this presentation tells you about how different components of blood are separated in blood bank for their maximum use to patients.

1. PRESENTED BY :-
MANTHAN
NIPS ,PGIMER

2.  In the past whole blood was the only preparation
 But it cause unnecessary administration of unwanted
cells or plasma constituents.
 A significant advance in transfusion medicine was
made when techniques became available for
separation of blood in a closed system and patient
could be administered specific replacement therapy.
 Hence whole blood can be utilized for preparation
of different components and thus can benefit
multiple patients.

3. This presentation will enable participants to
• Understand the basic principles and
procedure of Component Separation
• Know the different components that can
be prepared in a blood bank

4.  1926-THE BRITISH RED CROSS instituted the
firsthuman blood transfusion service in the
world.
 1940-the freeze dried plasma was developed.
 1950-Glycerol cryoprotectant for freezing red
blood cells.
 1950-Carl walter and W.P.Murphy introduced
the plastic bags for whole blood.
 1979-A new anticoagulant preservative CPDA-
1,which extend shelf life of whole and red
blood cells to35 days is introduced.

5.  In a statistical study it is seen that total no
of blood donation in
West Bengal- 713535 (Vol. 85.71%)
Bihar – 47863 (Vol. 22.74%)
Jharkhand --- 73238 (Vol. 33.13%)
Uttar Pradesh- 394699 (Vol. 17.3%)
Maharashtra- 377110 (Vol. 86.36%)

6. FULFIL CRITERIA
 Hb ≥ 12.5 gm/dl
 Weight
- 45 kg (350 ml)
- 55 kg (450 ml)
 No Aspirin < 3 days
 Single bold venipuncture, free flow of blood
 Collection - time < 10 minutes with frequent mixing

9.  Weighing balance
 Two pan balance
 Refrigerated centrifuge
 Laminar air flow bench
 Deep freezers (-40,-80oC)
 Platelet shaker/ incubator
 Refrigerated water bath
 Plasma expressor
 Tube sealer
Additional
 Sterile tubing welder
 Gamma irradiator
 Cell separator
(apheresis)

10. STANDARD
 Whole blood
 Packed Red Cells
 Platelets
PRP, RDP, SDP
 Fresh Frozen Plasma
(FFP)
 Cryoprecipitate
SPECIALIZED
 Saline-washed Red Cells
 Frozen Red Cells
 Leucodepleted products
 Irradiated products

11. Plasma Expresser

12. Dielectric Sealer

13. Multiple integratedblood bags

14. Quadruple Blood Bags

15. Whole blood
Red cells Plasma Platelets
(Fresh) frozen
plasma (F(FP)
Cryoprecipitate Cryo
supernatant
plasma (CSP)
F lX*
Immune
Globulin
Albumin
Fractionated
products
F Vlla*
F Vlll*
Granulocytes
* Now available as recombinant products

16. Preparation protocol
Counter
balancing
Weighing
Centrifugation
Expression

17. Centrifugation
Principle
Sediment of blood cells depend
on their size as well as the difference
of their density from that of the
surrounding fluid, viscosity of
medium, flexibility of the cells which
are temperature dependent

19. It is an old time, crude but cheaper method to separate
plasma from whole blood
 Steps
1. blood collected in a double or triple bag system.
2. Blood is kept hanging overnight at least for 12-16 hours.
There is clear plasma above and packed red cells below.
3. The supernatant plasma is expressed into the satellite
bag, leaving behind, 80-100 ml of plasma.
4. Plasma thus collected is called single donor plasma and
can be stored for 24-26 days at 4 - 6°C or for 1 year at
-20°C or below.

20. (b) Low speed refrigerated centrifugation:-
It is used for preparing Platelet Rich Plasma (PRP)
(c) High speed refrigerated centrifugation:-
It is used for preparing Platelet Concentrate (PC), Fresh
Frozen Plasma (FFP) and Cryoprecipitate (CP).
Effect of centrifugation
•High speed of centrifugation
causes much trauma to cells and
also the breakage of plastic
container.
•Second hard spin in PRP-PC
method causes platelet activation
as they are forced against the wall
of the container.

