SAMPLE COLLECTION
Dr. SYEDA AFSHAN (MBBS , MD)
(SR)
Dept of Biochemistry.
Historically, evidence of liquid biological
specimens collected on paper appears as far back
as 780 AD.
Their documented use for analytical processes,
however, dates only to the early 1900s.
In 1947, Joseph Kleiner invented the Evacutainer,
a glass vacuum tube for drawing blood.
Types of Blood specimens
BLOOD SAMPLE COLLECTION
- Specimens must be obtained using-
-proper phlebotomy techniques,
-collected in the proper container,
-correctly labeled
-promptly transported to the laboratory.
-It is the policy of the laboratory to reject samples when
there is failure to follow these guidelines.
Sample collection is done from:
⚫ Capillaries
⚫ veins
⚫ arteries
Capillary blood:
The capillaries are the very fine blood vessels between arteries and
veins to allow the blood deliveries of oxygen to the tissues. Blood
tests from fingers contain capillary blood.
Higher Potassium than Venous blood(due to tissue debris
contamination)
Venous blood:
Venous blood is deoxygenated blood which
travels from the peripheral vessels, through the
venous system into the right atrium.
Deoxygenated blood is then pumped by the right
ventricle to lungs via the pulmonary
artery which is divided in two branches, left and
right to the left and right lungs respectively.
Blood is oxygenated in lungs and returns to the left
atrium through pulmonary veins
Has 15-20 mg/dl less glucose than capillary
blood.
Arterial blood:
⚫ It is the oxygenated blood in the circulatory system found in the
lungs, the left chambers of the heart, and in the arteries. It is
bright red in color.
Universal precautions
⚫ Assume that all human blood is potentially infectious for
HIV, hepatitis, and other infectious agents.
⚫ Practice Universal Precautions using gloves, eye protection,
and lab coats.
⚫ Always use sterile, single-use, disposable supplies for sample
collection.
Sample collection Toolkit
⚫ It Includes-
⚫ Needle Disposable Container
⚫ cotton Gauge
⚫ 70%(v/v) Iso-propyl alcohol ⚫ Gloves
⚫ Syringes-19/20/21 G ⚫ Couch
⚫ Needles ⚫ Bathroom
⚫ Phlebotomy Chairs ⚫ Biohazard Kit
⚫ Tourniquet ⚫ Place For Privacy
⚫ Spill Kit ⚫ Refrigerator
⚫ Accident And Incident Kit
⚫ 1% Hypochlorite Solution
Syringes Needles
VACUTAINER NEEDLE CUTTER CYCLOMIXER
PRESERVATIVES FOR BLOOD
HEPARIN EDTA ACD
•Commonly used anticoagulant •Chelating agent, binding •Samples for
• Heteropolysaccharide divalent cations like Ca+2 & molecular
• Mucoitin polysulfuric acid Mg+2 diagnosis
Disadvantages •Preserves the cellular •Preserves
• high cost components of blood – form and
• Inhibit acid phosphatase activity haematology. function of
• Affects binding of calcium to EDTA cellular
•Disodium / dipotassium/ organelles
• Affects T3, T4 binding to tripotassium salt
their carrier proteins.
ANTI-COAGULANTS
SODIUM FLUORIDE SODIUM CITRATE SODIUM, POTASSIUM,
AMMONIUM, LITHIUM
OXALATES
• Weak anticoagulant •34-38g/L, 1 part to 9 parts of •Insoluble complexes with Ca+2
•Preservative for Glucose in blood ions
blood. • Chelates calcium •Potassium Oxalate 1-2g/L is
•Unsuitable for measurement of most widely used
Inhibits Ca
glycolysis enzymes
urease & interferes with other
tests.
Venipuncture routine procedure
● Review and accession test
● Approach, identify, and ● Clean and air dry site
prepare patient ● Anchor vein by making the
● Verify diet restrictions and skin taught using thumb and
latex sensitivity insert needle at an angle of
● Sanitize hands 30-40 degrees.
● Position patient, Examine both
arms to find the best vein.
Locate the puncture site; apply
the tourniquet, and ask the
patient to make a fist.
● Establish blood flow, release tourniquet and ask patient
to open fist.
● Fill, remove, and mix tubes order of draw.
● Place gauze, withdraw needle, activate safety feature, and
apply pressure.
● Discard needle and holder unit.
