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: dif icult to detect.
 Cephalic  The butter ly 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 lat 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 low
CLINICAL CHEMISTRY BSMDT 2A

 inger stick - the lateral side of the middle or ring Serum

inger 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 ibrinogen 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 ibrin clot incorporating the cells; this clot blood cells that can result in hemoglobin
consumes ibrinogen. release
CLINICAL CHEMISTRY BSMDT 2A

 Proper patient identi ication is the irst 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 irst 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 identi ication 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 arti icial or
 the specimen should be suf icient 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 dif iculty 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
CLINICAL CHEMISTRY BSMDT 2A

BLOOD COLLECTION: VENIPUNCTURE  For deep veins: 12 inch needle

 needle’s gauge is a numerical value that indicates
VENIPUNCTURE is performed for one of ive reasons: the diameter of its lumen.
 to obtain blood for diagnostic purposes.  the needle top is color coded to indicate the gauge
 to monitor blood components levels.  20g – yellow
 21g – green
 22g – black
VENIPUNCTURE METHODS:
 23g – blue
 syringe method
 when performing venipuncture, the needle is
 evacuated tube method
insterded into vein with the bevel facing upward.
Syringe method
Tube Stoppers
 open system used when small and fragile veins are
involved.  The different color of tube stoppers indicates the
type of additives contained within.
Evacuated tube method  The most frequently used additive is an
 comprises a double pointed needle anticoagulant which prevent from blood clotting.
(without anticoagulant the specimen will clot)
 plastic holder/adapter
 Some tubes also contain a polymer gel (gray/yellow)
 vacuum tubes
 polymer gel: will created physical barrier
between the liquid portion of the sample and the
BLOOD COLLECTION NEEDLES
cellular elements.
 ETS needle: both ends are pointed with one being
 The additives in the tubes are precisely proportioned
shorter than other, the long end is used into vein, the
to the amount of the blood that will till the tube.
short end is used to pierce the vacuum tubes
 It is critical completely ill each tube to ensure an
 needles come in variety of sizes: 1-12 inches
accurate ratio of blood to chemical additive.
 ROUTINE VENIPUNCTURE: 1 inch needle
CLINICAL CHEMISTRY BSMDT 2A

Otherwise, the test result may be inaccurate, or the Site selection and Preparation
specimen may be rejected.  Tourniquet: the most frequently used is a thin,
lexible strap applied the elbow to restrict blood low
ORDER OF DRAW and locate the vein
 the order of draw is determined by the additive  Thr tourniquet is secured in such a way that is easily
contained in the tube, not by the color of the stopper. removed by one hand.
 Sodium polyantholesulfunate: yellow  The tourniquet should not be left in place for more
 Sodium Citrate: blue/black than 1 minute as this can alter the blood's
 No additives: Red composition.
 No additives with gel: gold
 Lithium Heparin: green  Select the puncture site after then apply the
 EDTA: lavender tourniquet.
 Sodium Fluoride: gray  Venipuncture should be performed on the forearm's
large veins:
Patient Identi ication and Preparation  median cubital
 the phlebotomist must wash or sanitizer hands  cephalic
 always wear gloves.  basilic
 the patient has a right to know the procedure that  Palpate the vein with the tip of your index inger to
will be performed. determine its direction, depth, and size.
 Identify the patient. (The patient must state their  Select the veins that are large and easily accessible.
name.  Clean the puncture site with 10% isopropyl alcohol
 Adjust the patient position to ensure his/her (typically prepacked swabs.
comfort and safety in case patient becomes faint and  Circularly rub the alcohol swab away from the site.
falls.  Apply suf icient presswire to the puncture site to
remove all dirt.
 After cleansing the area, avoid touching it.
CLINICAL CHEMISTRY BSMDT 2A

 To anchor the vein, the dumb should be 1 to 2

inches below the puncture site. SPECIMEN LABELING
 Position the needle in the same direction as the vein  Patients full name
at a 15‒20-degree angle  Unique Identi ier
 The penetration of vein should be smooth motion to  Date and Time
decrease patient's discomfort. Slow insertion of  Phlebotomist Initial
needle is more painful in most patient.
 If needle is inserted to slowly, blood will leak out at Rejection of samples
the puncture site  Hemolysis: caused by a procedural error, such as
using a small bore or pulling back too hard on
 After collecting all tubes of blood and removing the plunger.
tourniquet, remove the inal tube holder, place a  Clotting: failure to mix or inadequate mixing of
cotton or gauze pad over the site, and withdraw the samples collected into an additive tube.
needle smoothly.  Quantity Not Suf icient: certain additive tubes
 Apply no pressure prior to removing the needle. must be illed up appropriately.
 Gently invert all tubes containing additives:  Wrong evacuated tube
 SPS/ Blood Culture: 8-10 x  Improper storage and handling: certain tests must
 Sodium citrate: 3-4 x be collected on ice: ammonia & ABC, and protected
 No additive: 0 from light: bilirubin
 Serum gel separator: 5 x  Improper labeling
 Lithium Heparin: 8-10 x
 EDTA: 8-10 x
 Sodium Fluoride: 8-10 x