• Blood is drawn from volunteer Blood donors and is
given to places such as hospitals and community Blood
• Most of the whole Blood collected is separated, before
it is sold, into Blood products (Blood components)
including red Blood cells, Blood platelets, Blood plasma
and other human Blood clotting agents.
• Blood products are transfused into patients needing
just that particular part (fraction) of the whole Blood
that is donated.
• A number of Blood donors also donate platelets by
apheresis (a procedure in which Blood is drawn from a
Blood donor and separated into its components, some of
which are retained, such as plasma or platelets, and the
remainder of the Blood is returned, by transfusion, to the
Blood donor; also called hemapheresis).
• Rather than using whole Blood transfusions, doctors are
increasingly using individual Blood components such as red
cells, white cells, platelets, and plasma. Plasma is
fractionated into an increasing number of Blood products,
including albumin, gamma globulins and clotting factors .
6. Blood components advantages
Whole Blood is a more likely carrier of transfusion transmitted
Keeping in mind the frequency of serious shortages of quality
Blood, it is considered imprudent to use whole Blood.
Most patients require only one particular component of whole
Blood. Better patient management is achieved by giving only the
desired and/or essential component.
Utilizing normal blood storage techniques, blood products have a
greater shelf life than whole Blood.
Blood filtration and other techniques help to make Blood safer; and,
Blood products can often be infused regardless of ABO Blood group.
8. Blood donors
• Age range of 18 to 60 years - in good health.
• not have undergone major surgery within 6 months of
• blood containing not less than 12.5% w/v of
haemoglobin. (13.3 % in case of male)
• if females, not be pregnant.
• The frequency of donations of whole blood shall not
exceed once every 3 months with a maximum volume
of 1.5 litres in any consecutive 12-month
9. Blood collection
• The blood is drawn aseptically through a closed system
into a suitable sterile container containing a specific
amount of Anticoagulant Citrate Dextrose Solution (ACD
Solution) or Anticoagulant Citrate Phosphate Dextrose
Solution (CPD Solution) which is placed before the
container is sterilised. The quantity of anticoagulant
solution should not exceed 22% v/v of the final volume of
the mixture. No antimicrobial preservative is added.
• With every container of blood, a separate sample mixed
with the appropriate quantity of anticoagulant solution, is
collected for compatibility and other tests; this small
container is firmly attached to the main container.
• Acid –Citrate- Dextrose (ACD)
– Sodium Acid citrate – 2.0 – 2.5g
– Dextrose - 3g
– Water for Inj - to 120 ml.
• Citrate prevents clotting by binding the calcium ions as unionised
• Trisodium citrate is not used now because of caramelisation of
dextrose during sterilisation and two solutions have to be
• Acid citrate produces a pH of about 5.- produces little or no
caramelisation – less likely to induce flaking of glass.
• Higher conc is preferred because it more effectively reduces the
formation of small clots.
• Dextrose delays hemolysis in –vitro and prolongs their life span
• Naturally occurring
• Made by the connective tissue surrounding blood
• Inhibits clotting in the circulatory system
• It is used in blood for transfusion when, large volumes
of blood must be given to one patient and
corresponding amounts of citrates would be harmful.
• It quickly loses its activity in vitro and normal
quantities are effective for about a day (ACD prolongs
the storage life to three weeks.)
• Expensive and continues its action after transfusion,
necessitating the administration of neutralising
substances such as potamine sulphate.
12. Disodium edetate
• Chelating agent that chelates divalent cation
• Preferred when preservation of blood platelets is
essential, although the stability of these seems to
depend much more on preventing contact with
glass surfaces; if plastic bags or silicon treated
bottles are used, ACD is almost as effective for
• The survival of RBC in dextrose – edetate
solutions in as good as in ACD.
• Two small additional samples are collected at the
time of withdrawal of blood.
• One is obtained by draining the withdrawal tube
– attached firmly with the main container for
testing compatibility with the recipient.
• When plastic bag is used, the collection tubing
itself is sealed blood filled, with special tool.
• Separate specimen – helps to prevent a possible
• Second larger sample: used as early as possible –
blood serological testing Grouping and Rh typing.