21.  450 ml of blood
 63 ml of anticoagulant solution.
 Hct-36-44%
 No components have been removed.
 Store at 1-6 oC
 Shelf life-
 Citrate-Phophate-Dextrose (CPD) - 21 days
 CPDA-1 (adenine) - 35 days
 SAG-M – 42 days
 Administer through standard blood filter (150-280
micron)
 Infuse within 4 hours of issue

22.  Drawbacks:
› After storage for >24 hours, platelets and WBC are non-
functional
› Factor V and VIII decrease with storage
› Fluid overload
 Indications:
› Acute blood loss > 25% TBV
› Exchange transfusion
Contraindication
- Risk of volume overload : Chronic anemia
Incipient cardiac failure

23. units with red blood cells and some
plasma
- with Anticoagulant ACD / CPD / CPDA
– 1
- Hct ~ 75 – 80 %

24. Principle:
RBCs are obtained by removal of supernatant plasma from
centrifuged whole blood.
Preparation:
 Centrifuge whole blood unit in refrigerated centrifuge containing the
parameters RPM-3850,Time-5 Min, Temp- 40C
 Express the supernatant plasma with the help of plasma expressor.
 Double seal the tubing between the primary and satellite bag.
 Check that the satellite bag has the same donor number as that on the
primary bag and cut the tubing between the two seal.
Advantages:
 Oxygen carrying capacity equal to that of whole blood in half the
volume.
 Significantly decrease levels of isoagglutinins, metabolites and
electrolytes.

25. Shelf life: (If CPDA1 anticoagulant) -35 days
Storage temp. : 40C (Range is 2-60C)
QC Requirements: PCV 80% (Range is 65-80%)
Volume: 250- 300 ml.
CONTENTS: Red cells- 65-80%
Plasma – 20-35%
Some platelets, white cells storage lesion by
products and anticoagulant preservative solutions.
Transfusion Criteria's: ABO/Rh specific and compatible
Indications: Restore oxygen carrying capacity symptomatic
anemia and surgical blood loss.
Effect: 1 unit RBCs should raise HCT -3%, Hb 1 g/dl

26. Since 1978 citrate-phosphate-dextrose with
adenine (CPDA-1) is used as blood
preservative for 35 days at 2-40C.

27. Action of ingredients of anticoagulant solution.
Citrate
Prevents coagulation by
chelating calcium
Sodium di-
phospate
Prevents fall in pH
Glucose
Supports ATP generation
by glycolytic pathways
Adenine
Synthesizes ATP, increases
level of ATP, extends the
self life of RBC to 42
days.

28. Action of ingredients of anticoagulant solution.
- Blood pH on day of collection is 7.5 and on 35th
day become 6.84.
- A fall in pH in the stored blood results in a
decrease in red cell 2, 3-DPG level, which
results in increase in hemoglobin-oxygen
affinity. CPDA-1 maintains adequate
levels of 2,3-DPG for 10 -14 days.
- During storage Na+ and K+ leak through the
red cell membrane rapidly. K+ loss is
greater than Na+ gain during storage.

29. Centrifugation method
- easiest and least cost
- least efficient
- reduce WBC only 70 - 80%
- reduce RBC volume ~ 20%
Leukocyte poor red blood cells

30. 30
Filtration method
easy, quick, but more expensive
high efficient
remove WBC more than 99.9%
( third generation )
little loss of RBC volume
Leukocyte depleted red blood cells

31. Indication
- Minimizes white cell immunization in patients
- Prevention of FNHTR (Febrile Non-Hemolytic Transfusion
Reaction )
- Reduces risk of CMV transfusion
Contraindication
- Not prevent graft –vs- host disease
Dosage
- same as Packed Red Cell
Administration
- same as Whole Blood

