© World Health Organization

WHO Technical Report Series, No. 961, 2011

Annex 5
Supplementary guidelines on good manufacturing
practices for heating, ventilation and air-
conditioning systems for non-sterile pharmaceutical
dosage forms

1. Introduction
2. Scope of document
3. Glossary
4. Protection
4.1 Products and personnel
4.2 Air filtration
4.3 Unidirectional airflow
4.4 Infiltration
4.5 Cross-contamination
4.6 Displacement concept (low pressure differential, high airflow)
4.7 Pressure differential concept (high pressure differential, low airflow)
4.8 Physical barrier concept
4.9 Temperature and relative humidity
5. Dust control
6. Protection of the environment
6.1 General
6.2 Dust in exhaust air
6.3 Vapour and fume removal
7. Design of HVAC systems and components
7.1 General
7.2 Air distribution
7.3 Recirculation system
7.4 Full fresh-air systems
7.5 Additional system components
8. Commissioning, qualification and maintenance
8.1 Commissioning
8.2 Qualification
8.3 Maintenance
9. Premises
References
Further reading

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1. Introduction
Heating, ventilation and air-conditioning (HVAC) play an important role in
ensuring the manufacture of quality pharmaceutical products. A well designed
HVAC system will also provide comfortable conditions for operators.
These guidelines mainly focus on recommendations for systems for
manufacturers of solid dosage forms. The guidelines also refer to other
systems or components which are not relevant to solid dosage form
manufacturing plants, but which may assist in providing a comparison
between the requirements for solid dosage-form plants and other systems.
HVAC system design influences architectural layouts with regard to items
such as airlock positions, doorways and lobbies. The architectural components
have an effect on room pressure differential cascades and cross-contamination
control. The prevention of contamination and cross-contamination is an
essential design consideration of the HVAC system. In view of these critical
aspects, the design of the HVAC system should be considered at the concept
design stage of a pharmaceutical manufacturing plant.
Temperature, relative humidity and ventilation should be appropriate and
should not adversely affect the quality of pharmaceutical products during
their manufacture and storage, or the accurate functioning of equipment.
This document aims to give guidance to pharmaceutical manufacturers
and inspectors of pharmaceutical manufacturing facilities on the design,
installation, qualification and maintenance of the HVAC systems.
These guidelines are intended to complement those provided in Good
manufacturing practices for pharmaceutical products (1) and should be read
in conjunction with the parent guide. The additional standards addressed by
the present guidelines should, therefore, be considered supplementary to
the general requirements set out in the parent guide.

2. Scope of document
These guidelines focus primarily on the design and good manufacturing
practices (GMP) requirements for HVAC systems for facilities for the
manufacture of solid dosage forms. Most of the system design principles
for facilities manufacturing solid dosage forms also apply to other facilities
such as those manufacturing liquids, creams and ointments. These guidelines
do not cover requirements for manufacturing sites for the production of
sterile pharmaceutical products. These guidelines do not cover the specific
requirements relating to facilities handling hazardous products. Guidelines
for hazardous product facilities are covered in a separate WHO guideline.
These guidelines are intended as a basic guide for use by pharmaceutical
manufacturers and GMP inspectors.

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 They are not intended to be prescriptive in specifying requirements and design
parameters. There are many parameters affecting a clean area condition and it
is, therefore, difficult to lay down the specific requirements for one particular
parameter in isolation.
Many pharmaceutical manufacturers have their own engineering design and
qualification standards and requirements may vary from one manufacturer
to the next. Design parameters and user requirements should, therefore, be
set realistically for each project, with a view to creating a cost-effective
design, yet still complying with all regulatory standards and ensuring that
product quality and safety are not compromised. The three primary aspects
addressed in this manual are the roles that the HVAC system plays in product
protection, personnel protection and environmental protection (Figure 1).
Cognisance should be taken of the products to be manufactured when
establishing system design parameters. A facility manufacturing multiple
different products may have more stringent design parameters with respect
to cross-contamination control, compared with a single product facility.
Figure 1
The guidelines address the various system criteria according
to the sequence set out in this diagram

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3. Glossary
The definitions given below apply to terms used in these guidelines. They
may have different meanings in other contexts.

acceptance criteria
Measurable terms under which a test result will be considered acceptable.

action limit
The action limit is reached when the acceptance criteria of a critical
parameter have been exceeded. Results outside these limits will require
specified action and investigation.

air changes per hour (ACPH)

The volume of air supplied to a room, in m3/hr, divided by the room volume, in m3.

air-handling unit (AHU)

The air-handling unit serves to condition the air and provide the required air
movement within a facility.

airlock
An enclosed space with two or more doors, which is interposed between
two or more rooms, e.g. of differing classes of cleanliness, for the purpose
of controlling the airflow between those rooms when they need to be
entered. An airlock is designed for and used by either people or goods
(PAL, personnel airlock; MAL, material airlock).

alert limit
The alert limit is reached when the normal operating range of a critical
parameter has been exceeded, indicating that corrective measures may need
to be taken to prevent the action limit being reached.

as-built
Condition where the installation is complete with all services connected and
functioning but with no production equipment, materials or personnel present.

at-rest
Condition where the installation is complete with equipment installed and
operating in a manner agreed upon by the customer and supplier, but with
no personnel present.

central air-conditioning unit (see air-handling unit)

change control
A formal system by which qualified representatives of appropriate
disciplines review proposed or actual changes that might affect a validated

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 status. The intent is to determine the need for action that would ensure that
the system is maintained in a validated state.
clean area (cleanroom)18
An area (or room or zone) with defined environmental control of particulate
and microbial contamination, constructed and used in such a way as to reduce
the introduction, generation and retention of contaminants within the area.
closed system
A system where the product or material is not exposed to the manufacturing
environment.
commissioning
Commissioning is the documented process of verifying that the equipment
and systems are installed according to specifications, placing the equipment
into active service and verifying its proper action. Commissioning takes
place at the conclusion of project construction but prior to validation.
containment
A process or device to contain product, dust or contaminants in one zone,
preventing it from escaping to another zone.
contamination
The undesired introduction of impurities of a chemical or microbial nature,
or of foreign matter, into or on to a starting material or intermediate, during
production, sampling, packaging or repackaging, storage or transport.
controlled area
An area within the facility in which specific environmental facility
conditions and procedures are defined, controlled, and monitored to prevent
degradation or cross-contamination of the product.
critical parameter or component
A processing parameter (such as temperature or relative humidity) that
affects the quality of a product, or a component that may have a direct
impact on the quality of the product.
critical quality attribute (CQA)
A physical, chemical, biological or microbiological property or characteristic
that should be within an appropriate limit, range or distribution to ensure
the desired product quality.

