LESSON 3: SPECIMEN PROCESSING

I. Learning Objectives
At the end of this chapter, the students should be able to:
A. Describe the different complications of venepuncture;
B. Enumerate the sites to be avoided in blood collection; and
C. Explain the procedures in specimen processing including chain of custody,
delivery time, and transport.

II. Concepts and Information from the PowerPoint

Complications of Venipuncture
A. Layers of Blood Vessels
Veins and Arteries:
 Tunica intima (endothelium)
 Tunica media- changes vessel diameter (vasodilation and vasoconstriction)
(smooth muscles and elastic tissue)
 Tunica externa (connective tissue covering)
Capillaries
 single layer of flattened endothelial cell
B. Late Local Complications
1. Hemoconcentration
- Is an increase in the number of formed elements in blood resulting from a
decrease or increase in plasma volume
Common causes:
- prolonged tourniquet application
- massaging or squeezing the site
- Long term IV therapy
Tests affected:
- Albumin, ammonia, Bilirubin, Calcium, Enzymes, Lipids, Potassium, Proteins,
Iron
2. Failure of blood to enter the syringe or vacutainer tube
- Incorrect bevel position (bevel down)
- Absence of vacuum
 - Not able to hit the vein:
Superficial puncture, rolling vein, lack of skill
- Went through the vein
- Collapsed vein, bevel resting on vein wall
3. Syncope (fainting)
- It is the transient loss of consciousness due to lack of oxygen in the brain and
results in the inability to stay in an upright position
- The needle should be removed immediately. Patient’s head should be lowered
between legs and patient should be instructed to breathe deeply.
C. Late Local Complications
1. Thrombosis
- Is an abnormal vascular condition in which thrombus develops within a blood
vessel of a body
2. Thrombophlebitis
- Inflammation of the vein accompanied by a clot which occurs as a result of
trauma at the vessel wall
3. Hematoma
-Occurs when there is leakage of blood from vessel into surrounding soft tissue.
Can be controlled with direct pressure.
Causes:
-Vein is fragile or too small for needle size
-The needle penetrates all the way through the vein
-The needle is partly inserted through the vein
-Needle is removed while tourniquet is still on
-Pressure is not adequately applied after puncture
D. Late General Complication
- Serum Hepatitis and AIDS. Acquired from contaminated needles and needle
stick.
 Sites to be Avoided in Blood Collection
A. Intravenous lines
- Use opposite arm.
- Stop IV for 2-5 minutes.
- If no other site available and IV cannot be stopped, a site below the IV line
should be sought. Initial 5 mL of blood drawn should be discarded.
- Increased glucose, chloride, potassium and sodium. Decrease in urea and
creatinine
B. Burned or scarred areas
- Select another site
C. Areas with hematoma
- Select another site
D. Thrombosed veins
- Select another site
E. Edematous arms/ sites
- Swelling caused by excess fluid trapped in your body's tissues
- Select another site
F. Partial or Radial mastectomy
- The risk is that the body’s ability to fight an infection in the affected limb is
compromised because of lymph node removal during mastectomy.
- Draw from opposite arm or select another site.
G. Cast/s on arm or leg
- Select another site

Anticoagulant: Action: Common Tests

Anticoagulant Action Common Tests

Citrate Combines with calcium in a non- Coagulation tests, Prothrombin time

ionized form (PT), Activated Partial
Thromboplastin Time (APTT)
Ethylenediamine Combines with calcium through CBC, Erythrocyte Sedimentation
tetraacetic acid chelation Rate, blood smears, HbA1C
(EDTA)

