Basic Introduction to Phlebotomy, the equipment, and anticoagulants required. To buy the whole presentation other books and contents, please message/email me. danandrewcruz.hva@heal.com

1. Phlebotomy- Blood collection,
and Anticoagulants
Dan Andrew Cruz,
RMT, PBMcT, DTA HIPAA Certified,

2. The study of the subject matter is important due to the bearing of the number of questions in the board exam

4. Basics in Blood Collection
● Primary duty of is to collect blood specimens for lab testing.
● Blood collection include arterial puncture, capillary puncture, and venipuncture
● Phlebotomist must be familiar with various types of equipment for selecting
appropriate collection devices.

5. Basics in Blood Collection: Equipment
● Tourniquet - a device used to restrict blood flow in a patient's arm to make it
easier to draw blood or perform other medical procedures.
○ Apply the tourniquet tightly enough to restrict venous flow but not so tightly
that it restricts arterial flow.
○ Tourniquets should not be left in place for more than a minute, as this can
change the composition of the blood.
○ They are made of stretchable materials like latex, nitrile, or vinyl and are
usually disposable.

6. Notes:
● Distance from Puncture site - 3-4 inches above or 7.5 to 10cm above
○ If closer to the site, the vein may collapse as blood is above the intended venipuncture
site. withdrawn. If too far above the site, it may be ineffective.
● But not tight enough to restrict arterial blood flow
● Should be applied < 1minute - Causes hemoconcentration
● Blood Pressure Cuff (Alternative) Maintain at patient’s Diastolic pressure OR
<40mmHg
● Potential to transmit MRSA

7. Antiseptics - used to prevent sepsis, which is the presence of microorganisms or
their toxic products within the bloodstream.
● Antiseptics prevent or inhibit the growth and development of microorganisms
but do not kill them.
● 70% isopropyl alcohol (isopropanol) is the most common
● since many patients are allergic to povidone–iodine.
● Benzalkonium chloride - 1 step antiseptic
Adhesives or Bandages

8. Gloves
● A new pair should be used per patient and should be a good fit for the hands
● Non-sterile, disposable, non-powdered (skin irritation)
● Types: Latex, Nitrile, Neoprene, Polyethylene and Vinyl

9. Sharps and needles disposal containers: Oftentimes colored red for easy
identification.
Biohazard bags: leak proof plastic bags that are commonly used to transport
blood and other specimens from the collection site to the laboratory.

10. Disinfectants - that are used to remove or kill microorganisms on surfaces and
instruments, typically corrosive and are not safe to use on human skin
● Bleach A 1:100 dilution is recommended for decontaminating nonporous
surfaces after cleaning up blood or other body fluid spills.
● When spills involve large amounts of blood or other body fluids or occur in the
laboratory, a 1:10 dilution.

11. Basics in Blood Collection: Equipment
Phlebotomy Needles: sterile, disposable, and designed for a single use only.
● They include multisample needles, hypodermic needles, and winged infusion
(butterfly) needles used with both the evacuated tube system and the syringe
system.
● Multisample needles are commonly enclosed in sealed twist off shields or
covers. Hypodermic needles and butterfly needles are typically sealed in sterile
pull-apart packages.

13. Needle Gauge: a measure of the thickness of a needle, with higher numbers
indicating a smaller diameter.
● Size of is chosen based on the size and condition of the patient's veins, type of
procedure being, and equipment being used.
● Most blood collection, needles gauges ranging from 20 to 23 are appropriate, a
21-gauge needle being the standard for most routine adult venipunctures.
● The speed of blood flow through a needle is generally faster through larger-
diameter needles.
Note: Using small gauge needles may cause blood cell rupture.

15. Basics in Blood Collection

16. Evacuated Tube System (ETS): most common and efficient method for collecting
blood samples
● Preferred by the Clinical and Laboratory Standards Institute (CLSI).
● A closed system that allows multiple tubes to be collected with a single
venipuncture,
● preventing the blood from being exposed to the air or contaminants.
● The ETS consists of a special blood-drawing needle, a tube holder, and various
types of evacuated tubes.

17. Multisample needles (ETS needles): are used in the evacuated tube system for
collecting multiple tubes of blood during a single venipuncture.
● They have a threaded portion in the middle and beveled points on each end.
The end that pierces the vein is longer and has a longer bevel. The shorter end
is used to penetrate the tube stopper.
● The needle is covered by a retractable sleeve that prevents leakage of blood
when the tube is removed.

18. Traditional ETS Needle
ETS Needle With Hub

19. Tube holder: a clear, disposable plastic cylinder used in the evacuated tube system
for collecting blood.
● It has a small threaded opening at one end (called the hub) where the needle is
screwed in, and a large opening at the other end where the collection tube is
placed.
● The large end has flanges or extensions on the sides to help with placement
and removal of the tube.

20. Evacuated Tubes: used to collect blood samples in the evacuated tube system
(ETS) and the syringe method.
● They are available from various manufacturers in different sizes and volumes,
ranging from 1.8 to 15 mL.
● The appropriate size of tube is chosen based on the age of the patient, the
amount of blood needed for the test, and the size and condition of the
patient's vein.
● Both plastic and glass tubes are available, with many laboratories choosing
plastic for safety reasons.

