Unique characteristic of Sterile products:

✓ All products must be sterile.

✓ All products must be free from pyrogenic (endotoxin)
contamination.
✓ Products should be isotonic.
✓ All products must be stable, not only chemically and
physically like all other dosage forms, but also ‘stable’
microbiologically
✓ Products must be compatible.
 Formulation of Parenteral:
1. Therapeutic agents
2. Added substances (Additives)
✓ Antimicrobials
✓ Antioxidants
✓ Buffers
✓ Bulking agents or tonicity modifiers
✓ Chelating agents
✓ Solubilizing agents
✓ Local anesthetics
✓ Stabilizers
[Link]
✓ Water
✓ Water miscible vehicles
✓ Non-aqueous vehicles
 Formulation of Parenteral

[Link] ingredients
✓ Antibiotics
✓ Insulin
✓ Anticancer
✓ Steroids
✓ Vaccines
✓ Antipyretic
✓ Analgesics
✓ Anti-inflammatory
✓ LVP’s like Dextrose, NaCl or combination etc….
 Vehicles/Solvents

Vehicles used must be:

✓ Pharmacologically inert
✓ Non-toxic and non-irritating
✓ Be Physically and chemically stable
✓ Must be pyrogen free
✓ Not affect activity of medicinal agent

Types of Vehicles
➢ Aqueous vehicles
➢ Water miscible vehicles
➢ Non aqueous vehicles
Aqueous vehicles:
Water is used as vehicle for majority of injections because
water is tolerated well by the body and is safest to
administer.

✓ Purified water (USP)

✓ Water for Injection (WFI)

✓ Sterile Water for Injection (SWFI)

✓ Bacteriostatic Water for Injection (BWFI)

✓ Sodium chloride injection (USP)

Types of water:

[Link] water (USP):

✓ Drinking water that has been passed through a purification process
such as distillation, ion exchange columns or reverse osmosis to
remove dissolved solids and microorganisms.
✓ Purified water, USP should not have more than 100 recoverable
bacterial colonies per mL.

✓ Purified water is used for cleaning equipment as well as a source of

water for steam generation.

✓ It is not suitable for preparing injection solutions.

2. Water for injection, USP (WFI)

✓ It is the water intended to be used in the manufacture of

injectable products, which are to be sterilized after their
preparation.

✓ It is the most frequently used solvent.

✓ It contains no added substances.

✓ It is NOT used for the dilution of packaged parenteral products.

✓ Although it is not required to be sterile, it should be free from

pyrogens.

✓ It must be clear, colorless and odorless.

✓ Prepared by distillation or reverse osmosis.

✓ The total number of dissolved solids must not exceed 10 ppm.

✓ Water for injection, USP should not have more than 50

recoverable bacterial colonies per mL
✓
✓ Unless it is used within 24 h of its collection, WFI should be
discarded or maintained sterile or stored in tight containers at:

1-25.0 ºC for small volumes

2-80.0 ºC for larger tanks

(below and above the range at which microbial growth occurs)

3. Sterile water for injection, USP
✓Sterile Water for Injection (USP) is a sterile, non-pyrogenic
preparation of water for injection

✓contains no antimicrobial agent or added buffer and is supplied

only in single-dose containers.

✓used to dilute or dissolve already-sterilized and packaged

injectable medications (dry powders of Na phenobarbitol &
Ampicillin Na).
✓It is added under aseptic conditions.

✓It must be pyrogen-free.

✓Does not contain antimicrobial agent.

✓It must be isotonic when intended for Intra-vascular.

Note: This water may contain a slightly greater amount of total

solids than water for injection due to the leaching of solids
from the glass-lined tanks during the sterilization process
 4. Bacteriostatic Water for Injection, USP

✓ Sterile, nonpyrogenic preparation of water for injection

containing 0.9 % of benzyl alcohol added as a preservative.
✓ It is supplied in a multiple-dose container from which repeated
withdrawals may be made to dilute or dissolve drugs for
injection. The pH is 5.7 (4.5 to 7.0).

