By

Noorul Alam.I

Pharmaceutical Products?

Pharmaceutical products -

are suitable for treatment or prevention of diseases in humans or animals,

or

can be used in or given to humans or animals to restore, change or affect physiological functions by

means of a pharmacological, immunological or

metabolic effect, or to make a medical diagnosis.

Blood Products?

Blood products are the whole blood or its components which are collected, processed and stored

which are suitable for treatment of diseases in humans

or animals.

Biopharmaceutical products purified from human blood, such as the blood clotting factor VIII used

to treat haemophiliacs.

What is blood?

Blood is a connective tissue

Humans contain 5 liters on average

8% of body weight is blood

Functions of Blood

Transportation

Regulation

Protection

INTRODUCTION

1628 - William Harvey discovers the circulation of blood.

1665 - The first recorded successful blood transfusion occurs in England: Physician Richard Lower keeps dog

alive by transfusing blood from other dogs.

1818 - British obstetrician James Blundell performs the first successful transfusion of human blood to a patient for

the treatment of postpartum hemorrhage

Blood Group by Landsteiner, 1901

1907 - Ludvig Hektoen suggests that the safety of transfusion might be improved by cross-matching blood

between donors and patients to exclude incompatible

mixtures.

Reuben Ottenberg performs the first blood transfusion using blood typing and cross-matching.

Sodium Citrate Hustin, 1915.

1st World War (19141918) stimulus.

1939-1940 - The Rh blood group system is discovered by Karl Landsteiner, Alexander Wiener, Philip Levine and R.E.

Stetson.

Whole Human Blood:

Whole Human Blood:

Blood with suitable anticoagulant.

Donor 1. Not suffering from transmitting disease due to blood transfusion like syphilis, serum jaundice, etc.

2. Not anemic - Hb count not < 12.5 (females)

not < 13.3 (males)

Collection:

Collected - from Median cubital Vein.

- in a sterile container

containing anticoagulant.

contd

Gently shaken during Collection Maximum of 420 ml is taken. Then sealed and cooled to 4-6C

r

SINGLE_BLOOD_BAG_HOLDER

contd

Blood Clotting: Classical Theory:

1. Prothrombin Thrombin.

2. Fibrinogen Fibrin

Thromboplastin + Ionized Calcium

Thrombin

ANTICOAGULANTS:

1. Citrates:

Most commonly used Acid citrate Dextrose (ACD) Composition:

Sodium acid citrate 2.0 to 2.5 G Dextrose - 3.0 G

WFI - 120 ml

Mechanism: Calcium ion --------------- unionized calcium citrate.

Trisodium citrate was also used due to alkaline pH cause caramellisation of dextrose during sterilization.

Dextrose delays haemolysis of erythrocytes (in vitro) & prolong their life after trans fusion. Synthesis ATP

2.Heparin:

Naturally occurring by mast cells. Occasionally used during large transfusion. Eg: cardiac surgery. As large amount of citrate is harmfull.

Expensive, needs neutralizing substance (protamine sulphate).

3.Disodium Edetate:

Chelates with Calcium divalent ion. Used when preservation of blood platelets is essential.

contd

Blood Groups:

ABO system

Antigen Aglutinogens Antibody - Agglutinins Rh system

Subtypes is blood group:

A1 Negative (A1 -ve)

A1 Positive (A1 +ve)

A1B Negative (A1B -ve)

A1B Positive (A1B +ve)

A2 Negative (A2 -ve)

A2 Positive (A2 +ve)

A2B Negative (A2B -ve)

contd

A2B Positive (A2B +ve)

B Negative (B -ve)

B Positive (B +ve)

B1 Positive (B1 +ve)

O Negative (O -ve)

O Positive (O +ve)

Storage:

Stored at 4-6C - Within half an hour from collection.

Deleterious reaction s:

Leucocytes disintegrates within few hours

Platelets disintegrates within few hours

RBCs shows fall in ATP and other organic Phospates.

- reduction in oxygen carrying capacity.

- increased fragility due to lipid loss from the membrane.

Fitness of blood for transfusion based on its appearance. Complete haemolysis sign of bacterial infection. Pseudomonas and other members of coli-aerogenes survive at refrigerator temp.

Uses:

Used during

Loss of blood hemorrhage,. Loss of vital constituents Shock,

Burns,

Uncontrolled Diarrhoea,

Vomitting,

Hemorrhage,etc.

