This document provides information about phlebotomy and blood specimen collection. It discusses that phlebotomy involves making an incision in a vein with a needle, which is known as a venipuncture. Blood can be obtained through skin punctures or venous sampling. The document outlines the different types of collection tubes used including those containing EDTA, sodium citrate, sodium fluoride, and clot activators. It provides the recommended order of draw and notes on proper inversion of tubes to mix additives. The venipuncture procedure and avoiding problematic sites are summarized. New technologies for vein finding and infant blood collection are also mentioned.

1. Phlebotomy
Lecturer : Kalit . M. Abdullah
MPH ,BMS,

2. Phlebotomy
• Phlebotomy is the process of making an
incision in a vein with a needle.
• The procedure is known as a venipuncture.
• A person who performs phlebotomy is called a
phlebotomist, although doctors, nurses & medical
laboratory scientists.

3. Blood Specimen Collection and
Processing:
•The first step in acquiring a quality lab. Test result for any patient
is the specimen collection procedure.
•Blood specimens are obtained through capillary skin punctures (finger,
toe, heel), arterial or venous sampling.
•Capillary / dermal /skin puncture, earlobe stick, finger stick, heel
stick (infants).
•Venous puncture (direct puncture of vein by

4. An Introduction Venipuncture:
•Venipuncture is the process of obtaining blood
samples from veins for laboratory testing, It is
probably the most common procedure in the
medical field, usually performed for the following
reasons:…

5. 1. To obtain blood samples in order to perform diagnostics.
2. To collect blood for later use should the patient’s condition requires
transfusions.
3. To remove blood that was found with excessive levels of erythrocytes or iron.
An Introduction Venipuncture:

7. EDTA salts (ethylenediaminetetraacetic
acid) are used to anticoagulated whole
blood for hematological investigations as
the cellular components of the blood are
particularly well preserved by EDTA.
EDTA K₂ / K₃
Mixing method
✔ 8-10 inversions
After the blood sample has been taken is extremely important to avoid micro clotting.
Classification
✔ Whole Blood

8. Clot Activator
In order to obtain serum samples from
plastic tubes, the tube must have a
coagulation activator added. As the
plastic surface alone is insufficient to
trigger the coagulation within an
acceptable time, plastic serum tubes
have silica particles added for this
purpose
≤ 1300 g for 10 minutes at 18-25°C
✔Serum analysis
Centrifuging
Clotting
✔ 60 minutes.

9. Clot
Activator
with Gel
During the centrifugation of tubes, an inert gel
separates the serum and the blood clot
preventing the contamination of the serum
from the separated cellular components.The
stability of the gel barrier is a distinct
advantage
during transport and storage.The blood to
additive ratio is 9
Clotting
✔ 30 minutes.
1300-2000 g for 10 minutes
Centrifuging
✔Serum analysis

10. Clotting
✔ 30 minutes.
For coagulation analyses different plasma specifications can
be obtained from the citrated blood.
✔ Platelet rich plasma:150-200 g for 5 minutes at 18-25°C"
✔ Platelet poor plasma:2000-2500 g for 10-15 minutes at 18-
25°C"
✔ Platelet free plasma:>3000 g for 15-30 minutes at 18-
25°C"
Centrifuging
Trisodium Citra
te
Trisodium citrate is used as an anticoagulant
for coagulation investigations. It works as an
anticoagulant by forming complexes with
metal ions such as calcium inhibiting the
coagulation cascade. Anticoagulation with
trisodium citrate is reversible.The blood to
additive ratio is 9:1.

11. Glucos
e
Glucose tubes are available in Sodium Fluoride,
Potassium
Oxalate. Glucose values in unpreserved blood samples
decrease quickly after collection as glucose is
metabolized
by the blood cells. The additives contained in
Fluoride/Oxalate tubes will stop enzymatic activity
at the glycolytic pathway.

12. Sodium & Lithium
For clinical chemistry are available with spray-
dried sodium hepari
nor lithium heparin additives. Heparin acts as
an anticoagulant as it
develops an antithrombin complex. This
complex inhibits thrombin
and the activated factor X and thus prevents
coagulatio

13. ESR
For ESR determination without the use of
sedimentation
pipettes. The blood is taken directly into tubes and
mixed
by inversion 8-10 times. Immediately before the tubes
are
placed in ESR stand for measurement; the tubes must
be
mixed again. After one or two hours the results are
read.
The measurement results achieved correspond to the
Westergren method

14. Venipuncture Procedure Steps:
1.Explain the procedure and purpose for the patient.
2.Assess the patient's physical disposition (i.e. diet,
exercise).
3.Position the patient - sitting or lying (NEVER allow the
patient to sit upright on a high stool or standing due to the
possibility of syncope).
4.Check the requisition form for requested tests, patient
information, and any special requirements.