32.  Obtained by apheresis from family
members for administration to cancer
patients.
 Contain 1.0 x 1010 granulocytes
 Pre-treatment with recombinant G-CSF and
dexamethasone can yield 4-8 x 1010
granulocytes
 Stored at 24o C
 Infuse within 24 hours of collection

33.  ANC <500
 Fever
 Documented infection (bacterial or
fungal) for 24-48 hours
 Unresponsive to appropriate antibiotics
 Reasonable hope of marrow recovery

34. It is prepared from the whole blood within six-hours of collection,
preferably stored at room temperature of 20-24°C.

35. 1. The blood is collected in-CPDA-1 double or triple bag
system.
2. The blood bags are weighed on a weighing balance and
bags weighing equal are placed opposite to each other in
the buckets of centrifuge.
3. Temperature of the centrifuge is adjusted between 20 -
24°C
4. The speed of the centrifuge is calculated according to the
radius of the arm of the centrifuge rotor.
5. The calculated speed for platelet preparation is 1750 rpm for
11 min. in PGI
6. After centrifugation, the bags are taken out from centrifuge
chamber with minimal disturbance and the PRP is expressed
from the primary bag into the satellite bag with the help of a
plasma expresser. After placing proper knots, labeling the
blood group and ensuring the screening status the satellite
bag is detached from the primary bag.

36. 7. The PRP for storage can be kept at 22-24°C in
a, platelet incubator with constant agitation for
a maximum of 48-72.hours (at our centre in
PGI).
8. As a part of internal quality control 1% of the
random components units over a month are
tested for pH and yield. For PRP the pH should
always be > 6.2 and yield 4.5 x 1010 / bag (Drugs
and Cosmetics Act) and 5.5 x 1010/ bag (AABB
Technical Manual) and one bag of PRP
generally raises the platelet count in the
recipient by 5000-10000 / µl.
Note: Aspirin and related analgesics affect the
platelet function, so the donor for platelets is
accepted after 3 days of ingestion of these
drugs.

37. This supplies the same amount of platelets as PRP, but in lesser
volume (40 - 50ml).
Principle:
Platelets are harvested from whole blood following ‘light spin‘
centrifugation. The platelets are concentrated by 'heavy spin'
centrifugation with subsequent removal of supernatant plasma.
Steps
1. Blood is collected in the triple bag system only
2. PRP is prepared by following the above mentioned steps
3. After detaching the satellite bags from the primary bag, the PRP
in again spun at 4000 - 5000 rpm (high spin) for 4 to 5 minutes.
4. A platelet button is formed at bottom and platelet poor plasma
is expressed from the 1st satellite bag to the 2nd satellite bags,
leaving behind 40 - 50 ml plasma for platelet button suspension.
5. The platelet concentrate in stored at 20°C-24°C for a maximum
of 3 days (at PGI) with constant agitation.

38. Shelf life: 3 days in platelet incubator & agitator.
24 hrs if no storage cabinet
Storage temp.: 20°C - 24°C
Q.C. Requirements: To be prepared within 8 hrs after
collection, pH should be 6.2 or more at the
end of storage time. Platelet
count > 5.5 x 1010 /unit.
Volume: 30 to 50 ml
Contents: Platelet - 5.5 x 1010 /bag
Plasma - 30 to 50 ml and some white
cells
Transfusion Criteria: ABO / Rh specific and compatible
Indications: Severe thrombocytopenia, qualitative
platelet defects
Effect: Increases in platelet count 10,000 / ul per
unit

39.  Dosage
 1 unit of PC / 10 kg B.W.
 Increment will be less in
- Hypersplenism
- DIC
- Septicemia
1 unit of PC  Platelet 5000-10,000 / ul

40.  Indications
Treatment of bleeding due to
 Thrombocytopenia
 Platelet Dysfunction
 Prevention of bleeding
 Contraindication
prophylaxis of bleeding in surgical patients

