Parenteral products:

These are sterile preparations containing one or more active ingredients intended for administration by
injection, infusion or implantation into the body.

Injections:

These are sterile solutions or suspensions of drugs in aqueous or oily vehicle meant for administration
into the body by means of a needle under or through one or more layers of skin.

Advantages of parenteral:

 Onset of action is fast.

 Drug can be given to unconscious patient.
 Drug action can be prolonged by modifying the formulation.
 Bioavailability is high.
 Free from first pass metabolism.
 Drugs which cannot be given orally, can be given by parenteral route.
 Used in emergency situation.

Disadvantages:

 It requires trained person for injecting the drug solution.

 It can cause pain and inflammation.
 Drug withdrawl is not possible.
 It is costly.
 There are chances of tissue damage.
 It may cause infection at site of injection.
 It may cause sepsis.
 It may cause thrombophlebitis.
 It may cause fluid overload.
 Disposal of needles, syringes or other infusion devices required extra efforts.

Routes of administration:
Intradermal injections:

 These are given between the layers of skin.

 Skin of left forearm is selected for this injection.
 0.1– 0.2 ml is given by this route.
 Diagnostic agents are given by this route.
  25 – 26 gauge needle is used.

Subcutaneous injections:

 These are given under the skin, into the subcutaneous tissues.
 1ml or less volume is given by this route.
 Injection is given into upper arm.
 Most convenient route.
 Solutions, emulsions and suspension are given by this route.

Intramuscular route:

 These injections are given into the muscular tissues.

 The muscles of the shoulder, thigh or buttock are selected for this injection.
 Volume upto 2ml can be given by this route.
 Aqueous or oily suspensions are given by this route.

Intravenous route:

 These injections are given into the vein and hence drug directly reaches to the blood.
 Volume from 1ml to 1000ml can be given by this route.
 19 – 20 gauge needle should be used.
 IV infusion of large volume parenteral is called Venoclysis and it is used to give electrolyte and
nutrients.
 Suspension and oily injections cannot be given by this route.

Intra- arterial route:

 These injections are directly given into the artery.

 It requires generally surgery.

Intracardiac route:

 These are given into the heart muscles or ventricle only in emergency.
 E.g. A stimulant in case of cardiac arrest

Intrathecal route:

 These are given into the subarachnoid space that surrounds the spinal cord.

Intra – articular route:

 These are given directly into the joints.

Intracerebral route:

 These are given into the cerebrum.

General requirements for parenteral dosage forms

Stability:

 The physical as well as chemical stability of parenteral product should be maintained during
storage.

Sterility:

 The formulation should be free from all type of microbes.

Free from pyrogens:

 The parenteral formulation should be free from pyrogens , hence test for pyrogens should be
done.

Free from foreign particles:

 The product should be free from all types of foreign particles. To ensure it, clarity test should be
done.

Isotonicity:

 The parenteral product should be isotonic with blood plasma or body fluids.

Specific gravity:

 The parenteral product meant for spinal cord should have gravity as same of spinal fluid.

Chemical impurity:

 The parenteral product should be free from any type of chemical impurity.

Types of parenteral products:

Injection:

Injections are sterile, pyrogen free solutions or dispersions (emulsion or suspension) of one or
more drugs in a suitable solvent. Injections are prepared using aqueous solvent if possible.
Injections which are formulated as dispersion (emulsion or suspension) should be stable so that
after shaking a homogenous dose of drug can be withdrawn. Single dose injections should not
have antimicrobial agent or preservative and injections having antimicrobial agent should not be
given by intracisternal, intrathecal, epidural route. Multidose injections have a suitable
antimicrobial agent in sufficient concentration. The multidose containers are equipped to ensure
protection of the contents after partial withdrawn. The contents of multidose preparations
generally should not exceed 30ml to minimize the risk of contamination due to multiple
penetrations of the closure. Example: Insulin injection

Infusions:
These are sterile, pyrogen free aqueous solutions or emulsions (O/W type) usually prepared to be
isotonic. These are prepared for administration in large volume (more than 100ml) and do not
contain any antimicrobial agent or preservative. Example: Dextrose solution

Powder for injections or sterile powders:

Drugs which are not stable in solution form are prepared as dry sterile solids which are dissolved in water
for injection just before administration. Sterile powders may be prepared by freeze-drying, sterile
recrystallization or spray drying methods. Example: Cefuroxime sterile powder

Concentrate for injections:

These are sterile, pyrogen free solutions which are diluted with prescribed volume of sterile water for
injection at the time of administration.

