BLOODBANKING

BLOOD
COLLECTION,
PRESERVATION AND
STORAGE
• INDIA- collection, processing, storage and preparation of blood components is
regulated under Food and Drugs Administration (FDA). Blood is regarded as “DRUG”.

• Three main types of donations-

1. Whole blood donation (one unit- 350ml whole blood in an anticoagulant solution).
2. Autologous donation (for subsequent transfusion to self)
3. Apheresis donation (separation and retention of desired product from whole blood and
return of remaining portion to donor)
eg.- plasmapheresis, red cell apheresis, plateletpheresis, leucapheresis or stem cell
collections.
Donor’s screening with complete demographic, medical history and physical
examination is done. Hemoglobin estimation and ABO Rh blood grouping is done
prior donation.
DEFERRAL CRITERIA
 Equipment and Materials
• Blood Containers: Polyvinyl chloride (PVC) plastic bags
-single, double or triple bags for collection of 350 ml or 450 ml blood.

• anticoagulant solutions:
-citrate-phosphate-dextrose (CPD) or citrate-phosphate-dextrose-adenine
(CPDA-1)
-volume :49 ml for 350 ml or 63 ml for 450 ml of blood (14 ml CPD or CPDA-
1 for 100 ml blood).

• Sphygmomanometer

• Automatic mixing of blood and weighing of blood bag machine

• Sterile cotton swabs and band-aids/bandages

• Methylated spirit, tincture of iodine, povidone-iodine solution (1 %) &

 METHOD OF DONOR BLOOD COLLECTION
• wash hands with soap and water and wear
sterile gloves.
• Inspect the bag for leakage or any other
defect. The anticoagulant solution must be
clear.
• Check the donor name, donation number on
the form bag and pilot tube.
• Place the bag on a balance which is below
the level of the arm.
• choose the site of venipuncture in the
antecubital area of the arm (area that is free of
any skin lesion/needle marks.)
• clean 4-5 cm area on proposed site of
venipuncture ,starting at the site of
venipuncture and moving outwards in a
concentric spiral way with methylated
spirit/alcohol. Apply 10% povidone-iodine
solution (betadine) or tincture of iodine in the
same way ,allow it to dry.
• Inflate blood pressure cuff to maintain pressure 50-60 mm of hg.
• Ask the donor to close the fist.
• Uncover the sterile needle and perform venipuncture immediately
• ask the donor to open and close hand or to squeeze a rubber ball.
the donor should be under constant observation throughout the
phlebotomy.
• Mix the blood and anticoagulant gently and periodically during
collection of blood. Mixing can be done by hand or blood
collection monitor -scale/mixer. The flow of blood should be
uninterrupted and constant
• once donation is complete clamp the tubing of the bag with
plastic clip , deflate the cuff or release the tourniquet.
• Place the sterile swab at the venipuncture site, apply light pressure
and withdraw the needle.
• Take the bag to the processing table, Loosen the plastic clip and apply light pressure on the bag to
transfer 5-6 ml of blood in the pilot tube.

• Seal the tube with di-electric tube sealer and separate the needle.

• Invert bag several times to mix blood thoroughly; then allow tubing to refill with anticoagulated
blood from the bag.

• Keep the blood bag at 2-6°C in the refrigerator immediately after collection. If platelets are to be
harvested, blood bag should be kept at 20-24°C until platelets are separated. Platelets should be
separated within 6-8 hours after the collection.

• The donor should remain on the bleeding couch for 8-10 minutes under observation.
STORAGE AND PRESERVATION
 ANTICOAGULANT PRESERVATIVE SOLUTION
• THE NEED OF THESE SOLUTION IS FOR :
1. Prevent coagulation
2. Preserve the life and function of red cell for long time
• Solutions are mainly :
• Acid Citrate Dextrose
• Citrate Phosphate Dextrose
• Citrate Phosphate Dextrose Adenine (CPDA 1)
 FUNCTION OF VARIOUS CHEMICAL USED IN
ANTICOGULANT PRESERVATIVE SOLUTION
• CITRATE –most commonly used. It acts BY CHELATING CALCIUM hence preventing clotting.
• Citric acid prevents the caramelization of glucose in the citrate dextrose solution during
autoclaving ,it is fairly weak tribasic hydroxy acid .Along with trisodium citrate, it gives an optimal
Ph .
• DEXTROSE- supports ATP generation by glycolytic pathway- cellular energy
• ADENINE-increase in the post transfusion survival of red cells to 35 days because of enhanced
ATP production
• MONOBASIC TRISODIUM PHOSPHATE- Maintains Ph during storage by adequating the
levels of 2,3-DPG
 SHELF LIFE