41. › Fresh Frozen Plasma
› Frozen Plasma :- Aged plasma
› Cryoremoved plasma
› Cryoprecipitate

42. Fresh Frozen Plasma

43.  Plasma along with anticoagulant preservative
 Volume ~ 250-300 ml
 Prepared from blood within 8 hrs of donation
 Maximum level of labile and non-labile clotting factors
(about 1 IU per ml) V & VIII, proteins C and S,
complement, and immunoglobulin.
 Good for 24 hours post thaw
Then it can be stored for 5 days as liquid plasma
(labile factors V and VIII decreased)
 Shelf life: 1 year

44. It is prepared from the whole blood collected in a CPDA-1 double or triple bag
system within 6-8 hours of its collection.
Principle:
Plasma is separated from cellular blood elements and frozen to preserve the
activity of labile coagulation factors.
Steps
1. Blood is collected in a CPDA-1 double or triple bag system.
2. The blood bags are weighed in a weighing balance and bags weighing equal
are placed opposite to each other in the buckets of the centrifuge.
3. The temperature of the centrifuge is adjusted at 4 - 6°C.
4. The bags are subjected to a high spin (3850 rpm) centrifugation for 5 min.
5. After the centrifuge stops, the blood bags are taken out with minimal
disturbance and supernatant plasma is expressed into the satellite bag, with
the help of a plasma expressor, leaving behind 80-90ml.
6. After labeling the group and ensuring screening status, the plasma is stored
at -20°C or below. The storage shelf life is one year at -20°C and 5 years at -70
to-80°C

45. Fresh Frozen Plasma contains coagulation factors and other
plasma protein (per unit or bag)
Volume - 200-250 ml
Factor VIII - 0.6 IU / ml
Factor IX - 0.9 IU / ml
Fibrinogen - 250-300 mg / bag
Proteins - Albumin, globulin, etc.
1 IU / kg of factor VIII or factor IX raises the factor VIII levels in
plasma by 2% and factor IX levels by 1% respectively.
Shelf life: One year
Storage temp.: -20°C or below
Q. C, Requirements: The entire proems of preparation and
freezing should be completed within 8 hrs after collection.
Volume: 50 to 200 ml

46.  Clinically significant deficiency of Factors II, V, X, XI
 DIC
 Plasma exchange
 Immunodeficiencies
 Massive transfusion of stored blood.
 Liver disease
 Urgent reversal of warfarin therapy
 Correction of known coagulation factor deficiencies for
which specific concentrates are unavailable
 Correction of microvascular bleeding in the presence of
elevated (> 1.5 times normal) PT or PTT
 Correction of microvascular bleeding secondary to
coagulation factor deficiency in patients transfused with
more than one blood volume and when PT and PTT cannot
be obtained in a timely fashion

47.  Dose
› 10-15 ml/kg B.W
› For warfarin reversal, 5-8 ml/kg of FFP
 Contraindication
› Volume expansion
› Immunoglobulin replacement
› Nutritional support
› Wound healing

48.  Precaution
› Acute allergic reaction are common
› Anaphylactic reaction may occur
› Hypovolemia alone is not an indication for use
 Dosage
Initial dose of 15 - 20 ml / kg B.W
 Administration
› Must be ABO compatible
› Infuse as soon as possible after thawing (
within 6 hrs )
› using standard blood administration set

49.  Plasma which separate from whole blood at
any time during storage.
 Contain all non-labile coagulation factors.
 Indication
 Treatment of stable coagulation deficiencies
 Contraindication
 same as FFP

50. Cryoprecipitate is the cold – insoluble portion of plasma that precipitates when
FFP is thawed between 1-60
C

51.  Cryoprecipitate contains precipitated proteins of plasma,
rich in factor VIII and fibrinogen, obtained from FFP
prepared within 6-8 hours of collection, subsequent
thawing at 4- 60 C and removal of supernatant.
 Also the advantage of Cryoprecipitate is that we can
administer large amount of factor VIII without overloading
the recipient, especially in pediatric patients.
 Principle: Coagulation factor VIII can be concentrated by
cryoprecipitation of freshly collected plasma.
Cryoprecipitation is accomplished by rapid freezing of
plasma and slow thawing at low temperature.