Implants:

These are sterile solid preparations having one or more drugs placed into the body subcutaneous or
intramuscular tissues by a minor surgery. These provide controlled release of the drug over a period of
time at the site of implantation. Example: Probuphine

Formulation of parenteral:

It consists of following:

Vehicles:

 Aqueous
 Non- aqueous

Additives:

 Solubilising agents
 Stablizers
 Buffers
 Preservatives
 Chelating agents
 Tonicity factors
 Suspending agents, emulsifiers and wetting agents

Vehicles:

These are added in parenteral preparations to dissolve or disperse drug and to deliver it to the site of
action. There are two types of vehicles which are used in preparation of parenteral:
Aqueous vehicle:

Water is used as vehicle for most of the preparations because it is well tolerated by the body and safest to
administer.

The aqueous vehicles used are:

 Water for Injection

 Sterile Water for Injection
 Bacteriostatic water for injection

Water for injection:

It must be free from pyrogen free but not necessary that it is sterile. It is obtained by deionsation and
distilled water. It should have not more than 1mg/100ml of total solids.

Sterile water for injection:

It is water for injection which must be pyrogen free and sterile. It is packed in single dose container
having a volume less than 1 litre. It does not contain any antimicrobial agent.

Bacteriostatic water for injection:

It is sterile water for injection having one or more antimicrobial agents. It is packed in multiple dose
container so that repetitive water withdrawn from container can be possible without contamination.
Volume does not exceed more than 5ml since bacteriostatic agents are toxic if used in large
concentrations.

Non – aqueous vehicle:

These are used due to following reasons:

 To increase the solubility of water insoluble drugs

 To protect the drugs which undergo hydrolysis.

The mostly used non- aqueous vehicle are:

Water miscible vehicles:

Water miscible solvents are used along with main solvent in parenteral preparation. These are used to
increase solubility of certain drugs in an aqueous vehicle or to decrease the hydrolysis of drugs.

Example:

 Ethyl alcohol in preparation of hydrocortisone injection.

 Propylene glycol in preparation of digoxin injection.

Water immiscible vehicles:

Fixed oils such as arachis oil, almond oil and sesame oil etc used as vehicle. These are used generally
when depot action of formulation is needed or drug is insoluble in water.

Example:

 Dimercaprol injection using arachis oil as vehicle.

Additives:

 These agents are used to increase the stability or quality of the product.
 These agents are used to increase solubility of drug in desired solvent.
 These agents are used to make formulation isotonic with blood plasma.
 These are used to maintain sterility in the formulation.
 These agents should be physically and chemically compatible with formulation.

Solubilising agents:

 These are the agents which increase solubility of the drug into solvent. The solubilizing agents
may be of two types:
 Surfactants
 Co-solvents

Surfactants: These agents increase the solubility of the drug into solvent by reducing the surface tension
of the drug. Example: Tween 80 (polyoxyethylene sorbitan monooleate), Tween 20 (polyoxyethylene
sorbitan monolaurate), Lacithin etc.

Co-solvents: These are the solvents which are used in conjunction with another solvent to dissolve the
drug. Example: Propylene glycol, glycerin, ethanol, polyethylene glycol, sorbitol etc.

Stabilizers:

 These are the agents which prevent oxidation and hydrolysis of the formulation.
 E.g. Ascorbic acid (0.02 – 0.1%), Thiourea (0.005%), Sodium metabisulfite ( 0.1 – 0.15%)

Buffers:

 These are the agents which resist change in pH of formulation or increase the solubility of drug in
solution.
 E.g. Acetic acid (1-2%), Citric acid (1-5%), Phosphoric acid (0.8-2%)

Preservatives:

 These are the agents which prevent the growth of microbes in the formulation. These are added in
multiple dose containers to prevent the contamination of content due to repeated withdrawn of the
content. Large volume, single dose container does not have preservative.
  E.g. Phenol (0.065 – 0.5%), Benzyl alcohol (0.5 – 10%), Thimerosal (0.001 – 0.2%)

Chelating agents:

 These agents are used to form chelates with the metallic ions present in the formulation an hence
prevent the drug degradation.
 Example: Oxidation of adrenaline by copper, iron etc which can be avoided by use of EDTA
(Ethylene diamine tetra acetic acid), disodium edetate etc. in concentration of 0.05%.

Suspending agents:

 These agents are used to increase the viscosity and to suspend the particles for a long time.
 E.g. Gelatin (2%), Methylcellulose (0.03 – 1.05%), Pectin (0.2%)

Emulsifiers:

 These are the agents which are used in sterile emulsions to make miscible.
 E.g. Lecithin (0.5 -2.3%), Polysorbate 80.