1
Note: Clean area standards, such as ISO 14644-1, provide details on how to classify air cleanliness by
means of particle concentrations, whereas the GMP standards provide a grading for air cleanliness
in terms of the condition (at-rest or operational), the permissible microbial concentrations, as well
as other factors such as gowning requirements. GMP and clean area standards should be used in
conjunction with each other to define and classify the different manufacturing environments.

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 cross-contamination
Contamination of a starting material, intermediate product or finished
product with another starting material or product during production.
design condition
Design condition relates to the specified range or accuracy of a controlled
variable used by the designer as a basis for determining the performance
requirements of an engineered system.
design qualification (DQ)
Design qualification is the documented check of planning documents and
technical specifications for conformity of the design with the process,
manufacturing, GMP and regulatory requirements.
direct impact system
A system that is expected to have a direct impact on product quality. These
systems are designed and commissioned in line with good engineering
practice (GEP) and, in addition, are subject to qualification practices.
exfiltration
Exfiltration is the egress of air from a controlled area to an external zone.
facility
The built environment within which the clean area installation and associated
controlled environments operate together with their supporting infrastructure.
good engineering practice (GEP)
Established engineering methods and standards that are applied throughout
the project life-cycle to deliver appropriate, cost-effective solutions.
hazardous substance or product
A product or substance that may present a substantial risk of injury to health
or to the environment
indirect impact system
This is a system that is not expected to have a direct impact on product
quality, but typically will support a direct impact system. These systems are
designed and commissioned according to GEP only.
infiltration
Infiltration is the ingress of air from an external zone into a controlled area.
installation qualification (IQ)
Installation qualification is documented verification that the premises,
HVAC system, supporting utilities and equipment have been built and
installed in compliance with their approved design specification.

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no-impact system
This is a system that will not have any impact, either directly or indirectly, on
product quality. These systems are designed and commissioned according
to GEP only.
non-critical parameter or component
A processing parameter or component within a system where the operation,
contact, data control, alarm or failure will have an indirect impact or no
impact on the quality of the product.
normal operating range
The range that the manufacturer selects as the acceptable values for a parameter
during normal operations. This range must be within the operating range.
operating limits
The minimum and/or maximum values that will ensure that product and
safety requirements are met.
operating range
Operating range is the range of validated critical parameters within which
acceptable products can be manufactured.
operational condition
This condition relates to carrying out room classification tests with the
normal production process with equipment in operation, and the normal
staff present in the room.
operational qualification (OQ)
Operational qualification is the documentary evidence to verify that the equipment
operates in accordance with its design specifications in its normal operating range
and performs as intended throughout all anticipated operating ranges.
oral solid dosage (OSD)
Usually refers to an OSD plant that manufactures medicinal products such
as tablets, capsules and powders to be taken orally.
pass-through-hatch (PTH) or pass box (PB)
A cabinet with two or more doors for passing equipment or product, whilst
maintaining the pressure cascade and segregation between two controlled
zones. A passive PTH has no air supply or extract. A dynamic PTH has an
air supply into the chamber.
performance qualification (PQ)
Performance qualification is the documented verification that the process and/
or the total process related to the system performs as intended throughout all
anticipated operating ranges.

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 point extraction
Air extraction to remove dust with the extraction point located as close as
possible to the source of the dust.

pressure cascade
A process whereby air flows from one area, which is maintained at a higher
pressure, to another area at a lower pressure.

qualification
Qualification is the planning, carrying out and recording of tests on
equipment and a system, which forms part of the validated process, to
demonstrate that it will perform as intended.

quality critical process parameter (CPP)

A process parameter which could have an impact on the critical quality
attribute.

relative humidity
The ratio of the actual water vapour pressure of the air to the saturated
water vapour pressure of the air at the same temperature expressed as
a percentage. More simply put, it is the ratio of the mass of moisture
in the air, relative to the mass at 100% moisture saturation, at a given
temperature.

standard operating procedure (SOP)

An authorized written procedure, giving instructions for performing
operations, not necessarily specific to a given product or material, but of a
more general nature (e.g. operation of equipment, maintenance and cleaning,
validation, cleaning of premises and environmental control, sampling and
inspection). Certain SOPs may be used to supplement product-specific
master and batch production documentation.

turbulent flow
Turbulent flow, or non-unidirectional airflow, is air distribution that is
introduced into the controlled space and then mixes with room air by means
of induction.

unidirectional airflow (UDAF)

Unidirectional airflow is a rectified airflow over the entire cross-sectional
area of a clean zone with a steady velocity and approximately parallel
streamlines (see also turbulent flow).

validation
The documented act of proving that any procedure, process, equipment,
material, activity or system actually leads to the expected results.

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 validation master plan (VMP)
Validation master plan is a high-level document which establishes an
umbrella validation plan for the entire project, and is used as guidance by
the project team for resource and technical planning (also referred to as
master qualification plan).

4. Protection
4.1 Products and personnel
4.1.1 Areas for the manufacture of pharmaceuticals, where pharmaceutical
starting materials and products, utensils, primary packing materials and
equipment are exposed to the environment, should be defined as “clean
areas”, “clean zones”, “controlled areas” or “cleanrooms”.
4.1.2 The achievement of a particular clean area condition depends on a
number of criteria that should be addressed at the design and qualification
stages. A suitable balance between the different criteria will be required in
order to create an efficient clean area.
4.1.3 Some of the basic criteria to be considered which affects room
cleanliness should include:
• building finishes and structure
• air filtration
• air change rate or flushing rate
• room pressure
• location of air terminals and directional airflow
• temperature
• relative humidity
• material flow
• personnel flow
• gowning procedures
• equipment movement
• process being carried out (open or closed system)
• outside air conditions
• occupancy
• type of product
• cleaning standard operating procedures (SOPs).
4.1.4 Air filtration and air change rates should be set to ensure that the
defined clean area condition is attained.
4.1.5 The air change rates should be determined by the manufacturer and
designer, taking into account the various critical parameters using a risk
based approach with due consideration of capital and running costs and