14. Blood products
• Whole Human Blood
• Conc Human RBC
• Fresh Human Plasma
• Dried Human Plasma
• Clotting factors
• Human fibrinogen & human thrombin
15. WHOLE HUMAN BLOOD I.P
• Whole Human Blood is blood drawn from
selected human donors and mixed with a
16. WHOLE HUMAN BLOOD I.P
• Description: Deep red fluid which, on standing,
separates into a lower layer of sedimented red cells
and a yellowish, almost clear, upper layer of plasma,
free from visible signs of haemolysis, with a greyish
layer between the two consisting of leucocytes and
• Storage: Store in colourless, transparent and sterile
containers into which it was originally drawn. The
containers should be provided with a hermetic,
contamination-proof closure. Store at a temperature
between 2° and 8°
• The label states
– The distinctive code number by reference to which the
details of the donor are available.
– The ABO group with the approved colour scheme for
different groups as specified by the appropriate national
• A yellow, AB white; B Red; O Light Blue
– The Rh group.
– The total volume of fluid, the proportion of blood, and the
nature and volume of anticoagulant solution.
– The date on which the blood was withdrawn.
– The expiry date which should not exceed 21 days from the
date of withdrawal of blood
18. WHOLE HUMAN BLOOD
• The blood is cooled to 4 to 6° immediately after withdrawal
from the donor and is maintained at that temperature
throughout its period of 21 days, except for short periods, not
exceeding 30 minutes, which are necessary during
transportation or testing.
• Even at this temperature
– The leucocytes disintegrate in few hours and the platelets
in few days.
– The RBC shows a fall in ATP and other organic phosphates,
a reduction in oxygen carrying capacity.
19. Blood Haemolysis
• Blood underwent haemolysis is unfit for use.
• Complete haemolysis especially if it occurs rapidly is
a symptom of bacterial infection.
• But the absence of haemolysis is not confirmation of
freedom from bacteria.
• Certain psychrophillic bacteria, especially
pseudomonas and members of coli aerogens group,
can grow in blood at refrigerater temperature,
without causing haemolysis.
• Many organisms that can contaminate blood can use
Citrate as its sole source of carbon and as expected
this can leads to clot formation.
22. How long can Blood Components
• Red cells: 35 days to maximum 42 days
• Platelets: 05 days with continuous agitation
• Plasma: 12 months at -18°C or 24 hours
23. Concentrated human red blood corpuscles B.P
• This product is made by removing most of the citrated plasma
from whole blood that is not more than a fortnight old and
has been allowed to stand or has been centrifuged to deposit
• The quantity of fluid removed must not less than 40% of the
total volume. Certain amount of plasma is left so that the cell
preparation is not too viscous for administration.
• The blood from which the concentrate is prepared meets the
pharmacopoeia requirements for whole human blood and
must not be more than 14 days old.
• The haemoglobin concentration of the final preparation is not
less than 15.5% w/v.
• Since there is a risk of bacterial contamination, the product
must be used within 12 hours.
24. Concentrated human red blood
• Storage: 4 – 6°C
• Blood from each donor contributing to the
concentrate must be grouped and
compatibility-tested against the blood of the
• Labeling: Similar to Whole human blood, but
include in addition a statement of the date
and time of removal of plasma.
• Uses: Principally in the treatment of anemia.
• Platelets are prepared from whole blood collected in
anticoagulant solution and filtered to remove
• Platelets are suspended in a small amount of the
original plasma. A unit contains at least 55 x 109
platelets suspended in 50-55 mL of plasma.
• Trace amounts of red blood cells can be present in
• Platelets may also be obtained by apheresis.
• The primary role of platelets is to prevent
bleeding of injured blood vessel walls by
forming an aggregate at the site of injury.
• Platelets also participate in blood coagulation,
inflammation and wound healing.
• The transfusion of platelets to a patient with
thrombocytopenia or bleeding should produce
a rise in the platelet count and control
27. Fresh frozen plasma
• Plasma prepared from whole human blood,
with in a few hours of its collection from the
donor stored in the frozen state, preferrably in
• Fresh frozen plasma contains a normal
concentration of fibrinogen and the labile
coagulation factors VIII and V.
• Used as a source of Clotting factor VIII.
• Fresh frozen plasma contains the clotting
factors that are necessary for hemostasis.
28. Frozen Plasma
• Frozen plasma contains stable coagulation
factors such as Factor IX and fibrinogen in
concentrations similar to FFP, but reduced
amounts of Factor V and VIII.
• The indications are the same as for FFP, except
that FP should not be used to treat
coagulation factor deficiencies of Factor V and
29. Dried human plasma
• Transfusion material with long life and stability
during storage. Can store up to 5 years.
