Sterile Pharmaceutical Products Development

Are dosage forms of therapeutic agents

Are dosage forms of therapeutic agents

that are free of viable M.O. as possess high

that are free of viable M.O. as possess high

level of purity

level of purity

 including:

(Parenteral, ophthalmic, and irrigating

solutions).



Sterile products used in pharmaceutical

industry for product development,

production, control and packaging.

Objective:

Objective:

 increase of a therapeutic effect in a

patient.

Parenteral preparations:

Parenteral preparations:

Like I.V., Intra-spinal, I.M., S.C., intradermal.



I.V. and intra-spinal route: 

I.V. and intra-spinal route: 

given in the form of

aq. solution. rather than suspension 

(to avoid

(to avoid

blockage due to P.S.

blockage due to P.S.

, but it should be sterile

since (

the danger come from sensitivity of

the danger come from sensitivity of

nerve tissues and blood capillaries for any

nerve tissues and blood capillaries for any

contamination

contamination

).



I.M., S.C., or intradermally:

I.M., S.C., or intradermally:

Given in the form of solution, suspension or

emulsion but care must be taken to 

avoid

avoid

undue tissue irritation and mild local irritation.

undue tissue irritation and mild local irritation.

Notes:

Notes:

1.

Solvent systems suitable for sterile products

Solvent systems suitable for sterile products

limited for the products with little or no

limited for the products with little or no

tissue irritation like water.

tissue irritation like water.

2.

Physical and chemical contaminants 

Physical and chemical contaminants 

cause

cause

irritation to body tissues, since small

irritation to body tissues, since small

quantities of these may cause 

quantities of these may cause 

degradation

degradation

of the products (chemical changes)

of the products (chemical changes)

particularly during heating period when

particularly during heating period when

thermal sterilization is employed.

thermal sterilization is employed.

Most important vehicle for sterile products is 

water

water

(vehicle for all natural body fluids).

(sterilization can be done by distillation or reverse

(sterilization can be done by distillation or reverse

osmosis)

osmosis)

To remove dissociated and undissociated organic

To remove dissociated and undissociated organic

and inorganic substances present in water (like

and inorganic substances present in water (like

ionic content and pyrogens)

ionic content and pyrogens)

Tests for purity of water:

Tests for purity of water:

1- Gravimetric method

1- Gravimetric method

 (measure total solid

contents of dissociated and undissociated organic

and inorganic substances).

2- Electrolyte measurement for conductivity

2- Electrolyte measurement for conductivity

(conductivity indicate presence of microbes).



Used in sterile pharmaceutical products 

because

because

of hydrolytic reactions and solubility factors

of hydrolytic reactions and solubility factors

.



Properties of non-aq. solvents:

Properties of non-aq. solvents:

 non-toxic, non-

irritating, non-sensitizing and no adverse effect

on the ingredients of formulations.



Evaluation for such solvents

Evaluation for such solvents

(physical properties):

density, viscosity, miscibility, polarity, stability,

solvent activity and toxicity.

Example:

Example:

solvents miscible with water 

solvents miscible with water 

(PEG 400 &

600, glycerin and propylene glycol)

solvents immiscible with water 

solvents immiscible with water 

(fixed oils).

Requirements:

Requirements:



Physical and chemical purity

Physical and chemical purity

(free of microbial and

pyrogenic contaminants) 

of solutes

of solutes

should be

determined.



Containers for storage of solutes

Containers for storage of solutes

should be designed to

prevent contamination especially after open.

Added substance

Added substance

(enhance stability of products)

(solubilizers, antioxidants, chelating agents, buffers, tonicity

(solubilizers, antioxidants, chelating agents, buffers, tonicity

contributors, antibacterial agents, antifungal agents, hydrolysis

contributors, antibacterial agents, antifungal agents, hydrolysis

inhibitors and antifoaming agents)

inhibitors and antifoaming agents)

Properties:

Properties:

1- should be non adversely effect on the products.

1- should be non adversely effect on the products.

2- not interfere with therapeutic efficacy nor assay of active

2- not interfere with therapeutic efficacy nor assay of active

ingredients

ingredients

3- non-toxic

3- non-toxic

1.

Antibacterial agents

Antibacterial agents

-

(in formulation of multiple dose vial) 

(in formulation of multiple dose vial) 

at

at

Bacteriostatic conc.

Bacteriostatic conc.

-

 In formulations to be sterilized or made by

 In formulations to be sterilized or made by

aseptic manipulation.

aseptic manipulation.

2.

Antioxidants

Antioxidants

Protect therapeutic agents from oxidation

particularly under accelerated conditions of

thermal sterilization. 

They act in many ways:

They act in many ways:

A- 

A- 

Reducing agents

Reducing agents

B- 

B- 

Blocking agents (oxidative chain reaction)

Blocking agents (oxidative chain reaction)

C-

C-

Synergist compounds (increase antioxidants).