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 energy usage. Primarily the air change rate should be set to a level that will
achieve the required clean area condition.
4.1.6 Air change rates are normally determined by the following
considerations (could normally vary between 6 and 20 air changes per hour):
• area condition required: whether a specific room cleanliness condition
is in fact required and whether the room condition is rated for an “at
rest” condition or an “operational” condition (air change rate should be
selected on need rather than tradition)
• the product characteristics (e.g. odours, hygroscopicity, etc)
• the quality and filtration of the supply air
• particulates generated by the manufacturing process
• particulates generated by the operators
• configuration of the room and air supply and extract locations
• sufficient air to achieve containment effect and to clean up the area
• sufficient air to cope with the room heat load
• sufficient air to balance extract rates
• sufficient air to maintain the required room pressure.
4.1.7 If a cleanroom classification is specified the manufacturer should
state whether this is achieved under “as-built” (Figure 2), “at-rest” (Figure 3)
or “operational” (Figure 4) conditions.
4.1.8 Room classification tests in the “as-built” condition should be
carried out on the bare room, in the absence of any equipment or personnel.
4.1.9 Room classification tests in the “at-rest” condition should be carried
out with the equipment operating where relevant, but without any operators.
Because of the amounts of dust usually generated in a solid dosage facility,
the clean area classifications would be rated for the “at-rest” condition.
4.1.10 Room classification tests in the “operational” condition are
normally carried out during the normal production process with equipment
operating, and the normal number of personnel present in the room.
Generally a room that is tested for an “operational” condition should be
able to be cleaned up to the “at-rest” clean area classification after a short
clean-up time. The clean-up time should be determined through validation
and is generally of the order of 20 minutes.
4.1.11 Materials and products should be protected from contamination
and cross-contamination during all stages of manufacture (see also section
4.5 for cross-contamination control).
Note: contaminants may result from inappropriate premises (e.g. poor design,
layout or finishing), poor cleaning procedures, contaminants brought in by
personnel, poor manufacturing process and a poor HVAC system.

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Figure 2
“As-built” condition

Figure 3
“At-rest” condition

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Figure 4
“Operational” condition

4.1.12 Airborne contaminants should be controlled through effective

ventilation and filtration.
4.1.13 External contaminants should be removed by effective
filtration of the supply air (see Figure 5 for an example of a shell-like
building layout to enhance containment and protection from external
contaminants).
4.1.14 Internal contaminants should be controlled by dilution and
flushing of contaminants in the room, or by displacement airflow (See
Figures 6 and 7 for examples of methods for the flushing of airborne
contaminants).
4.1.15 Airborne particulates and the degree of filtration should be
considered critical parameters with reference to the level of product
protection required.
4.1.16 Personnel should not be a source of contamination.
4.1.17 The level of protection and air cleanliness for different areas should
be determined according to the product being manufactured, the process
being used and the product’s susceptibility to degradation (Table 1).

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Figure 5
Shell-like containment control concept

4.2 Air filtration

Note: The degree to which air is filtered plays an important role in the
prevention of contamination and the control of cross-contamination.
4.2.1 The type of filters required for different applications depends on
the quality of the ambient air and the return air (where applicable) and
also on the air change rates. Table 2 gives the recommended filtration
levels for different levels of protection in a pharmaceutical facility.
Manufacturers should determine and prove the appropriate use of
filters.
4.2.2 Filter classes should always be linked to the standard test method
because referring to actual filter efficiencies can be very misleading (as
different test methods each result in a different value for the same filter).
(Referring to filter classifications such as an 85% filter or a 5 μm filter are not
valid classifications and should not be used, as this can lead to the incorrect
filter being installed. Only the EN 779 and EN 1822 classifications, as per
the table below, should be used.)

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Figure 6
Turbulent dilution of dirty air

Low-level extract is ideal for dust suppression purposes, but is not essential. (Low-level extract is
essential for Grade A, B & C areas.)

Figure 7
Unidirectional displacement of dirty air

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Table 1
Examples of levels of protection (based on ISPE oral solid dosage (OSD)
Guideline criteria)
Level Condition Example of area
Level 1 General Area with normal housekeeping and maintenance
where there is no potential for product
contamination, e.g. warehousing.
Level 2 Protected Area in which steps are taken to protect the
pharmaceutical starting material or product from
direct or indirect contamination or degradation, e.g.
secondary packing, warehousing, first stage change
rooms.
Level 3 Controlled Area in which specific environmental conditions
are defined, controlled and monitored to prevent
contamination or degradation of the pharmaceutical
starting material or product, e.g. where product,
starting materials and components are exposed to
the room environment; plus equipment wash and
storage areas for equipment product contact parts.

Table 2
Levels of protection and recommended filtration
Level of protection Recommended filtration
Level 1 Primary filters only (e.g. EN 779 G4 filters)
Level 2 Protected areas operating on 100% outside air: primary plus
secondary filters (e.g. EN 779 G4 plus F8 or F9 filters)
Level 3 Production facility operating on recirculated plus ambient
air, where potential for cross-contamination exists: Primary
plus secondary plus tertiary filters (e.g. EN 779 G4 plus F8
plus EN 1822 H13 filters) (for full fresh air system, without
recirculation, G4 and F8 or F9 filters are acceptable)

Note: The filter classifications referred to above relate to the EN 1822 and
EN 779 test standards (EN 779 relates to filter classes G1 to F9 and EN
1822 relates to filter classes E10 to U17). Refer to Figure 8 for comparative
classifications of other filter standards.
4.2.3 In selecting filters, the manufacturer should have considered
other factors, such as particularly contaminated ambient conditions, local
regulations and specific product requirements. Good pre-filtration extends
the life of the more expensive filters downstream.
4.2.4 Materials for components of an HVAC system should be selected
with care so that they do not become a source of contamination. Any
component with the potential for liberating particulate or microbial
contamination into the air stream should be located upstream of the final
filters.

