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COLLECTION AND PREPARATION OF BLOOD COMPONENTS

I. INTRODUCTION

Established phlebotomy guidelines are to be followed. Be sure to adjust the balance or vacuum assist device so that only 500 ml of blood is collected. Blood can be collected into any of the following FDA-approved anticoagulant or anticoagulant-preservative systems:1-7

Citrate-phosphate-dextrose (CPD) anticoagulant
Citrate-phosphate-dextrose-adenine (CPDA1)
anticoagulant
CPD/AS-1 anticoagulant/preservative system
CP2D/AS-3 anticoagulant/preservative system
CPD/AS-5 anticoagulant/preservative system

Red blood cells prepared in anticoagulant/preservative systems have hematocrit values of approximately 55 V%. If the red blood cells are collected and stored in anticoagulant/preservative systems, they must be centrifuged to prepare a red blood cell concentrate with a hematocrit of 75+5 V% prior to glycerolization.

The ACP215 is equipped with a washing bowl with a 325 ml capacity. Collection of 500 ml of blood will not result in red cell spillage during the deglycerolization cycle and a reduced freeze-thaw-wash (FTW) recovery.

PROCESSING OF FRESH WHOLE BLOOD

Within 8 hours of collection and storage at room temperature, a red blood cell concentrate is prepared as follows:

Centrifuge the fresh whole blood unit at 1615 X g for 4 minutes. Be sure the brake on the centrifuge is turned to the OFF position (no brake). Remove all the platelet rich plasma to an integrally attached transfer pack. Store the red blood cells in the original collection bag at 4 C for 3 to 6 days (inclusive) prior to freezing.

Ideally, a non-anticoagulated blood sample for serum should be collected from the venipuncture site after the unit of whole blood is collected. The serum is transferred to four (4) 2 ml polyethylene cryogenic vials for freezing and future testing. If sera is not available, prepare four 2 ml polyethylene cryogenic vials of plasma within 24 hours of collection for freezing for future testing of the plasma.

PROCESSING OF STORED WHOLE BLOOD, ADDITIVE-PRESERVED RED BLOOD CELLS OR RED CELL CONCENTRATES WITH UNKNOWN HEMATOCRIT VALUES

Centrifuge the stored whole blood, additive-preserved red blood cells or red cell concentrates with unknown hematocrit values at 1615 X g for 4 minutes. Be sure the brake on the centrifuge is turned to the OFF position (no brake). Remove all the supernatant to an integrally attached transfer pack. Store the red blood cells in the original collection bag at 4 C for 3 to 6 days (inclusive) prior to freezing.

Ideally, a non-anticoagulated blood sample for serum should be collected from the venipuncture site after the unit of whole blood is collected. The serum is transferred to four (4) 2 ml polyethylene cryogenic vials for freezing and future testing.

TABLE 1

SPEED AND LENGTH OF TIME FOR PROCESSING BLOOD COMPONENTS

1. Fresh whole blood spun at 1615 X g for 4 minutes in a 22 C refrigerate centrifuge (no brake) to prepare platelet-rich plasma (PRP) and a red blood cell concentrate with a hematocrit value of 75 + 5 V%.

2. Liquid preserved whole blood, additive-preserved red blood cells, or red cell concentrates with unknown hematocrit values, spun at 1615 X g for 4 minutes in a 22 C refrigerated centrifuge (no brake) to prepare a red blood cell concentrate with a hematocrit value of 75 + 5 V%.

3. Glycerolized red cells spun at 1248 X g for 10 minutes in a 22 C refrigerated centrifuge (no brake) to prepare a glycerolized red blood cell concentrate with a hematocrit value of 60 + 5 V%.

CALCULATIONS

RCF = 28.38 X R X (RPM/1000)2

OR

RCF = RELATIVE CENTRIFUGAL FORCE (X g)
R = RADIUS IN INCHES
RPM = REVOLUTIONS PER MINUTE

     
RPM REQUIRED FOR

CENTRIFUGE

ROTOR

RADIUS

1615 X g

1248 X g

SORVALL RC-3BFOR RC-3C

HG-4L

9.09 inches

2500

2200

H4000

9.09 inches

2500

2200

H6000A

10.25 inches

2356

2070

BECKMAN J6-B

JS5.2

8.90 inches

2527

2220

JS4.2

10.05 inches

2380

2093

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