Synergist compounds (increase antioxidants).

D-

D-

 Chelating agents (complex with catalysts).

 Chelating agents (complex with catalysts).

3.

Buffers

Buffers

(like Acetate, citrate and phosphate)

(like Acetate, citrate and phosphate)

Maintain required pH 

Maintain required pH 

since 

since 

change in pH

change in pH

significant 

significant 

alteration in the rate of degredative

alteration in the rate of degredative

reactions

reactions

Change in pH result from:

Change in pH result from:

a)

Dissolving glass constituents in the product.

Dissolving glass constituents in the product.

b)

Release of constituents from rubber closure or

Release of constituents from rubber closure or

plastic components in contact with the

plastic components in contact with the

product.

product.

c)

Dissolving of gases and vapors from the

Dissolving of gases and vapors from the

airspace in the container and diffusion

airspace in the container and diffusion

throughout the plastic or rubber component.

throughout the plastic or rubber component.

4.

Tonicity contributors

Tonicity contributors

Compounds that contribute to the isotonicty

of a product

 to reduce pain of injection in

areas with nerve endings.

(Buffers serve as tonicity contributors as well

(Buffers serve as tonicity contributors as well

as stabilizers for the pH).

as stabilizers for the pH).

Methods of determination of isotonicty:

Methods of determination of isotonicty:

1-

1-

 Freezing point depression

 Freezing point depression

2-

2-

 Permeability of a living semipermeable

 Permeability of a living semipermeable

membrane into the blood stream.

membrane into the blood stream.

General notes:

General notes:

1.

Importance appear from 

direct contact

direct contact

with the products.

with the products.

2.

But 

no container 

no container 

presently available is

totally nonreactive

totally nonreactive

, particularly with aq.

solutions.

3.

Physical characteristics

Physical characteristics

 are the primary

consideration 

in selection of a protective

in selection of a protective

container.

container.

Plastic containers for medical field consist of:

1.

Thermoplastic polymer

2.

Plasticizers

3.

Fillers

4.

Antioxidants

Mainly used because 

(light in weight, low

(light in weight, low

toxicity, non-breakable, low in additives and

toxicity, non-breakable, low in additives and

low reactivity with the products)

low reactivity with the products)

Glass is a preferred containers for injections.

Glass is a preferred containers for injections.

Two glass types available: (soda-lime and borosilicate).

Two glass types available: (soda-lime and borosilicate).

Physical characteristics:

Physical characteristics:

1- 

1- 

Protection from UV-light

Protection from UV-light

 by using amber glass

 by using amber glass

containers (made from iron oxide).

containers (made from iron oxide).

2- 

2- 

Sufficient physical strength

Sufficient physical strength

 to withstand high pressure

 to withstand high pressure

during autoclaving, shipping, processing and storage.

during autoclaving, shipping, processing and storage.

3- 

3- 

Thermal expansion 

Thermal expansion 

to withstand thermal shocks

to withstand thermal shocks

during washing and sterilization.

during washing and sterilization.

4- 

4- 

Transparency

Transparency

 to facilitate inspection.

 to facilitate inspection.

5- 

5- 

Uniform physical dimensions 

Uniform physical dimensions 

to facilitate handling by

to facilitate handling by

machines in automatic operations.

machines in automatic operations.

-

Seal openings in vials, bottles

Seal openings in vials, bottles

-

Provide soft and elastic enough to permit entry

Provide soft and elastic enough to permit entry

and withdrawal

and withdrawal

 of a hypodermic needle

without loss of integrity of the sealed

container.

-

Non-reactive

Non-reactive

 with the products in contact.

Compatibility problems:

a)

Leaching of ingredients from rubber

 with

subsequent reaction with the product.

b)

Removal of ingredients from the product 

by

sorption or by vapor transfer through the

closure.

Convey of products from container into the body

or from one container to another, thus compatibility

between product and device is evaluated.

Examples:

Examples:

1.

hypodermic needles (stainless-steel)

2.

plastic irrigating solution bottles

3.

plastic ophthalmic dropping bottles

4.

transfer needles

5.

transfer set (I.V. catheter from silicone rubber

and nylon)



All device components must be visible clean

All device components must be visible clean

and fluid path through the device must meet the

and fluid path through the device must meet the

same rigid standards for cleanliness as the

same rigid standards for cleanliness as the

product.

product.

Ophthalmic preparations

Ophthalmic preparations

Instilled into the eye and are similar to

Instilled into the eye and are similar to

parenterals.

parenterals.

Characteristics:

Characteristics:

 1-

 1-

Stable, 

Stable, 

2-

2-

high purity of

high purity of

ingredients as well as freedom from physical

ingredients as well as freedom from physical

(particles), chemicals and microbial

(particles), chemicals and microbial

contaminants. 

contaminants. 