Wetting agents:

 These are the agents which are used to prevent the interfacial tension between solid and liquid
phase.
 E.g. Propylene glycol ( 0.2 – 50%), Polysorbate 80 ( 0.04 – 0.4%)

Tonicity factors:

 These are the agents which are used to make preparation isotonic with blood plasma or other
tissue fluid.
 E.g. Lactose (0.14 – 5%), Mannitol (0.4 – 2.5%), Sorbitol (2%), sodium chloride (0.9%)

Importance of isotonicity in parenteral:

Isotonicity means two solutions have same osmotic pressure across the semi-permeable
membrane. A parenteral preparation is said to be isotonic if it has same osmotic pressure as that
of blood plasma. A parenteral preparation should be isotonic otherwise it may cause crenation or
haemolysis of the RBC.

When a hypertonic (Having high solute concentration) solution is injected then it causes
shrinkage of RBC present in blood plasma leading to crenation of the RBC due to flow of fluid
from RBC into the solution.
 When hypotonic solution (having low solute concentration) is injected then it causes swelling
of RBC leading to haemolysis of the RBC due to flow of fluid from solution into the RBC.

Various agents having property to maintain isotonicity are used in formulation of parenteral
products like sodium chloride, dextrose, Lactose etc. 0.9% w/v solution of sodium chloride is
used as isotonic preparation.

Method of preparation of parenteral or processing:

Steps of preparation of parenteral:

 Cleaning of container, closure and equipment:

 Sterilization of equipment
 Compounding of material
 Filtration of solutions
 Filling the preparation into final containers
 Sealing of container
 Sterilization of final container

Cleaning of container, closure and equipment:

 New unused containers can be cleaned only by rinsing.

 Previously used containers should be cleaned by using detergents followed by final rinsing with
water for injection.
 Containers should be cleaned after disassembling if possible.
 Containers should be used only for one type of product which minimizes the contamination.
 For large tanks, pipes and equipment which can be isolated and placed in a process unit, a special
cleaning technique called CIP (cleaning in place) is used.
 Alternative hot and cold water treatment should be used for cleaning the new containers and final
washing should be done using Water for injection.
 It can be done using rotary rinser and conveyor type rinser.
 Rubber closures should be washed with hot solution of 0.5% sodium pyrophosphate in water,
then washed with water followed by rinsing with water for injection.

Sterilization of containers:

 Containers and equipment are sterilized by autoclaving at 115°c to 116°c for 30 minutes and by
hot air oven at 160°c for 2 hours.

Compounding of material:

 All material should be compounded according to the standard given in IP and method of mixing
and method of preparation should be decided before compounding.
  The parenteral preparation should be compounded under clean conditions but not essential that
compounding area should be aseptic.

Filtration of preparation;

 The compounded solution is filtered through bacteria proof filters, seitz filters, filter candle, and
sintered glass filers.
 The objective of filtration is clarification and sterilization.
 Clarification is removal of microbes of at least 0.3µm and sterilization is removal of microbes
less than 0.3µm size.
 After filtration the solution must be protected from environmental contamination until it is sealed
in final container.
 It can be done by feeding the filtrate from collecting vessel to the filling machine through sterile
hose connections.
 A secondary in –line filter is included near the outlet to pick any particulate material from lines
and equipment.

Filling in final container:

 The filtered solution is filled into final container like ampoule, vials and transfusion bottles which
are previously cleaned and dried.
 Ampoules are used for filling single dose while vials are used for filling multidose.
 Bottles are used for filling transfusion fluids.
 On small scale, filling is done manually by using hypodermic needles and syringes.
 On large scale filling is done by automatic filling machines.
 Solutions are filled into final containers by gravity filling, vacuum filling and pressure filling.
 The sterile powder is filled into final container by individual weighing or by automatic and semi-
automatic equipment.
 During filling of ampoule, it should be considered that it should be filled below the neck of the
ampoule and solution should not touch the neck of ampoule.
Sealing the containers:

 Sealing should be done in aseptic area adjacent to the filling area.

Sealing is of two types:

Tip sealing;

 It is done by melting the sufficient glass at the tip of ampoule neck to form a bead and close the
opening.

Pull sealing:

 It is done by heating the neck of a rotating ampoule below the tip and then pulling the tip away to
form a small capillary to being melted closed.
 Pull sealing is slower process but it is more reliable than tip sealing.

Sterilization of final container:

 The parenteral preparation after sealing should be sterilized in final container.

