The document provides guidelines for blood sample collection procedures. It discusses introducing oneself to the patient, checking identification, selecting a vein, using the proper order and techniques for drawing blood into tubes, labeling the tubes, and applying pressure after withdrawing the needle. Precautions are outlined such as universal safety protocols, proper disposal of sharps, and cleaning up spills. The document also provides information on collecting samples from infants and the indications for rejecting specimens.

1. BLOOD SAMPLE
COLLECTION PROCEDURES
PRESENTED BY :
NIDHI SINGH
NURSING OFFICER
ERA’S MEDICAL COLLEGE AND
HOSPITAL LUCKNOW
GUIDE BY:
DR.ANJALATCHI
VICE PRINCIPAL
ERA’S MEDICAL COLLEGE AND
HOSPITAL LUCKNOW

2. INTRODUCTION
 A blood sample is taken by a physician, in a laboratory or
hospital and in many cases the blood sample is sent on
to analysis. It takes about 5 minutes to take a blood
sample. Blood samples are taken in the arm, hand,
finger or ear, depending on the analysis to be made.

3. COLLECTIO OF BLOOD

4. BLOOD SPECIMEN COLLECTION PROCESS

5. DEFINITION OF BLOOD SAMPLE COLLECTION

6. Indications for blood sampling and
blood collection

7. • capillary sampling (i.e. finger or heel-pricks or, rarely, an ear lobe
puncture) for analysis of capillary blood specimens for all ages;
examples include testing of iron levels before blood donation,
blood glucose monitoring, and rapid tests for HIV, malaria and
syphilis.
•Blood collection is used to obtain blood from donors for various
therapeutic purposes.

9. CONTENT:
 Introduction
 Definition
 Indication
 Preparation of patient.
 preparation of environment .
 preparation of articles .
 procedure .
 After care .
 records and report .
 checklist .

10. TYPE OF SAMPLE COLLECTION

11. Lab requsition form contain

12. ROUTINE VENIPUNCTURE GUIDELINES
MATERIALS
 1. Safety Needles, 22g or less
 2. Butterfly needles. 21g or less
 3. Syringes
 4. Vacutainer tube holder
 5. Transfer Device
 6. Blood Collection Tubes.
 The vacuum tubes are designed to draw a predetermined volume of blood.
 Tubes with different additives are used for collecting blood specimens for specific types of
tests.
 The color of cap is used to identify these additives.
 7. Tourniquets. Single use, disposable, latex-free tourniquets
 8. Antiseptic. Individually packaged 70% isopropyl alcohol wipes.
 9. 2×2 Gauze
 10. Sharps Disposal Container. An OSHA acceptable, puncture proof container
marked “Biohazardous”.
 11. Bandages or tape

13. SAFETY
 1. Observe universal (standard) safety precautions.
 2. Observe all applicable isolation procedures.
 3. PPE’s will be worn at all times.
 4. Wash hands in warm, running water with a appropriate hand washing
product,
 5. If hands are not visibly contaminated a commercial foaming hand
wash product may be used before and after each patient collection.
 6. Gloves are to be worn during all phlebotomies, and changed between
patient collections.
 7. Palpation of phlebotomy site may be performed without gloves
providing the skin is not broken.

14. CONTINUED
 8. A lab coat or gown must be worn during blood collection procedures.
 9. Needles and hubs are single use and are disposed of in an appropriate
‘sharps’ container as one unit.
 10. Needles are never recapped, removed, broken, or bent after phlebotomy
procedure.
 11. Gloves are to be discarded in the appropriate container immediately after
the phlebotomy procedure.
 12. All other items used for the procedure must be disposed of according to
proper biohazardous waste disposal policy.
 13. Contaminated surfaces must be cleaned with freshly prepared 10% bleach
solution. All surfaces are cleaned daily with bleach.
 14. In the case of an accidental needlestick, immediately wash the area with an
antibacterial soap, express blood from the wound, and contact your supervisor.

15. PREPRATION OF PATIENT
 Check consult .
 Identify and prepare the patient condition .
 Select the site.
 Perform hand hygiene and put on gloves.
 Disinfect the entry site.

16. PREPRATION OF ENVIRONMENT
 Provide calm environment .
 Wear gloves and a lab coat or gown when handling
blood/body fluids.
 Dispose of needles immediately upon removal from the
patient's vein.
 Clean up any blood spills with a disinfectant such as
freshly made 10% bleach.
 If you stick yourself with a contaminated needle:

17. PREPRATION OF ARTICLE

19. PROCEDURE OF
BLOOD COLLECTION

20. PROCEDURE OF BLOOD COLLECTION
 Introduce yourself to the patient, and ask the patient to state
their full name.
 Check that the laboratory form matches the patient's identity
.
 Ask whether the patent has allergies, phobias or has ever
fainted during previous injections or blood draws.
 If the patient is anxious or afraid, reassure the person and
ask what would make them more comfortable.
 Make the patient comfortable in a supine position (if possible).
 Place a clean paper or towel under the patient's arm.
 Discuss the test to be performed (see Annex F) and obtain
verbal consent. The patient has a right to refuse a test at any
time before the blood sampling, so it is important to ensure
that the patient has understood the procedure.