3-

3-

 Also free from pyrogen

 Also free from pyrogen

(although not absorbed systemically from eye)

(although not absorbed systemically from eye)

but considered microbiologically active.

but considered microbiologically active.

Requirements:

Requirements:

 Buffers (isotonicty) and

 Buffers (isotonicty) and

antioxidants (stabilizers).

antioxidants (stabilizers).

Water for injection

Water for injection

Prepared by distillation and Reverse osmosis

Prepared by distillation and Reverse osmosis

approved by USP.

approved by USP.

Specifications for still:

Specifications for still:

1.

Prepurification of water 

Prepurification of water 

by 

by 

(deionization or filtration)

(deionization or filtration)

To improve the quality of distillate

To improve the quality of distillate

and 

and 

reduce the frequency of

reduce the frequency of

required cleaning due to insoluble scale in boiler.

required cleaning due to insoluble scale in boiler.

2.

Removal of entrained contaminants from vapor before

Removal of entrained contaminants from vapor before

condensed

condensed

(by passage through an efficient baffle system).

(by passage through an efficient baffle system).

3.

Ejection of volatile constituents from top of the system before

Ejection of volatile constituents from top of the system before

vapor is cooled 

vapor is cooled 

To prevent from redissolve and appear in the

To prevent from redissolve and appear in the

condensate.

condensate.

4.

Construction of all surfaces that contact with vapor and

Construction of all surfaces that contact with vapor and

condensate

condensate

(from pure tin, 304 stainless steel, or borosilicate

(from pure tin, 304 stainless steel, or borosilicate

glass) 

glass) 

To prevent even small traces from dissolve

To prevent even small traces from dissolve

Rinsing new containers

Rinsing new containers

Cleaning new glasswares without detergent

Cleaning new glasswares without detergent

treatment, the cycle is essentially rinsing process to

treatment, the cycle is essentially rinsing process to

loose debris by 

loose debris by 

(hot clean steam and cold treatment

(hot clean steam and cold treatment

and final rinses with filtered WFI), 

and final rinses with filtered WFI), 

this should be done

this should be done

by using machines for:

by using machines for:

1- Containers: inverted in spindles in the front of

1- Containers: inverted in spindles in the front of

machine and carried through a series of rinses in one

machine and carried through a series of rinses in one

rotation.

rotation.

2- Ampules and containers with constricted opening

2- Ampules and containers with constricted opening

makes water drainage incomplete thus a blast of clean

makes water drainage incomplete thus a blast of clean

air to blow out remaining water.

air to blow out remaining water.



After cleaning containers, are often removed from

After cleaning containers, are often removed from

the rinser and placed in clean stainless boxes for

the rinser and placed in clean stainless boxes for

sterilization under the protection of HEPA-filtered

sterilization under the protection of HEPA-filtered

airflow.

airflow.

Cleaning Rubber and Plastic

Cleaning Rubber and Plastic

Components

Components

Rubber and plastic materials accumulate

Rubber and plastic materials accumulate

surface debris accumulated from surface

surface debris accumulated from surface

molding operation and from handling

molding operation and from handling

(attracted and held on surface by

(attracted and held on surface by

electrostatic forces)

electrostatic forces)

Washed by mechanical agitation in a tank

Washed by mechanical agitation in a tank

of hot detergent solution (0.5% sodium

of hot detergent solution (0.5% sodium

pyrophosphate)  followed by a thorough

pyrophosphate)  followed by a thorough

water rinses , the final rinses being WFI.

water rinses , the final rinses being WFI.

Filtration of solutions

Filtration of solutions

Primary objectives of filtering solutions are

Primary objectives of filtering solutions are

Clarification and sterilization

Clarification and sterilization

Liquids may be subdivided from a bulk

container to individual dose containers more

easily and uniformly than a solid.

Mechanical subdivision:

Mechanical subdivision:

1.

Mobile, low density liquid

Mobile, low density liquid

achieved by

achieved by

with light-duty machinery.

with light-duty machinery.

2.

Viscous, sticky or high density liquids

Viscous, sticky or high density liquids

requires much more rugged machines to

requires much more rugged machines to

withstand the pressure required to

withstand the pressure required to

dispense them.

dispense them.



Filling machines should have parts:

Filling machines should have parts:

1.

Through which liquid flows easily for cleaning and

Through which liquid flows easily for cleaning and

sterilization.

sterilization.

2.

Constructed of non-reactive materials such as

Constructed of non-reactive materials such as

borosilicate glass or stainless steel.

borosilicate glass or stainless steel.

Example:

Example:

 Syringes made from stainless steel when

the pressures required for delivery of viscous liquids

or large volumes while they are unsafe for glass

syringes.

