HEMATOLOGY II

Laboratory Evaluation of Hemostasis

LABORATORY
SPECIMEN COLLECTION
EVALUATION
OF SPECIMEN MANAGEMENT

HEMOSTASIS
PRIMARY HEMOSTASIS

SECONDARY HEMOSTASIS
 A. HEMOSTASIS
• involves the interaction of vasoconstriction,
platelet adhesion and aggregation, and
coagulation enzyme activation to stop
bleeding.
• the key cellular elements of hemostasis are
the cells of the vascular intima,
extravascular tissue factor (TF)–bearing
cells, and platelets.

• Overview
 A. Purpose
1. Provide a blood component to a patient
that will provide maximum benefit while
minimizing potential for harm.
2. This requires a systematic process that is
fairly rigid
3. Regulated in the US by:
a) CLIA (Clinical Laboratory Improvement
Amendments, 1988)
b) AABB (formerly “American Association of
Blood Banks”)
c) College of American Pathologists (CAP)
d) FDA

• Overview
 • Most hemostasis laboratory procedures are
performed on venous whole blood
collected by venipuncture and mixed 9:1
with a 3.2% solution of sodium citrate
anticoagulant. The specimen is maintained
as well-mixed whole blood for platelet
function testing or centrifuged to provide
platelet-poor plasma (PPP) for other
procedures.

• SPECIMEN COLLECTION
 • Patients need not fast, but they should
avoid vigorous activities and should rest
quietly for 30 minutes prior to collection
for hemostasis testing.
• There are numerous drugs that affect the
outcomes of coagulation tests.
• aspirin suppresses most platelet
function, and Coumadin (warfarin)
reduces the activities of factor II
(prothrombin), factor VII, factor IX,
factor X, protein C, protein S, and
protein Z and prolongs the
• SPECIMEN COLLECTION prothrombin time (PT) test.
 • Hemostasis Specimen Collection Tubes
• Most hemostasis specimens are collected
in plastic blue-stopper (blue-top, blue-
closure) sterile evacuated blood collection
tubes containing a measured volume of
0.105 to 0.109 M (3.2%) buffered sodium
citrate anticoagulant.

• SPECIMEN COLLECTION
 • Hemostasis Specimen Collection
Protocol
• Laboratory directors typically prefer
evacuated blood collection tube systems
for hemostasis blood collections; however,
many directors may require syringe
collection and initial “discard tubes” in
special circumstances.

• SPECIMEN COLLECTION
 • Hemostasis Specimen Collection
Protocol
• Laboratory directors typically prefer
evacuated blood collection tube systems
for hemostasis blood collections; however,
many directors may require syringe
collection and initial “discard tubes” in
special circumstances.

• SPECIMEN COLLECTION
 • Hemostasis Specimen Collection
Protocol
• If the hemostasis specimen is part of a
series of tubes to be filled from a single
venipuncture site, it must be collected first
or immediately after a nonadditive tube.
• The ratio of whole blood to anticoagulant
must be 9 parts blood to 1 part
anticoagulant.
• When specimens are collected using
winged-needle butterfly sets, the
phlebotomist must compensate for the
• SPECIMEN COLLECTION internal volume of the tubing, which is
usually 12 inches long and contains
approximately 0.5 mL of air.
 • Hemostasis Specimen Collection
Protocol
• Clotted specimens are useless for
hemostasis testing, even if the clot is
small.
• Excessive specimen agitation causes
hemolysis (RBC rupture),procoagulant
activation, and platelet activation.
• Excess needle manipulation may promote
the release of procoagulant substances
from the skin and connective tissue, which
contaminate the specimen and cause
• SPECIMEN COLLECTION clotting factor activation.
• During blood collection, the phlebotomist
must remove the tourniquet within 1
minute of its application to avoid blood
stasis
 • Hemostasis Specimen Collection
Protocol
• Clotted specimens are useless for
hemostasis testing, even if the clot is
small.
• Excessive specimen agitation causes
hemolysis (RBC rupture),procoagulant
activation, and platelet activation.
• Excess needle manipulation may promote
the release of procoagulant substances
from the skin and connective tissue, which
contaminate the specimen and cause
• SPECIMEN COLLECTION clotting factor activation.
• During blood collection, the phlebotomist
must remove the tourniquet within 1
minute of its application to avoid blood
stasis
 • Hemostasis Specimen Collection
Protocol
• Selection of Needles for Hemostasis Specimens
• Whether evacuated collection tubes or
syringes are used, the bore of the needle
should be sufficient to prevent hemolysis
and activation of platelets and plasma
procoagulants.

• SPECIMEN COLLECTION
 • Hemostasis Specimen Collection
Protocol
• Selection of Needles for Hemostasis Specimens
• Whether evacuated collection tubes or
syringes are used, the bore of the needle
should be sufficient to prevent hemolysis
and activation of platelets and plasma
procoagulants.

• SPECIMEN COLLECTION
 • Hemostasis Specimen Collection
Protocol
• Specimen Collection Using Capillary
Puncture
• Several near-patient testing (point-of-
care) coagulometers generate PT
results from a specimen consisting of
10 to 50 mL of whole blood.
• Anticoagulants Used for Hemostasis
Specimens
• Sodium Citrate (Primary Hemostasis
Anticoagulant)
• The anticoagulant used for hemostasis
• SPECIMEN COLLECTION testing is buffered 3.2% (0.105 to 0.109
M) sodium citrate, molecular weight 294.1
Daltons. Sodium citrate binds calcium
ions to prevent coagulation, and the buffer
stabilizes specimen pH as long as the tube
 • Hemostasis Specimen Collection
Protocol
• Adjustment of Sodium Citrate Volume for
Elevated Hematocrits
• The 9:1 blood-to-anticoagulant ratio is
effective, provided the patient’s hematocrit
is 55% or less. In polycythemia, the
decrease in plasma volume relative to
whole blood unacceptably raises the
anticoagulant-to-plasma ratio, which
causes falsely prolonged results for clot-
based coagulation tests.

• SPECIMEN COLLECTION
 • Hemostasis Specimen Collection
Protocol
• Adjustment of Sodium Citrate Volume for
Elevated Hematocrits
• The 9:1 blood-to-anticoagulant ratio is
effective, provided the patient’s hematocrit
is 55% or less. In polycythemia, the
decrease in plasma volume relative to
whole blood unacceptably raises the
anticoagulant-to-plasma ratio, which
causes falsely prolonged results for clot-
based coagulation tests.

• SPECIMEN COLLECTION
 HEMOSTASIS SPECIMEN
MANAGEMENT
• Hemostasis Specimen Storage
Temperature

• SPECIMEN COLLECTION
 HEMOSTASIS SPECIMEN
MANAGEMENT
• Hemostasis Specimen Storage
Temperature

• SPECIMEN COLLECTION