• BLOOD STORAGE AT 2-6 ˚C IN ACD SOLUTION – 21 DAYS

• CPD SOLUTION -21 DAYS
• CPDA -1 SOLUTION – 35 DAYS BECAUSE OF ENHANCED ATP PRODUCTION AND
BETTER PRESERVATION OF 2,3 –DPG .
• CPDA 1 SOLUTION IS MOST COMMONLY USED.
 RED CELL PRESERVATION
 The goal of blood preservation to provide viable and functional blood component
for patient requiring BT. More than 70% of red cells should remain viable in
circulation 24 hr after transfusion of stored blood in CPDA 1 for 35 days.

 The loss of red cell viability is correlated with the lesion “ lesion of storage “ due
to various biochemical changes:
 Decrease in Ph
 Build up of lactic acid
 Decrease in glucose consumption
 Decrease in ATP level
 Low 2,3 DPG levels
 ADDITIVE SYSTEM
The new blood collection system has a primary bag containing a standard anticoagulant CPD and
a satellite bag containing a additive solution. Blood is collected in primary bag .After the plasma is
removed from the whole blood into another empty satellite bag ,the additive solution is added to
red cells .
• IT provides nutrients to red cell and extend the red cell storage to 42 days and to harvest
maximum amount of plasma. These should be added to red cells within 72 hr since phlebotomy
 FIRST GENERATION ADDITIVE SOLUTION:
• The first basic additive solution of saline ,adenine ,and glucose was developed by hogman. It
also permitted 5 week storage of PRC but without the adverse effect associated with the high
Ph of preservative solution.

• ADDITIVE SOLUTIONS ARE-

SAG –M is standard RBC storage system in many countries. (100 ml to the packed RBCs prepared
from a 450-ml blood collection)
• Adsol – AS1
• Nutricel- AS3
• Optisol- AS5
All of these variants appear to be equivalent ,permit 42 days of storage with recovery of about 80% of cell
with around 0.3% haemolysis at the end of storage period.
These additives allow for harvesting more plasma and platelets from the unit.
 SECOND GENERATION ADDITIVE SOLUTION

• The potential for 7-8 weeks of storage

• Better recovery
• Reduction in membrane loss
• Improve 2,3-DPG and/or ATP concentration for all stored RBCs.
• Solutions are-
1. PAGGS –consisting of phosphate ,adenine, glucose, guanosine and saline.
2. PAGGG –M :- sodium chloride replaced by sodium gluconate so as to take advantages of “chloride
shift” to maintain red cells 2,3 DPG
3. Erythosol 1 and 2 take advantages of chloride shift to increase the intracellular Ph, while at the same
time ,using a more alkaline final suspending solution and additional phosphate to maintain ATP.
4. EAS 81.
STORAGE TEMPERATURE FOR BLOOD
• Blood always must be stored at a temperature of 2-60 C . main reason
for storing blood at this temperature:-
1. To keep the rate of glycolysis at lower level so dextrose is not used
quickly
2. To minimize the proliferation of any bacterial contamination
3. To minimize the rate of diffusion of electrolytes across the cell
membrane
The lower limit of +2 0 c is also very important because red cells
are very sensitive for freezing it cause red cell to haemolyse.
 HEPARIN
• Heparin prevent coagulation by inactivating the prophylactic activity of thrombin .
• Dose of heparin for anticoagulation is 0.5-2.0 IU/ml blood.
• Heparinized blood should be used within 24 hours. The effect of heparin can be neutralised
with protamine sulphate.
 REJUVENATE SOLUTIONS

