Sterile Product

Manufacturing

Introduction
To give an overview of the principles involved in the
manufacture of sterile products
The overall objective is to produce product that has a
high assurance of sterility (and which meets all other
quality parameters)
This presentation:
Summarises the general approach
Gives a framework for other detailed guides on specific aspects of

sterilisation & sterile manufacturing

Illustrates the underlying principles
Provides advice and gives recommendations.

General Principles of Sterile

Manufacturing

Moist Heat Sterilization

Dry Heat Sterilization
Aseptic Processing
Environmental Monitoring
Ethylene Oxide Sterilization
Sterile Filtration
Water systems validation
Sterility testing
Radiation Sterilization
Visual Inspection

Fundamentals

Sterility is the absence of living organisms

This is an absolute definition

The probability of achieving sterility depends on the overall process

It is generally accepted that a terminally sterilized product should have a probability

of non-sterility of less than 10-6 (i.e., a lower probability than one in a million of
having a non-sterile unit)

This is often expressed as an SAL Sterility Assurance Level of 106

This is a worst-case figure (with a challenge more resistant than product bioburden).
Real confidence levels are generally very much higher

A figure that has sometimes been quoted for aseptically filled product is probability of
non-sterility of less than 10-3. However, this is harder to analyse as contamination
does not follow a clear statistical distribution. Potential contamination sources are not
randomly distributed.

Why Validate and Control?

The test for sterility cannot confirm that the whole
batch is sterile
It is performed on a sample from a batch and has

statistical limitations
It can miss contamination if only a proportion of units
are non-sterile

It is thus necessary to recognize and understand

every aspect that could lead to loss of sterility
assurance
Such conditions should be prevented by the
application of carefully designed barriers and/or
control measures.

Development Validation and

Control

It is important that the product and process are designed to

maximise sterility assurance

Wherever possible, the product should be developed to withstand

sterilization in the final container

Once the product design is defined, a suitable production process

must be developed

This is installed and validated

The process must then be tightly controlled to assure reliability and

consistency.

Product Design Considerations

Facility Design

Cleaning and disinfection of the

Facility

Water

Categories of Water

Gases and Vacuum

Equipment (1)

Equipment (2)
Equipment Sterilization and handling

Sterilization must follow a validated procedure

Aseptic processes designed to minimise aseptic

assembly and intervention

Unavoidable aseptic assembly needs clear &
precise procedures
Aseptic assembly must be simulated (worst-case)
in media fill simulation trials

Sterilization In Place is a good method where

possible must be validated.

Personnel
Training - personnel appropriately trained for sterile processing,
including assessment and documentation:

Basic GMP
Fundamentals of microbiology
Personal hygiene, health and cleanliness
Behaviour and aseptic working techniques
Gowning and entry procedures
Cleaning and disinfection
Sterilization procedures, validation and routine operation
Emergency procedures to protect product quality (e.g. loss of HVAC System, loss of
power, equipment interventions etc.)

Personnel participating in aseptic processing must have practical

training in aseptic techniques before doing aseptic manipulations
They must have participated in a successful media fill run.

Gowning and Aseptic Technique

Gowning
Personnel must correctly wear appropriate clean room garments
Detailed, easily understood, gowning procedure (preferably illustrated)

Aseptic Techniques
Personnel in the aseptic manufacturing area, must understand the
principles of aseptic procedures
They must only be considered qualified after appropriate training,
working under supervision and demonstration of competence
The supervisor should observe technique & correct as necessary
All personnel directly involved in aseptic processing must participate in
a media fill at least once per year

Glove disinfection
Sterile disinfectants must be available (e.g., alcohol based)
Glove disinfection must be reasonably frequent, defined in SOP.

Environmental Monitoring (1)

The scope of environmental monitoring
includes:
Non-viable particulates,
Viable (microbial) counts
Differential pressures
Temperatures
Humidities
Air flows

Environmental Monitoring (2)

Monitoring During Room Qualification

Operational Qualification (OQ) at rest conditions to verify operation
Performance Qualification (PQ) in worst case operational conditions
Action levels should meet USP or Euro GMP as applicable
Alert levels tight enough to detect deterioration, but not so tight that
they become meaningless due to frequent transgression
PQ must cover a sufficient period to establish consistency
Routine Monitoring
Ensures area remains satisfactory. Results should be within alert level
Results above alert levels need review and perhaps corrective actions
Above action levels, must trigger appropriate actions (described in
guide),
Results must be assessed for trends so that progressive or sudden
changes in the results may be observed. This should be reviewed
regularly.

Environmental Monitoring (3)

Deviation Reports and Failure Investigations

Recommended Methods for Routine Monitoring

Monitoring Plan

The data must be analysed

Where necessary further investigations initiated
Possible contamination sources to be assessed and, eliminated
Outcome and detail must be reported

Physical measurements of the air supply

Physical and microbiological monitoring of the environment
Particles (viable and non-viable) in the air
Micro-organisms settling out of the air
Micro-organisms contaminating surfaces
Presence of micro-organisms on the hands and garments

Defined monitoring plans: tests, locations, alert/action levels & frequencies

May contain details of water, compressed gas clean steam testing
A review of environmental data is a requirement for batch release.