52.  Cryoprecipitate contains (1 unit)
Volume - 10-20 ml
Factor VIII-C - 80-120 IU
Factor VIII R: Ag - high levels
Factor VIII vWF - high level
Fibrinogen - 150-200mg / bag
Factor XIII - 20-30% of original level

53. Shelf life: Frozen - 1 year
Thawed - 6 hours
Storage temp: Frozen - -20°C or less
Q. C. Requirements: Thaw at 37°C
Factor VIII: C-80 units/bag
Volume: 10 to 20'ml
Contents: Factor VIII: C - 80 to 150 units/bag
Fibrinogen - 150 to 250 mg/bag
Factor Xlll - 20 - 30% of whole blood
von Willebrand factor - 40-70% of whole
blood
Transfusion Criteria:ABO compatibility not required
Indications: Correction of factor VIII deficiency
(Hemophilia A, von Willebrand disease).

54.  Indication
› Quantitative and Qualitative Fibrinogen
Deficiency : DIC
› von Willebrand Disease
› Factor XIII deficiency
› Uremic Coagulopathy
› Fibrin Glue
› Factor VIII ( haemophilia A )

55.  Administration
› Dose of Cryo is based on the desired target level
of the specific factor to be replaced
› ABO compatible if possible no compatibility
testing required
› After thawing & pooling, infuse as soon as
possible through blood admin. Set
› must be infused within 6 hours of thawing

56. Heavy spin,4o
C(within 8 hrs)
Fresh Whole Blood
Packed Red Cells
Stored in 1- 6o
C
Fresh Plasma
Freeze -80o
C immediately
Stored at < -18o
C

57. Fresh Whole Blood
Packed Red Cells
Light spin, 22o
C(within 8 hrs)
Platelet Rich Plasma
Platelet Concentrate Fresh Plasma
Store at 22o
C Freeze(FFP)
Heavy spin,22o
C

58. Thaw at 4o
C & heavy spin
Fresh Frozen Plasma
Cryoprecipitate
-Refrozen within 1 hr
-Store at < - 18
o
C
Cryoremoved Plasma
Freeze -80o
C immediately
Stored at < -18o
C

59.  Red blood cells: 1-6°C
 Platelets: 22-24°C, with continuous agitation
 Plasma:
FFP:  -18°C (after thawing ~ 1-6°C for 24 hours)
FP:  -18°C (after thawing ~ 1-6°C for 24 hours)
CSP:  -18°C (after thawing ~ 1-6°C for 24 hours)
 Cryoprecipitate:  -18°C (after thawing ~ 20°C for 4 hours)

60.  Red cells: 42 days, collected in
CP2D/AS-3 35 days,
collected in CPDA-1
 Platelets: 5 days with continuous
agitation
 Cryo: 12 months at -18°C or
4 hours after thawing
 Plasma: 12 months at -18°C or
(FFP/FP/CSP) 24 hours after
thawing

61.  The word apheresis is derived from a
Greek word which means separation.
Apheresis also known as hemapheresis is
removal of whole blood from donor /
patient, separation into components,
retention of desired or unwanted
components and reinfusion of remaining
constituents to the donor or patient.

63.  Citrate toxicity may occur in the form of
numbness and tingling sensation around
the mouth if the amount of citrate
infused exceeds the body's ability to
metabolize it. This problem can be
solved by decreasing the rate Of infusion
of anti-coagulant or by giving
exogenous calcium to the donor.
 The other side effects are similar to that
of normal blood donation.