These solutions having Phosphate, Inosine, Glucose, Pyruvate, Adenine increase the level of 2,3 DPG
and ATP in stored red cell can be added at any time between 3 days post collection and 3 days after
expiration of red cell .
The solution is added directly to red cell mixed and incubated at 37 degree for 1 hour .
The rejuvenated cells should be transfused within 24 hour after washing or they are glycerolyzed for
keeping red cell in frozen state to improve the quality of red cells.
 RED CELL FREEZING
• Frozen red cell are primarily used for autologous transfusion and storage of rare blood group, freezing
damage red cells due to intracellular ice formation and hypertonicity.
• Glycerol is most commonly used cryoprotective agent that are less than 6 days old.
• Glycerol limit the ice formation and provide liquid phase and avoid excessive hypertonicity.
• The freezing and storage temperature depend on the concentration of glycerol.

Two concertation's are used to freeze red cells.

1. HIGH CONCENTRATION GLYCEROL (40% WEIGHT IN VOLUME) .
2. LOW CONCENTRATION GLYCEROL (20% WEIGHT IN VOLUME).
 METHOD OF FREEZING AND PRESERVATION OF
RED CELL IN FROZEN STATE

1. Glycerolised red cell having final concentration of 40% W/V of glycerol can be
frozen at -80 degree over a period of 30 min using mechanical refrigeration .Then it
can be preserved at -60 to -65 degree C for 10 years.
2. Glycerolised red cells having final concentration of 20% w/v of glycerol can be
frozen at -196 0 c using liquid nitrogen for 2 -3 min and can be preserved in the gas
phase of liquid nitrogen at -120 degree C for 3 years.
 HIGH GLYCEROL SOLUTION
• Solution consist of 6.2 M glycerol solution that contains-
.Glycerol 57g %
.Na lactate 1.6g %
.KCL 0.03g%
.Monobasic disodium phosphate 25m eq/ l
 FREEZING PROCEDURE
1. Blood is collected in CPDA 1 double pack. The pack is centrifuged at 3000 rpm for 7 min and plasma is
removed.

2. 12% glycerol solution equal to half the volume of the packed cells is added to it, mixed and allowed to
equilibrate at room temperature for 10 min.

3. 60% glycerol solution equal to half the volume of packed cell is added to the pack and mixed ,allow to
equilibrate for `10 min.

4. Dispense into labelled 10 ml sterile plastic /glass tubes(15x100 mm)

5. Freeze at -400 C ,keep it in lowest part of freezer and switch freezer to rapid freeze or place in top of
liquid nitrogen tank for 5-10 min.

6. When frozen , tubes may be stored at -200 c

 LOW GLYCEROL SOLUTIONS
• This consist of
Glycerol – 35g%
Mannitol -2.88%
NaCl – 0.65 g
Procedure
1. Whole blood in CPD is centrifuged at 3000 rpm for 7 min and plasma is removed and freezed.
2. Low glycerol solution equal in volume to the red cells is added with vigorous shaking.
3. The glycerolised red cells are transferred to polyolefin plastic bag and kept in aluminium container
which is then placed upright in a bath of liquid nitrogen at -196 0c ,and then stored at -120 0c in liquid
nitrogen vapour.
 THAWING AND DEGLYCEROLATION
• Frozen red cells are thawed in water bath at 37 degree for about 10 min.

• Glycerol must be properly removed from the thawed cells to avoid haemolysis in-vivo or in-vitro.

• Washing the glycerolized cell in hypertonic (12% saline ) solution to less hypertonic solution
finally to isotonic electrolyte solution containing glucose (0.2% dextrose).

• Glycerol contents must be reduced to 1-2% otherwise they will haemolyse on contact with plasma.

• Washing can be caried out either by continuous flow washing in the haemonetic blood processor or
by in Fenwal Elutramatic system.
 INDICATION OF USE OF FROZEN RED CELL
1. Freezing of rare blood group enables long term storage and supply on a regional and
national basis.