21. PROCEDURE
 1. Identify the patient, two forms of active identification are required.
 Ask the patient to state their name and date of birth.
 This information must match the requisition.
 2. Reassure the patient that the minimum amount of blood required for
testing will be drawn.
 3. Verify that any diet or time restrictions have been met.
 4. Order of Draw
 The following order of draw is the approved order as established by CLSI.
 This order of draw should be followed whenever multiple tubes are drawn
during a single venipuncture.
 This is to prevent cross contamination by the tube additives that could
lead to erroneous results.

22. ORDER OF BLOOD DRAW FOR SAMPLE
COLLECTION
 1. Blood Culture
 2. Light Blue Top (plasma): 3.2% sodium citrate. These tubes are
used for coagulation tests and need to be completely filled to ensure the
proper ratio of blood to anticoagulant.
 3. Red Top (serum): Plain and gel. Used for chemistry and reference
tests.
 4. Green Top (plasma): With and without gel, contains lithium heparin.
These tubes are used primarily for chemistry tests.
 5. Lavender or Pink Top (plasma): Contains EDTA. Used primarily for
hematology and blood bank testing.
 6. Gray Top (plasma): Contains sodium fluoride/potassium oxalate.
Used by chemistry for glucose testing.
 7. Yellow Top (plasma and cells): Contains ACD solution A or B. Used
for Genetics testing.

23. Order of Draw
1) Blood culture tube
2) Citrate tube
3) Plain tube
4) Heparin tube
5) EDTA tube
6) Fluoride tube

24. CONTINUED
 NOTE: When using a winged blood collection set for venipuncture and a
coagulation tube is the first tube needed, first draw a discard tube (plain
red top or light blue top). The discard tube does not need to be filled
completely.
 1. Assemble the necessary equipment appropriate to the patient’s
physical characteristics.
 2. Wash hands and put on gloves.
 3. Position the patient with the arm extended to form a straight-line form
shoulder to wrist.
 4. Do not attempt a venipuncture more than twice. Notify your
supervisor or patient’s physician if unsuccessful.
 5. Select the appropriate vein for venipuncture.

25. 5. Select the appropriate vein for venipuncture
 .
 The larger median cubital, basilic and cephalic veins are most frequently used, but
other may be necessary and will become more prominent if the patient closes his fist
tightly.
 At no time may phlebotomists perform venipuncture on an artery.
 It is not recommended that blood be drawn from the feet .The Providers permission
is required to draw from this site.
 Extensive scarring or healed burn areas should be avoided
 Specimens should not be obtained from the arm on the same side as a mastectomy.
 Avoid areas of hematoma.
 If an IV is in place, samples may be obtained below but NEVER above the IV site.
 Do not obtain specimens from an arm having a cannula, fistula, or vascular graft.
 Allow 10-15 minutes after a transfusion is completed before obtaining a blood
sample.

26.  6. Apply the tourniquet 3-4 inches above the collection site.
 Never leave the tourniquet on for over 1 minute.
 If a tourniquet is used for preliminary vein selection, release
it and reapply after two minutes.
 7. Clean the puncture site by making a smooth circular pass
over the site with the 70%
alcohol pad, moving in an outward spiral from the zone of
penetration.
 Allow the skin to dry before proceeding.
 Do not touch the puncture site after cleaning.

27. 8. Perform the venipuncture
 1. Attach the appropriate needle to the hub by removing the plastic cap over the
small end of the needle and inserting into the hub, twisting it tight.
 2. Remove plastic cap over needle and hold bevel up.
 3. Pull the skin tight with your thumb or index finger just below the puncture site.
 4. Holding the needle in line with the vein, use a quick, small thrust to penetrate the
skin and enter the vein in one smooth motion.
 5. Holding the hub securely, insert the first vacutainer tube following proper order of
draw into the large end of the hub penetrating the stopper. Blood should flow into
the evacuated tube.
 6. After blood starts to flow, release the tourniquet and ask the patient to open his
or her hand.
 7. When blood flow stops, remove the tube by holding the hub securely and pulling
the tube off the needle.

28. CONTINUED s
 8. Gently invert each tube
 Light blue top- invert 3-4 times
 Red and gold tops invert 5 times.
 All other tubes containing an additive should be gently
inverted 8-10 times.
 9. DO NOT SHAKE OR MIX VIGOROUSLY. If multiple
tubes are needed, follow the proper order of draw

29. Phlebotomy

30.  9. Place a gauze pad over the puncture site and remove the needle.
 10. Activate the safety device and properly dispose of the vacutainer holder with
needle attached into a sharps container.
 11. Immediately apply slight pressure to the gauxe pad over the venipuncture
site..
 Ask the patient to apply pressure for at least 2 minutes.
 When bleeding stops, apply a fresh bandage, gauze or tape.
 12. Tubes must be positively identified after filling with a firmly attached patient
label.
 The label must include the patient first and last names, DOB,, collection date and
time and collectors initials.
 If no patient labels are available, manually label the tubes with the required
information. All labels must include two identifiers .
 The tube must be labeled before leaving the patient.
 13. Observe special handling requirements
 Some test specimens require special handling for accurate results.
 Refer to the specific test in the online directory for handling and storage
requirements.

