because it
preserves the cell morphology and inhibits
platelet aggregation better than any other
anticoagulant.
v Sodium citrate
• Sodium citrate is used for coagulation tests
because it does the best job of preserving
the coagulation factors.
• Mechanism: Binding or chelating calcium
v Heparin
• Heparinized plasma is used for a number
of chemistry tests, especially stat tests and
in other rapid-response situations when a
fast turnaround time (TAT) is needed.
• Faster TAT is possible due to eliminated
time that would normally be required for a
specimen to clot before serum could be
obtained.
• Mechanism: Inhibiting thrombin formation
• There are three heparin formulations:
ammonium, lithium, and sodium heparin
with the second causing the least
interference in chemistry testing and is the
most widely used anticoagulant for both
plasma and whole-blood chemistry tests.
v Potassium oxalate
• Potassium oxalate is the most widely used.
• Mechanism: Precipitating calcium
• It is added to tubes containing glucose
preservatives to provide plasma for
glucose testing.
o Special-Use Anticoagulants
v Acid Citrate Dextrose (ACD)
• ACD is available in two formulations for
immunohematology tests such as DNA
testing and human leukocyte antigen (HLA)
phenotyping.
• Mechanism: Binding calcium, with little
effect on cells and platelets

5. UNIT IV
BLOOD COLLECTION, EQUIPMENT, ADDITIVES, AND ORDER OF DRAW
• Dextrose acts as an RBC nutrient and
preservative by maintaining its viability
v Citrate Phosphate Dextrose (CPD)
• CPD is used for blood donation.
• Mechanism: Citrate chelates calcium,
phosphate stabilizes the pH level, and
dextrose provides the cells with energy and
helps keep them alive.
v Sodium Polyanethol Sulfonate (SPS)
• SPS is used for blood culture because it also
reduces the action of a protein, called
complement, which destroys bacteria.
• It also slows down phagocytosis and
reduces the activity of certain antibiotics.
• Mechanism: Binding calcium.
o Antiglycolytic Agents
v It is a substance that prevents glycolysis, the
breakdown of glucose by blood cells.
v If not prevented, the glucose concentration
decreased at a rate of 10 mg/dL per hour.
v The most common is sodium fluoride that
preserves glucose for up to three days and also
inhibits the growth of bacteria.
v It is commonly used in combination with the
anticoagulant potassium oxalate to provide
plasma specimens for rapid-response
situations.
o Clot Activators
v Clot activators are substances that enhance
coagulation for serum specimens.
v They contain substances that provide more
surface for platelet activation such as:
• Glass (silica) particles that can be found in
serum-separator tubes (SST); which causes
clotting within 15 to 30 minutes.
• Inert clays such as Celite that are used with
some point-of-care coagulation systems.
• Thrombin clotting factors.
o Thixotropic Gel Separator
v Thixotropic gel is an inert (nonreacting)
synthetic substance initially contained in or near
the bottom of certain tubes.
v Its density is between that of the cells and the
serum or plasma.
v During centrifugation, the cell changes in
viscosity and moves to a position between the
cells and the serum or plasma, forming a
physical barrier.
v The separation prevents the cells from
continuing to metabolize substances such as
glucose in the serum or plasma.
o Trace Element-Free Tubes
v Trace element-free tubes are free of trace
element contamination as possible; for they are
used for trace element tests, toxicology studies,
and nutrient determination.
v Tests such as measure substances present in
small quantities that those elements found in the
glass, plastic, or stopper may leach into the
specimen and falsely elevate results.
v It contains EDTA, heparin, or no additive to
meet various test requirements.
Order of Draw
o Carryover/Cross-Contamination
v It is the transfer of an additive from one tube to
the next.
v It occurs when blood in an additive tube touches
the needle during ETS blood collection or when
blood is transferred from a syringe into ETS
tubes.
v Blood remaining on or within the needle can be
transferred to the next tube drawn or filled,
contaminating that tube with the additive from
the previous tube and possibly affecting the
results
o Tissue Thromboplastin Contamination
v Tissue thromboplastin is a substance in tissue
fluid that activates the extrinsic coagulation
pathway and can interfere with coagulation
tests.
v It is picked up by the first tube during ETS
collection, or it is mixed with blood collected in a
syringe.
v Although it does not pose a significant problem
for prothrombin time (PT) and partial
thromboplastin time (PTT) unless the draw is
difficult and involves a lot of needle
manipulation, it may compromise results of
other coagulation tests.
v An extra discard tube will be used to remove the
tissue fluid from the needle and is then thrown
away.
o Microbial Contamination
v Blood cultures detect microorganisms in the
blood and require a special site-cleaning
measures prior to collection to prevent
contamination by microorganisms on the skin.
v Blood culture bottles are sterile and collected
first because the sterility of the site is optimal
and to prevent microbial contamination of the
needle from unsterile tops of other tubes.

6. UNIT IV
BLOOD COLLECTION, EQUIPMENT, ADDITIVES, AND ORDER OF DRAW
o Order, Color, Additive, Inversions, Test/s
REFERENCES:
McCall, R. E., & Tankersley, C. M. (2012). Phlebotomy
Essentials (5th ed.). Lippincott Williams & Wilkins.
Order Color Additive Inversions Test/s
1 Yellow Sodium
polyanethol
sulfonate
(SPS)
3-4 Blood culture
2 Light Blue Sodium
citrate (1:9)
3-4 Coagulation
3 Black Sodium
citrate (1:4)
3-4 ESR
4 Red None 0 Chemistry
Blood bank
Serology/
Immunology
5 Gold Thixotropic
gel and clot
activator
5 Serology
6 Orange Thrombin 5-6 Chemistry
tests for
patients with
anticoagulant
therapy
7 Green/Light
Green
Heparin 8 Clinical
chemistry
Arterial blood
gas (ABG)
8 Lavender/
Pink
EDTA 8 Hematology
9 Yellow CPD 8 Human
leukocyte
antigen (HLA)
10 Gray Potassium
oxalate
8 Glucose
tolerance test