● Label tubes.
● Check patient’s arm and apply bandage.
● Dispose of used materials
● Thank patient, remove gloves, and sanitize hands
● Transport specimens to lab.
Time of collection
• For fasting samples
1) a minimum of 8-10 hrs fasting is required for glucose
2)10-12 hrs fasting for lipid profile(no exercise,no
smoking,no alcohol consumption,no fatty meal).
• The post prandial sample should be collected exactly after 2
hrs from the beginning of meal.
• It is important to mention time to hormonal assays (to know
the circadian variation in hormonal levels)
GTT
ORAL GTT - FASTING AND 2HR
Gestational GTT - FASTING , 1HR , 2HR
75 gm of GLUCOSE TO be administered
after the withdrawal of fasting sample
Few important Variations
1. Circadian variations are seen with Cortisol (7 am),
potassium is more in afternoon.
2. GTT performed in morning is more accurate than
Afternoon sampling due to influence of insulin levels
and other metabolic changes
3. Cyclical variations (menstrual) - seen with
Hormones (estrogens, progesterone ,LH, FSH, etc) &
cholesterol decreases during ovulation.
PHYSIOLOGICAL VARIABLES DURING
BLOOD COLLECTION
□ Posture
□ Prolonged bed rest
□ Exercise
□ Physical training
□ Circadian variation
□ Menstrual cycle
PRE-ANALYTICAL ERRORS:
● Incorrect pt. Preparation
● Improper collection tube
● Improper specimen
● Insufficient mixing of sample
● Inadequate Clotting
● Improper centrifugation
● Improper Transportation
● Incorrect labelling
HEMOLYSIS
CAUSES: Too small a needle, Shaking Vigorously, Presence of
excess anticoagulant,Centrifuging at high speeds before clotting of
blood has occurred,Freezing/Thawing of blood, Unclean tubes or
presence of water in tubes.
TESTS AFFECTED:
● [Link]
● [Link]
● SGOT
● [Link]
● [Link] Phosphatase
REJECTION CRITERIA
• QUANTITY INSUFFICIENT
• CLOTTED SAMPLE
• HEMOLYSED SAMPLE
• INCORRECT ORDER OF DRAW-Intermixing
• INCORRECT SAMPLE DELIVERY([Link] ice packs for
ABG sample transport)
• IMPROPER LABELLING
Preanalytic considerations
⚫ Phlebotomist be able to recognize problem sites,
⚫ identify procedural error risks,
⚫ address certain patient conditions([Link]/PAD)
⚫ Handle patient complications.
⚫ Problem sites:
⚫ Burns, scars, and tattoos
⚫ Damaged veins
⚫ Edema
⚫ Hematoma
⚫ mastectomy
Patient conditions and complications
⚫ Allergies to supplies or equipment
⚫ Excessive bleeding
⚫ Fainting
⚫ Nausea or vomiting
⚫ Obese patients
⚫ Pain
⚫ Petechiae
⚫ seizures/convulsions
Arterial blood sampling
⚫ Sample is collected from an artery, primarily to determine arterial blood gases.
⚫ should only be performed by health workers who have demonstrated proficiency after
formal training.
⚫ The sample can be obtained either through a catheter placed in an artery, or by using a
needle and syringe to puncture an artery.
Choice of site
The first choice is the radial artery
brachial artery
femoral artery
o may be harder to locate, because they are less superficial than the radial artery
o have poor collateral circulation;
o are surrounded by structures that could be damaged by faulty technique.
a container with crushed ice for transportation of the sample to the
laboratory (if the analysis is not done at the point of care) where
applicable, local anesthetic and an additional single-use sterile syringe
and needle.
Holding the syringe and needle like a dart, use the index finger to
locate the pulse again, inform the patient that the skin is about to be
pierced then insert the needle at a 45 degree angle, approximately 1 cm
distal to (i.e. away from) the index finger, to avoid contaminating the
area where the needle enters the skin.
COMPLICATIONS
• Arteriospasm
• Hematoma
• Nerve damage
• Fainting or a vasovagal response
•Other problems - drop in blood pressure, complaints of feeling faint, sweating
or pallor that may precede a loss of consciousness.
Why this is important?
CO2 present in blood has higher
concentration than in atmosphere , so
there is passive diffusion if not transported
in ICE box , which may lead to deranged
Blood Gas Values.
