BLOOD COLLECTION:

ROUTINE VENIPUNCTURE AND SPECIMEN

HANDLING

VENIPUNCTURE PROCEDURE

The venipuncture procedure is complex, requiring both knowledge and skill to perform.
Each phlebotomist generally establishes a routine that is comfortable for her or him.

Phlebotomists are considered to have occupational exposure to blood borne pathogens.

The performance of routine vascular access procedures by skilled phlebotomists
requires, at a minimum, the use of gloves to prevent contact with blood. Laboratory
coats or work smocks are not typically needed as personal protective equipment during
routine venipuncture, but an employer must assess the workplace to determine whether
certain tasks, workplace situations, or employee skill levels may result in an employee's
need for laboratory coats or other personal protective equipment to prevent contact with
blood. It is an employer's responsibility to provide, clean, repair, replace, and/or dispose
of personal protective equipment/clothing. As part of presenting a professional
appearance, an institutional dress code may include wearing of a laboratory coat or
smock.

Several essential steps are required for every successful collection procedure:

1. Patient comfort. Is the seating comfortable and has the patient been seated for at
least 5 minutes to avoid being rushed or confused?
2. Carry out hand hygiene before and after each patient procedure, before putting
on and after removing gloves.
3. Identify the patient using two different identifiers, asking open ended questions
such as, "What is your name?" and "What is your date of birth?"
4. Assess the patient's physical disposition (i.e. diet, exercise, stress, basal state).
5. Check the requisition form for requested tests, patient information, and any
special requirements.
6. Label the collection tubes at the bedside or drawing area.
7. Select a suitable site for venipuncture.
8. Prepare the equipment, the patient and the puncture site.
9. Perform the venipuncture, collecting the sample(s) in the appropriate
container(s).
10. Recognize complications associated with the phlebotomy procedure.
11. Assess the need for sample recollection and/or rejection.
12. Promptly send the specimens with the requisition to the laboratory.
ORDER FORM / REQUISITION

A requisition form must accompany each sample submitted to the laboratory. This
requisition form must contain the proper information in order to process the specimen.
The essential elements of the requisition form are:

 Patient's surname, first name, and middle initial.

 Patient's ID number.
 Patient's date of birth and sex.
 Requesting physician's complete name.
 Source of specimen. This information must be given when requesting microbiology,
cytology, fluid analysis, or other testing where analysis and reporting is site specific.
 Date and time of collection.
 Initials of phlebotomist.
 Indicating the test(s) requested.

An example of a simple requisition form with the essential elements is shown below:
LABELING THE SAMPLE

A properly labeled sample is essential so that the results of the test match the patient.
The key elements in labeling are:

 Patient's surname, first and middle.

 Patient's ID number.
 NOTE: Both of the above MUST match the same on the requisition form.
 Date, time and initials of the phlebotomist must be on the label of EACH tube.

Automated systems may include labels with bar codes.

Examples of labeled collection tubes are shown below:

EQUIPMENT:

THE FOLLOWING ARE NEEDED FOR ROUTINE VENIPUNCTURE:

 Evacuated Collection Tubes - The tubes are designed to fill with a predetermined
volume of blood by vacuum. The rubber stoppers are color coded according to the
additive that the tube contains. Various sizes are available. Blood should NEVER be
poured from one tube to another since the tubes can have different additives or coatings
(see illustrations at end).
 Needles - The gauge number indicates the bore size: the larger the gauge number, the
smaller the needle bore. Needles are available for evacuated systems and for use with a
syringe, single draw or butterfly system.
 Holder/Adapter - use with the evacuated collection system.
 Tourniquet - Wipe off with alcohol and replace frequently.
 Alcohol Wipes - 70% isopropyl alcohol.
 Povidone-iodine wipes/swabs - Used if blood culture is to be drawn.
 Gauze sponges - for application on the site from which the needle is withdrawn.
 Adhesive bandages / tape - protects the venipuncture site after collection.
 Needle disposal unit - needles should NEVER be broken, bent, or recapped. Needles
should be placed in a proper disposal unit IMMEDIATELY after their use.
 Gloves - can be made of latex, rubber, vinyl, etc.; worn to protect the patient and the
phlebotomist.
 Syringes - may be used in place of the evacuated collection tube for special
circumstances.
ORDER OF DRAW

Blood collection tubes must be drawn in a specific order to avoid cross-contamination of

additives between tubes. The recommended order of draw for plastic collection tubes is:

1. First - blood culture bottle or tube (yellow or yellow-black top)

2. Second - coagulation tube (light blue top). If just a routine coagulation assay is the only
test ordered, then a single light blue top tube may be drawn. If there is a concern
regarding contamination by tissue fluids or thromboplastins, then one may draw a non-
additive tube first, and then the light blue top tube.
3. Third - non-additive tube (red top)
4. Last draw - additive tubes in this order:
1. SST (red-gray or gold top). Contains a gel separator and clot activator.
2. Sodium heparin (dark green top)
3. PST (light green top). Contains lithium heparin anticoagulant and a gel separator.
4. EDTA (lavender top)
5. ACDA or ACDB (pale yellow top). Contains acid citrate dextrose.
6. Oxalate/fluoride (light gray top)

NOTE:Tubes with additives must be thoroughly mixed. Erroneous test results may be
obtained when the blood is not thoroughly mixed with the additive.

PROCEDURAL ISSUES

PATIENT RELATIONS AND IDENTIFICATION:

The phlebotomist's role requires a professional, courteous, and understanding manner in

all contacts with the patient. Greet the patient and identify yourself and indicate the
procedure that will take place. Effective communication - both verbal and nonverbal - is
essential.

Proper patient identification MANDATORY. If an inpatient is able to respond, ask for a

full name and always check the armband or bracelet for confirmation. DO NOT DRAW
BLOOD IF THE ARMBAND OR BRACELET IS MISSING. For an inpatient the nursing
staff can be contacted to aid in identification prior to proceeding.
An outpatient must provide identification other than the verbal statement of a name.
Using the requisition for reference, ask a patient to provide additional information such
as a surname or birthdate. A government issued photo identification card such as a
driver's license can aid in resolving identification issues.

If possible, speak with the patient during the process. The patient who is at ease will be
less focused on the procedure. Always thank the patient and excuse yourself
courteously when finished.

PATIENT'S BILL OF RIGHTS:

The Patient's Bill of Rights has been adopted by many hospitals as declared by the Joint
Commission on Accreditation of Healthcare Organizations (JCAHO). The basic patient
rights endorsed by the JCAHO follow in condensed form are given below.

The patient has the right to:

 Impartial access to treatment or accommodations that are available or medically

indicated, regardless of race, creed, sex, national origin, or sources of payment
for care.
 Considerate, respectful care.
 Confidentiality of all communications and other records pertaining to the patient's
care.
 Expect that any discussion or consultation involving the patient's case will be
conducted discretely and that individuals not directly involved in the case will not
be present without patient permission.
 Expect reasonable safety congruent with the hospital practices and environment.
 Know the identity and professional status of individuals providing service and to
know which physician or other practitioner is primarily responsible for his or her
care.
 Obtain from the practitioner complete and current information about diagnosis,
treatment, and any known prognosis, in terms the patient can reasonably be
expected to understand.
 Reasonable informed participation in decisions involving the patient's health care.
The patient shall be informed if the hospital proposes to engage in or perform
human experimentation or other research/educational profits affecting his or her
care or treatment. The patient has the right to refuse participation in such activity.
 Consult a specialist at the patient's own request and expense.
 Refuse treatment to the extent permitted by law.
 Regardless of the source of payment, request and receive an itemized and
detailed explanation of the total bill for services rendered in the hospital.
 Be informed of the hospital rules and regulations regarding patient conduct.
VENIPUNCTURE SITE SELECTION:

Although the larger and fuller median cubital and cephalic veins of the arm are used
most frequently, the basilic vein on the dorsum of the arm or dorsal hand veins are also
acceptable for venipuncture. Foot veins are a last resort because of the higher
probability of complications.

Certain areas are to be avoided when choosing a site:

 Extensive scars from burns and surgery - it is difficult to puncture the scar tissue
and obtain a specimen.
 The upper extremity on the side of a previous mastectomy - test results may be
affected because of lymphedema.
 Hematoma - may cause erroneous test results. If another site is not available,
collect the specimen distal to the hematoma.
 Intravenous therapy (IV) / blood transfusions - fluid may dilute the specimen, so
collect from the opposite arm if possible. Otherwise, satisfactory samples may be
drawn below the IV by following these procedures:
o Turn off the IV for at least 2 minutes before venipuncture.
o Apply the tourniquet below the IV site. Select a vein other than the one
with the IV.
o Perform the venipuncture. Draw 5 ml of blood and discard before drawing
the specimen tubes for testing.
 Lines - Drawing from an intravenous line may avoid a difficult venipuncture, but
introduces problems. The line must be flushed first. When using a syringe
inserted into the line, blood must be withdrawn slowly to avoid hemolysis.
 Cannula/fistula/heparin lock - hospitals have special policies regarding these
devices. In general, blood should not be drawn from an arm with a fistula or
cannula without consulting the attending physician.
 Edematous extremities - tissue fluid accumulation alters test results.