 For thermostable substances, the final containers can be sterilized by autoclaving at 115°c - 116°c
for 30 minutes or 121°c for 20 minutes and by hot air oven at 160°c for 2 hours.
 Thermolabile substances are sterilized by bacteria- proof filters and it may also contain a suitable
bacteriostatic agent to prevent the growth of micro- organism but should not be used if dose is
exceeding 15ml and used by IV route.

Production facilities for parenetral products:

The production of parenteral products requires special precautions and facilities in order to
maintain sterility and freedom from any particulate matter. Hence the production area of
parenteral products can be divided into main five sections:

1. Clean up area
2. Preparation area
3. Aseptic area
4. Quarantine area
 5. Finishing and packaging area

Clean up area:

It is not an aseptic area. The area and environment of the room should be free from dust, fibres
and microorganisms. This area should be constructed in such a way that it can withstand
moisture, steam and detergent. Ceiling and walls can be coated with epoxy resins to prevent the
accumulation of dust and microorganisms. Exhaust fans should be fitted in this area. This area
should be kept clean to eliminate the chances of entry of contamination into the aseptic area. The
containers and closures are washed and cleaned in this area.

Preparation area:

Various ingredients of the formulation are mixed and prepare for filling into the final container
into this area. It is not essential that this area is aseptic. Cabins and counters should be made up
of stainless steel and filled in such a way that no space is left for accumulation of dust. Ceiling,
walls and floor should be properly cleaned and painted to eliminate the chances of
contamination.

Aseptic area:

The parenetral formulation is filtered, filled into final container and sealed in this area. The air ij
aseptic area should be free from any type of contamination, fibres or microorganisms which is
achieved by use of HEPA (High efficiency particulate air) filters which can remove 99.97%
0.5µm or larger particles from the air. HEPA filters are fitted into the laminar air flow system in
which air free from dust and fibres move in uniform velocity. Ultra violet lamps are also fitted to
maintain sterility. The entry of personnel in this area should be through air lock system. The
trained personnel should only be allowed to enter in this area. The personnel entering in this area
should be neat, clean, reliable and wear sterile clothing. There should be minimum movement
into this area. The environment of this area should be class 100 room (means less than 100
particles per cubic feet of 0.5µm or larger size should be present).

Quarantine area:

The preparations after filtering, filling into final container and sealing are kept in this area. Then
random samples of preparations are sent to quality control department for evaluation. After
passing the evaluation, the preparations are transferred to finishing and packaging area.

Finishing and packaging area:

The parenteral products are properly lebelled and packed in this area. Proper packaging is
required to protect the product from physical damage during transportation, handling and
storage. This area is not aseptic.

Formulation of Injections:
These are sterile solutions or suspensions of drugs in aqueous or oily vehicle meant for administration
into the body by means of a needle under or through one or more layers of skin.

The formulation of injections is same as that of formulation of parenteral products hence refer
formulation of parenteral products.

Formulation of sterile powders or lyophilized products:

Drugs which are not stable in solution form are prepared as dry sterile solids which are dissolved in water
for injection just before administration. Example: Cefuroxime sterile powder. Sterile water for injection is
supplied with dry powders to make solution or suspension for injection. It can be given by IV or IM route
but suspension is not given by IV route.

Methods of preparation of sterile powder:

Sterile recrystallization:

The drug is dissolved in a solvent and obtained solution is sterilized through membrane filter having pore
size 0.22µ. A sterile anti-solvent is then added to crystallize the drug particles followed by filtration and
drying aseptically.

Lyophilization:

It is a process of separating a solid substance from solution by freezing the solvent and evaporating the
ice under vacuum. In this, drug solution is sterile filtered into the sterile trays, which are placed in freeze
dryer at temperature -50°c and then dried by vacuum to separate the drug powder.

Spray drying:

The solution of drug is sprayed into a dry chamber where it comes in contact with a hot steam of a sterile
gas (80-100°c).

Formulation:

Fillers or bulking agents:

These are used to make the sufficient structure of the cake after drying or to make up the volume of the
cake. These are used when the dose of the drug is too small or when it is less than 2%. Example:
Mannitol, glycine, glucose, sucrose, dextran etc.

Buffering agents:

These are the agents which are used to resist the change in pH of the product. Example: sodium citrate,
sodium phosphate, sodium hydroxide etc.

Collapse temperature modifiers:

These are the agents which increase the collapse temperature. During lyophilization process, the primary
drying temperature should be below than collapse temperature otherwise drying timing increases.
Example: Dextran, Ficoll, gelatin etc.