Note:

Note:

Sterile solutions of low potency dispense in

Sterile solutions of low potency dispense in

large volume (1L) don’t require precision of small

large volume (1L) don’t require precision of small

vol. of potent injectables. So bottles of solutions are

vol. of potent injectables. So bottles of solutions are

filled by gravity, pressure or vacuum filling devices.

filled by gravity, pressure or vacuum filling devices.

Sterile solids (such as antibiotics) 

Sterile solids (such as antibiotics) 

are

more

difficult to subdivide accurately precisely into

individual dose containers than are liquids.



The rate of flow of solids tend to be slow and

The rate of flow of solids tend to be slow and

irregular,

irregular,

particularly if finely powdered while

small, granular particles flow most evenly

small, granular particles flow most evenly

.



Most containers 

Most containers 

even

with large opening must

be used; even so, 

have slow filling rate and a

have slow filling rate and a

risk of spillage

risk of spillage

. The 

tolerance

tolerance

 for the content

of such containers must be 

relatively

relatively

large

large

.



Sterile products evaluated for:

1.

Pyrogen test on WFI

2.

Glass tests on containers

3.

Identity test on rubber closure

4.

Microbial load test to determine No. and

type of M.O. present.

5.

Conductivity measurement

6.

Conformation of volume of fill

7.

Leaker test

8.

Clarity test

9.

Sterility test

Intended to detect incompletely sealed ampoules

Intended to detect incompletely sealed ampoules

(having capillary pores or tiny cracks)

(having capillary pores or tiny cracks)

M.O. or dangerous contaminants enter ampoules or contents may

M.O. or dangerous contaminants enter ampoules or contents may

leak to the outside and spoil the appearance of package.

leak to the outside and spoil the appearance of package.

Detection of leaks by:

Detection of leaks by:

1.

Spark tester probe

Spark tester probe

(moving from liquid layer into air space).

(moving from liquid layer into air space).

Thus blue spark occur if airspace is evacuated.

Thus blue spark occur if airspace is evacuated.

2.

Dying method 

Dying method 

(by applying –ve pressure and submerged

(by applying –ve pressure and submerged

ampoule in 0.5-1% methylene blue dye bath during autoclave

ampoule in 0.5-1% methylene blue dye bath during autoclave

cycle – adv. Of leaker detection and sterilization)

cycle – adv. Of leaker detection and sterilization)

so if dye

so if dye

penetrate then leak is present.

penetrate then leak is present.

Note:

Note:

capillaries of 15 Micron or smaller may not be detected.

capillaries of 15 Micron or smaller may not be detected.

Subjective evaluation of the observer.

Subjective evaluation of the observer.



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The presence of pyrogenic substances in parenteral

The presence of pyrogenic substances in parenteral

preparations is determined by:

preparations is determined by:

In vivo test: 

In vivo test: 

a qualitative biologic test based on fever

response in rabbits.

In vitro test: 

In vitro test: 

utilizing gelling property of the LAL. In the

presence of pyrogenic endotoxins from gram –ve

bacteria, a firm gel is formed within 60 min. when

incubated at 37°C.

Note:

Note:

A- 

A- 

LAL test is 5-10 times more sensitive than rabbit

LAL test is 5-10 times more sensitive than rabbit

test.

test.

B – 

B – 

Greater danger present from injection of large volume

Greater danger present from injection of large volume

solutions containing pyrogens than from small volumes.

solutions containing pyrogens than from small volumes.

C- 

C- 

Pyrogenic effect is less with I.M. injection than with I.V.

Pyrogenic effect is less with I.M. injection than with I.V.

Packaging of sterile products to convey for the

user the quality, purity and reliability of the

product and representing:

1.

Dignified

2.

Neat

3.

Attractiveness

4.

Accurate and completely provide with

information for its use.

5.

Protect the package against physical

damage during shipping, handling and

storage and from UV radiation (for light

sensitive substances).

Packaging requirements for injection

Packaging requirements for injection

according to USP:

according to USP:

1.

The volume of an injection for single-dose container

The volume of an injection for single-dose container

should provide the amount specified at one time

should provide the amount specified at one time

(to prevent later time use later exposure to

contamination).

2.

Preparations intended for intraspinal, intracisternal

Preparations intended for intraspinal, intracisternal

or peridural should be packaged only in single-

or peridural should be packaged only in single-

dose containers 

dose containers 

(because of sensitivity of nerve

tissues to irritation from added substances- such as

antibacterial agents- ).

3.

No multiple dose container shall contain a volume

No multiple dose container shall contain a volume

of injection more than is sufficient to permit

of injection more than is sufficient to permit

withdrawal of 30 ml

withdrawal of 30 ml

 (because larger volumes would

provide for the withdrawal of more doses thereby

increasing the potential for contamination).