PROCEDURE FOR VEIN SELECTION:

 Palpate and trace the path of veins with the index finger. Arteries pulsate, are
most elastic, and have a thick wall. Thrombosed veins lack resilience, feel cord-
like, and roll easily.
 If superficial veins are not readily apparent, you can force blood into the vein by
massaging the arm from wrist to elbow, tap the site with index and second finger,
apply a warm, damp washcloth to the site for 5 minutes, or lower the extremity
over the bedside to allow the veins to fill.
PERFORMANCE OF A VENIPUNCTURE:

 Approach the patient in a friendly, calm manner. Provide for their comfort as
much as possible, and gain the patient's cooperation.
 Identify the patient correctly.
 Properly fill out appropriate requisition forms, indicating the test(s) ordered.
 Verify the patient's condition. Fasting, dietary restrictions, medications, timing,
and medical treatment are all of concern and should be noted on the lab
requisition.
 Check for any allergies to antiseptics, adhesives, or latex by observing for
armbands and/or by asking the patient.
 Position the patient. The patient should either sit in a chair, lie down or sit up in
bed. Hyperextend the patient's arm.
 Apply the tourniquet 3-4 inches above the selected puncture site. Do not place
too tightly or leave on more than 2 minutes (and no more than a minute to avoid
increasing risk for hemoconcentration). Wait 2 minutes before reapplying the
tourniquet.
 The patient should make a fist without pumping the hand.
 Select the venipuncture site.
 Prepare the patient's arm using an alcohol prep. Cleanse in a circular fashion,
beginning at the site and working outward. Allow to air dry.
 Grasp the patient's arm firmly using your thumb to draw the skin taut and anchor
the vein. The needle should form a 15 to 30 degree angle with the surface of the
arm. Swiftly insert the needle through the skin and into the lumen of the vein.
Avoid trauma and excessive probing.

 When the last tube to be drawn is filling, remove the tourniquet.

 Remove the needle from the patient's arm using a swift backward motion.
 Press down on the gauze once the needle is out of the arm, applying adequate
pressure to avoid formation of a hematoma.
 Dispose of contaminated materials/supplies in designated containers.
 Mix and label all appropriate tubes at the patient bedside.
 Deliver specimens promptly to the laboratory.
PHLEBOTOMY PROCEDURE ILLUSTRATED:

 Patient identification
 Filling out the requisition
 Equipment
 Apply tourniquet and palpate for vein
 Sterilize the site
 Insert needle
 Drawing the specimen
 Drawing the specimen
 Releasing the tourniquet
 Applying pressure over the vein
 Applying bandage
 Disposing needle into sharps
 labeling the specimens

PERFORMANCE OF A FINGERSTICK:

 Follow the procedure as outlined above for greeting and identifying the patient. As
always, properly fill out appropriate requisition forms, indicating the test(s) ordered.
 Verify the patient's condition. Fasting, dietary restrictions, medications, timing, and
medical treatment are all of concern and should be noted on the lab requisition.
 Position the patient. The patient should either sit in a chair, lie down or sit up in bed.
Hyperextend the patient's arm.
 The best locations for fingersticks are the 3rd (middle) and 4th (ring) fingers of the non-
dominant hand. Do not use the tip of the finger or the center of the finger. Avoid the side
of the finger where there is less soft tissue, where vessels and nerves are located, and
where the bone is closer to the surface. The 2nd (index) finger tends to have thicker,
callused skin. The fifth finger tends to have less soft tissue overlying the bone. Avoid
puncturing a finger that is cold or cyanotic, swollen, scarred, or covered with a rash.
 Using a sterile lancet, make a skin puncture just off the center of the finger pad. The
puncture should be made perpendicular to the ridges of the fingerprint so that the drop of
blood does not run down the ridges.
 Wipe away the first drop of blood, which tends to contain excess tissue fluid.
 Collect drops of blood into the collection device by gently massaging the finger. Avoid
excessive pressure that may squeeze tissue fluid into the drop of blood.
 Cap, rotate and invert the collection device to mix the blood collected.
  Have the patient hold a small gauze pad over the puncture site for a couple of minutes to
stop the bleeding.
 Dispose of contaminated materials/supplies in designated containers.
 Label all appropriate tubes at the patient bedside.
 Deliver specimens promptly to the laboratory.

