SOP FOR EDTA-PLASMA SPECIMEN COLLECTION AS RECOMMENDED
BY HUPO PLASMA PROTEOME PROJECT SPECIMENS COMMITTEE AND
APPROVED AT 5th HUPO WORLD CONGRESS WORKSHOP (Oct, 2006)
Note to HUPO: The following are general procedures recommended by Pfizer for
collection of plasma/serum specimens in clinical laboratories equipped with low speed
centrifuges. Individual biomarker assays may have better performance with optimized
temperature and centrifugation conditions. HUPO note: if you are concerned about
platelet activation, be wary of the 4C step and especially the re-warming of the specimen.
Biomarker Human Serum & Plasma Collection Standard Operating Procedure with
EDTA-Plasma Tube
Overview
The measurement of biomarkers from blood samples collected in clinical and
methodology studies can provide useful insights to our understanding of physiology and
pathophysiology. While not one blood collection matrix and procedure can produce
samples appropriate for the measurement of all blood based markers, many biomarkers
can be accurately measured in blood collected in lavender-top K2 EDTA tubes for
plasma, or red-top tubes (containing no anticoagulant, preservative, or separator) for
serum. BD Vacutainer® venous blood collection tubes with spray-coated K2 EDTA are
routinely used for whole blood hematology and immunohematology testing, and
preferred over K3 EDTA tubes (which have a liquid additive). The BD Vacutainer®
plastic serum tubes with spray-coated silica are the tubes of choice for serum. Following
centrifugation, 10 ml blood collections produce 4-5 mL of serum or plasma.
http://www.bd.com/vacutainer/products/venous/plus_plastic_tubes_docs.asp
Lavender-top K2 EDTA plasma and red-top serum BD Vacutainer® venous blood
collection tubes can be stored at room temperature for extended periods of time.
Expiration dates are provided and should be adhered to.
Materials and Equipment
 A refrigerated (2-8 ºC) centrifuge capable of spinning at 1500-2000g that will
hold 16 x 100 mm tubes
 10 mL lavender-top K2 EDTA BD Vacutainer® venous blood collection tubes and
10 mL red-top serum BD Vacutainer® venous blood collection tubes
 Disposable transfer pipettes
 VWR Low Temperature 5 ml Freezer vials, Sterile (Product # 16001-104). Serum
and plasma from the BD collection tubes are transferred to these secondary tubes
for freezing and storage.
 Freezer and cryobox for storing freezer vials. Ultra-cold freezers at or below -70
ºC are preferred.
Finally, personnel performing these tasks should be receive Blood Borne Pathogen
training and annual recertification from their Environmental Health and Safety
Departments, and adhere to adhere to all relevant guidelines regarding the safe handling
and disposal of human blood products, including but not limited to wearing safety
glasses, gloves, and lab coats.
Instructions for Obtaining Plasma for Biomarker Research Purposes
1. Pre-chill 10 mL lavender-top K2 EDTA BD Vacutainer® venous blood collection
tubes on wet ice for at least 5 minutes.
2. Follow standard phlebotomy practices and procedures, including wearing gloves
during venipuncture and when handling blood collection tube and secondary tube
to minimize exposure hazard. Consult product inserts for details of collection
procedures, and order of collection when also collecting samples for other
purposes.
3. Immediately after allowing the lavender-top Vacutainer® tube to completely fill,
gently invert the tube 8 times, and return the tube to the wet ice bath
4. Within 30 minutes of collection, centrifuge at 1500 – 2000g for 10 min in a
refrigerated centrifuge (2-8 ºC).
5. Within 30 min of centrifugation carefully transfer the plasma into a 5 ml VWR
freezer vial. Do not attempt to retrieve the last 0.2 ml of plasma.
6. Plasma samples should be immediately frozen at -70˚C to -80˚C, and shipped on
dry ice to a designated facility.
7. Deviations from the prescribed collection protocol should be reported to the
designated coordinator.
Instructions for Obtaining Serum for Biomarker Research Purposes
1. Follow standard phlebotomy practices and procedures, including wearing gloves
during venipuncture and when handling blood collection tube and secondary tube
to minimize exposure hazard. Consult product inserts for details of collection
procedures, and order of collection when also collecting samples for other
purposes.
2. After allowing the red-top Vacutainer® tube to completely fill, invert 4-6 times
and store upright at room temperature until clotted (approximately 30-45 min).
3. Centrifuge at 1500 – 2000g for 10 min in a refrigerated centrifuge (2-8 ºC). Room
temperature centrifugation is also acceptable for serum collections.
4. Within 30 min of centrifugation carefully transfer the serum into a 5 ml VWR
freezer vial. Do not attempt to retrieve the last 0.2 ml of serum.
5. Serum samples should be immediately frozen at -70˚C to -80˚C, and shipped on
dry ice to a designated facility.
6. Deviations from the prescribed collection protocol should be reported to the
designated coordinator.
Provided by:
Gregory M. Preston, Ph.D.
Pfizer Global Research & Development
Groton/New London Laboratories
Translational Biomarkers, Discovery
Email: gregory.m.preston@pfizer.com