Tonicity modifiers:

These are the agents which make formulation isotonic with blood plasma. Example: Dextrose, sodium
chloride etc.

Antimicrobial agents:

These are the agents which prevent the growth of microbes in formulation. These are generally added in
multi-dose containers. Example: Ethyl and methyl paraben, phenol, cresol etc.

Solubilizing agents:

 These are the agents which increase solubility of the drug into solvent. The solubilizing agents
may be of two types:
 Surfactants
 Co-solvents

Surfactants: These agents increase the solubility of the drug into solvent by reducing the surface tension
of the drug. Example: Tween 80 (polyoxyethylene sorbitan monooleate), Tween 20 (polyoxyethylene
sorbitan monolaurate), Lacithin etc.

Co-solvents: These are the solvents which are used in conjunction with another solvent to dissolve the
drug. Example: Propylene glycol, glycerin, ethanol, polyethylene glycol, sorbitol etc.

Complexing agent:

 These agents are used to form Complex with the metallic ions present in the formulation an hence
prevent the drug degradation.
 Example: EDTA (Ethylene diamine tetra acetic acid), disodium edetate etc. in concentration of
0.05%.

Lyophilization and lyophilized products:

Lyophilization is a process in which water is frozen, followed by removal from the sample, initially by
sublimation and then by desorption. In this process, moisture content of the product is decrease to such an
extent that it does not cause microbial growth. This process has following steps:

Freezing:

This step involves the freezing of sample solution at -50°c.

Primary drying:

In this, frozen ice is sublimed and dry and structurally an intact product is formed. This is the most time
consuming step in this process.
Secondary drying:

In this step, after primary drying the residual moisture content is reduced by continuing the drying at slow
rate.

Formulation of small volume parenteral:

These are the parenteral preparations administered in less than 100ml. Formulation of small volume
parenteral is same as that of injections hence refer formulation of injections.

Types of small volume parenteral:

 Ampules
 Vials
 Prefilled syringes

Ampules:

 These are single dose containing small volume parenteral.

 These are glass containers with an elongated neck that must be broken off.
 Most of the ampules are weakened around the neck for easy breaking off the ampules and some
ampules have colored bend around the neck.
 Whole formulation in ampule is used at once.

Vials:

 These are multi dose containing small volume parenteral

 These are made up of glass or plastic and sealed with rubber stopper.
 A needle is used to add contents and withdraw the formulation in vials.
 A portion of formulation is withdrawn from vial at one time and remaining portion in vial can be
used again.
 Vials may be single dose containing and multi dose containing.
 Multi dose containing vials have preservative to prevent the bacterial contamination.

Prefilled syringes:

These are of two types:

 Cartidge type package is a single syringe and needle unit which is placed in a special holder
before use and needle and syringe unit is discarded after use but holder can be reused.
 Other type of package consists of a glass tube closed at both ends with rubber stoppers. The
prefilled tube is placed in special designed syringe that has a needle attached to it and after use all
unit is discarded.

Formulation of large volume parenterals:

 These are the parenteral preparations administered in more than 100ml.

  These are only single dose preparations.

Formulation of large volume parenteral is same as that of formulation of injections except the
antimicrobial agents which are not added in large volume parenterals.

Types:

 Hyperlimentation fluid or total parenteral nutrition (TPN)

 Cardioplegic fluid
 Dialysis fluid
 Irrigating fluid

TPN:

 IV administration of calories, nitrogen and other nutrients in sufficient amount to produce tissue
synthesis is called TPN and fluid which is used for this purpose is called hyperlimentation fluid.
 These products are generally injected to avoid multiple injections of nutritional requirements of
the patient by IV route.
 TPN is generally used in pre- operative and post- operative conditions.
 E.g. Dextrose solution 25%

Cardioplegic fluid:

 It is the solution by which the ischemic myocardium is protected from cell death which is
achieved by reducing myocardium metabolism through a reduction in cardiac work load.

Irrigation fluid:

 The fluid which is sued to rinse a body cavity is called irrigation fluid.
 E.g. 0.9% sodium chloride irrigation fluid which is sued during surgical procedures for rinsing
the tissues,, body cavity, wounds or irrigation of a special tube called catheter.

Dialysis fluid:

 Dialysis fluid is the solution which is used in dialysis process and dialysis process is the process
by which waste material from the body is removed out and it is carried out when kidney don’t
work properly.
These are of two types:
 Haemodialysis fluid
 Intraperitoneal fluid
 SVP LVP
Volume is 100 ml or less Volume is more than 100ml

Routes of administration are IV, IM, SC IV route

May be single dose or multi dose Single dose

Technique is vein puncture Venoclysis

Needle used is 20 -22 guage 18 -1 9 gauge

Preservative is used in case of multi dose No Preservative

Buffers are used No buffer

E.g. Solution, emulsion and suspension Solution, O/W type emulsion.