Bioburden and Components

Active Ingredients, Excipients, Additives

All ingredients should have appropriate biological specifications
Any limitations to sterilization must be defined
Description of origin (e.g. virological / prion risk)
Materials Used in the Process
Where appropriate, determine bioburden (e.g., ion exchange materials)
Primary Packaging Components
Container and the closure and cleaning / sterilization to be clearly
specified
Steps such as siliconization may need monitoring
If cleaning/sterilization is by supplier, same exigencies apply
Container-closure integrity
The integrity must be validated
Simulate, where appropriate: stress from processing
Method appropriate to container/closure system

Weighing, Compounding and

Sterilization

Weighing and compounding must be carried out in suitably classified rooms

Vessels must be cleaned, and sterilized or sanitised as appropriate and

stored dry in a way to prevent microbial contamination

Storage of pre-sterilization intermediates to be controlled & time limited

Following aspects to be considered:

Pre-filtration bioburden (filter sterilized material)
Pre-sterilization bioburden
Appropriate in-process controls

Sterilization of product and product contact materials

Selection of a suitable sterilization protocol must be based on SAL
Method must also consider the stability of the product
Validation always required
Change control is vital; even apparently minor change must be assessed

Terminal Sterilization

Steam Sterilization

By far the most common method for aqueous-based pharmaceuticals

Preferred cycle is the Pharm Eur reference cycle is 15 minutes at 121C
The sterilization cycle chosen must be compatible with product stability
Sterilization parameters clearly defined
In conjunction with other controls, the required SAL must be demonstrated
Validation to confirm sterilization conditions consistently throughout the load

Sterilization by Ionizing Radiation

Common for medical devices, but not for pharmaceuticals.

Pharm. Eur. reference condition, 25 KiloGray (kGy), has been widely accepted. Other

conditions may be used if validated and accepted by the regulator

Important to consider susceptibility of the product to radiation damage

Dry Heat Sterilization

Lower antimicrobial efficacy than moist heat, thus higher temperatures and/or longer

exposures. Pharm Eur reference cycle is 2 hours @ 160C

Rarely used for terminal sterilization of pharmaceuticals; in rare cases heat resistant
non-aqueous products may be terminally sterilized.

Sterilization of Items for Aseptic Fill

(1)

Steam Sterilization

Widely used, but careful validation needed particularly complex items

Broadly similar to terminal steam sterilization, but two aspects are critical

Quality of saturated steam

Removal of air and subsequent steam penetration

Sterilization by Ionizing Radiation

Dry Heat Sterilization/Depyrogenation

May be used for temperature sensitive primary packaging or components

Used for disposables for sterile areas and sterility testing areas
Validation includes dosimetry, - correct, even, irradiation of the items
Sterilization/ depyrogenation of heat resistant primary packaging materials
Pharm Eur notes that temperatures in excess of 220 oC have been frequently used,

the USP suggests 250 15 oC

Validation must include endotoxin challenge studies
Dry heat may be used to sterilize non-aqueous preparations (e.g. Ointment bases) at
lower temperature/time relationships, without depyrogenation.

Sterilization of Items for Aseptic Fill

(2)
Ethylene Oxide Sterilization
Quite widely used to sterilize heat labile components
European Pharmacopoeia and the European GMP guide indicate
that this method should only be used where there is no suitable
alternative
Hazardous - toxic, potentially carcinogenic, flammable,
potentially explosive
Generally conducted by specialized contractors
There are strict regulatory limits on maximum permissible
product residues
Bulk packs for sterilization must be gas permeable, but sealed
against microbial ingress
Sterilization must consider packaging, load pattern, gas
penetration (ethylene oxide & water vapour), bulk pack integrity
Validation and routine monitoring must include Biological
indicators.

Sterilization by Filtration (Liquids)

Principle:

Contaminating organisms are not killed, but are retained on the filters. Any faults in

the filter structure, may compromise this

Validation includes:

Validation should address:

Retention of bacterial challenge: B. diminuta at 107 per cm2

This is correlated with an integrity test value
Filter suitability - toxicity, extractables, shedding of particles
Adsorption of product
Compatibility with product solvents
The required filter size and suitability of the filtration equipment
Retention of B.diminuta in the actual product under process conditions
Parameters for the physical integrity test

Routine Filtration

Conducted in line with the validated parameters

Check integrity testing, process time, differential pressure, flow rates, sterilization and

reuse of filters.

Performance Qualification of Aseptic

Manufacturing
Based on simulating the risk of contamination in all
aseptic operations
For a new process, a minimum of three consecutive
satisfactory media filling trials
For aqueous liquid products, simulation trials use a
liquid microbiological medium
For solid dosage forms, a powder placebo is used,
followed by aseptic reconstitution into a liquid
microbiological medium
The following slide gives a general overview....

Aseptic Process Simulation

(Media Fill Trial)
Media Fill Trials (MFTs)

All process stages simulated as closely as possible

Particularly interventions and manual manipulations
Must follow routine procedures and include all interventions
Regular interventions simulated with the same frequency as actual
process
In each case, the worst-case eventuality must be covered
Process must be successfully validated before product filling is
permitted
Revalidation by media fill must be conducted every half year (each line)

Manufacturing Environment

Microbiological monitoring must be performed during the trial

Filling Conditions and Equipment

All according to routine operating conditions and at normal times of day

Containers must be passed through all stages.

Thank You
Any Questions