ANTICOAGULANTS & OTHER

PRESERVATIVESBy

Soumya Das

INTRODUCTION

Anticoagulant and preservative solutions allow red blood cells to be stored and transported, enabling modern blood banking.

DEFINITIONS Stored Blood – Citrated blood which has

been kept for hours or days at temp between 2-8’C & to which nothing has been added to inhibit deterioration of RBC.

Preserved Blood – Blood to which some additional substance has been added to check disintegration of RBC.

HISTORY 1893 - Idea of using Citrate for human transfusion

proposed. 1914-15 – 1st RBC Storage Soln (mixture of Citrate &

Glucose was prepared, by Rous & Turner. March, 1914 – 1st Citrated Blood was given to Human,

done by Hustin. 1918 – Oswald H. Robertson first use of cold blood storage

or "banked blood”, also called as Father of Blood Bank. 1943 - Loutit and Mollison during the Second World War

introduced acidified citrate dextrose (ACD) solution. 1957 - Gibson et al, preservative of citrate-phosphate-

dextrose (CPD) introduced. 1978 - Citrate-Phosphate-Dextrose with Adenine (CPDA-1)

preservative was developed. 1980s – Additive Soln were introduced.

PRINCIPLE Basic Principle of these Soln is to

prevent the loss of viability of RBC & increase the capacity to off load Oxygen when given to the recipient.

ATP Metabolism of ATP provides energy for

red cells. It is the Predictor of viability of RBC. It is

suggested that the exposure of Phosphatidyl serine on the outer membrane of stored RBC, hastens the signal for Red Cell Aging.

During Component shelf life, ATP levels drop, but when ATP-reduced red cells are transfused, ATP is regenerated and normal energy metabolism is restored.

2,3 BISPHOSPHOGLYCERATE (2,3 BPG)

Oxygen transport is dependent on 2,3 BPG, as it affects the ability of Hb to release bound oxygen.

On storage, 2,3 BPG levels drops (can drop upto 0 in just 2 weeks) the affinity of Hb for Oxygen increases proportinately. So transfused cells can’t deliver Oxygen readily to tissue.

Once in circulation stored RBC regenerate 2,3 BPG which returns to ½ in 4hrs & to normal in 24hrs.

PRESERVATIVES

Liquid Frozen

CPD – Citrate-Phospate-Dextrose

Glycerol

CP2D – Citrate-Phospate-Dextrose-Dextrose

CPDA-1 – Citrate-Phosphate-Dextrose-Adenine

ACD-A - Acid-Citate-Dextrose formula A

ACD-B - Acid-Citate-Dextrose formula B

AS-1 ( Adsol) AS-3 (Nuticel)

AS-5 (Optisol)

PrimaryAdditive

ACID-CITRATE-GLUCOSE SOLUTIONS (ACD)

Character – a colourless or faintly yellow, clear liquid, practically free from particles. pH – 4.5 - 5.5. Shelf life of Whole Blood – 21 days. >70% transfused cells viable after 24hrs.

Constituents ACD-A ACD-B

Sodium citrate

22.0g 13.2g

Citric acid monohydrat

e

8.0g 4.8g

Glucose 24.5g 14.7g

Distilled Water

1ltr 1ltr

Vol used for 100ml of

blood

15.0ml 25.0ml

Ratio (mL soln to Blood)

1.5:10 2.5:10

CITRATE-PHOSPATE-DEXTROSE-ADENINE SOLUTION

pH – 5.3-5.9 Character – a colourless or faintly yellow, clear liquid, practically free from particles. Ratio (ml soln to blood) – 1.4:10, 14ml is added to each 100ml of blood.

Composition

CPD CPDA-1 CP2D

Trisodium citrate

26.30g

26.30g 26.30g

Citric acid

3.27g 3.27g 3.27g

Sodium dihydrogen phosphate

2.22g 2.22g 2.22g

Dextrose 25.5g 31.8g 44.0g

Adenine - 0.275g -

Water 1l 1l 1l

FDA- Approved shelf life(days)

21 21 35

FUNCTION OF EACH COMPONENT Citrate ( Calcium chelator) –1. prevents coagulation of blood 2. retards gylcolysis. Dextrose –1. Improves red cell viability. 2. provides energy for ATP synthesis.3. decreases rate of hydrolysis of

phosphorus.

