SECTION: BLOOD BANK

COPPER SULFATE HGB TEST

 RBCs specific gravity = 1.0971
 Copper Sulfate = 1.053
 Principle:
- A drop of blood, allowed to fall into a copper sulfate solution of specific
gravity 1.053, becomes encased in a sac of copper proteinate, which
prevents dispersion of fluid for 15 seconds but not later than 2 mins
- If specific gravity of blood is higher than the solution, the drop will sink or
else it will remain suspended for some time.

>1.053 = 12.5 mg/dL Hgb

<1.053 = <12.5 mg/dL Hgb (Anemia)

SELECTION OF METHODOLOGY
 Weigh processed blood components to ensure they met specified volume
requirement.
 WB – 405ml to 495ml in 63ml anticoagulant
 PRBC – 230ml to 330ml
 PC – 50ml to 70ml
 FFP – 150ml to 250ml
 CRYO – less than or equal to 15ml
 CRYOSUP – 190ml to 210ml

 Terumo Blood Bag

Mother Bag – AG = 42g
Satellite Bag = 27g
 Fresenius Kabi
Mother Bag – AG = 44g
Satellite Bag = 32g

LABELLING
 Checking system
 Barcode Label
 Segment Number
 MBD Code
STORAGE AND TRANSPORT CONDITIONS:

WHOLE BLOOD AND RED CELLLS

CONDITION TEMPERATURE RANGE STORAGE TIME

Transport: Pre – Processed +20C to +24C Less than 8 hours
Storage: Pre/Post-
+2C to +6C Approx. 35 days
Processing
Transport: Processed Blood +2C to +10C Less than 24 hours

PLATELET CONCENTRATE

Required Temperature:
+20C to +24C
Processing to start within 8 hours of donation
Constant agitation during storage
Shelf life: 5 days

FRESH FROZEN PLASMA (FFP) AND CRYOPRECITATE

Separated from whole blood within 6 to 8 hours of collection
Rapidly Frozen

STORAGE IN RELATION TO TEMPERATURE

Product Storage Temperature Maximum Storage Life

FFP -65C or colder 7 years
FFP or Cryo -40C to -64C 2 years
FFP or Cryo -30C to -39C 1 year
FFP or Cryo -25C to -29C 6 months
FFP or Cryo -20C to -24C 3 months
WHOLE BLOOD
 Life span and storage
o CPD-A1: 35 days at +2C to +6C
o SAG-M: 42 days at +2C to +6C
 Volume: 450ml  10% volume excluding the anticoagulant (405-495 mL)
 There should be no hemolysis: (upon expiry: <0.8% of red cell mass)
 Frequency: 1% of all units collected with a min. of 4 units/month

PLATELET CONCENTRATE
 Frequency: 1% of all units collected with a min. of 4 units/month
 Note: Platelet concentrates must demonstrate “Platelet ribbon” & “Swirling
phenomenon”

Blood unit/s collected more than 15 minutes should not be processed into
components other than PRBC

How to do QC of Platelet Count:

1. Strip pilot tube and mix platelet concentrate or knead the platelet concentrate
to get a good representative sample of the unit
2. Seal about 1-2 inch. Cut pilot tube and place sample in a clean test tube
3. Run sample in Hema Analyzer
4. Get the platelet count and compute

Computation:
Platelet ct. of PC = Platelet ct. (expressed in 10^9/L) x Total volume of Platelet
Concentrate in Liter

Example:
 Platelet count = 1050 x 10^9/L
 Volume of PC = 60 mL = .060 L

Platelet Count of PC = 1.050 x .060

= 63 x 10^9/L or 6.3 x 10^10/L

PLATELET APHERESIS
 Volume: 300ml
 Content:
 Platelet: 3.0 x 10 11
 Plasma: 300ml
  WBC
 Expiration:
 Closed System: 5 days
 Open System: 4 hours
 Dosage Effect: 30,000 – 60,000 per unit

FRESH FROZEN PLASMA

 Life span and storage: -30C or colder for 12 months
 Volume: 150-250ml
 Content: All coagulation factors
: 400mg fibrinogen
 Factor VIIIc: 70% of the value of the freshly collected plasma unit

FRESH COMPONENTS STORAGE TIME

STORAGE TEMPERATURE MAXIMUM STORAGE TIME

-65C or below 7 years
-40C to -64C 24 months
-30C to -39C 12 months
-25C to -29C 6 months
-20C to -24C 3 months
BLOOD TYPING
(3-5% RCS Preparation)

Prepare RCS
 1,000 uL NSS + 50 uL PRBC of Patient
 Mix

TEST PROCEDURE:
1. Dispense 10 uL of Patient’s RCS on A, B, D and control wells
2. Dispense 10 uL of A1 and B cells + 40 uL Patient’s plasma
3. Centrifuge for 5 mins. (No incubation required)

COOMB’S TEST

Prepare RCS
 1,000 uL NSS + 50 uL PRBC of Patient
 Mix

TEST PROCEDURE:
1. 50 uL Surgiscreen cells (I,II,III) + 40 uL Patient’s Plasma (IAT)
2. 10 uL Patient’s RCS + 40 uL Patient’s plasma + 50 uL BLISS (Autocontrol)
3. Dispense 10 uL of Patient’s RCS only (No incubation required) (DAT)
4. Incubate for 15mins.
5. Centrifuge for 5 mins.

REMINDERS:
! Examine cassettes for discrepancies before use
! Handle BLISS careful to avoid contamination
! No need for centrifugation of newly delivered cassettes
! Put red cell reagents in refrigerator when not in use

WORKING PROCEDURES IN ORTHO WORKSTATION

CROSSMATCHING
(3-5% RCS Preparation)

Prepare RCS
 1,000 uL NSS + 50 uL PRBC of Donor
  Mix

TEST PROCEDURE:
1. 10 uL Donor’s RCS + 40 uL Patient’s Plasma + 50 uL BLISS (MAJOR)
2. 10 uL Patient’s RCS + 40 uL Donor’s Plasma + 50 uL BLISS (MINOR)
3. Incubate for 15 mins. (if along with AB screening, do 15 mins.)
4. Centrifuge for 5 mins.

AUTOCONTROL

Prepare RCS
 1,000 uL NSS + 50 uL PRBC of Patient
 Mix

TEST PROCEDURE:
1. 10 uL Patient’s RCS + 40 uL of Patient’s plasma + 50 uL BLISS
2. Incubate for 15 mins. (if along with Ab screening, do 15 mins.)
3. Centrifuge for 5 mins.

ANTIBODY SCREENING
(0.8% PREP)

TEST PROCEDURE:
1. 50 uL Surgiscreen cells (I,II,III) + 40 uL Patient’s Plasma
2. Incubate for 15 mins.
3. Centrifuge for 5 mins.

DONOR’S ANTIBODY SCREEN PROCEDURE

(USE ONLY 1 WELL)

1. 25uL (I) + 25uL (III) SURGISCREEN

CELLS
2. Add 40uL DONOR’S PLASMA
3. INCUBATE for 15 mins.
4. CENTRIFUGE for 5 mins.