Blood Component Separation

PURPOSE
The Immunohaematology Laboratory (IHL) separates Whole Blood into components
to maximize its usage.
EQUIPMENT
Tube sealer
Laminar flow
Refrigerated centrifuge
Plasma expresser
Double pan weighing balance
SUPPLIES
Triple bags
Manuals of all equipment for reference regarding use and maintenance of each
equipment
SPECIAL SAFETY PRECAUTIONS
Follow universal precautions when working in a laboratory environment with
potentially infectious agents.
PROCEDURE
Preliminary Statement
It is the responsibility of the component room technician to separate
components from whole blood collected in multiple bags. The packed cells
unit should be within the range of 235-350mls, Fresh Frozen Plasma 200260mls, Platelets and Cryoprecipitate 60-80mls.
Preparation of Red Blood Cells (RBC) and Fresh Frozen Plasma(FFP)
1. Turn centrifuge on and ensure it is set to 4C
2. Remove whole blood from 2-8C unclear storage area or donor area.
Process all whole blood units within 6hrs of collection.
3. Resuspend all whole blood units before placing into centrifuge cups,
massage blood out of blood bag ports.
4. Place whole blood bags on cups, balance centrifuge cups two at a time
using balance scale.
5. Repeat with remaining bags and close centrifuge lid.
6. Enter Program 3 on centrifuges (7mins at 4200RPM) press enter.
7. Remove whole blood bags gently from centrifuge cup.
8. Place blood bag on plasma expresser and release spring clamp on
expresser.
9. Break the seal of the tube connecting to the satellite bag.
10. Express plasma into satellite bag, leaving 50-60mls of plasma on red
blood cells.
11. Seal plasma tubing and red cell tubing. Separate after ensuring unit
number is on both bags.

12. It is noted to label plasma prepared within 6hrs of collection Fresh

Frozen Plasma. FFP expires 1year after collection date.
13. It is noted that plasma prepared after 6hrs of collection time must be
discarded.
Preparation of Red Blood Cells, Fresh Frozen Plasma and Platelet Concentrate
1. Process the blood collected within 6 hours.
2. Turn on centrifuge and ensure it is set at20C.
3. It is noted that whole blood units for platelet preparation must remain
at room temperature. If whole blood has been refrigerated or placed on
ice during shipment, platelets cannot be made
4. Resuspend whole blood units before placing into centrifuge cups and
massage blood out of blood bag ports
5. Place whole blood units into centrifuge cups, balance centrifuge cups
two at a time using balance scale
6. Verify temperature at 20C.Place two balanced centrifuge cups in
centrifuge in opposite position from each other. Repeat with remaining
bags and close centrifuge lid.
7. Enter Program 4 on centrifuge (5mins at 2600RPM)
8. Remove blood bags gently from centrifuge and place on plasma
expresser and release spring clamp on expresser.
9. Temporarily seal off or clamp off plasma satellite bag. Break the seal
of the tube connecting to the satellite bag.
10. Express plasma into platelet satellite bag leaving 50-60mls plasma on
the red cells.
11. Seal off red cell bag and place in uncleared blood bank refrigerator
after verifying bar code
12. Place satellite bags back into centrifuge at 20C and close lid
13. Enter Program 3 on centrifuge (7mins at4200RPM)
14. Gently remove bags from centrifuge and place on plasma expresser
15. Remove temporary seal or clamp from plasma satellite bag.
16. Express the plasma into empty plasma bag leaving 50-60mls plasma
on the platelet concentrates.
17. Note: It is better to leave less plasma on platelets and add additional
plasma back into platelet until you attain correct volume. NEVER
remove additional plasma from platelet after you have removed it from
plasma expresser. Resuspended platelets would be lost back into
plasma bags.
18. Seal the tubing of the plasma bag leaving an approximate 6 inch
segment.
19. Verify bar code number on both plasma and platelet bags.
20. Separate plasma bag and place in uncleared plasma freezer.
21. Seal tubing of platelet bag leaving an approximate 6 inch segment.
Allow platelet concentrate to rest on table for 1 hr.
22. Gently resuspend platelet concentrate using a circular motion.

23. Place platelet in 18-24C incubators on the uncleared shelf. Expiration

date is midnight of fifth day after collection.
Preparation of Cryoprecipitated AHF(cryo)
1. Prepare FFP from triple bag system temporarily sealing tubing
between FFP and satellite bag.
2. Store FFP in -18C or colder freezer.
3. Cryo can be prepared at time from FFP during the 12 month storage
period, but cryo expiration date is 1 year from date whole blood was
collected.
4. Remove FFP from storage freezer. Thaw FFP in 2-8C refrigerators for
approx. 6-10 hrs until slushy consistency is reached.
5. Centrifuge plasma in 1-6C refrigerated centrifuge at 4200RPM for
10mins (program 1) after balancing centrifuge cups.
6. Remove plasma from centrifuge and place on plasma expresser.
7. Express plasma into plasma bag, leaving 10-25mls plasma on cryo
8. Seal the tubing on the cryo and plasma bags and verify accuracy of bar
code numbers.
9. Separate cryo and FFP. Place cryo in appropriate -18C or colder
refrigerator.
10. Discard cryo reduced plasma.
11. Use Delphyn Software System to transform units (FFP to cryo). Refer
to Delphyn Processing Module, Section B. pg. 7 to 11.
Decontamination and withdrawing of Blood and its Components
1. Blood components are rejected due to incompatible grounds, it will
then be withdrawn. When withdrawing Blood and/or its components,
the blood must first be decontaminated by injecting five (5) ml
concentrated bleach directly into the bag of blood (should try not burst
bag). After which the decontaminated bag of blood is withdrawn using
the MOH facility. All Blood, and thus their components, that are
rejected based on rejection criteria, are withdrawn.
2. Withdraw units using Delphyn Software System. Refer to Delphyn
Processing Module, Section E. pg. 16 to 20.
Quality Control of blood component preparation
Whole blood:
Volume : 450-500ml 10 % of body volume excluding anticoagulant
Hb minimum 12.5g/dL
Sterility : no growth
Packed Cell:
Volume : 280ml 50ml, frequency of control 1% of all units
Sterility : no growth
Platelet concentrates:
Storage : 18-22C

Temperature should be recorded at least every 12Hrs during storage.

Volume > 40ml.
Macroscopic appearance : no visible platelets aggregates
Sterility : no growth

Fresh Frozen Plasma

Volume: 200-260ml
No leakage after pressure in plasma extractor, before freezing and after
thawing
Macroscopic : no abnormal color or visible clots
AHF Cryoprecipitate
Volume : 50-80 mls
Macroscopic : homogenous
Sterility: no growth
Delphyn System
Enter results into Delphyn Software System. Refer to Delphyn Donor Module,
Section E. pg. 24 to 36.
1.0 REFERENCES
Technical Manual of American Association of Blood Banks 13 th edition 1999 Pgs
26, 168-170, 172-173,177, 716-717, 723-726.
WHO Model Standard Operating Procedures, 2002.
Introduction to Transfusion Medicine - Zarin Bharucha & D.M. Chouhan 1 st
edition 1990 Pg 124-125.
Circular of information for the use of human blood and components.
Delphyn Donor Module, Section E. pg. 24 to 36.
2.0 RELATED DOCUMENTS
IHL-200-F01A Component Register
IHL-200-F01B Blood and Components Destruction Log
IHL-200-F01C Components Quality Control
Delphyn Donor Module, Section E. pg. 24 to 36.
3.0 APPENDICES
IHL-200-F01A Component Register
IHL-200-F01B Blood and Components Destruction Log
IHL-200-F01C Components Quality Control