CONT. Citric acid – 1. Acidic pH counteracts the marked rise of pH

that occurs when blood is cooled to 4⁰C.2. One of the earliest anticoagulants, not used

these days.3. Acidic pH doesn’t maintain 2,3 BPG levels. Sodium dihydrogen phosphate - more

alkaline pH and better maintains 2,3 BPG levels.

VOLUME USED The volume of each anticoagulant soln

used for 450ml of whole blood is 63ml., and for 500ml is 70ml, according to AABB standard guidelines.

If smaller quantities of blood are to be drawn, then the volume of anticoagulant is reduced proportionately.

RED CELL ADDITIVE SOLN The conversion of Whole blood into

components requires removal of a significant fraction of both plasma and red cell preservative from the solution of RBC.

At this stage, blood has been effectively anticoagulated so the presence of CPD is no longer required by the Red cell concentrate remaining in the primary collection bag.

So, to provide nutrient support and fluid for normal flow, Additive soln are added.

CONTENTS OF ADDITIVE SOLN(MG/100ML)Constituent AS-1 (Adsol) AS-5

(Optisol)AS-3

(Nutricel)

SAGM

Dextrose 2200 900 1100

Adenine 27 30 30

Monobasic sodium phosphate

0 0 276

Mannitol 750 525 0

Sodium chloride

900 877 410

Sodium citrate

0 0 588

Citic acid 0 0 42

FUNCTION OF EACH COMPONENT OF ADDITIVE SOLN. Saline - Fluid in which Red Cells are

suspended to provide the desired flow rate cond.

Glucose/Dextrose – Basic nutrients for glycolysis.

Adenine – support ATP levels Mannitol – to prevent Haemolysis. The 2,3 BPG level is reduced by 90% at

42 days. The loss of 2,3 BPG can be prevented by incoporating bicarbonate and a CO2 trap into the system.

VOLUME USED The volume of each anticoagulant soln

used for 450ml of whole blood is 100ml, and for 500ml is 111ml, according to AABB Standard guidelines

FROZEN STORAGE OF ERYTHROCYTES Preservation of RBC by freezing retards

the deleterious biochemical changes that occur during liquid storage.

By this method RBC can be effectively stored as long as for 21yrs.

Substance used: 1. Glycerol – most commonly used. 2. Hydroxyethyl starch 3. Dextran.

Technique – Slow Freezing method in which the RBC are equilibrated with 40-50% Glycerol and cooled to -80 to -120’C using mechanical refrigeration.

Frozen RBC must be thawed and the glycerol is removed gradually by washing in glycerol soln of decreasing conc. To prevent osmotic hemolysis.

Such thawed and washed RBC must be used within 24hrs because processing breaks the closed system and introduces the possibility of bacterial contamination.

CORD BLOOD STORAGE SOLN USED In a Bag of 200 mL fill, 25 mL of Citrate

Phosphate Dextrose (CPD) anticoagulant used for 150 mL of cord blood

ANTICOAGULANTS IN LABORATORY STUDIES

Anticoagulants

Conc Properties Used Tube

EDTA (Ethylene diamine tetraacetic acid)

1 -2mg/ml

o Most Powerful Calcium Chelator.oBest for preservation of Cell Morphology.oPlatelet clumping inhibited.oCalcium is bound in an unionized, but still complex is formed

All CELL COUNTS & SMEARS.

Double Oxalate3 parts of Ammonium oxalate + 2 parts of Potassium oxalate.

2mg in 0.1ml

o forms insoluble Ca oxalate compound which ppt.o never used for transfusion purpose.

Hb Estimation, PCV, WBC & RBC, Prothrombin Time, Wintrobe ESR

Trisodium

Citrate

1:9 Coagulation Studies

1:4 Westergen ESR

Heparin 0.1-0.2mg/cc

o an inhibitor of thromboplastin form Anti-thrombino Inhibit the action of Thrombin on Fibrinogen.

• Osmotic fragility test• Electrolyte estimation in biochemistry.• Can be used as an anticoagulant in blood bank.

ALL TUBESTube Use Image

Vaccum Blood Collection Tube with Gel & Clot Activator

Serum-separating tubes,,used in medical clinical chemistry tests requiring bloodserum.

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