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Preservation and Storage of Blood

The first anticoagulant preservative was introduced by Rous and Turner in 1916. It consisted of a citrate-glucose solution in which blood from rabbits was stored for two weeks, which prevented anaemia when transfused in another rabbit who had suffered from blood loss. Rous Turner's solution as used for storage of human blood during the First World War (Mollison 1987). The next important development occurred in 1943 during the Second World War when acidifieditrate dextrose (ACD) solution was introduced for clinical use by Loutit and Mollison.In 1957 Gibson et al developed an improved preservative of citrate-phosphate-dextrose (CPD), which was less acidic than ACD and maintained 2,3-diphosphoglycerate (2,3-DPG) level better than in ACD solution. CPD eventually replaced ACD and became commonly used preservative for storage of blood/red cells in liquid form. Shelf-life of blood stored in CPD at 2-4 °C was 21 day. In 1978 citrate-phosphate-dextrose with adenine (CPDA-1) preservative was developed. The addition of adenine improved the synthesis of adenosine triphosphate (ATP) in the stored blood,which prolonged the storage of blood/red cells at 2-4 °C to 35 days.

Approved Preservative Solutions
Compositions of Preservatives/Anticoagulants

	ACD	CPD	CP2D	CPDA-1
Trisodium citrate (g)	22.0	26.30	26.35	26.35
Citric acid (g)	8.0	3.27	3.27	3.27
Dextrose (g)	24.5	25.50	51.10	31.90
Monobasic sodium phosphate (g)	-	2.22	2.22	2.22
Adenine (g)	-	-	-	0.27
Distilled water (ml)	1000	1000	1000	1000
Preservative (ml) / 100ml blood	15	14	14	14
Preservative (ml) / 450 ml blood	67.5	63	63	63
Initial pH of preservative	5.0	5.6	5.6	5.6
On first day pH of blood in bag	7.0	7.2	7.4	7.3
Storage time (days) at 2-6 °C	21	21	21	3

Action of ingredients of anticoagulant solution
Glucose _ supports ATP generation by glycolytic pathways.
Adenine _ synthesizes ATP, increases level of ATP, extends the shelf-life of red cells to 42 days.
Citrate _ prevents coagulation by chelating calcium.
Sodium di-phosphate prevents fall in pH

Red Cells Preservation
The goal of blood preservation is to provide viable and functional blood components for patients requiring blood transfusion. More than 70% of red cells should remain viable in circulation 24hours after transfusion of stored blood in CPDA-1 for 35 days. The blood is stored at 2-6 °C to maintain the optimal viability.
The loss of red cells viability is correlated with the "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

Decrease in pH
When blood is stored at 2-6 °C, glycosis is reduced but does not stop. Giycosis results in the production of lactate, with subsequent decrease in pH. Whole blood collected in CPD has a pH7.20 on day 0 and 6.84 on day 21. Preservative solutions provide buffering capability to minimize pH changes and optimize the storage period.

Loss of Adenosine Triphosphate (ATP)
ATP is associated with the red cells viability. Loss of ATP causes increase in cellular rigidity and decrease in red cell membrane integrity and deformability. A decrease in ATP allows the leak of Na+ and K+ through red cell membrane at levels exceeding those normally seen in vivo. The ATP level in CPDA-1 red cells at day 35 is 45 % (±12) of the initial level.

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