Note: If the patient will receive more than 5mL of parenteral

preparation, BWFI is NOT the vehicle of choice. Why?
5. Sodium Chloride Injection, USP:

✓ Is a sterile isotonic solution of NaCl in water for injection.

✓ Contains no antimicrobial agent.

✓ May be used as a sterile vehicle in solutions or

suspensions of drugs for parenteral administration.
2. Water miscible vehicles
Solvent which are miscible with water primarily to effect solubility of
drugs and / or reduce hydrolysis
✓ Dioxolanes, Dimethylacetamide, Butylene glycol
✓ PEG 400, 600, propylene glycol, glycerin, Ethyl alcohol
Eg., Cardiac glycoside injection

3. Non aqueous vehicles

The most important group of non aqueous vehicles are the fixed
oils.
The USP provides specifications for such vehicles, indicating that
the fixed oils must be
✓ vegetable origin so that they will be metabolized,
✓ will be liquid at room temperature
✓ will not become rancid readily.
• The USP also specifies limits for the degree of unsaturation and
free fatty acid content.

• The oils most commonly used are corn oils, cotton seed oil, peanut
oil and sesame oil.

• The fixed oils for injection must not contain mineral oils or solid
paraffins as these would not be metabolized by the body and,
therefore, might eventually cause tissue reactions and even
tumors.

• Fixed oils are used particularly as vehicles for certain hormone

preparations.

• The label must state the name of the vehicle so that the user may
beware in case of known sensitivity other reactions to it.

Examples: progesterone, testosterone, deoxycorticicosterone) and

vitamin (e.g., Vitamin K, Vitamin E) preparations.
 Antioxidants
• Many drugs are liable to oxidative degradation in aqueous
solution.
• Parenteral products containing such drugs therefore the
addition of a suitable antioxidant.
Function of antioxidants
1. Antioxidant (Reducing agent): are preferentially oxidized
and gradually used up and protect drugs prone to oxidation

2. Antioxidants (Blocking agent): act by inhibiting an oxidative

chain reaction in which they are not usually consumed and
protect drugs prone to oxidation

Note: Those product in which oxygen enters into a degradative

reaction, an antioxidant effect can be achieved by displacing
oxygen from contact with the product.
by replacement of the air from the container of the parenteral
product with an inert gas like nitrogen or carbon dioxide.
Suitable antioxidants (reducing agent) for aqueous injections
include :
✓ Sodium sulphite
✓ Sodium metabisulphite
✓ Sodium thiosulphate
✓ ascorbic acid

Suitable antioxidants (Blocking agents) For oily injections

include:

✓ Tocopherol
✓ butylated hydroxytoulene
✓ butylated hydroxyanisole
✓ propyl gallate
 Buffers
• The solubility and stability of medicaments in solution are
greatly influenced by the pH of the solution.

• Additionally, consideration of pH is important in case of

parenteral products since a highly alkaline pH (above 9) can
cause tissue necrosis while an acidic pH ( below 3) can result
in extreme pain at the site of injection.

• The ideal pH of a parenteral product is 7.4, the pH of blood.

• Buffers are usually added for adjusting the pH of parenteral
products.
• The most common buffers used are
✓ Acetate buffers
✓ Phosphate buffers
✓ Citrate buffers
✓ Glutamate buffers
Bulking agent or Tonicity modifiers
The osmotic pressure of blood is approximately 300 milliOsmoles/L
and ideally any sterile solution would be formulated to have the
same osmolarity.
examples
✓ 0.9% w/v Sodium Chloride iv solution has an osmolarity of
308 mOsmole/L
✓ 5% w/v Dextrose iv solution has an osmolarity of 280
mOsmol/L.

• Methods to determine Tonicity: Freezing point depression method

• Intravenous solutions that have larger osmolarity values

(hypertonic) or smaller osmolarity values (hypotonic) may cause
damage to red blood cells, pain, and tissue irritation.