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Figure 8
Comparison of filter test standards

230
 4.2.5 Where possible ventilation dampers, filters and other services should
be designed and positioned so that they are accessible from outside the
manufacturing areas (service voids or service corridors) for maintenance
purposes.
4.2.6 Directional airflow within production or primary packing areas
should assist in preventing contamination. Airflows should be planned in
conjunction with operator locations, so as to minimize contamination of the
product by the operator and also to protect the operator from dust inhalation.
4.2.7 HVAC air distribution components should be designed, installed
and located to prevent contaminants generated within the room from being
spread.
4.2.8 Supply air diffusers should be selected with care taking consideration
of, e.g. room requirements and positions of equipment and operators in the
room. Supply air diffusers of the high induction type (e.g. those typically
used for office-type air-conditioning) should where possible not be used
in clean areas where dust is liberated. Air diffusers should be of the non-
induction type, introducing air with the least amount of induction so as to
maximize the flushing effect. In rooms where the process results in high
dust liberation; perforated plates or low induction swirl diffusers with
low level extract or return should be used (to contain the dust at the lower
level of the room) (see Figures 9–11 for illustrations of the three types of
diffuser). In cases where dust liberation is low, ceiling return air grilles may
be acceptable.
4.2.9 Induction and certain swirl diffusers induce room air vertically
up to the diffuser to mix with the supply air. These diffusers create good
dilution of contaminants in the room and may be used in rooms where there
is low dust liberation. However, if used in rooms where excessive dust is
generated, the distribution of dust in the room could be hazardous for the
operators in the room.

4.3 Unidirectional airflow

4.3.1 Unidirectional airflow (UDAF) should be used for weighing booths
or sampling booths to provide operator and product protection and should
also have a slight air in-flow from the room to enhance containment. Dust
containment at the weigh booth should be demonstrated by smoke airflow
pattern tests, or other appropriate tests. UDAF can also be used to provide
protection of other dusty processes.
4.3.2 Sampling of materials such as starting materials, primary packaging
materials and products, should be carried out in the same environmental
conditions that are required for the further processing of the product.

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Figure 9
Induction diffuser

Figure 10
Perforated plate diffuser

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Figure 11
Swirl diffuser

4.3.3 In a weighing booth situation, the aim of the UDAF is to provide

dust containment and operator protection.
Example: In Figure 12 the dust generated at the weighing station is
immediately extracted through the perforated worktop, thus protecting the
operator from dust inhalation, but at the same time protecting the product
from contamination by the operator by means of the vertical unidirectional
airflow stream.
4.3.4 The unidirectional flow velocity should be such that it does not
disrupt the sensitivity of balances in weighing areas. Where necessary the
velocity may be reduced to prevent inaccuracies during weighing, provided
that sufficient airflow is maintained to provide containment. Conventional
unidirectional airflow systems, where a Grade A condition is required, have
a guidance airflow velocity of 0.36 to 0.54 m/s. However, in a weigh booth
or sampling booth a lower velocity can be used as a Grade A condition is
not required. It is often necessary to reduce velocities to a lower level in
order not to influence balance readings. The airflow velocity and directional
flow should still ensure product containment. For this type of application
it is sometimes better to refer to the unit as an airflow protection booth
(APB) rather than a UDAF, in order to avoid confusion, with a Grade A
requirement.
4.3.5 The position in which the operator stands relative to the source of
dust liberation and airflow should be determined to ensure that the operator
is not in the path of an airflow that could lead to contamination of the
product (Figure 13).

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Figure 12
Operator protection at weighing station

4.3.6 Once the system has been designed and qualified with a specific
layout for operators and processes, this should be maintained in accordance
with an SOP.
4.3.7 There should be no obstructions in the path of a unidirectional flow
air stream that may cause the operator to be exposed to dust.
Figure 14 illustrates the incorrect use of a weighing scale which has a solid
back. The back of the weighing scale should not block the return air path
as this causes air to rise vertically, resulting in a hazardous situation for the
operator.
Figure 15 illustrates a situation where an open bin is placed below a vertical
unidirectional flow distributor. The downward airflow should be prevented
from entering the bin, and then being forced to rise again, as this would
carry dust up towards the operator’s face. In such an occurrence it may be
necessary to add a partial cover over the bin to limit the entry of air. Point
extraction could also be used but this can result in the excessive loss of
product.
Figure 16 shows that a solid worktop can sometimes cause deflection of
the vertical unidirectional airflow resulting in a flow reversal. A possible
solution would be to have a 100 mm gap between the back of the table and
the wall, with the air being extracted here.

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Figure 13
Operator protection by horizontal airflow

4.3.8 The manufacturer should select either vertical or horizontal

unidirectional flow (Figure 17) and an appropriate airflow pattern to provide
the best protection for the particular application.
4.3.9 Return or exhaust air grilles in rooms or at weigh or sampling booths
should preferably be of the perforated grille types, which are easy to clean.
Return/exhaust air filters can either be installed at the room terminal or in
the air-handling unit. Maintenance and cleaning of filters and ducts should
be addressed to ensure constant airflow.

4.4 Infiltration
4.4.1 Air infiltration of unfiltered air into a pharmaceutical plant should
not be a source of contamination.

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Figure 14
Operator subject to powder inhalation due to obstruction

Figure 15
Operator subject to powder contamination due to airflow reversal in bin

236
Figure 16
Operator subject to powder inhalation due to worktop obstruction

4.4.2 Manufacturing facilities should normally be maintained at a positive

pressure relative to the outside, to limit the ingress of contaminants. Where
facilities are to be maintained at negative pressures relative to the ambient
pressure, special precautions should be taken. Refer to the WHO guideline
for hazardous products, for further guidance on negative pressure facilities.
4.4.3 The location of the negative pressure facility should be carefully
considered with reference to the areas surrounding it, particular attention
being given to ensuring that the building structure is well sealed.
4.4.4 Negative pressure zones should, as far as possible, be encapsulated
by surrounding areas with clean air supplies, so that only clean air can
infiltrate into the controlled zone.

4.5 Cross-contamination
4.5.1 Where different products are manufactured at the same time, in
different areas or cubicles, in a multiproduct OSD manufacturing site,
measures should be taken to ensure that dust cannot move from one cubicle
to another.
4.5.2 Correct directional air movement and a pressure cascade system can
assist in preventing cross-contamination. The pressure cascade should be
such that the direction of airflow is from the clean corridor into the cubicles,
resulting in dust containment.
4.5.3 The corridor should be maintained at a higher pressure than the
cubicles, and the cubicles at a higher pressure than atmospheric pressure.