Blood Products:

Blood

Plasma

Immunoglobin

(Gamma globulin) Serum

Clotting factor

Fibrinogen Thrombin

Fibrin (foam)

RBC

CONCENTRATED RBC:

Preparation - plasma removed by centrifugation of

whole blood (not>fortnight

old).

Thorough aseptic precautions carried out.

Used within 12 hrs, Hb not < 15.5% Cells matched with recipients plasma

Uses:

To treat chronic anaemia, (whole blood may overtax) Exchange transfusions in infants.

Plasmapheresis

DRIED HUMAN PLASMA:

Disadvantage of whole blood:

Short life (three weeks)

Need refrigeration

Should be compatible with recipient

Advantage of plasma:

Five years if properly stored.

Can be stored at RT 20C (light protected)

Given to recipient of any blood group.

Used as substitute for whole blood.

Preparation: Two major problems:

(A) transmission of Viral Jaundice:

1) Infective Jaundice incubation period 5 weeks 2) Homologous serum Jaundice incubation period 20 weeks.

B) Neutralization of plasma agglutinins:

Occasionaly the plasma agglutinins cause haemolysis of

recipient cell.

ratio of mixing plasma from different groups. (cross

neutralizing)

9 of A : 9 of O : 2 of B : or AB

Preliminary freezing:

centrifuged at -18C Preliminary Drying:

placed in drying chamber with high vacuum. Condensing coil at

-50C, small heater provide latent heat of evoporation. Duration -2 days

Secondary Drying:

dried by vacuum desiccation over phosphorous pentoxide.

Duration 1 day.

contd

Storage: At 20 C and protected from light , moisture, and Oxygen.

Expiry date 5 years

Fitness of which is shown by the solubility when reconstituted with

WFI, NaCl injection, or Solution contationing 2.5% of

dextrose, 0.45% NaCl , eqivalent to original volume of plasma.

- must dissolve in 10 mins at RT.

Labelling: Label should state,

1. Name and % of anticagulant and added substance.

2. WFI required

3. Protein content

4. content must be used within 3 hrs after reconstitution

Uses: Used as the alternative to the Whole blood

Treatment of Burns and scalds.

Restore blood volume ( Emergency)

In fibrinogen deficiency: dried plasma reconstituted to 1/3 rd or th of

original vol.

Dried residue of plasma ( 400ml) - 1 g fibrinogen.

contd

FRESH FROZEN PLASMA:

Preparation: Freshly collected blood - Centrifuged stored in frozen state (-30 C)

If it is not pooled shold be labelled and used according to stated blood group.

During usage thawed (45 mins) not exceeding 37C

Use: Labile clotting factors present

So source of factor VIII to treat minor haemorrhage

Fractionation of Plasma:

Protein seperation method salting out ( dialysis required)

- Not suitable for plasma frationation

Cohnss technique use of organic solvent (Ethanol or Ether).

Advantages of organic solvent:

Volatility- so removed easily

Salt in low conc to improve resolution

Bactericidal activity

Being liquid easy to add aseptically.

Human Fibrinogen:

Fibrinogen is the soluble constituent of plasma. With thrombin it forms as fibrin

Preparation

Fractionation of Plasma

Precipitate is collected by centrifugation

Dissolved in citrate-saline and freeze dried

Air in the container is removed by nitrogen.

Citrate prevents spontaneous clotting when reconstituted with saline

Fibrinogen dissolves slowly but froth badly, so agitation should be limited to rocking.

Must be used immediately after reconstitution (within 3 hrs).

USE: To treat fibrinogen deficiency. More often used along with Thrombin

Ether 11 %

Plasma Fibrinogen

0 C pH 7.7

Human Thrombin

Ether 11 %

Plasma Fibrinogen

0 C pH 7.7

Ether 10 %

Supernatant 1 Prothrombin

0 C pH 5.35

Thrombin is the enzyme converts fibrinogen to fibrin

Prothrombin is obtained by fractionation and washed with water.

Dissolved in citrate saline.

It is converted to thrombin by

Adjusting the pH to 7 and

Adding Thromboplastin and calcium citrate.

Solution is filtered and Freeze dried.

Air in container is replaced with nitrogen.

USE:

* Fibrinogen + thrombin = Fibrin

* Fibrin used in surgery to suture severed nerves

* Assist adhesion of skin graft.

* As Haemostat..

