4.
Haematoma (blood accumulation under the skin) –
Phlebotomy – CC2
Whole blood – Chemistry (HgA1C)
Serum – Chemistry (we are not just clotting the blood; it
is all trapping the impurities/ unnecessary elements in
the blood; creating the fibrin mesh)
INDICATIONS:
Diagnostic:
- Obtain blood samples for analysis.
- (e.g., systematic problems – Fe anemia, glucose
DM, INR, infections, cholesterol. Immunology,
liver enzymes/functions).
Therapeutic:
- Treat Polycythemia Vera (elevated RBC volume
aka hematocrit). Myeloproliferative (bone
marrow uncontrollable production of elements
of blood) disorders abnormal increases of RBC,
WBC, and platelets.
- Treat hemochromatosis (dangerously high iron
levels). One of the causes of liver damage; does
not manifest in the early stage of life.
- Donation for transfusion.
CONTRAINDICATIONS:
- Low oxygen levels in blood (hypoxemia).
RISKS:
1. Infection
- Negligible if sterile environment, proper
use/disposal of needles, and proper
management of samples.
2. Hitting a nerve or artery (arterial stab) – very close
proximity to the artery in our basilic vein (antecubital).
If you hit the artery, since it is oxygenated blood, it will
move your plunger.
- Remove the needle and apply pressure.
- Press the wound area with cotton stay for
around 1-2 minutes and put pressure to avoid
hematoma and swelling.
SIDE EFFECTS
1. Some pain, possible bruising.
2. Fainting and lightheaded (vaso-vagal) – Sit erect the
patients in case of fainting there is protection.
3. Excessive bleeding – Get cotton, tape, and put
pressure, check from time to time.
through and through (cross stitch) of the vein.
5. Iron deficiency anemia (in therapeutic phlebotomy) –
especially in children who looked pale.
ALTERNATIVES
1. No real alternative to phlebotomy, however, there are
various sites on the body that could be used.
- Last option is your foot.
- Primarily your antecubital area (antecubital,
central vein, cephalic and basilic)
2. See method
3. Never attempt more than twice:
• Refer patient back.
PROCEDURE – think action & rationale.
At every step know: WHAT are you doing? And WHY are
you doing it?
Patient identification -> Labelling of tubes
*****WASH HANDS*****
Equipment: Sterile Tray with:
• Pair of gloves
• Tourniquet
• Alcohol wipes
• Gauze
• VACUTAINER barrel and Needle
• Blood bottles (color coded according to additive
e.g., anticoagulant or preservative).
RULES OF ASEPSIS
STEP 2: CHECK THE PATIENT DETAILS
• Ask full name, DOB, Gender, and compare with
blood request form.
• Check blood form has been signed by the
requesting doctor.
• If special requirements, check patient has
complied, e.g., fasting.
• Have you had blood taken before? (Preferred
vein).
PUT GLOVES On.
• Ensure patient is in a relaxed position.
FIND A SUITABLE VEIN (Palpation: bouncy, large, and
superficial.
• 90% used – Anterior Cubital Fossa – Median
cubital vein, Cephalic, Basilic.
• Back of hand – Cephalic (housemans) vein. Artery – femoral 90 degrees; Radial 40 degrees.
• Feet, Central line, Peripheral Venous line,
Femoral stab (groin harder to disinfect).

• Amount drawn depends on indication (see

request form
• However, 5-25 ml is enough.

• Apply gauze to the puncture site for 1 minute,

with some pressure. But according to the
research cotton can cause platelet clump.
• Remove gloves and wash hands.

WASH HANDS
Vacuum Tubes
RED TOP
ADDITIVE: None (glass); Clot Activator (plastic)
MODE OF ACTION: Blood clots, and the serum is separated by
centrifugation.
USES:
1. Serum Chemistries
2. Immunology and Serology
3. Blood Bank (Crossmatch)
********Glass (no inversion) ********Plastic (5 inversions)

GOLD TOP
ADDITIVE: Gel and Clot Activator
MODE OF ACTION: The serum separator tube (SST) contains a gel
at the bottom to separate blood from serum during centrifugation.
USES:
1. Serum Chemistries
*********5 inversions**********