21. ● Evacuated tubes fill with blood automatically because there is a vacuum
(negative pressure) in them.
● The tubes are designed to draw a precise volume of blood, as indicated on the
label, by creating a vacuum that removes a specific amount of air and creates
negative pressure.
● The vacuum in the tube must be maintained until the desired volume of blood is
collected,

22. ● Improper storage, opening the tube, dropping the tube, advancing the tube too
far onto the needle before venipuncture, or if the needle bevel becomes
partially out of the skin during venipuncture decreases vacuum.
● If the vacuum is lost prematurely or the tube is removed before the vacuum is
exhausted, or if blood flow is stopped during the draw, the result is an
underfilled tube called a partial draw or "short draw."
● Test results may be compromised in partially filled tubes that contain additives
because the ratio of blood to additive has been altered.

24. Additive tubes are those that have substances other than the tube stopper placed
within them.
● Additives serve specific purposes,
○ preventing clotting or
○ preserving certain components of the blood.
● The presence or absence of additives in the tube will determine whether the
collected blood will clot or not.
● If the additive prevents clotting, the resulting specimen is called a whole-blood
specimen, which can be used for testing as is, or centrifuged to separate the
cells from the fluid portion called plasma for testing.
● If the additive causes the blood to clot, the specimen must be centrifuged to
obtain the fluid portion called serum.

25. Nonadditive tubes - which do not contain any substances to prevent clotting, are
rare and are usually only used for discarding or clearing purposes.
● A few glass nonadditive red-top tubes are still available, but most of these are
being phased out due to safety concerns.
● Nonadditive tubes produce serum samples because the blood collected in them
will naturally clot when there is nothing to inhibit the process.
Most tubes used for collecting blood samples contain additives to prevent the blood
from clotting.

27. ● Citrated tubes with different
capacity of blood.
● Markers for the min and to the
maximum allowable limit of blood
to anticoagulant ratio to be
achieved.
● Ideal: 9:1 (blood to anticoagulant)

28. Serum - comes
from coagulated
blood.
Plasma - comes
from
uncoagulated
blood

29. ETS Tubes with volume markings
ETS Tubes With Different
colored stopper

32. Winged infusion set - is a medical device used for collecting blood or administering
fluids.
● Consists of a small, flexible tube with a needle at one end and a connector for a
syringe or evacuated tube system at the other end.
● It is often used on small or hard-to-reach veins, such as those found in the
hands or in elderly or pediatric patients.
● It is generally easier to use and more precise than a needle and syringe alone.
● Multisample Luer adapters can also be used with winged infusion sets.

34. Syringe - a medical device used to inject liquids into or withdraw fluids from the
body.
● It consists of a barrel with graduated markings, a plunger that fits snugly inside
the barrel, and a needle attached to the barrel.
● The barrel is marked with graduations indicating the volume of fluid in the
syringe. The plunger is used to draw fluids into the syringe or expel them from
the barrel.
● Used for patients with small or difficult veins.

35. Note:
● Blood collected in a syringe must be transferred into ETS tubes.
● Blood transfer by poking the syringe needle through the tube stopper or by
removing the tube stopper and ejecting blood from the syringe into the tube is
considered unsafe.
● Use a syringe transfer device allows the safe transfer of blood into the tubes
without using the syringe needle or removing the tube stopper.
○ The device is similar to an ETS tube holder but has a permanently attached needle
inside.

36. Note:
● Transfer devices reduces any chance for
accidental needlesticks pricks, confines any
aerosol or spraying of the specimen to the
phlebotomist
● Removing the stopper/ cap can generate
specimen aerosolization.

37. ADVANTAGES OF EVACUATED TUBE
METHOD
● Large amount of blood can be obtained
● Blood can be transported and stored
for future use
● Additional and repeated tests can be
done
● Fastest method of collecting sample
which requires various anticoagulation
requirements
● Blood collected is ideal for chemistry
and other serological tests
DISADVANTAGES OF EVACUATED TUBE
METHOD
● Requires more time and skill on the
part of the phlebotomist
● Requires more equipment
● More complications may arise
● Difficult to do on infants, children, and
obese individuals

38. Basics in Blood Collection: Anticoagulants
Anticoagulants: substances that prevent blood from clotting, their mechanisms and uses are
as ff:
● Binds or precipitate calcium, or inhibit the production of thrombin.
● Used to ensure that the blood sample remains in a liquid form for testing.
● Often added to tubes used for collecting blood samples to prevent clotting, particularly if
the test requires whole blood or plasma.
Note: Important to mix anticoagulant-treated blood immediately after collection to prevent the
formation of small clots, and to mix gently to avoid breaking red blood cells.
Note: Never shake or otherwise vigorously mix a specimen aka Jostling, as this can cause
hemolysis, which makes most specimens unsuitable for testing.

39. Parameters Affected by Hemolysis
Increased:
● “KLA6MP ITC2”
● K+
● LDH (150x)
● ACP
● ALP
● Aldolase
● ALT
● AST
● Albumin
● Mg2+
● Phosphorus
● Iron
● Total protein
● Affects bilirubin levels
● ESR
Decreased:
● Creatinine,
● Alkaline phosphatase (ALP),
● Potassium,
● Lipase (due to inhibition)
● packed cell volume (PCV),
● Mean corpuscular volume (MCV).
● Cell count
● Hematocrit (HCT),
● Activated PTT
Legend:
Hematology
Clinical Chemistry

40. ● There are various types of anticoagulants
that are used for different tests.
○ Some of the most commonly used
anticoagulants include
○ EDTA,
○ citrates,
○ heparin,
○ oxalates.
● Using the correct anticoagulant is
important to ensure the accuracy and
reliability of the test results.