Fluoride Forms weakly dissociated Glucose, Lactate

calcium compounds; prevents
glycolysis. Glucose is stable for
24 to 48 hours

Heparin Inhibits or neutralization of Routine biochemistry, Ammonia,

thrombin Blood gas, Blood Alcohol

Specimen Processing: Delivery Time, Transport and Chain of Custody

A. Specimen Processing
- Laboratory scientist must ascertain if sample is acceptable for further
processing.
Criteria:
Volume considerations
Proper anticoagulants or preservatives
Proper handling, timing and transportation
- Unless whole blood analysis is being performed, the sample is then
centrifuged and the serum or plasma is separated from the cells.
- Once processed, laboratory scientist notes the presence of hemolysis, icterus
or lipemia in serum or plasma
- Samples should be analyzed within 4 hours. If >4 hours, refrigerate at 4°C for
up to 8 hours
B. Routine Handling
- Additive tubes must be mixed immediately after collection by inverting 3-8 times
depending on additive type.
- Gentle inversions to avoid hemolysis
- If tubes are not thoroughly mixed immediately, microclots can form in
anticoagulant tubes.
C. Transporting Specimen
- According to Clinical Laboratory Standards Institute (CLSI) and Occupational
Health and Safety Administration (OSHA) guidelines, blood specimen tubes must
be transported in a plastic bag with biohazard logo and a liquid-tight closure
- Blood specimen tubes must be transported carefully to prevent breakage and
protect specimen integrity
- Agitation and rough handling can cause hemolysis
D. Delivery Time Limits
STAT specimen- transported, processed and tested immediately
Routine blood specimen- delivered to laboratory within 45 minutes and
centrifuged within 1 hour of arrival
- The maximum time limit for serum or plasma separation according to CLSI
standards is 2 hours after collection
- Prompt separation is essential to minimize effects of metabolic processes
such as glycolysis. Aldosterone, calcitonin, enzymes and phosphorus are also
affected.
E. Chain of Custody
Chain of custody- a special documentation protocol required for specimens
collected for forensic or legal reasons
- Anyone participating in the COC of the specimen can be summoned to
participate in a related legal proceeding
- A special form accompanies the specimen from collection up to reporting. It
identifies and is signed by:
o Individual submitting to test
o Witness (if applicable)
o All who will handle the specimen (collection, processing, testing and
results)
- Date and time of receipt and condition of the specimen should be
documented
F. Processing Safety
- Large laboratories typically have a specific area (usually called central
processing) where specimens are received, evaluated for suitability, identified,
logged and sorted by department.
- All procedures involving blood must be performed in a manner that minimizes
splashing, spraying, splattering, and generation of droplets.
 - OSHA regulations require use of PPE (gloves, fully closed fluid-resistant lab
coats, protective face gear such as mask and goggles)
G. Centrifugation
- Unless the test is performed on whole blood, a blood specimen must be
centrifuged to separate the serum or plasma from the cells.
- Heparinized and other anticoagulated specimens can be centrifuged right away
- Specimens for tests that require serum must be completely clotted before
centrifugation. Incomplete clotting - latent fibrin formation that can interfere with
testing.
- After centrifugation, specimens should be visually checked. The presence of
hemolysis, icterus and lipemia or any abnormality, should be noted.
H. Aliquot Preparation
- Aliquot (portion of a specimen)- created when multiple tests are ordered on a
single specimen
- Aliquot tubes should be labeled with the same information as the specimen
tube.
- Serum and plasma or plasma obtained from different types of anticoagulants
must not be mixed together.
- Aliquot preparation has an inherent risk of error, and each specimen must be
carefully matched with the corresponding aliquot tube to avoid misidentified
samples.

III. Assignment

Make a comprehensive list of the process or rules/guidelines to be followed for

blood collection and blood sample acceptability and rejection in the clinical
chemistry section. (Give at least 10)

IV. Discussion/Generalization

Give a summary or generalization of what you have learned from Lesson 3:

Specimen Processing. Minimum of 5 sentences.

V. Quiz with Answers

Choose the best answer among the given choices.

1. This is swelling caused by excess fluid trapped in the body's tissues.

Thrombosis

Edema

Hematoma

Mastectomy

2. The following are causes of immediate local complications except:

loss of consciousness due to lack of oxygen in the brain

the vein moves or rolls out of the way when punctured

leakage of blood from vessel into surrounding soft tissue

absence of vacuum in blood collection tubes

3. This layer of blood vessel is made up of connective tissues.

Tunica media

Tunica intima

Tunica serosa

Tunica adventitia

4. This anticoagulant forms weakly dissociated calcium compounds and prevent

glycolysis.

Sodium Polyanethol Sulfonate

Sodium Heparin
 Ethylenediamine tetraacetic acid

Sodium Fluoride

5. Routine blood specimen for whole blood analysis should be centrifuged within.

2 hours

3 hours

not centrifuged

1 hour

6. This is the area usually found in large laboratories where specimens are
received, evaluated, identified and sorted.

Central processing

Main processing

Capital processing

Central operations

7. The maximum time limit for serum and plasma separation according to CLSI
standards is

5 hours

4 hours

2 hours

3 hours

8. Collecting blood from intravenous lines may cause these changes on analytes
of the specimen except:
 decreased sodium

decreased creatinine

increased glucose

increased potassium

9. This is a special documentation protocol required for specimens collected for

forensic or legal reasons.

Chain of Command

Chain of Control

Chain of Custody

Chain of Care

10. This anticoagulant combines with calcium through chelation.

Sodium fluoride

Ethylenediamine tetraacetic acid

Sodium citrate

Sodium heparin

11. All of these are late local complications in blood collection except:

increased number of formed elements in blood

leakage of blood from vessel into surrounding tissue

vascular condition in which thrombus develops within the blood vessel

 inflammation of the vein accompanied by a clot

12. A phlebotomist should not collect blood from the arm on the side of a
mastectomy because

it is painful for the patient

the affected limb's ability to fight infection is compromised

the blood collected will be contaminated with excess fluid trapped in body's
tissues

finding a suitable vein is harder

13. In mixing additive tubes you must gently invert the specimen to avoid

icterus

hemolysis

lipemia

contamination

14. This is created when multiple tests are ordered on a single specimen.

Fraction

Specimen

Aliquot

Sample

15. If samples cannot be processed or analyzed within 4 hours it should be

refrigerated at

4 degrees Celsius for up to 8 hours

 4 degrees Celsius for up to 10 hours

4 degrees Celsius for up to 12 hours

2 degrees Celsius for up to 8 hours

VI. References

✘ Bishop, M. L., Fody, E. P., & Schoeff, L. E. (2013). Clinical chemistry: principles,
techniques, and correlations. 7th ed. Philadelphia: Wolters Kluwer
Health/Hippincott Williams & Wilkins

✘ McPherson, R. A., Pincus, M. R., & Henry, J. B. (2007). Henry's clinical

diagnosis and management by laboratory methods. Philadelphia: Saunders
Elsevier

✘ Rodriguez, M.T. (2019). Clinical Chemistry Reviewer Handbook