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Figure 17
Diagram indicating horizontal and vertical unidirectional flow

4.5.4 Containment can normally be achieved by application of the

displacement concept (low pressure differential, high airflow), or the
pressure differential concept (high pressure differential, low airflow), or the
physical barrier concept.
4.5.5 The pressure cascade regime and the direction of airflow should be
appropriate to the product and processing method used.

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 4.5.6 Highly potent products should be manufactured under a pressure
cascade regime that is negative relative to atmospheric pressure.
4.5.7 The pressure cascade for each facility should be individually
assessed according to the product handled and level of protection required.
4.5.8 Building structure should be given special attention to accommodate
the pressure cascade design.
4.5.9 Ceilings and walls, close fitting doors and sealed light fittings should
be in place, to limit ingress or egress of air.

4.6 Displacement concept (low pressure differential, high airflow)

Note: This method of containment is not the preferred method, as the
measurement and monitoring of airflow velocities in doorways is difficult.
This concept is commonly found in production processes where large
amounts of dust are generated.
4.6.1 Under this concept the air should be supplied to the corridor,
flow through the doorway, and be extracted from the back of the cubicle.
Normally the cubicle door should be closed and the air should enter the
cubicle through a door grille, although the concept can be applied to an
opening without a door.
4.6.2 The velocity should be high enough to prevent turbulence within the
doorway resulting in dust escaping.
4.6.3 This displacement airflow should be calculated as the product of
the door area and the velocity, which generally results in fairly large air
quantities.
Note: Although this method of containment may still exist on older facilities,
it is not the preferred method, as the measurement and monitoring of doorway
velocities is difficult. In addition, simultaneously maintaining the correct
room pressure and the correct room air change rate is often not achieved.

4.7 Pressure differential concept (high pressure differential,

low airflow)
Note: The pressure differential concept may normally be used in zones where
little or no dust is being generated. It may be used alone or in combination
with other containment control techniques and concepts, such as a double
door airlock.
4.7.1 The high pressure differential between the clean and less clean
zones should be generated by leakage through the gaps of the closed doors
to the cubicle.

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 4.7.2 The pressure differential should be of sufficient magnitude to ensure
containment and prevention of flow reversal, but should not be so high as to
create turbulence problems.
4.7.3 In considering room pressure differentials, transient variations, such
as machine extract systems, should be taken into consideration.
4.7.4 A pressure differential of 15 Pa is often used for achieving
containment between two adjacent zones, but pressure differentials of
between 5 Pa and 20 Pa may be acceptable. Where the design pressure
differential is too low and tolerances are at opposite extremities, a flow
reversal can take place. For example, where a control tolerance of ± 3 Pa
is specified, the implications of rooms being operated at the upper and
lower tolerances should be evaluated. It is important to select pressures and
tolerances such that a flow reversal is unlikely to occur.
4.7.5 The pressure differential between adjacent rooms could be
considered a critical parameter, depending on the outcome of risk analysis.
The limits for the pressure differential between adjacent areas should be
such that there is no risk of overlap in the acceptable operating range, e.g.
5 Pa to 15 Pa in one room and 15 Pa to 30 Pa in an adjacent room, resulting
in the failure of the pressure cascade, where the first room is at the maximum
pressure limit and the second room is at its minimum pressure limit.
4.7.6 Low pressure differentials may be acceptable when airlocks
(pressure sinks or pressure bubbles) are used to segregate areas.
4.7.7 The effect of room pressure tolerances are illustrated in Figure 18.
4.7.8 The pressure control and monitoring devices used should be
calibrated and qualified. Compliance with specifications should be regularly
verified and the results recorded. Pressure control devices should be linked
to an alarm system set according to the levels determined by a risk analysis.
4.7.9 Manual control systems, where used, should be set up during
commissioning, with set point marked, and should not change unless other
system conditions change.
4.7.10 Airlocks can be important components in setting up and maintaining
pressure cascade systems and also to limit cross-contamination.
4.7.11 Airlocks with different pressure cascade regimes include the
cascade airlock, sink airlock and bubble airlock (Figures 19–21):
• cascade airlock: higher pressure on one side of the airlock and lower
pressure on the other;
• sink airlock: lower pressure inside the airlock and higher pressure on both
outer sides;

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Figure 18
Examples of pressure cascades

Figure 19
Example of cascade airlock
(In most cases the internal pressure of the airlock is not critical. The pressure differential
between the two outer sides is the important criteria.)

241
Figure 20
Example of sink airlock

Figure 21
Example of bubble airlock

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• bubble airlock: higher pressure inside the airlock and lower pressure on
both outer sides.
Note: The diagrams above and the differential pressures shown here are
for illustration purposes only. Pressures indicated in these examples are
absolute pressures, whereas the local pressure indication would most likely
be pressure differential from room to room.
4.7.12 Doors should open to the high pressure side, so that room pressure
assists in holding the door closed and in addition be provided with self-
closers. Should the doors open to the low pressure side, the door closer
springs should be sufficient to hold the door closed and prevent the pressure
differential from pushing the door open. There should be a method to
indicate if both doors to airlocks are open at the same time, or alternatively
these should be interlocked. The determination of which doors should be
interlocked should be the subject of a risk assessment study.
4.7.13 Central dust extraction systems should be interlocked with the
appropriate air-handling systems, to ensure that they operate simultaneously.
4.7.14 Room pressure differential between adjacent cubicles, which are
linked by common dust extraction ducting, should be avoided.
4.7.15 Air should not flow through the dust extraction ducting or return
air ducting from the room with the higher pressure to the room with the
lower pressure (this would normally occur only if extract or return systems
were inoperative). Systems should be designed to prevent dust flowing back
in the opposite direction in the event of component failure or airflow failure.
4.7.16 Adequate room pressure differential indication should be provided
so that each critical room pressure can be traced back to ambient pressure
(by summation of the room pressure differentials), in order to determine the
room actual absolute pressure. Room pressure indication gauges should have
a range and graduation scale which enables the reading to an accuracy, as
appropriate; normal operating range, alert and action limits should be defined
and displayed at the point of indication. A colour coding gauge may be helpful.
Room pressure indication may be either analogue or digital, and may be
represented as either pressure differentials or absolute pressures. Which
ever system is used any out-of-specification condition should be easily
identifiable.
4.7.17 Material pass-through-hatches (PTH) or pass boxes (PB) can also
be used for separating two different zones. PTHs fall into two categories,
namely a dynamic PTH or a passive PTH. Dynamic PTHs have an air
supply to or extraction from them, and can then be used as bubble, sink or
cascade PTHs.