Containers for parenteral:

Glass containers:

 Glass is the preferred material for containers for injectable products.

 It is principally composed of silicon dioxide tetrahedron which is modified using oxides like
sodium, potassium, calcium etc.

Types of glass:

1. Type1: Highly resistant borosilicate glass

2. Type2: Treated soda lime glass
3. Type3: Soda lime glass
4. Type4: General purpose glass

 Generally type1 glass is used for most of the sterile products.

 But type2 and Type 3 glass can also be used when product has non aqueous vehicle.
 The protection of light sensitive drugs can be done by use of amber colored glass which is
achieved by iron oxides.
 But iron oxides leach out in the product hence amber colored container should not be used in case
of product which has iron catalyzed chemical.
 The glass should have sufficient mechanical strength to withstand high pressure during
autoclaving and abuse during processing.
Sterile products made by glass:

Ampules:

 These are single dose containing formulations.

 Generally Type 1 glass is used.

Package type Type of formulation Minimum quality of

glass that can be used
Ampule Aqueous injections of any pH Type1
Aqueous injections of pH less Type 2
than 7
Non- aqueous injections Type 3

Vials:

These are used for multi dose parenteral products and provided with closure followed by aluminum seal
to ensure the air tight packing.

Package type Type of formulation Minimum quality of

glass that can be used
Vials Aqueous injections of any pH Type1
Aqueous injections of pH less Type 2
than 7
Non- aqueous injections Type 3

Plastic containers:

Advantages:

 Light in weight
 Non breakable
 Has low toxicity
 Low reactivity with products

Disadvantages:

 Tissue toxicity can occur

 Reactivity due to sorption
 Leaching

Plastic is of two types:

Thermoplastic :
 1. Polyethylene
2. LDPE (Low density polyethylene)
3. HDPE (High density polyethylene)
4. Polypropylene
5. Polyvinyl chloride
6. Polystyrene
7. Polyamide (Nylon)

Thermosets:

1. Melamine
2. Phenol formaldehyde
3. Urea formaldehyde

 The two plastics have more interest in parenteral field that is polypropylene and copolymer
polyethylene and polypropylene.
 Polypropylene is mostly used plastic because it has high tensile strength, high melting point
(165°c) and low permeability to gases and water vapors.

Plastic Use
Polyethylene For ophthalmic solutions
PVC For intravenous solution bags
Polyolefins For making bottles for parenteral soultion
HDPE Disposable syringes
Polypropylene Packing of dialysis fluid
Polycarbonate Disposable syringes
LDPE Irrigation solution bags
Acrylonitrile IV Infusion

Rubber:

 Rubber closures are mostly used to seal the opening of cartridges, vials, bottles and to provide
soft and elastic permit to enter and withdrawn of a needle without loss of integrity of container.
 Principle unit of rubber closure is latex and vulcanizing agent, accelerator, activator, filler etc are
added.
 Closures should be non- reactive with the product.
 Physical properties of rubber are elasticity, hardness, porosity.
 Sometimes plastic or lacquer coating is also done to rubber to prevent the sorption, vapor transfer
and to provide complete barrier as desired.
FILLING AND SEALING OF PARENTERAL:

Filling of liquids:

Liquids can be easily and uniformly filled from the bulk container into the individual dose
container.

Filling of small volume:

Small volumes are generally filled into the ampoules or vials. For filling of small volumes of
liquid generally hypodermic needles and syringes are used which deliver the liquid into the
container by the stroke of plunger of the syringe.

Filling of large volume:

Sterile solutions of low potency are dispensed in large volume. For filling of large volumes of
sterile solutions generally gravity filling, pressure filling and vacuum filling are used.

Gravity filling:

In this, the reservoir of sterile solution is kept above the filling line having a connection between
the reservoir and shut-off device at the filling line. The shut-off device operated manually and it
stops the entry of liquid after filling of container or bottles to the graduations on the container.

Pressure filling:

In this, the filling of sterile solution is assisted by the pressure. This is equipped with a overflow
tube connected to a receiver to stop excess filling of the container.