31. Blood collection tubes:
 Serum separating tubes (SST)
 Plasma separating tubes (PST)

32. Top Color Additives Principle Uses
Lavender EDTA
Dose= 1to2g/l of
blood
-The strongest anti-coagulant
-Ca+2 chelating agent
- Hematology
- Blood bank
Light Blue Sodium Citrate
2g/l
Ca+2 chelating agent - PT
- APTT:
Green Sodium Heparin
or Lithium
Heparin
Heparin binds to Thrombin and inhibits the
second step in the coagulation cascade
(Prothrombin Thrombin)
Fibrinogen Fibrin
Enzymes
Hormones
Electrolytes (Na+, K+, Mg+,
Cl)
Heparin
Plasma Separating Tubes (PST)

33. Top Color Additives Principle Uses
Gray -Sodium Fluoride
2g/l
-Potassium Oxalate
Glycolysis inhibitor
Anti-Coagulant
Glucose tests
Top Tubes Additives Principle Uses
Red ------
Sometimes it has gel or
silicon at the bottom of tube
to reduce hemolysis
Enhancing the
formation of blood
clot
Serology
-Antibodies
-Hormones
-Drugs
Virology
Chemistry
Blood cross matching before
blood transfusion
Serum Separating Tubes (SST)

36. VENIPUNCTURE PROCEDURE USING A SYRINGE:
 1. Place a sheathed needle or butterfly on the syringe.
 2. Remove the cap and turn the bevel up.
 3. Pull the skin tight with your thumb or index finger just below the puncture
site.
 4. Holding the needle in line with the vein, use a quick, small thrust to penetrate
the skin and vein in one motion.
 5. Draw the desired amount of blood by pulling back slowly on the syringe
stopper. Release the tourniquet.
 6. Place a gauze pad over the puncture site and quickly remove the needle.
 7. Immediately apply pressure. Ask the patient to apply pressure to the gauze
for at least 2 minutes.
 8. When bleeding stops, apply a fresh bandage, gauze or tape.
 9. Transfer blood drawn into the appropriate tubes as soon as possible using a
Blood Transfer Device, as a delay could cause improper coagulation.
 10. Gently invert tubes containing an additive 5-8 times.
 11. Dispose of the syringe and needle as a unit into an appropriate sharps
container.

37. INFANT/CHILD PHLEBOTOMY
 1. Confirm the patient’s identification
 2. Secure patient to Papoose apparatus for stabilization if
child is unable to sit upright on their own.
 3. Assemble the required supplies
 4. Select the collection site and proceed as routine
phlebotomy. If the child is old enough, collect blood as in
an adult.

38. SPECIMEN REJECTION CRITERIA
Specimen rejection criteria:
Specimen improperly labeled or unlabeled
Specimen improperly collected or preserved
Specimen submitted without properly completed request
form
 contaminated form
Improperly volume & leakage sample
Absurd blood sample -: High electrolyte level
Hemolyzed sample (show tubes)

39. AFTER CARE
 Leave the dressing strip in place for 30 minutes to an
hour.
 For the Rest of the Day .
 Avoid tight or restrictive clothing on the arm
 If bleeding occurs, apply firm pressure with your fingers
directly over the needle site
 Avoid strenuous use of the arm eg sports or heavy lifting
 If you have discomfort or swelling of the arm, elevate
your arm and place ice over the site for 10 minutes.
 Rest the arm as much as possible.

41. RECORD AND REPORTS
 Checked vital sign and recorded on time .
 First - blood culture bottle or tube (yellow or yellow-black
top)
 Second - coagulation tube (light blue top).
 Third - non-additive tube (red top)
 Last draw - additive tubes in this order:
 SST (red-gray or gold top). Contains a gel separator and
clot activator.
 Sodium heparin (dark green top)
 PST (dark green green top with gold rim). Contains lithium
heparin anticoagulant and a gel separator.
 EDTA (lavender top)
 Oxalate/fluoride (light gray top) or other additives

43. 1. Never do mouth pipetting.
2. Barrier protection such as gloves, masks,
goggles and apron must be available,
3. Phlebotomists must change gloves and
adequately dispose of them between
drawing blood from different patient.
4. Frequent hands washing whenever gloves
are changed.

44. 6. Facial barrier protection used for
spattering of blood or body fluid.
7. Avoid using syringes whenever
possible and
dispose of needle in white coloured
container
8. Make a habit of keeping your hands
away from your mouth, nose, eyes and
other mucous membrane inoculation.

45. 11.Decontaminate all surfaces and reusable device
after use.
12.Before centrifuging tubes, inspect them for
cracks.
13.Use biohazard disposal techniques. Eg. Red
bag.
14.Never leave a discarded tube or infected
material unattended and unlabeled.
15.All employees must be vaccinated with
hepatitis B vaccine.

46. Biohazard Flammable Material Hazard
Toxic Material Hazard
Ionizing Radiation Hazard

47. THANK YOU