FINGERSTICK PROCEDURE ILLUSTRATED:

 Equipment
 Proper location on finger
 Puncture with lancet
 Drop of blood
 Wipe first drop
 Collecting the specimen
 Specimen container

ADDITIONAL CONSIDERATIONS:

To prevent a hematoma:

 Puncture only the uppermost wall of the vein

 Remove the tourniquet before removing the needle
 Use the major superficial veins
 Make sure the needle fully penetrates the upper most wall of the vein. (Partial
penetration may allow blood to leak into the soft tissue surrounding the vein by
way of the needle bevel)
 Apply pressure to the venipuncture site

To prevent hemolysis (which can interfere with many tests):

 Mix tubes with anticoagulant additives gently 5-10 times

 Avoid drawing blood from a hematoma
 Avoid drawing the plunger back too forcefully, if using a needle and syringe, or
too small a needle, and avoid frothing of the sample
 Make sure the venipuncture site is dry
 Avoid a probing, traumatic venipuncture
 Avoid prolonged tourniquet application or fist clenching.
Indwelling Lines or Catheters:

 Potential source of test error

 Most lines are flushed with a solution of heparin to reduce the risk of thrombosis
 Discard a sample at least three times the volume of the line before a specimen is
obtained for analysis

Hemoconcentration: An increased concentration of larger molecules and formed

elements in the blood may be due to several factors:

 Prolonged tourniquet application (no more than 1 minute)

 Massaging, squeezing, or probing a site
 Long-term IV therapy
 Sclerosed or occluded veins

Prolonged Tourniquet Application:

 Primary effect is hemoconcentration of non-filterable elements (i.e. proteins). The

hydrostatic pressure causes some water and filterable elements to leave the
extracellular space.
 Significant increases can be found in total protein, aspartate aminotransferase
(AST), total lipids, cholesterol, iron
 Affects packed cell volume and other cellular elements
 Hemolysis may occur, with pseudohyperkalemia.

Patient Preparation Factors:

 Therapeutic Drug Monitoring: different pharmacologic agents have patterns of

administration, body distribution, metabolism, and elimination that affect the drug
concentration as measured in the blood. Many drugs will have "peak" and
"trough" levels that vary according to dosage levels and intervals. Check for
timing instructions for drawing the appropriate samples.
 Effects of Exercise: Muscular activity has both transient and longer lasting
effects. The creatine kinase (CK), aspartate aminotransferase (AST), lactate
dehydrogenase (LDH), and platelet count may increase.
 Stress: May cause transient elevation in white blood cells (WBC's) and elevated
adrenal hormone values (cortisol and catecholamines). Anxiety that results in
hyperventilation may cause acid-base imbalances, and increased lactate.
 Diurnal Rhythms: Diurnal rhythms are body fluid and analyte fluctuations during
the day. For example, serum cortisol levels are highest in early morning but are
decreased in the afternoon. Serum iron levels tend to drop during the day. You
must check the timing of these variations for the desired collection point.
  Posture: Postural changes (supine to sitting etc.) are known to vary lab results of
some analytes. Certain larger molecules are not filterable into the tissue,
therefore they are more concentrated in the blood. Enzymes, proteins, lipids,
iron, and calcium are significantly increased with changes in position.
 Other Factors: Age, gender, and pregnancy have an influence on laboratory
testing. Normal reference ranges are often noted according to age.

REASONS FOR CANCELING A LABORATORY TEST

A test that has been ordered may be cancelled due to problems unrelated to drawing the
specimen, and these are the most common causes for cancellations:

 Duplicate test request

 Incorrect test ordered
 Test no longer needed

A test may be cancelled due to a technical problem in the specimen collection process:

 Hemolysis of the specimen

 Clotted specimen
 Quantity of specimen not sufficient
 Collection of specimen in incorrect tube
 Contaminated specimen
 Identification of the specimen is suspect
 Delay in transport - specimen too old

SAFETY AND INFECTION CONTROL

Because of contacts with sick patients and their specimens, it is important to follow
safety and infection control procedures.