• Osmolarity adjustment is made usually by using sodium chloride,

glucose, glycerine, dextrose, sorbitol or mannitol etc.
Chelating agents: Edetate disodium, Edetate calcium
disodium, Edetate tetrasodium

Local Anesthetics: Procaine HCL, Benzyl alcohol

Suspending agent: Gelatin, Methyl cellulose, Pectin, PEG

4000 etc

Solubilizing agents: Used to increase solubility of slightly

soluble drugs
They acts by any one of the following:
1. Solubilizers, 2. emulsifiers or 3. wetting agents.
Examples:
Dimethylacetamide, Ethyl alcohol, Glycerin, PEG –400,
polysorbate
 Stabilizers
• The drugs in the form of solutions are more liable to deteriorate due
to oxidation and hydrolysis.

• The stabilizers are added in the formulation to prevent this.

• The oxidation can be prevented by adding a suitable antioxidant,

such as, thiourea, ascorbic acid, sodium metabisulphite, or the
product is sealed in an atmosphere of nitrogen or carbon dioxide.

• Hydrolysis can be prevented by using a non-aqueous vehicle or by

adjusting the pH of the preparation.
Examples of stabilizers: Creatinine, Glycine, Niacinamide, Sodium
saccharin
Chemical stability of medicaments in parenterals.

✓ Parenterals are hermetically (air-tight) sealed container and hence

medicament is protected against oxidation due to oxygen present in
the air.

✓ Parenterals use high purity materials hence metal impurities such

as Fe, Cu, are less and this lead to better stability.

✓ Parenterals contain minimum excipients, in most of the cases

none other than vehicles, as a result stability problems due to drug
excipients interaction are not encountered, e.g., surface active agent
catalysed decomposition.
 Sterilization of Parenteral
Sterilization: Elimination of vegetative and sporal form of microorganism

.
 Dry heat sterilization
Principle:
The killing of microorganisms by heat is a function of the time-
Tankertanker Design

temperature combination used. If the temperature is increased then the time

required for killing all the bacteria will be decreased.
Condition: Cycles recommended as per BP 1988 are:
A minimum of 1800C for not less than 30 minutes. A
minimum of 1700C for not less than 1 hour.
A minimum of 1600C for not less than 2 minutes.
Mechanism of killing the bacteria:
The vital constituents of cells such as proteins (enzymes) and
nucleic acids are denatured by oxidation.

2
3
 Dry heat sterilization
Applications

✓Dry heat is used to sterilize Glass ware ( e.g. test tubes,

Pasteur- pippettes, petridishes, flasks, glass syringes etc )
✓Porcelain and metal equipment such as forceps, scalpels,
scissors etc.
✓Fats, oils and greasy materials (like petroleum jelly) those
are impermeable to moisture.
Limitation of Dry heat

✓Aqueous solution cannot be sterilized by dry heat because

the water will evaporate if the container is kept open. If the
container is kept close then it may burst due to pressure
developed by the steam.
✓Powders that cannot withstand the high temperature are not
sterilized by dry heat method. Tankertanker Design
 Moist heat sterilization

Autoclave

MHS

Heating with
Tyndalization
bactericide

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5
Autoclave
Principle:
Mechanism of killing of microorganisms:
Bacterial death by moist heat is due to denaturation and coagulation of
essential protein molecules (enzymes) and cell constituents.

Conditions:
The USP and BP recommended the following condition:
Pressure: : 15 lb/square inch (psi)
Temperature : 1210C
Time : 15 minutes
The following combinations of temperature and holding time are normally
employed for sterilizing by heat in autoclave:
Holding Temp (°C) Holding Time (Min)
115 to 118 30
121 to 124 15
126 to 129 10
134 to 138 3
Advantages of moist heat sterilization(Autoclave)

✓ High heat content plus rapid heat transfer.