2. Storage of blood for patient with antibodies against high frequency antigens.

3. Storage of blood for autotransfusion, specially in patients with rare blood group.

4. Prevention of non haemolytic febrile transfusion reaction in patient sensitise to leucocytes

and platelet.

5. Prevention of sensitization against HLA antigen in potential recipient of tissue transplant.

 PLATELET PRESERVATION
• The preservation of viable and functional platelet depend on
following factors
1. Temperature-22-240 c
2. Ph –above 6
3. PLASTIC BAG- platelet stored in bag made of polyvinyl
chloride with plasticizer di(2 ethyl hexyl)phthalate have shelf
life of 3 days .New plastic bags made of polyolefin with no
plasticizer and thin walled PVC with tri-(-2- ethyl hexyl)
trimellate plasticizer maintain Ph and function about 7 days .
4. The pooled platelet can be stored for 4 hour at 22 -24 O C
before they are transfused .
 FRESH FROZEN PLASMA

• Shelf life of FFP is 12 month at -18 C or

lower.
• After thaw FFP can be stored at 2-6 C
for 12 hrs before transfusion
• SINGLE DONOR PLASMA - shelf life of SDP is 5 years at -18 0c or lower
• Thawed plasma can be stored at 2-60 c for 5 days before transfusion.
• CRYOPRECIPITATE - can be stored for 12 month at -18 0 c or lower . thawed
cryoprecipitate can be stored for 6 hrs at 2-60 c and pooled cryoprecipitate kept at 2-60 c should
be used within 4 hrs.
• GRANULOCYTES - shelf life of these 24 hrs at 22 – 24 0c .
STORAGE OF BLOOD DURING
TRANSPORTATION
• Blood should be packed in cold boxes
surrounded by icepacks ,making sure that the
blood never touches the ice packs directly.
• Temp range kept within 2 -8 C during
transportation.
• IN HOSPITAL COLLECTION- If ambient temp is
more than 25 C or if there is possibility that the
blood will not be transfused immediately then
also transport into a cold box.
 SHIPPING OF PLATELET

• Should use well insulated container with no ice ,to maintain the temp b/w 22 -24 C.
• Shipping of frozen component - FFP and Cryoprecipitate must be shipped at -18
C or below. Dry ice is used to maintain the frozen state.
PRODUCT DESCRIPTION/CONTENTS STORAGE SHELF LIFE EFFECTS
WHOLE BLOOD RED CELLS All components of donor blood plus 2-6 C CPD- 21 days Increase HCT BY 3 % and
anticoagulant CPD, CPDA-1 whole CPDA- 35 days Hb by 1gm/dl
blood with 200-250ml plasma Open system- 24 hr.
removed, final volume- 300ml. With AS additive- 42 days
AS 100 ml additive added and most
plasma removed final volume= 350 ml
RED CELLS- LEUCOCYTES Modified by centrifugation removing 2-6 C CPD- 21 days
REDUCED buffy coat or filtration to remove >70% CPDA- 35 days
leucocytes Open system- 24 hr.

RED CELLS WASHED Red cells washed with normal saline 2-6 C 24 hrs
RED CELLS FROZEN- Red cells frozen with glycerol -65 (high glycerol) 10 yr
DEGLYCEROL
-120 (low glycerol) 3 yr

-deglycerolized at 2-6 C 24 hrs

PLATELETS >5.5 X 1010 20-24 with agitation 3-5 days Increase platelets by 5-10
In 40-70 ml of plasma 4hrs after pooling x109 /L

GRANULOCYTES About 1x1010 in 200- 250 ml plasma 20-24 without agitation 24 hrs

FFP 200-250 ml with anticoagulant Frozen -18 C 1 Yr Increase Fibrinogen by 10

Thawed 2-6 C 12 hrs mg/dl
CRYOPRECIPITATE 80- 120 Units factor VIII Frozen -18 C 1 yr Increase Fibrinogen by 10
40-70% von willebrand factor Thawed 2-6 C 6 hrs mg/dl
after pooling 4 hrs
THANKYO
U