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4.8 Physical barrier concept
4.8.1 Where appropriate, an impervious barrier to prevent cross-
contamination between two zones, such as closed systems, pumped or
vacuum transfer of materials, should be used.

4.9 Temperature and relative humidity

4.9.1 Where appropriate, temperature and relative humidity should be
controlled, monitored and recorded, where relevant, to ensure compliance
with requirements pertinent to the materials and products and provide a
comfortable environment for the operator where necessary.
4.9.2 Maximum and minimum room temperatures and relative humidity
should be appropriate. Alert and action limits on temperatures and
humidities should be set, as appropriate.
4.9.3 The operating band, or tolerance, between the acceptable minimum
and maximum temperatures should not be made too close. Tight control
tolerances may be difficult to achieve and can also add unnecessary
installation and running costs.
4.9.4 Cubicles, or suites, in which products requiring low relative humidity
are processed, should have well-sealed walls and ceilings and should also
be separated from adjacent areas with higher relative humidity by means of
suitable airlocks.
4.9.5 Precautions should be taken to prevent moisture migration that
increases the load on the HVAC system.
4.9.6 Humidity control should be achieved by removing moisture from
the air, or adding moisture to the air, as relevant.
4.9.7 Dehumidification (moisture removal) may be achieved by means of
either refrigerated dehumidifiers or chemical dehumidifiers.
4.9.8 Appropriate cooling media for dehumidification such as low
temperature chilled water/glycol mixture or refrigerant should be used.
4.9.9 Humidifiers should be avoided if possible as they may become a
source of contamination (e.g. microbiological growth). Where humidification
is required, this should be achieved by appropriate means such as the injection
of steam into the air stream. A product-contamination assessment should be
done to determine whether pure or clean steam is required for the purposes
of humidification.
4.9.10 Where steam humidifiers are used, chemicals such as corrosion
inhibitors or chelating agents, which could have a detrimental effect on

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 the product, should not be added to the boiler system. Only appropriate
additives should be added to the boiler system.
4.9.11 Humidification systems should be well drained. No condensate
should accumulate in air-handling systems.
4.9.12 Other humidification appliances such as evaporative systems,
atomizers and water mist sprays, should not be used because of the potential
risk of microbial contamination.
4.9.13 Duct material in the vicinity of the humidifier should not add
contaminants to air that will not be removed by filtration further downstream.
4.6.14 Air filters should not be installed immediately downstream of
humidifiers, as moisture on the filters could lead to bacterial growth.
4.9.15 Cold surfaces should be insulated to prevent condensation within
the clean area or on air-handling components.
4.9.16 When specifying relative humidity, the associated temperature
should also be specified.
4.9.17 Chemical driers using silica gel or lithium chloride are acceptable,
provided that they do not become sources of contamination.

5. Dust control
5.1 Wherever possible, dust or vapour contamination should be removed
at source. Point-of-use extraction, i.e. as close as possible to the point where
the dust is generated, should be employed. Spot ventilation or capture hoods
may be used as appropriate.
5.2 Point-of-use extraction should be either in the form of a fixed high
velocity extraction point or an articulated arm with movable hood or a fixed
extraction hood.
5.3 Dust extraction ducting should be designed with sufficient transfer
velocity to ensure that dust is carried away, and does not settle in the ducting.
Periodic checks should be performed to ensure that there is no build up of
the dust in the ducting.
5.4 The required transfer velocity should be determined: it is dependent on
the density of the dust (the denser the dust, the higher the transfer velocity
should be, e.g. 15–20 m/s).
5.5 Airflow direction should be carefully chosen, to ensure that the
operator does not contaminate the product, and also so that the operator is
not put at risk by the product.

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 5.6 Point extraction alone is usually not sufficient to capture all of the
contaminants, and general directional airflow should be used to assist in
removing dust and vapours from the room.
5.7 Typically, in a room operating with turbulent airflow, the air should be
introduced from ceiling diffusers, located at the door entry side of the room
and extracted from the rear of the room at low level to help give a flushing
effect in the room. Correct flushing of the rooms may be verified by airflow
visualization smoke tests.
5.8 When dealing with particularly harmful products, additional steps,
such as handling the products in glove boxes or using barrier isolator
technology, should be used.

6. Protection of the environment

6.1 General
6.1.1 It should be noted that protection of the environment is not addressed
in this guideline, and discharges into the atmosphere should be compliant
with relevant local and national environmental legislation and standards.
6.1.2 Dust, vapours and fumes could be possible sources of contamination;
therefore, care should be taken when deciding on the location of the inlet
and exhaust points relative to one other.

6.2 Dust in exhaust air

6.2.1 Exhaust air discharge points on pharmaceutical equipment and
facilities, such as from fluid bed driers and tablet-coating equipment, and
exhaust air from dust extraction systems, carry heavy dust loads and should be
provided with adequate filtration to prevent contamination of the ambient air.
6.2.2 Where the powders are not highly potent, final filters on a dust
exhaust system should be fine dust filters with a filter classification of F9
according to EN 779 filter standards.
6.2.3 Where reverse-pulse dust collectors are used for removing dust from
dust extraction systems, they should usually be equipped with cartridge
filters containing a compressed air lance, and be capable of continuous
operation without interrupting the airflow.
6.2.4 Alternative types of dust collectors (such as those operating with a
mechanical shaker, requiring that the fan be switched off when the mechanical
shaker is activated) should be used in such a manner that there is no risk
of cross-contamination. There should be no disruption of airflow during a
production run as the loss of airflow could disrupt the pressure cascade.

246
 6.2.5 Mechanical-shaker dust collectors should not be used for applications
where continuous airflow is required, in order to avoid unacceptable
fluctuations in room pressures, except in the case where room pressures are
automatically controlled.
6.2.6 When wet scrubbers are used, the dust-slurry should be removed by
a suitable means, e.g. a drainage system or waste removal contractor.
6.2.7 The quality of the exhaust air should be determined to see whether the
filtration efficiency is adequate with all types of dust collectors and wet scrubbers.
6.2.8 Where necessary, additional filtration may be provided downstream
of the dust collector.