15. Venipuncture Procedure Steps:
5. Select a suitable site for venipuncture.
6. Prepare the equipment, the patient and the puncture site.
7. Perform the venipuncture.
8. Collect the sample in the appropriate container.
9. While the tube fills, remove the tourniquet.
10. Label the collection tubes at the drawing area.
11. Immediately send the specimens with the requisition to the

16. Order of draw:
• Blood collection tubes must be drawn in a specific order to
avoid cross-contamination of additives between tubes.
• A Vacutainer blood collection tube is a sterile glass or plastic
tube with a closure that is evacuated to create a vacuum inside
the tube facilitating the draw of a predetermined volume of
liquid.
• The recommended order of draw for plastic vacutainer tubes is:
……..

17. Order of draw Stopper color Additives Laboratory Use
1 Plain Tube Red Top Tube no additive
2 EDTA Tube Lavender Top Tube EDTA*
3 Heparin Tube Green Top Tube Sodium Heparin
4 ESR Tube Black Top Tube Sodium citrate
5 Glucose Tube Grey Top Tube Sodium fluoride
6 PT Tube Blue Tube Sodium citrate
Drug levels,
blood bank
CBC*,HbA1c
Plasma chemistry
,ammonia level
Westergren
sedimentation rate
glucose tolerance
testing, alcohol level
PT*,PTT*
7 Serum Tube Yellow Top Tube
ACD* Whole blood
SPS* determination
8 Thrombin Tube Orange Top Tube Thrombin Chemical testing
9 Gel Yellow tube Serum Gel Tube
contains a clot
activator and serum
gel separator
various laboratory
tests
EDTA=Ethylene diamine tetraacetic acid, ACD= Acid citrate dextrose, CBC=Complete blood
count,PT=Prothrombin times, PTT=Partial thrombinplastin times, SPS=sodium polyanethol sulfonate

18. Notes:
• Additives preserve the specimen, prevent deterioration and
coagulation, or block action of certain enzymes in blood cells.
• After the tube has been filled with blood, immediately invert the
tube several times to prevent coagulation.
• Tubes with anticoagulants should be gently and completely
inverted (end over end) 4 to 6 times after collection. This process
ensures complete mixing of anticoagulants with the blood
sample and prevents clot formation.
• Complete filling of this tube is essential to obtain accurate
results.Some evacuate tubes contains mark line , you must fill
the tube into this mark line.

19. Specimens may be rejected for the
following reasons (Unacceptable
Specimens):
1. There is a patient name or file number discrepancy
between specimen label and request form.
2. There is no patient name or other unique identifier on
specimen.
3. Specimen is too old when received.
4. There is apparently no specimen in container.
5. The expiration date of the transport medium has been
exceeded.

20. Avoid performing a venipuncture on
1. Arm on side of mastectomy. If drawn here, the test results
could be inaccurate because of lymph edema.
2. Scarred or burned areas. Performing a venipuncture at these
sites is more difficult due to the scar tissue.
3. Arm in which blood is being transfusion / IV cannula. The
fluid in the IV could dilute the specimen.
3
2
1

21. 4.A hematoma (A hematoma is an abnormal collection of blood
outside of a blood vessel. It occurs because the wall of a blood
vessel wall, artery, vein, or capillary, has been damaged and blood
has leaked into tissues ) If drawn here, could cause incorrect test
results.
5.Edematous (Edema is swelling caused by fluid retention)
should be avoid because the accumulated fluid could alter test
results.

22. New vein finder devices :
• Devices that utilize laser/infrared technology for Vein Viewer (vein finders(.
Veinlite AccuVein VeinViewer Flex

23. New Device for Infant Blood Collection:
Innovac Quick-Draw

24. New Device for Infant Blood Collection:
•Automatic sample mixing reduces repeat collections
due to clots and hemolysis.
•Collect even the smallest drops from the puncture
site with no squeezing, scooping, or stopping to mix
the sample blood immediately.

25. Practical Part
•Trying to drawing of blood sample
from your colleagues.

26. Any
Questions ??!!!