METHODS OF COMPONENT SEPARATION INCLUDING TIME RESTRICTIONS

Donors taking anti-platelet drugs (e.g. Aspirin), donors who have consumed alcohol during the preceding 12 hours and donors who give history of bleeding tendencies should NOT be selected for preparation of Platelet Concentrates
Donors selected for component making should be given a serial number. This is done after consulting Blood Donor Register and giving the next serial Number. Write this number on (1) Donor Form; (2) Blood Bags, including all satellite bags; and (3) two clean glass tubes selected as 'Pilot Tubes'
If phlebotomy is traumatic or if blood collection takes longer than 10 minutes, record this on the donor form.
If phlebotomy is traumatic or if blood collection takes longer than 10 minutes, record this on the donor form Collection bags from which Platelet Concentrates are to be prepared should NOT be refrigerated. Keep such bags between 20 and 24 degrees Celsius (either in the platelet incubator or in a room where air-conditioner is on)

Before starting component separation make appropriate entries in the "Component Log Book". Make sure that component separation is completed with six hours after blood collection.

A. When blood is collected in a "double bag system" the two bags are to be separated into (1) Whole Blood and (2) Platelet Concentrate (PLC) Use the refrigerated centrifuge machine after balancing the opposing buckets carefully.
Temp. r.p.m. r.c.f. Time (min)

PLATELET CONCENTRATES
 
A. When blood is collected in a "double bag system" the two bags are to be separated into (1) Whole Blood and (2) Platelet Concentrate (PLC)
Use the refrigerated centrifuge machine after balancing the opposing buckets carefully.
Temp. rpm rcf time (min)

FIRST SPIN : 22 1219 321 20

[The main bag has RBCs at the bottom and Platelet-Rich-Plasma (PRP) above]
Break the connecting seal between the main bag and satellite bag after placing the main bag on the 'plasma extractor'. Place the satellite bag on weighing balance. Release the "presser" part of the extractor and allow supernatant plasma (PRP) to drain into the satellite bag. Stop this process if red cells start entering the connecting tube. Use two plastic clamps to shut the connection between the bags. Keep both bags in the refrigerated centrifuge machine for second spin.

SECOND SPIN : Temp.: 22; r.p.m.: 3600; r.c.f.: 2797; Time: 12 min.

[The satellite bag has, now, platelets (with few RBCs) at the bottom and a clear supernatant plasma]
After the second spin keep the satellite bag on the extractor, main bag on the table, release the clamps on the tubing, allow the presser to force supernatant plasma back into the main bag till only about 50 ml is left in the satellite bag and stop draining put a clamp, use tube sealer and RECHECK serial numbers on both bags before separating the two bags.
Store main bag in the refrigerator and allow the satellite bag (containing Platelet Concentrate: PLC) to remain standing for 1 hour, at the end of which keep the bag on platelet agitator.
B. When blood is collected in a "triple bag system" the three bags are to be separated into
(1) RBC Concentrate (Packed Cells), (2) Platelet Concentrates (PLC) and (3) Fresh-Frozen-Plasma (FFP).

FIRST SPIN : as above
After the first spin open the connection between the main bag and the first satellite bag. Keep the main bag on the plasma extractor and allow PRP to drain into the first satellite bag. Seal the connecting tubing at least in three places and separate the main bag from the two satellite bags.
The main collecting bag now contains RBC Concentrate. Keep this in the "Quarantine Fridge" until all "quality control tests" are completed.
Keep the remaining "two bag system" in the refrigerated centrifuge machine after carefully balancing the opposite buckets for the second spin.
SECOND SPIN : as above.
[The first satellite bag has now, platelets (with a few RBCs) at the bottom and a clear supernatant plasma] Open the connection between the second and the third satellite bags. Keep the second bag on the plasma extractor and drain the supernatant plasma into the third bag, leaving about 50 ml in the second bag for later suspending the platelets. Seal the connecting tubes at least at three places and separate the bags after ensuring that both the bags are properly numbered for identification. The third bag containing clear plasma is to be kept in the deep freezer (having minus 30 degrees temperature).The second bag containing PLC is to be left standing for 1 hour and then kept on "platelet agitator".

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