6.3 Vapour and fume removal

6.3.1 Vapour should be extracted at the point of generation. When planning
the system for the extraction of residual vapours, the density of the vapour
should be taken into account. If the vapour is lighter than air, the extract
grilles should be at a high level, or possibly at both high and low levels.
6.3.2 The systems for fume, dust and effluent control should be designed,
installed and operated in such a manner that they do not become possible
sources of contamination or cross-contamination, e.g. an exhaust-air
discharge point located close to the HVAC system fresh air inlet.
6.3.3 Fumes should be removed by means of wet scrubbers or dry
chemical scrubbers (deep-bed scrubbers).
6.3.4 Wet scrubbers for fume removal normally require the addition of
various chemicals to the water to increase the adsorption efficiency.
6.3.5 Deep-bed scrubbers should be designed with activated carbon filters
or granular chemical adsorption media. The chemical media for deep-bed
scrubbers should be specific to the effluent being treated.
6.3.6 The type and quantity of the vapours to be removed should be
known to enable the appropriate filter media, as well as the volume of media
required to be determined.

7. Design of HVAC systems and components

7.1 General
7.1.1 The required degree of air cleanliness in most OSD manufacturing
facilities can normally be achieved without the use of high-efficiency
particulate air (HEPA) filters, provided the air is not recirculated or

247
 in the case of a single-product facility. Many open product zones of
OSD form facilities are capable of meeting ISO 14644-1 Class 8 or
Grade D, “at-rest” condition, measured against particle sizes of 0.5 ìm
and 5 ìm, but cleanliness may not necessarily be classified as such by
manufacturers.
A risk assessment should be carried out to determine the cleanroom
conditions required and the extent of validation required.
7.1.2 There are two basic concepts of air delivery to pharmaceutical
production facilities: a recirculation system, and a full fresh air system
(100% outside air supply). For recirculation systems the amount of fresh air
should not be determined arbitrarily on a percentage basis, but, for example,
by the following criteria:
• sufficient fresh air to compensate for leakage from the facility and loss
through exhaust air systems;
• sufficient fresh air to comply with national building regulations; and29
• sufficient fresh air for odour control.
7.1.3 Where automated monitoring systems are used, these should be
capable of indicating any out-of-specification condition without delay by
means of an alarm or similar system. Sophisticated computer-based data
monitoring systems may be installed, which can aide with planning of
preventive maintenance and can also provide trend logging.
(This type of system is commonly referred to as a building management
system (BMS), building automation system (BAS) or system control and
data acquisition (SCADA) system.) If these systems are used for critical
decision-making, they should be validated.
7.1.4 Failure of a supply air fan, return air fan, exhaust air fan or dust
extract system fan can cause a system imbalance, resulting in a pressure
cascade malfunction with a resultant airflow reversal.
7.1.5 All critical alarms should be easily identifiable and visible and/or
audible to relevant personnel.
7.1.6 Appropriate alarm systems should be in place to alert personnel if a
critical fan fails. A fan interlock failure matrix should be set up, such that if
a fan serving a high pressure zone fails, then any fans serving surrounding
lower pressure areas should automatically stop, to prevent an airflow
reversal and possible cross-contamination.

2
Depending on occupant density, between 1 and ACPH will often satisfy occupancy requirements.

248
7.2 Air distribution
7.2.1 The positioning of supply and extract grilles should be such as to
provide effective room flushing. Low-level return or exhaust air grilles are
usually preferred. However, where this is not possible, a higher air change
rate may be needed to achieve a specified clean area condition, e.g. where
ceiling return air grilles are used.
7.2.2 There may be alternative locations for return air. For example,
referring to Figure 22, Room 1 (low-level return air) and Room 2 (ceiling
return air). The airflow diagram in Figure 22 is an example of a typical
system with a lower clean area condition.

Figure 22
Air-handling system with high-efficiency particulate air filters in air-handling unit

The airflow schematics of the two systems (Figures 22 and 23) indicate
air-handling units with return air or recirculated air, having a percentage
of fresh air added. Depending on product characteristics and dust loading
it is sometimes preferable to fit filters on return air outlets or in return air
ducting.

249
 Figure 23 is a schematic diagram of an air-handling system serving
rooms with horizontal unidirectional flow, vertical unidirectional flow and
turbulent flow, for rooms 3, 4 and 5, respectively.

Figure 23
Horizontal unidirectional flow, vertical unidirectional flow and turbulent flow

7.3 Recirculation system

7.3.1 There should be no risk of contamination or cross-contamination
(including by fumes and volatiles) due to recirculation of air.
7.3.2 Depending on the airborne contaminants in the return-air system
it may be acceptable to use recirculated air, provided that HEPA filters
are installed in the supply air stream (or return air stream) to remove
contaminants and thus prevent cross-contamination. The HEPA filters for
this application should have an EN 1822 classification of H13.
7.3.3 HEPA filters may not be required where the air-handling system
is serving a single product facility and there is evidence that cross-
contamination would not be possible.
7.3.4 Recirculation of air from areas where pharmaceutical dust is not
generated such as secondary packing, may not require HEPA filters in the
system.
7.3.5 HEPA filters may be located in the air-handling unit or placed
terminally. Where HEPA filters are terminally mounted they should

250
 preferably not be connected to the ducting by means of flexible ducting.
Due to the high air pressure required for the terminal filter, this connection
should preferably be a rigid duct connection. Where flexible ducting is
used, it should be as short as possible and properly fixed to withstand duct
pressure.
7.3.6 Air containing dust from highly toxic processes and/or solvents or
flammable vapours should never be recirculated to the HVAC system.

7.4 Full fresh-air systems

Figure 24 indicates a system operating on 100% fresh air and would
normally be used in a facility dealing with toxic products or solvents, where
recirculation of air with contaminants should be avoided.
7.4.1 The required degree of filtration of the exhaust air depends on the
exhaust air contaminants and local environmental regulations. HEPA filters
in the exhaust system would normally only be required when handling
hazardous materials.

Figure 24
Full fresh-air system

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Figure 25
Full fresh-air system with energy recovery

7.4.2 Energy-recovery wheels if used in multiproduct facilities should

have been subjected to a risk assessment to determine if there is any
risk of cross-contamination. When such wheels are used they should
not become a source of possible contamination (see Figure 25). Note:
Alternatives to the energy-recovery wheels, such as crossover plate heat
exchangers and water-coil heat exchangers, may be used in multiproduct
facilities.