✓ Destroys micro-organism more efficiently than dry heat.
✓ It can be used for a large number of injections, ophthalmic
solutions, irrigants, dialysis fluids etc.
✓ It rapidly penetrates porous materials and is therefore very suitable
for sterilizing surgical dressings and materials.
✓ The process is adaptable for plastic containers and some other
special dosage forms.
Disadvantages of moist heat sterilization
✓ It is not suitable for anhydrous materials such as powders and oils.
✓ It cannot be used for thermo labile substances.
✓ It does not destroy pyrogen’s.

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7
 Heating with a bactericide
This method is used for sterilizing aqueous solutions that are thermo
labile to withstand normal autoclaving temperature.
Condition:
Temperature: 98 to 1000 C boiling water
Time: 30 minutes
Bactericides: Bactericide that is compatible with the
product, container and closure:
e.g. For injection
Chlorocresol 0.2%(w/v)
Phenylmercuric nitrate (PMN) 0.002%(w/v)
Phenylmercuric acetate (PMA) 0.002%(w/v)
e.g. For eye drop
Thiomersal 0.01%(w/v)
Chlorhexidine acetate 0.01%(w/v)
PMA and PMN 0.002%(w/v)
Benzalkonium chloride (BAC) 0.01%(w/v)

In the following cases this method can be used:

✓ Injection fluids in the final container (i.e. terminal sterilization).
✓ Eye drops.
 Limitation of Bactericide with

✓ Solution of medicaments is not suitable for intrathecal, intra-

atrial, peridural, intra-peritoneal, intra-cisternal and any
other route of injection giving access to the cerebro-spinal
fluid and for intracardiac and intra-ocularly.
✓ Large volume parenteral fluids having a volume greater
than 15 ml is not recommended to use bactericide
because, though the concentration of the bactericide in
the solution is less but the total amount entering into the
system may be considerable due to large volume. e.g.
bactericides are not used in saline solutions.

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9
 Gaseous sterilization
This process involves exposure of materials to sterilize gasses
such as

✓ Ethylene oxide(large scale of industrial and medical applications)

✓ Formaldehyde
✓ Glutaraldehyde
✓ Propylene oxide

Mechanism of action: Ethylene oxide acts by alkylation.

Factors affecting gaseous sterilization

✓ Concentration of sterilize gasses .
✓ Humidity
✓ Temperature
✓ Time (Duration of exposure)
✓ physical nature and permeability of the load
✓ atmospheric preconditioning of the load before sterilization
 STERILIZATION BY RADIATION
Radiation can be divided into two groups:

1. Electromagnetic waves:
✓ Infra-red radiation (IR)
✓ Ultraviolet radiation (UV)
✓ X-rays
✓ Gamma rays
2. Streams of particulate matter
✓ alpha radiation
✓ beta radiation

Radiation Wave length Energy

UV-radiation 190 to 370 nm 5 eV
Gamma radiation 1 to 10-4 nm 1.3 MeV*
High velocity electrons 4MeV
 Ultraviolet radiation
Dose of sterilizing radiation:10 to 60 microwatts/cm2

✓ reduce the populations of vegetative cells by 90% in a short

period.

Application:
• Surface sterilization,
• Sterilization of clean air and water in thin layers.
Disadvantages:
• It has very poor penetration power.
• UV-light sterilization is not absolutely reliable because
DNA may get repaired in some favorable condition
3
2
Ionizing Radiation
Mode of action of ionizing radiation:
ionization and where water is present free radical formation. Free
radicals are powerful oxidizing (OH, HO2) and reducing (H) agents,
which are capable of damaging essential molecules (enzymes and
DNAs) in living cells. This results in cell death.
Gamma irradiation

Source: radioactive isotope of Cobalt 60 Co, is used as

a source of gamma emission.

