1) Blood components like packed red cells, platelet concentrates and fresh frozen plasma can be prepared by separating whole blood into its components using centrifugation and expressors. 2) Optimal storage conditions and times allow individual components to be stored and transfused separately as needed rather than transfusing whole blood. 3) The document outlines the equipment, procedures and quality indicators for preparing the main blood components from a single donor to benefit multiple recipients.

1. Blood component preparation
Dr. Akshay Agarwal
MD Pathology Resident
drakshay90@gmail.com

2. Index
• Need for separation
• Types of blood components
• Equipments
• Precautions
• Preparation of
– Packed red cells
– Platelet rich plasma
– Fresh frozen plasma

3. Need for separation
• Optimal survival of each constituents.
– In whole blood stored at 2-6°C, platelets stay
viable for 1 day and factor V & VIII decrease.
– While after separation, platelets stay upto 5 days
and factor V and VIII can be stored as FFP for 1 yr
at -30°C
• Transfusion of only required component thus
avoids the use of unnecessary component
which could be contraindicated in a patient.
• One donor can save several patients.

4. Blood
components
Cellular
PC, Platelets,
others
Plasma
FFP,
Cryoprecipitate
Plasma derivatives
Albumin 5%, 25%,
Factor VIII,
coagulation
factors

5. Equipments
• Freezer: – 40°C and -70°C
• Blood bank Refrigerator 2-6°C
• Refrigerated centrifuge with swing out head and oval
cups
• Laminar flow
• Weighing scale
• Dielectric sealer or aluminium clips & sealer
• Stripper
• Water bath 37°C or plasma defroster (Microwave)
• Platelet agitator
• Sterile docking device

6. Precautions
A. In collection of blood:
1. Proper selection of donor
2. Clean aseptic site of venipuncture
3. Uninterrupted flow of blood
4. Collect correct amount of blood
5. Automatic mixing of blood during collection
6. Separate platelets within 6-8 hours
7. triple pack and quad pack system

7. Precautions
B. In centrifugation
1. Balancing & use of rubber discs
2. Broad side of the bag faces the outside wall of
the cup
3. Correct speed and time to be maintained
4. Look for any abnormal vibration

8. Whole Blood
• 450 ml +/- 45 ml or350 ml +/- 35 ml of donor
blood and anticoagulant solution
• Minimum 70% of transfused red cells should
survive in the recipient’s circulation 24 hours
after transfusion.
• No functional platelets and no labile
coagulation factors V and VIII

9. A unit of blood collected from a single donor in a
sterile container with an appropriate
anticoagulant.(CPD-A1)
• •Shelf life; 35 days at 2–6 C°
• •Volume ; 450ml +- 10%
• •One unit whole blood contain ~ 200ml RBC
• •Indications ;
• –Massive transfusion
• –Exchange transfusion
• •Must be ABO identical , cross match compatible

10. Preparation of Packed Red Cells
• Red cells have a higher specific gravity than
plasma, thus settle in the lower portion of the
bag.
• Types of preparation:
– Sediment red cells: PCV- 60-70%; 30% plasma and
all original leukocytes and platelets.
– Centrigued red cells: PCV- 70-80%; 15%
– Red cells with additive (Adsol or SAG-M): PCV -50-
60%; minimum plasma and all leukocytes and
platelets.

11. A unit of RBC prepared after removing 2/3 of plasma
• •Volume; 250 - 300 ml
• •Shelf life; 35 days (CPDA1)
• •Storage temperature; 2–6 C°
• •Indications ;
• oSymptomatic anaemia
• oBleeding during surgery
• oSevere haemorrhage for patients with surgical, complications
• otrauma
• ocancer
• • Must be ABO compatible and cross match compatible
• •1 Unit raises ~ 1.5 g/dL of Hb in an adult

12. Steps in separation
1st
• Store the bag at 2-6°C till processed
2nd
• Place bag in buckets of refrigerated centrifuge and balance
the opposite bags
• Centrifuge at 5000 x g for 5 mins at 2-6°C
3rd
• Express approx 3/4th of the plasma in the satellite bag
• Keep red cell at 2-6°C and plasma at -30°C

13. RCC Quality Indicators
• •Volume 250 – 300 mL
• •Haematocrit 55 – 75 %
• •Haemoglobin >45 g/dL
• •pH 6.4 – 7.4
• •Haemolysis < 0.8 %
• •Sterility Sterile
• •Residual leukocyte count < 2x109/Unit