• Blood older than 21 days not normally
considered suitable for transfusion, but
plasma proteins are more stable.
• Normal procedure is to aspire plasma from
sedimented red cells and to use it for
30. Dried human plasma
• The plasma of some donors contains haemagglutinating
antibodies of the ABO system which on transfusion into
patients of certain blood groups may cause intravascular
• This risk can be avoided by neutralisation of haemagglutinins
with soluble blood group substances present in the plasma of
other donors of appropriate blood groups.
• Plasma is pooled from donations of blood selected so that,
the normal distribution of ABO groups is represented; thus a
ten-donor pool contains plasma from about equal numbers of
A and O donations, together with atleast one B or AB
31. Dried human plasma
• Difficult to filter sterilize a large volume of plasma.
• Thus it is necessary to test each of the ten-donor
pool for sterility.
• Pools that have passed sterility tests can be
redistributed to transfusion bottles and can be freeze
• It is reconstituted to its original vol. with water for
inj.at R.T, and dissolves compleately in less than 10
minutes to give a cloudy soln.
• Protein content is not less than 45g/l
32. Dried human plasma
• Other than general requirements of label, it should state,
– The names and percentages of anticoagulants and other
– The quantity of water for injections required to
reconstitute to the original volume.
– The protein content of the reconstituted liquid
– The contents must not be used for more than 3 hours after
– Under dry conditions, below 20°, protected from light
• Uses and advantages compared to whole blood
– For restoration of plasma volume.
– It can be given to patients of any blood group.
– Properly stored it keeps well for five years.
33. Dried human serum, BP
• Blood collected as per the guidance for whole human blood,
without addition of anticoagulant is allowed to clot, the
seperated fluid pooled, bottled and freeze dried.
• The ratio of blood groups in the pool, and the pool size are as
for dried human plasma, and all other requirements are
identical for those for this product, with the exception that
protein content must be less than 65g/l, and the test for
coagulation with calcium chloride does not apply.
• Use is same as that of dried plasma, except that it cannot be
used as a source of fibrinogen.
34. • It was in 1666, Richard Lower carried out the first successful
blood transfusion between two animals. But it took another
150 years before blood transfusion between humans began.
• In the beginning of the 20th century Dr Karl Landsteiner
discovered four main blood groups; A, B, AB and O. This break
through was made just in time for the world War when blood
was used to save the lives of soldiers.
• "Dried plasma" was developed and first used in WWII. A large
project was begun in August of the year 1940 to collect blood
in New York City hospitals for the export of plasma to Britain.
Dr. Charles Drew was appointed medical supervisor of the
"Plasma for Britain" project.
35. • Dried plasma package
came in two tin cans. One
bottle contained enough
distilled water to
completely reconstitute the
dried plasma contained
within the other bottle. In
about three minutes, the
plasma would be ready to
use and could stay fresh for
around four hours.
36. Fractionation of plasma
• Second world war demanded a transfusion material with a long
life (Than blood) and more stability during storage (Unlike liquid
• E J Cohn and his colleagues in the USA investigate the separation
of Albumin from plasma.
• About 60% of the plasma protein is albumin.
• It plays an important role in the maintenance of high osmotic
pressure in the blood vessels.
• A very successful solvent precipitation technique was developed
by which, other proteins as well as albumin were separated from
• Some of them like such a fibrinogen, prothrombin and
gammaglobulin are found to be therapeutically useful.
37. An ideal fractionation method
• Must not alter biological properties of the
fractions, nor their solubilities.
• Must be possible to carry out Aseptically,
ideally the conditions discouraging bacterial
• Any additive must be harmless or easily
removable after the use.
38. Different possible approaches
• Salting out – not suitable for plasma due to
high concentration of proteins are needed.
• Dialysis is difficult to perform aseptically.
• Cohn’s technique is the use of an Organic
solvent (Alcohol) to reduce the solubility of
the proteins and was given flexibility by
alternations of pH and ionic strength.
39. Advantages of Solvent over salt as a
• Because of its volatility, it can be removed easily during freeze
drying of the final product.
• Salt can be used in lower concentrations to improve
• It helps to control contaminants because of its bacteriostatic
• Being a liquid, it is easy to add aseptically.
• On the other hand, it is necessary to keep the temperature
very low (0 to -5°C) to prevent solvent denaturation of the
• In some places like UK ether is preferred over alcohol, since it
is claimed to cause less protein denaturation.