PROTECT YOURSELF
  Practice universal precautions:
o Wear gloves and a lab coat or gown when handling blood/body fluids.
o Change gloves after each patient or when contaminated.
o Wash hands frequently.
o Dispose of items in appropriate containers.
 Dispose of needles immediately upon removal from the patient's vein. Do not
bend, break, recap, or resheath needles to avoid accidental needle puncture or
splashing of contents.
 Clean up any blood spills with a disinfectant such as freshly made 10% bleach.
 If you stick yourself with a contaminated needle:
o Remove your gloves and dispose of them properly.
o Squeeze puncture site to promote bleeding.
o Wash the area well with soap and water.
o Record the patient's name and ID number.
o Follow institution's guidelines regarding treatment and follow-up.
o NOTE: The use of prophylactic zidovudine following blood exposure to
HIV has shown effectiveness (about 79%) in preventing seroconversion

PROTECT THE PATIENT

 Place blood collection equipment away from patients, especially children and
psychiatric patients.
 Practice hygiene for the patient's protection. When wearing gloves, change them
between each patient and wash your hands frequently. Always wear a clean lab
coat or gown.

TROUBLESHOOTING GUIDELINES:
IF AN INCOMPLETE COLLECTION OR NO BLOOD IS OBTAINED:

 Change the position of the needle. Move it forward (it may not be in the lumen)

 or move it backward (it may have penetrated too far).

 Adjust the angle (the bevel may be against the vein wall).

 Loosen the tourniquet. It may be obstructing blood flow.

  Try another tube. Use a smaller tube with less vacuum. There may be no
vacuum in the tube being used.
 Re-anchor the vein. Veins sometimes roll away from the point of the needle and
puncture site.
 Have the patient make a fist and flex the arm, which helps engorge muscles to fill
veins.
 Pre-warm the region of the vein to reduce vasoconstriction and increase blood
flow.
 Have the patient drink fluids if dehydrated.

IF BLOOD STOPS FLOWING INTO THE TUBE:

 The vein may have collapsed; resecure the tourniquet to increase venous filling.
If this is not successful, remove the needle, take care of the puncture site, and
redraw.

 The needle may have pulled out of the vein when switching tubes. Hold
equipment firmly and place fingers against patient's arm, using the flange for
leverage when withdrawing and inserting tubes.

PROBLEMS OTHER THAN AN INCOMPLETE COLLECTION:

  A hematoma forms under the skin adjacent to the puncture site - release the
tourniquet immediately and withdraw the needle. Apply firm pressure.

Hematoma formation is a problem in older patients.

 The blood is bright red (arterial) rather than venous. Apply firm pressure for more
than 5 minutes.

BLOOD COLLECTION ON BABIES:

 The recommended location for blood collection on a newborn baby or infant is the heel.
The diagram below indicates in green the proper area to use for heel punctures for blood
collection:

 Prewarming the infant's heel (42 C for 3 to 5 minutes) is important to obtain capillary
blood gas samples and warming also greatly increases the flow of blood for collection of
other specimens. However, do not use too high a temperature warmer, because baby's
skin is thin and susceptible to thermal injury.
 Clean the site to be punctured with an alcohol sponge. Dry the cleaned area with a dry
cotton sponge. Hold the baby's foot firmly to avoid sudden movement.
 Using a sterile blood lancet, puncture the side of the heel in the appropriate regions
shown above in green. Do not use the central portion of the heel because you might
injure the underlying bone, which is close to the skin surface. Do not use a previous
puncture site. Make the cut across the heelprint lines so that a drop of blood can well up
and not run down along the lines.
 Wipe away the first drop of blood with a piece of clean, dry cotton. Since newborns do
not often bleed immediately, use gentle pressure to produce a rounded drop of blood.
Do not use excessive pressure or heavy massaging because the blood may become
diluted with tissue fluid.
 Fill the capillary tube(s) or micro collection device(s) as needed.
 When finished, elevate the heel, place a piece of clean, dry cotton on the puncture site,
and hold it in place until the bleeding has stopped.
 Be sure to dispose of the lancet in the appropriate sharps container. Dispose of
contaminated materials in appropriate waste receptacles. Remove your gloves and wash
your hands.

HEELSTICK PROCEDURE ILLUSTRATED:

  Heelstick on baby

PEDIATRIC PHLEBOTOMY:

 Children, particularly under the age of 10, may experience pain and anxiety during the
phlebotomy procedure.
 A variety of techniques can be employed to reduce pain and anxiety. Effective methods
use distraction. These may include listening to music or a story, watching a video,
playing with a toy, having a parent provide distraction with talk or touch, using flash
cards, and squeezing a rubber ball. (Uman et al, 2013)