14. Leukocyte-reduced Blood Components
• Leukocytes can cause:
– NHFTR
– HLA alloimmunization
– CMV, EBV, HTLV-1
– TR-ALI
– TR- GVHD
– TR-Immunosuppresion
– Cytokines produced even in stored bags

15. Preparation of Platelet Concentrate
• Random donor platelet – prepared from
450ml of whole blood
• Single donor platelet – prepared by apheresis

16. • Random donor platelet concentrate, prepared from
one unit of blood collected from one donor
• •Volume 50ml
• •Storage temperature 22 - 24 C° (Agitation)
• •Shelf life 3 – 5 days
• •Adult Dose 1 unit /10kg (4 – 6 units)
• •Peads Dose 10 – 15 ml/Kg
• •ABO & Rh compatible
• •Cross match not necessary
• •Manual unit raises 5000 – 10,000 PLT in an adult

17. Procedure
1st
• Collect in triple pack system and store at room temperature at 20-22°C for
less than 6 hours.
2nd
• Place bag in buckets of refrigerated centrifuge and balance the opposite bags
• Centrifuge at 2000 x g for 3 mins at 20-24°C
3rd
• Express approx 4/5th of the platelet rich plasma in the satellite bag
4th
• Centrifuge the bag with PRP and another satellite bag at 20-40°C at 5000 xg
for 5 mins.
5th
• Express supernatant platelet poor plasma into another empty bag and leave
approx 50 ml of plasma.
6th
• Store platelet at 20-22°C under constant agitation.

18. Precautions
• Agitation during storage helps in:
– Exchange of gases
– Maintenance of pH
– Reduce formation of platelet aggregates
• pH should never fall below 6 because:
– Change in shape of platelets from disc to sphere
– Pseudopod formation
– Release of platelet granules

19. PLT Conc. Quality Indicators
• Platelet concentrate
• •Volume 45 – 55 mL
• •Swirling Present
• •Platelet count >55x109/Unit
• •Residual leukocyte count <1x109/Unit
• •Residual red cell count <1x109/Unit
• •pH 6.4 – 7.4
• •Sterility Sterile

20. Fresh Frozen Plasma (FFP)
• Plasma removed from cells within 6-8 hours of collection is rapidly frozen
to below
• –30C° temperature.
• •Before transfusion it is necessary thaw at 37 C°
• •Once thawed there is rapid deterioration of clotting factors, so use
immediately after thawing
• •Volume 200ml
• •Shelf life 1 year
• •Storage temp –30C°
• •Dose 12 - 15 ml/kg
• •Indications;
• –Single or multiple clotting factor deficiency
• –Severe liver disease
• –Massive transfusion
• –DIC

21. Preparation of fresh frozen plasma
1st
• Collect in triple pack system and store at 4°C or in air-conditioned room till
processed but not for more than 6-8 hours.
2nd
• Place bag in buckets of refrigerated centrifuge and balance the opposite bags
• Centrifuge at 5000 x g for 5 mins at 4°C
3rd
• Express approx 4/5th of the plasma in the satellite bag
4th
• Label the plasma bag and is rapidly frozen within 6-8 hours and the freezing
process should not be more than 1 hour at -70°C
5th
• Can be stored at -30°C or below for 1 year.
6th
• Thaw it in a plasma defroster (Microwave) or place in a plastic over wrap and
put it in a 37°C circulating water bath.

22. FFP Quality Indicators
• Fresh Frozen Plasma
• –Volume 180 – 200 mL
• –Factor VIII level > 0.7 IU/mL
• –Fibrinogen level >300 mg/Unit
• –Sterility Sterile

23. Precautions
• During blood collection:
– Flow should be continuous
– 450ml bag should not take more than 8 mins to fill
• During processing:
– Freezing the plasma as soon as it is separated
– Maintaining the storage temperature
• During thawing:
– The entry ports of the bag should remain above the
water
– Should be administered within 12 hours if kept at 2-
6°C.

24. Donor bed

25. Blood bag

26. Collected blood bag

27. Blood samples collected in appropriate
tubes for testing

28. Stripping and discarding of needle

29. Primary bag with satellite bags

30. cryocentrifuge

31. expressor

32. Platelet agitator