Dose: 1.25 MeV, Some users take the adequate

dose as 2.5 Mrad (Mega radiation unit)

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3
 Sterilization by Filtration
✓ Thermo-labile solutions.
✓ Filter preparation should aseptically filled in the sterile containers which
are then sealed.

Tankertanker Design
 Mechanism of filtration

The filters are thought to function by one or usually a combination

of the following:
✓ Sieving/screening/surface filteration,
✓ Entrapment/Deep-bed filtration
✓ Electrostatic attraction/ Adsorption

Tankertanker Design
 Types of filter
Membrane Sintered/fritted Ceramic
filter(0.22µm) glass filters.
✓ Finely
✓ Cellulose-derivative powdered
(acetate or nitrate) borosilicate
✓ sterilized by glass filled in
autoclaving or by disc and heated
ethylene oxide gas till in fused.
✓ Suitable for
sterilizing aqueous
and oily solutions.
✓ Not for organic
solvents such as
alcohol, chloroform
etc
 SEITZ FILTER
They are made of asbestos pad.

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7
 Sterile Bulk powder
Some drugs are difficult to sterilize terminally (i.e. when the total product
is ready within the container and package), the raw materials (i.e. bulk
drugs) of those products are required to be sterile.

Generally, the manufacture of a sterile bulk substance usually

includes the following steps:

✓ Conversion of non-sterile drug substance to the sterile form by

dissolving in an (organic or aqueous) solvent, sterilization of the
solution by filtration and collection in a sterilized reactor
(crystallizer).
✓ Aseptic precipitation or crystallization of the sterile drug
substance in the sterile reactor.
✓ Aseptic isolation of the sterile substance by centrifugation or
filtration.
✓ Aseptic drying (spray drying, lyophilization), milling and blending
of the sterile substance.
Tankertanker
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8
Summary Of Industrial Sterilization
Sterilization Sterilizable articles Non-
process / sterilizable
Conditions
articles
Moist heat or Autoclave Anything heat stable, Only when steam sterilization
15 lb/sqinch, 121°C, 15 • Most types of solid and liquid media with or is not applicable
mins without carbohydrate, gelatin
• Distilled water, saline solution.
• Rubber tubing and rubber stoppers
• Discarded cultures and contaminated media
prior to washing.
• Laboratory aprons coats.
• Canned foods.

Heating with Aqueous preparation unsuitable for higher • Intrathecal, intra-atrial,

bacteriostatic agents, temperature. peridural, peritoneal, intra-
100°C for 30 mins • Injection fluid cysternal, intracardiac,
• Eye drops intraocular injection
• Large volume parenteral
Tyndallization Gelatin, milk and carbohydrate
100°C, 20 mins, 3
consecutive days
Pasteurization Holder Beer and milk
method: 63 to 660C for
30 mins or Flash method:
720C for 20 sec.
Dry heat sterilization 1600C for • Heat stable nonaqueous products • Aqueous solution
2 hours and powders. • Surgical dressings
• Glass wares- test tubes, pipettes,
Petri-dishes and flasks.
• Forceps, scalpels, scissors, throat
swabs and glass syringes.
• Fats, oils, greasy materials (e.g.
petroleum jelly)
IR radiation Metal and surgical instruments Glass Same as dry heat
1800C or 2000C under vacuum syringes. sterilization
Ionizing radiation (gamma • Pre-packed disposable plastic Aqueous solution
radiation from Co-60 or Cs- syringes, catheters, plastic tubes in
137 or energized electrons saline sets etc.
from electron accelerator.), • Packaging material made of
Dose = 2.5 Mrads Aluminium and plastic strips.
UV-radiation 253.7 nm Air sterilization, surface sterilization
Source: Mercury vapor lamp
Filtration • Serum, physiological salt solution Suspensions and
0.22 nm pore size containing NaHCO3 viscous solutions, oils
• Enzymes, bacterial toxins etc.
• Solutions of antibiotics
Gas sterilization Disposable syringes, hypodermic
needles, pre-packed materials etc