41. Human fibrinogen
• Soluble constituent of plasma, that on addition of thrombin, is
converted to fibrin.
• After collection from plasma by fractionation, the precipitate is
collected by centrifugation. Dissolved in citrate saline, and freeze
• The air in the container is replaced by nitrogen.
• The citrate prevents spontaneous clotting when the material is
• Fibrinogen dissolves slowly and froths badly if shaken, and the
solid stabilised foam is very slow to disperse. Hence, agitation
should be limited to rocking.
• Solution should be used as early as possible, not later than three
hours of preparation.
• Use: In the treatment of fibrinogen deficiency. Often it is used in
conjugation with thrombin.
42. Human Thrombin
• It is the enzyme that converts fibrinogen to fibrin.
• The prothrombin obtained from the fractionation of
plasma is washed with distilled water and dissolved
in citrate saline.
• It is converted to thrombin by adjustment to pH 7
and adding thromboplastin and calcium ions.
• Solution is filtered and freeze dried.
• Air in the container is replaced with nitrogen.
• Reconstituted with saline when required.
43. Uses of Human thrombin
• The fibrin clot produced when, thrombin is mixed
with fibrinogen is used in surgery to suture severed
nerves and to assist adhesion of skin grafts.
• The mixture clots at a rate that depends on the
amount of thrombin present, and therefore if
necessary, it can be kept fluid long enough, for
adjustments.Eg. Skin grafts to made.
• The clot also act as a haemostat..
• Since it is well tolerated by human body, new cells
penetrate it rapidly.
44. Human fibrin foam
• This is a sponge like mass of human fibrin.
• Prepared by whipping a solution of fibrinogen in to a
froth by mechanical means.
• The product is poured in to trays and freeze dried.
• Cut in to pieces of convenient size and sterilised by
dry heat at 130°C for three hours.
• Storage conditions are similar to fibrinogen and
• No need to store under nitrogen.
• Along with thrombin, it is used as a haemostat
• Thrombin along with the rough surface
provided by the sponge promote blood
• The foam can be left in-situ, where it will be
absorbed because it is entirely of human
46. Human Normal immunoglobulin
• Obtained from the globulins fraction in the
fractionation of plasma
Globulins (Beta and
0°C,pH5, Ionic strength 0.01 Beta glubulins
-3.5°C, pH 6.75, ionic strength 0.025 Gamma globulins
The ionic strength are critical.
47. Human normal immunoglobulin
• Administered by dissolving in a suitable solvent, usualy 0.8%
• 0.01% thimersal is added as a preservative.
• Solution is sterilised by filtartion, packed in single dose
containers and stored at 4 - 6°C, protecting from light.
• Normally pools of not less than 1500 donations are used to
ensure a satisfactory representation of the various types of
• When it is for the production of antivaccinia and tetanus
immunoglobulin, the blood is of recently immunised donors
and pool size will be relatively smaller.
49. Human plasma protein fraction
• Solution of some proteins from liquid plasma.
• Contains albumin and certain globulins that retain their
solubility on heating.
• Prepared by fractionating pooled citrated plasma and is similar
to the fraction shown as crude albumin.
• Sodium caprylate or acetyl tryptophan is added as a stabiliser.
This allows the preparation to be heated for several hours at a
low temperature without significant denaturation of proteins.
• Sodium chloride is added to make the solution approximately
isotonic. Sterilised by filteration.
• Asceptically transferred to MRC bottles and then heated to 60
0.5 °C for ten hours to kill the viruses of infective Hepatitis
and homologous serum jaundice.
50. Human plasma protein fraction contd..
• No antibiotic or bactericide is allowed to control
contamination since, bactericide in a LVP (Large
volume Parentral) can cause toxicity and an antibiotic
may produce a risk of sensitization.
• The fractionation process involves concentration of
• This process may results in concentration of sodium
citrate, which leads to toxicity. The concentration of
sodium citrate in the final preparation is limited to
51. Human plasma protein fraction
• The protein content is not less than 4.3%w/v. The
product exerts a colloidal osmotic pressure
approximately equivalent to that of pooled liquid
plasma containing 5.2 % w/v of protein.
• Stored b/w 5 - 20°C, and protected from light.
• Since the product does not containig fibrinogen, the
product appears clear.
• Dried human plasma protein fraction is prepared by
freeze-drying human plasma protein fraction.
• Use: same as dried plasma.