. Index Need for separation Types of blood components Equipments Precautions Preparation of — Packed red cells — Platelet rich plasma — Fresh frozen plasmaNeed for separation * Optimal survival of each constituents. — In whole blood stored at 2-6°C, platelets stay viable for 1 day and factor V & VIII decrease. — While after separation, platelets stay upto 5 days and factor V and VIII can be stored as FFP for 1 yr at -30°C * Transfusion of only required component thus avoids the use of unnecessary component which could be contraindicated in a patient. * One donor can save several patients.Blood components Cellular Plasma Plasma derivatives PC, Platelets, others FER; Cryoprecipitate Albumin 5%, 25%, Factor VIII, coagulation factorsEquipments Freezer: — 40°C and -70°C Blood bank Refrigerator 2-6°C Refrigerated centrifuge with swing out head and oval cups Laminar flow Weighing scale Dielectric sealer or aluminium clips & sealer Stripper Water bath 37°C or plasma defroster (Microwave) Platelet agitator Sterile docking devicePrecautions A. In collection of blood: I 2 3. 4. 5 6 7. Proper selection of donor . Clean aseptic site of venipuncture Uninterrupted flow of blood Collect correct amount of blood . Automatic mixing of blood during collection . Separate platelets within 6-8 hours triple pack and quad pack systemPrecautions B. In centrifugation 1. 2. Balancing & use of rubber discs Broad side of the bag faces the outside wall of the cup . Correct speed and time to be maintained . Look for any abnormal vibrationWhole Blood * 450 ml +/- 45 ml or350 ml +/- 35 ml of donor blood and anticoagulant solution * Minimum 70% of transfused red cells should survive in the recipient’s circulation 24 hours after transfusion. * No functional platelets and no labile coagulation factors V and VIIIA unit of blood collected from a single donor ina sterile container with an appropriate anticoagulant.(CPD-A1) Shelf life; 35 days at 2-6 C° ¢Volume ; 450ml +- 10% *One unit whole blood contain ~ 200ml RBC e|ndications ; —Massive transfusion —Exchange transfusion *Must be ABO identical , cross match compatiblePreparation of Packed Red Cells * Red cells have a higher specific gravity than plasma, thus settle in the lower portion of the bag. * Types of preparation: — Sediment red cells: PCV- 60-70%; 30% plasma and all original leukocytes and platelets. — Centrigued red cells: PCV- 70-80%; 15% — Red cells with additive (Adsol or SAG-M): PCV -50- 60%; minimum plasma and all leukocytes and platelets.A unit of RBC prepared after removing 2/3 of plasma *Volume; 250 - 300 ml Shelf life; 35 days (CPDA1) «Storage temperature; 2-6 C° *Indications ; oSymptamatic anaemia oBleeding during surgery oSevere haemorrhage for patients with surgical, complications otrauma ocancer * Must be ABO compatible and cross match compatible #1 Unit raises ~ 1.5 g/dL of Hb in an adultSteps in separation * Store the bag at 2-6°C till processed } * Place bag in buckets of refrigerated centrifuge and balance the opposite bags * Centrifuge at 5000 x g for S mins at 2-6°C Express approx 3/4'” of the plasma in the satellite bag Keep red cell at 2-6°C and plasma at -30°CRCC Quality Indicators #Volume 250 — 300 mL *Haematocrit 55-75 % Haemoglobin >45 g/dL *pH6.4—7.4 *Haemolysis < 0.8 % Sterility Sterile *Residual leukocyte count < 2x109/UnitLeukocyte-reduced Blood Components * Leukocytes can cause: — NHFTR — HLA alloimmunization — CMV, EBV, HTLV-1 —TR-ALI — TR- GVHD — TR-Immunosuppresion — Cytokines produced even in stored bagsPreparation of Platelet Concentrate * Random donor platelet — prepared from 450ml of whole blood * Single donor platelet — prepared by apheresisRandom donor platelet concentrate, prepared from one unit of blood collected from one donor *Volume 50ml *Storage temperature 22 - 24 C° (Agitation) Shelf life 3-5 days *Adult Dose 1 unit /10kg (4-6 units) *Peads Dose 10-15 ml/Kg *ABO & Rh compatible *Cross match not necessary Manual unit raises 5000 — 10,000 PLT in an adult* Collect in triple pack system and store at room temperature at 20-22°C for less than 6 hours. Place bag in buckets of refrigerated centrifuge and balance the opposite bags * Centrifuge at 2000 x g for 3 mins at 20-24°C * Express approx 4/5" of the platelet rich plasma in the satellite bag | * Centrifuge the bag with PRP and another satellite bag at 20-40°C at 5000 xg for 5 mins. Express supernatant platelet poor plasma into another empty bag and leave approx 50 ml of plasma. * Store platelet at 20-22°C under constant agitation,Precautions * Agitation during storage helps in: — Exchange of gases — Maintenance of pH — Reduce formation of platelet aggregates * pH should never fall below 6 because: — Change in shape of platelets from disc to sphere — Pseudopod formation — Release of platelet granulesPLT Conc. Quality Indicators Platelet concentrate #Volume 45 — 55 mL *Swirling Present Platelet count >55x109/Unit Residual leukocyte count <1x109/Unit *Residual red cell count <1x109/Unit pH 6.4-7.4 *Sterility SterileFresh Frozen Plasma (FFP) Plasma removed from cells within 6-8 hours of collection is rapidly frozen to below —30C° temperature. *Before transfusion it is necessary thaw at 37 C° *Once thawed there is rapid deterioration of clotting factors, so use immediately after thawing *Volume 200m! *Shelf life 1 year *Storage temp —30C° *Dose 12 - 15 ml/kg Indications; —Single or multiple clotting factor deficiency —Severe liver disease —Massive transfusion =dICPreparation of fresh frozen plasma * Collect in triple pack system and store at 4°C or in air-conditioned room till | processed but not for more than 6-8 hours. Place bag in buckets of refrigerated centrifuge and balance the opposite bags vgs) © Centrifuge at 5000 xg for 5 mins at 4°C * Express approx 4/5" of the plasma in the satellite bag | * Label the plasma bag and is rapidly frozen within 6-8 hours and the freezing process should not be more than 1 hour at -70°C * Can be stored at -30°C or below for 1 year. * Thaw it in a plasma defroster (Microwave) or place ina plastic over wrap and put it in a 37°C circulating water bath.FFP Quality Indicators Fresh Frozen Plasma —Volume 180 — 200 mL —Factor VIII level > 0.7 IU/mL —Fibrinogen level >300 mg/Unit Sterility SterilePrecautions * During blood collection: — Flow should be continuous — 450ml bag should not take more than 8 mins to fill * During processing: — Freezing the plasma as soon as it is separated — Maintaining the storage temperature * During thawing: — The entry ports of the bag should remain above the water — Should be administered within 12 hours if kept at 2- 6°C.Donor bed i T I= = ;*Blood bagCollected blood bag (0 NatBlood samples collected in appropriate tubes for testingStripping and discarding of needlePrimary bag with satellite bagscryocentrifuge _expressorPlatelet agitator _Thank You