Vacuum filling:

It is generally used for large liquid volumes because it can be made automated. In this, a vacuum
is created in the bottle when a nozzle gasket makes a seal against the lip of the bottle to be filled.
Then the vacuum draws the liquid from the reservoir through delivery tube into the bottle to be
filled. When the liquid reaches to the adjustable overflow tube then seal is loosened and vacuum
is released.

NOTE: The filling is done always in slight excess to avoid spoilage that occurs during
administration due to adherence to the wall of bottle and retention on the syringe. The excess
volume is given in USP.

Filling of emulsion and suspension:

Due to high viscosity of suspension and emulsion these are required specially designed filling
equipments. To obtain adequate or uniform flow rate, high pressure and bottle with wide mouth
are required. Sometimes, jacketed reservoir tanks are used to increase the temperature of liquid
to decrease the viscosity. Generally emulsions and suspensions are stirred continuously in
reservoir tank during filling for uniform distribution of drug in every filled container or bottle.

Filling of solids:

Sterile solids are more difficult to fill into the individual container as compared to sterile
solutions. The flow rate of solid from the reservoir into the final container tends to be slow and
non-uniform especially if solids are very fine. Containers having wide mouth are used for filling
sterile solids because flow rate is slow and risk of loss is present due to falling of the solids.

Sterile solids can be filled into the final container by individual weighing. A scoop (like spatula)
can be used that holds a volume approx. equal to the weight required, but finally weighed on the
balance. This is a slow process.

When sterile solids are generally free-flowing then machines may be used for delivery of solid
into the final container. These machines measure and deliver a volume of sterile solid approx.
equal to weight required.

Another type of machine used for delivery of free flowing sterile solids uses augar in the funnel
shaped hopper and shape & size of the augar can be adjusted to deliver a regulated volume of the
solid into the container.

In another machine, a adjustable cavity in rim of filling wheel is filled by the vacuum as the
wheel passes under the hopper. The contents are held by the vacuum until the cavity is inverted
over the container when a jet of sterile air discharges the sterile solids. This machine dispenses
dry solids which flow less freely.

Sealing of ampoules:

Sealing should be done in aseptic area adjacent to the filling area.

Sealing is of two types:

Tip sealing;

It is done by melting the sufficient glass at the tip of ampoule neck to form a bead and close the
opening.

Pull sealing:

 It is done by heating the neck of a rotating ampoule below the tip and then pulling the tip
away to form a small capillary to being melted closed.
 Pull sealing is slower process but it is more reliable than tip sealing.
  Powder ampoule or other having wide mouth opening should be sealed using pull-
sealing.

Sealing bottles, cartridges and vials:

Glass or plastic vials and bottles are sealed by closing the opening with a rubber closure. The
closure must be inserted as soon as possible after filling to avoid contamination of the content of
container. Closures are inserted mechanically by high speed processing. When closures are
inserted by machines then surfaces of closure should be halogenated or coated with silicone to
reduce the friction. Rubber closures are held in position by means of the aluminium caps. The
closure can not be removed without destroying the aluminium cap or it is tamperproof. It may be
single aluminium cap/ double or triple aluminium caps. Single aluminium cap can be inserted
using hand crimper and double or triple aluminium cap required greater force for crimping.

Filling and sealing of infusion bottles:

Infusion bottles are filled and sealed by blow fill and seal technology (BFS). In this technology,
the container is formed, filled and sealed in one continuous, automated process. The basic
concept of this technology is that a container is formed, filled and sealed in one process without
human interference.

The BFS involves following steps:

Parison extrusion:

Pharmaceutically grade plastic resin is vertically heat extruded through a circular throat to form a
hanging tube called parison.

Container moulding:

Extruded tube is then enclosed within a two part mould and tube is cut above the mould.

Container filling:

The mould is transferred to filling zone, where filling needles are lowered and filled the mould

Container sealing:

Filling needles are removed and secondary top mould seals the container.

Container discharge:

The product is discharged to non-sterile area for labeling, packaging and distribution.
Evaluation of parenteral:

1. Sterility testing:

It is done to determine that particular lot of material is sterile.

Principle:

It is based on principle that if bacteria or fungi are kept in a medium which provide favourable conditions
for growth, the organism will grow and their presence can be detected.