Human Fibrin Foam:

It is a sponge like mass of human fibrin.

Preparation:

Whipping a solution of fibrinogen into a froth by mechanical means

Thrombin is added to it.

It forms fibrin clot.

Product is poured into tray and freeze dried, Then cut into pieces

Sterilized by Dry Heat at 130 C for 3 hrs.

Use:

Haemostat in surgery. (A piece is dipped into thrombin and applied

to the bleeding area)

Foam can be left in situ

Human Normal Immunoglobulin Injection:

Ether 11 %

Plasma Fibrinogen

0 C pH 7.7

Ether 10 %

Supernatant 1 Prothrombin

0 C pH 5.35

Ether 18.5 %

Supernatant 2 Globulin

-3.5 C pH 5.5 Ionic Conc. 0.035

Ether 9 %

Globulin Beta Globulin

0 C pH 5 Ionic Conc. 0.01

Ether 18.5 %

Supernatant Gamma Globulin

-3.5 C pH 6.75 Ionic Conc. 0.025

Ionic strengths are critical

Immunoglobulins are dissolved in suitable solvent (0.8% Na Cl)

Preservative is added to it.(0.01% thiomersol)

Sterilized by filtration

Packed in single dose container

Stored at 4 to 6 C with protection from light.

Pool of NLT 1500 donation used to get various type of antibodies

Antivaccina and antitetanus immunoglobulins obtained from recently

immunized donors

USE:

To prevent or attenuate Measles, Rubella and infectious Hepatitis.

Hypogammaglobulinaemia

Patient susceptible to bacterial infection (Gram +ve)

PLASMA SUBSTITUTES

Properties of an Ideal Plasma Substitute.

1.Same colloidal pressure as whole blood.

2.Viscosity similar to plasma.

3. Molecule do not easily diffuse through the capillary wall (. Molecular wt)

4. Low rate of excretion or destruction by the blood.

5. Complete elimination from the body.

6. Freedom from toxicity (No renal impairment)

7. Freedom from antigenecity, Pyrogenecity .

8. Isotonicity.

9. Highly stable in liquid form at normal condition and sterilization, transport

and storage.

10. Ease of preparation, ready availability and low cost.

Gum Saline (offficial I BP- 1932)

- Injection of sodium chloride and Acacia

- 6% Acacia in 0.9 % NaCl solution.

- Not metobolised but stored in different organs.

Polyvinylpyrolidone

- Synthetic colloid, For treatment of shock.

- Susceptible for Carcinogenicity. (in 1950 s withdrawn)

DEXTRAN

Polysaccharide produced by

Leuconostoc mesenteroides (Bacterium)

dextran-sucrase

n sucrose n(glucose H2O) + Fructose (used by organisms)

Natural Dextran

consists of approximately 200 000 glucose unit (Mol. Wt 5 million)

Drawbacks with Mol. Wt > 250 000:

Viscous solution difficult to administer.

Cause Renal damage & allergic Xns

Interfere with blood matching and sedimentation tests by causing

rouleaux formation.

So Size reduced by

1. Acid Hydrolysis ( Adjusted to pH 2 and heated to 90 C)

2. Thermal degradation

Heated at 160 C, under pressure.

In presence of

Sodium sulphite prevent oxidative degradation

Calcium carbonate neutralise acidity

3. Ultrasonic disintegration

4. seeding the fermenter

Drawbacks with Mol. Wt < 60 000:

Drawbacks with Mol. Wt < 60 000:

Rapidly excreated in urine

Pass into tissue fluids causing an adverse osmotic pressure

Mol. Wt : 60 000 100 000

Rreadily excreted in

urine

Lost into tissue fluids

Lost into tissue fluids

Acceptable range Causes allergy and renal

damage

Interfere with tests

60 000 100 000 250 000

DEXTRAN 110 Injection

- Solution of the polymer in Dextrose Injection 5 g/l, or in Sodium

Chloride Injection

- Sterilized either by autoclaving or filtration.

- Dextran 110 Injection in 5 g/l dextrose Store Below 25 - 5 Years

- Dextran 110 Injection in NaCl Injection Store Up to 40 - 5 Years.

- Emergency restoration of blood volume.

Dextran 40 Injection

- Avg. molecular weight of 40,000

- Small molecular weight dextran of this size has a specific action

in diminishing red cell aggregation

- Below 25 - 5 Years

- Method of preparation is similar to Dextran 110.

Questions

please