GREEN TOP
ADDITIVE: Sodium heparin or lithium heparin
MODE OF ACTION: Inactivates thrombin and thromboplastin.
USES:
1. Lithium level (use of sodium heparin).
2. Ammonia level (use of sodium/lithium heparin)
3. Plasma chemistries
**************8 inversions*************** • SOURCES:
- Di-Na+ salt of EDTA (powder)
LIGHT GREEN TOP (PST) - Versene or Versenate
ADDITIVE: Gel and Lithium heparin - Di-K+ salt of EDTA (liquid)
MODE OF ACTION: - Sequestrene
1. Anticougulates with lithium heparin.
2. Plasma is separated with Plasma Separator Tube gel at the SPECIAL CONSIDERATIONS
bottom of the tube. - Blood smear should be performed no more than 4 hours.
USES: - (cause red cell shrinkage)
1. Plasma chemistries
*************8 inversions ***************
LIGHT BLUE TOP
HEPARIN ADDITIVE: Buffered 3.2% (0.105M) Sodium Citrate (glass); 3.2%
• Acts as anti-thromboplastin and anti-thrombin. (0.109M) Na Citrate (plastic)
• Best anticoagulant of choice for minimal hemolysis. MODE OF ACTION: Forms calcium salts, in a non-ionized form of
• ALSO USED FOR: soluble complex.
• ESR, Hct, OFT, HDN cases. USES:
• Open heart surgery & transfusion 1. Coagulation tests (PT and PTT).
2. full draw required (3-4 inversions)
DISADVANTAGES
• Very expensive. CLEAR WITH LIGHT BLUE STOPPER
• Not ideal for smears (Cause changes in WBC and ADDITIVE: Citrate, Theophylline, Adenosine, Dipyridamole (CTAD)
platelets). USES:
• Can cause a bluish background. 1. Selected platelet function assays.
• Cannot be used for coagulation studies. 2. Routine coagulation determination.
**********3-4 inversions ******************
SPECIAL CONSIDERATIONS
• Blood films should be prepared immediately. YELLOW TOP
• ESR should be done within 2 hours. ADDITIVE: Sodium Polyanethol Sulfonate (SPS); ACD
• If the test be done in 2-3 hours, the specimen must be MODE OF ACTION: Complement inactivation; stabilize bacterial
refrigerated. growth.
USES:
• RBC swells within 6-24 hours at RT.
1. SPS: Blood culture (8 inversions)
2. ACD: HLA tissue typing, paternity testing, DNA studies (8
inversions)
ROYAL BLUE TOP
ADDITIVE: Disodium EDTA (plastic); Clot activator (plastic)
GRAY TOP
MODE OF ACTION: Designed to contain no contaminating metals.
ADDITIVE: Na+ fluoride; K oxalate
USES: Trace element testing; Toxicology
MODE OF ACTION: Antiglycolytic agent which preserves glucose
************ 8 inversions ***************
up to 5 days.
USES: Glucose, requires full draw (may cause hemolysis if short
LAVENDER TOP
draw).
ADDITIVE: Liquid Tri potassium EDTA (glass); Dipotassium EDTA
************8 inversions ****************
(plastic)
ADDITIVE: Na+ fluoride/ Disodium EDTA (8 inversions)
MODE OF ACTION: Chelates calcium.
ADDITIVE: Sodium fluoride (Serum tube) (8 inversions)
USES:
1. Hematology (CBC)
FLUORIDE
2. Blood Bank (crossmatch)
• Form a weakly dissociated Ca++ component.
3. Requires full draw – invert 8 times to prevent clotting and
platelet clumping. • With preservative action
- Interferes glycolysis.
EDTA
• Prevents clotting by chelation.
• Prevents calcium from ionizing.
ORANGE TOP
ADDITIVE: Thrombin-based clot activator with gel for serum
separation.
MODE OF ACTION: Activates fibrinogen to fibrin.
USES: STAT serum chemistries
********* 5-6 inversions *************

TAN TOP
ADDITIVE: Tripotassium EDTA (plastic)
USES: Lead determinations
***********8 inversions************

WHITE TOP
ADDITIVE: Dipotassium EDTA and gel for plasma separation.
USES: Molecular Diagnostic Test Methods (e.g., PCR)
*************** 8 inversions *************

PINK TOP
ADDITIVE: Spray-coated dipotassium EDTA (plastic)
USES: Whole blood hematology determinations;
immunohematology testings; cross-matching (AABB)
**********8 inversions**************

CLEAR TO WITH RED/LIGHT GRAY STOPPER

ADDITIVE: None (plastic)
USES: For use as a discard tube or secondary specimen tube.