Steps in sterility testing:

1. Selection of sample size

2. Selection of amount of product to be tested
3. Method of testing
4. Observation and results

Selection of sample size (containers):

Number of items in the batch Minimum number of items to be tested

Injectable preparations

Not more than 100 containers 10 % or 4 containers maximum

Between 100 – 500 containers 10 containers
More than 500 containers 2 % or 20 containers

Selection of amount of product to be tested:

Quantity of each container Minimum quantity to be used

For liquids
Less than 1ml Whole contents of container
1 – 4ml Half of the content
4 – 20ml 2ml
20 – 100ml 10% of the content

For solids
Less than 50mg Whole content of container
50 – 200mg Half content of the container
200mg or more 100mg

Methods of testing:

1. Membrane filtration method

2. Direct inoculation method
Membrane filtration method:

Sample is filtered through a membrane filter of 0.45µm size

Then membrane is divided into two halves

The first half is placed in 100ml of soyabean- digest media for growth of fungi at 20°c - 25°c for 7days.

Other half is placed in 100ml of fluid thioglycollate media for bacterial growth at 30°c - 35°c for 7 days

Then observe the growth

Direct inoculation method:

In this method, specified amount of sample is aseptically removed from the container

Transferred it into the culture media

Mix the sample with media and incubate for 14 days

Observe the growth

Interpretation:

Observe the growth

No evidence Evidence of
of growth growth

Pass the test

Re-testing is
done
 Evidence of No evidence of
growth growth

Isolate and
Passes the test
identify the
organism

If organism is If organism is
same as found in different as found
first testing in first testing

Fail the test Re-testing using

double number of
samples

Evidence of growth No evidence of growth

Fail the test Passes the test

Pyrogen testing:

Pyrogens are the endotoxins of the gram negative bacteria, thermostable, soluble in water,
polysaccharides, pass through bacteria proof filters and cause rise in body temperature of human.

There are two methods for pyrogen testing:

1. In-vivo testing (Rabbit test)

2. In-vitro testing (LAL test)

Rabbit test:

Principle:
It involves the measurement of rise in body temperature of rabbit when a IV injection of sample to be
tested is injected.

Material required:

 Thermometer, glass wares, syringes, needles, retaining boxes for rabbits.

 Three healthy adult rabbits of same sex.

Following rabbits should not use:

 A rabbit having weight less than 1.5kg

 A rabbit having temperature more than 39.8°c
 A rabbit showing temperature variation more than 0.2°c between two readings.

Method:

The test is performed in AC room.

The sample to be tested is dissolved in pyrogen free saline solution.

Warm the sample solution to 38.5°c before injection.

The volume to be injected is not less than 0.5ml or not more than 10ml per kg body weight

Thermometer is inserted into the rectum of the rabbit

Two normal temperature readings are taken an interval of 30 minutes before injecting the sample
Solution
Mean of two reading is calculated and noted as initial reading
The sample solution is injected into the ear vein of rabbit in doseppof 0.5 – 10ml/kg.

Record the temperature of each rabbit after 30 minutes of injecting the sample solution

Record the temperature of each rabbit upto 3 hours an interval of 30 minutes.

The difference between the initial temperature and maximum temperature recorded after injection is noted
as response.

Interpretation:
 Final reading – Initial reading = Response

Sum of response of group of Sum of response of group of 3

3 rabbits is less than 1.4°c rabbits is more than 1.4°c

Or Or

Response of every rabbit is Response of every rabbit is more

less than 0.6°c than 0.6°c

Passes the pyrogen test

Re-testing is done by taking another 5
rabbits

Sum of response of group of 8 Sum of response of group of

rabbits is less than 3.7°c 8 rabbits is more than 3.7°c

OR OR

Response of individual 3 Response of individual 3

rabbits among 8 is less than rabbits among 8 is more
0.6°c than 0.6°c

Passes the test Fail the tesT

In- vitro test (LAL Test):

Principle:

It involves the use of LAL reagent (from horse shoe crab) which shows coagulation in presence of
polysaccharides and pyrogens are also lipopolysaccharides. If pyrogens will be present it will show
coagulation.

Method:

 0.1ml sample is with 0.1ml LAL reagent in a depyrogenated glass plate.

 Plate is incubated at 37°c for 1 hour.
 Observe the plate under microscope.
  Presence of coagulation shows presence of pyrogens.
 If coagulation not occur it means pyrogens are absent.

Leakage testing:

It is done to check that the container in which parenteral preparation is filled is hermatically sealed.

Method:

 To check leakage, the container is immersed in 1% methylene blue solution in a vacuum chamber
under negative pressure and vacuum is released.
 If colored solution entered into the container, it shows presence of leakage into the container.

Clarity testing:

It is done to check the presence of any foreign particle or impurity in the container.

Method:

 To perform the clarity test, an instrument having black and white background and light source is
used.
 Dark colored preparations are seen against white background and light colored preparations are
seen against dark background.
 If any impurity or particle is present it will detected visually and preparation is rejected.