CLINICAL CHEMISTRY BSMDT 2A

Specimen Collection and Handling

The process of specimen collection, handling, and ❖ dilation of the veins

processing remains one of the primary areas of ❖ making for easier detection.
preanalytic errors.
The gauge of the needle
Types of Samples • Is inversely related to the size of the needle
• Phlebotomy, or venipuncture, is the act of obtaining • The larger the number, the smaller the needle bore
a blood sample from a vein using a needle attached and length.
to a collection device or a stoppered evacuated tube.
Routine venipuncture
Different volume sides of collection tubes: • uses a 23- or 21-gauge needle.
❖ 10 ml is the larger
❖ 150 ul is for pedriatric Winged infusion set
• May be used whenever the veins are fragile, small, or
Site of the vein: difficult to detect.
• Cephalic • The butterfly needle is attached to a piece of tubing,
• Basilic which is then attached to a hub or barrel.
• Medial antecubital vein of the arm (the most
frequent site for venipuncture) Capillary Puncture
• venipuncture, blood samples can be collected using
Tourniquet a capillary puncture technique that involves:
❖ made of pliable nonlatex rubber flat band or tubing ❖ heel stick - the outer area of the bottom of the
is wrapped around the arm, causing: foot for infants
❖ cessation of blood flow
CLINICAL CHEMISTRY BSMDT 2A

❖ finger stick - the lateral side of the middle or ring Serum

finger for individuals 1 year and older • The remaining liquid is called serum rather than
• A sharp lancet device is used to pierce the skin plasma
• An appropriate capillary or microtainer tube is • Most testing in the clinical chemistry laboratory is
used for sample collection. performed on either plasma or serum.
• The major difference between plasma and serum is
Analytic testing of blood that serum does not contain fibrinogen and some
• involves the use of whole blood, serum, or plasma. potassium is released from platelets (i.e., potassium
levels are slightly higher in serum than in plasma).
Whole blood - Plasma • It is important that serum samples be allowed to
• as the name implies, contains the liquid portion of completely clot (≈30 minutes) in an upright position
the blood, called plasma, and its cellular at room temperature before being centrifuged.
components (red blood cells, white blood cells, and
platelets). Centrifugation
• The collection of whole blood requires the vacuum • Centrifugation of the sample accelerates the
tube to contain an anticoagulant. process of separating the liquid portion and
• Complete mixing of the blood immediately following cellular portion.
venipuncture is necessary to ensure the • Specimens should be centrifuged according to
anticoagulant adequately inhibits the specimen from recommendations by the tube manufacturer or
clotting. test protocol
• As the tube of whole blood settles, the cells fall • Usually approximately 10 minutes at an RCF of
toward the bottom, leaving a clear yellow 1000 to 2000 g
supernatant on top, which is the plasma. • but should avoid having hemolysis
• If a tube does not contain an anticoagulant, the blood ❖ Hemolysis: the mechanical destruction of red
forms a fibrin clot incorporating the cells; this clot blood cells that can result in hemoglobin
consumes fibrinogen. release
CLINICAL CHEMISTRY BSMDT 2A

• Proper patient identification is the first step in • Use of proper anticoagulants or preservatives
sample collection. ❖ The specimen should be collected in the correct
evacuated tube
Steps that cannot be stressed strongly enough: • The timing is clearly indicated and appropriate for
• using the proper collection tube timed testing
• avoiding prolonged tourniquet application • Thr specimen is intact and has been properly
• drawing tubes in the proper order transported
• proper labeling of tubes ❖ Specimen sould be on ice, within a reasonable
period and protected from light
Antiseptics must be used when drawing blood: • Once the sample is processed, the laboratorian
❖ Isopropyl alcohol wipes - used for cleaning and should note the presence of any serum or plasma
disinfecting the collection site; however, characteristics such as:
❖ Chlorhexidine - used as the disinfectant in such ❖ hemolysis and icterus (increased bilirubin
cases. pigment)
❖ presence of turbidity often associated with
Sample Processing lipemia (increased lipids)
• The first processed, in the clinical chemistry
laboratory is correctly matching the blood collection • Many analytes are stable at room temperature
tube(s) with the appropriate test requisition and between 24 to 72 hours.
patient identification labels. • If testing is not to be performed within 8 hours, it is
• The laboratorian must also ascertain if the sample is recommended that serum and/or plasma be
acceptable for further processing. refrigerated between 2°C and 8°C.
• The criteria used depends on the test involved but • It is important to avoid exposing samples that are
usually include volume considerations light sensitive, such as bilirubin, to artificial or
❖ the specimen should be sufficient volume for ultraviolet light for extended periods of time.
testing needs
CLINICAL CHEMISTRY BSMDT 2A

• Separated serum and/or plasma may be frozen at Lipemia

−20°C and stored for longer periods without • Lipemia results when the lipid levels of the patient
deleterious effects on the results. are elevated and
• Visualized as a creamy or milky appearance to the
Hemolysis serum or plasma upon centrifugation.
• Hemolysis can be visually observed in a centrifuged • Lipemia can cause a volume displacement for some
patient sample as a red color due to the release of analytes, such as electrolytes, as well as interference
hemoglobin. in light- scattering methodologies due to the
• There are patient conditions where this may occur in turbidity present.
vitro such as: hemolytic anemia
• Also, hemolysis can be present due to preanalytic Icterus
collection variables such as: • Icterus is a deep yellow or golden appearance of the
❖ inappropriate needle gauge serum or plasma due to increased bilirubin levels,
❖ venipuncture site selection (small veins) and may cause spectral interference on certain
❖ venous trauma or difficulty in specimen analyzers in the chemistry lab.
collection.
Variables Affecting Select Chemistry Analytes
• Along with the release of hemoglobin, other
intracellular components may be released which Physiological factors
may impact patient values for these analytes, such • Samples requiring fasting: Glucose
as: • Analyte changes based on diurnal variation: Cortisol
❖ Potassium
❖ Phosphate Patient preparation factors
❖ Lactate dehydrogenase • Fasting: 8-12 hours fasting
CLINICAL CHEMISTRY BSMDT 2A

Collection and sample processing factors

• Venipuncture technique:
❖ Needle selection
❖ Site selection to decrease opportunity of
hemolysis
• Tube selection:
❖ appropriate sample tube
❖ inversion following collection
❖ appropriate clotting time
• Tourniquet use: prolonged use affects analytes
• Specimen transport:
❖ Protection from light = bilirubin
❖ Collect and store on ice = ABG and Ammonia