7.4.3 The potential for air leakage between the supply air and exhaust air
as it passes through the wheel should be prevented. The relative pressures
between supply and exhaust air systems should be such that the exhaust air
system operates at a lower pressure than the supply system.

7.5 Additional system components

7.5.1 A schematic diagram of the airflow for a typical system serving a
low relative humidity suite is represented in Figure 26. Air can be dried
with a chemical drier (e.g. a rotating desiccant wheel which is continuously
regenerated by means of passing hot air through one segment of the wheel).
Alternative methods of drying air are also available.

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Figure 26
Air-handling system with chemical drying

7.5.2 The figure illustrates the chemical drier handling part of the fresh
air/return air mixture on a bypass flow. The location of the chemical drier
should be considered in the design phase. The practice of locating the
complete chemical drier unit in the production cubicle is not recommended
as this could be a source of contamination or cross-contamination. Examples
of appropriate locations for the drying wheel could include:
— full flow of fresh/return air;
— partial handling of fresh/return air (bypass airflow);
— return air only;
— fresh air only; or
— pre-cooled air with any of the above alternatives.
7.5.3 Possible additional components that may be required in air handling
should be considered depending on the climatic conditions and locations.
These may include items such as:
— frost coils on fresh air inlets in very cold climates to preheat the air;
— reheaters for humidity control
— automatic air volume control devices
— sound attenuators

253
 — snow eliminators to prevent snow entering air inlets and blocking
airflow
— dust eliminators on air inlets in arid and dusty locations
— moisture eliminators in humid areas with high rainfall
— fresh air precooling coils for very hot or humid climates.

8. Commissioning, qualification and maintenance

8.1 Commissioning
8.1.1 Commissioning should include the setting up, balancing, adjustment
and testing of the entire HVAC system, to ensure that it meets all the
requirements, as specified in the user requirement specification (URS), and
capacities as specified by the designer or developer. The commissioning
plan should start at the early stages of a project so that it can be integrated
with qualification and verification procedures.
8.1.2 The installation records of the system should provide documented
evidence of all measured capacities of the system.
8.1.3 Acceptance criteria should be set for all system parameters. The
measured data should fall within the acceptance criteria.
8.1.4 Acceptable tolerances for all system parameters should be specified
prior to commencing the physical installation.
8.1.5 Training should be provided to personnel after installation of the
system, and should include operation and maintenance.
8.1.6 Commissioning should be a precursor to system qualification and
process validation.

8.2 Qualification
8.2.1 Validation is a many-faceted and extensive activity and is beyond
the scope of these guidelines (2) (see also Figure 27).
A risk-based approach should be used to identify the extent to which the
HVAC system requires qualification and verification. The basic concepts of
qualification of HVAC systems are set out below.
8.2.2 The qualification of the HVAC system should be described in a
validation master plan (VMP).
8.2.3 It should define the nature and extent of testing and the test
procedures and protocols to be followed.

254
Figure 27
Qualification is a part of validation

8.2.4 Stages of the qualification of the HVAC system should include DQ,
IQ, OQ and PQ.
8.2.5 Critical and non-critical parameters should be determined by means
of a risk analysis for all HVAC installation components, subsystems and
controls.
8.2.6 Any parameter that may affect the quality of the pharmaceutical
product, or a direct impact component, should be considered a critical
parameter.
8.2.7 All critical parameters should be included in the qualification
process. Note: A realistic approach to differentiating between critical and
noncritical parameters is required, to avoid making the validation process
unnecessarily complex.

Example:
• The relative humidity of the room where the product is exposed should
be considered a critical parameter when a humidity-sensitive product is
being manufactured. The humidity sensors and the humidity monitoring
system should, therefore, be qualified. The heat transfer system, chemical
drier or steam humidifier, which is producing the humidity controlled
air, is further removed from the product and may not require operational
qualification.
• A room cleanliness condition is a critical parameter and, therefore, the
room air change rates and HEPA filters should be critical parameters and
require qualification. Items such as the fan generating the airflow and the
primary and secondary filters are non-critical parameters, and may not
require operational qualification.
8.2.8 Non-critical systems and components should be subject to GEP and
may not necessarily require qualification.

255
 8.2.9 A change control procedure should be followed when changes are
planned to the direct impact HVAC system, its components and controls
that may affect critical parameters.
8.2.10 The design condition, normal operating ranges, operating range
and alert and action limits should be defined and be realistic.
8.2.11 Out-of-limit results (e.g. action limit deviations) should be
recorded and their impact should be investigated.
8.2.12 The relationships between design conditions, normal operating
range and validated acceptance criteria (also known as proven acceptable
range) are given in Figure 28.

Figure 28
System operating ranges

8.2.13 For a pharmaceutical facility, based on a risk assessment, some

of the typical HVAC system parameters that should be qualified may
include:
— temperature
— relative humidity
— supply air quantities for all diffusers
— return air or exhaust air quantities
— room air change rates
— room pressures (pressure differentials)
— room airflow patterns
— unidirectional flow velocities
— containment system velocities
— HEPA filter penetration tests

256
 — room particle counts
— room clean-up rates
— microbiological air and surface counts where appropriate
— operation of de-dusting
— warning/alarm systems where applicable.
8.2.14 The maximum time interval between tests should be defined by
the manufacturer. The type of facility under test and the product level of
protection should be considered. Table 3 gives various tests that can be
carried out. The required tests and intervals between testing should be
determined through risk assessment.

Table 3
Tests to demonstrate compliance
Test parameter Test procedure

Particle count test Dust particle counts to be carried out and result printouts
(Verification of cleanliness) produced.
No. of readings and positions of tests to be in accordance
with ISO 14644-1 Annex B5

Air pressure difference Log of pressure differential readings to be produced

(To verify non cross- or critical plants should be logged daily, preferably
contamination) continuously. A 15 Pa pressure differential between
different zones is recommended.
In accordance with ISO 14644-3 Annex B5

Airflow volume Airflow readings for supply air and return air grilles to be
(To verify air change rates) measured and air change rates to be calculated.
In accordance with ISO 14644-3 Annex B13

Airflow velocity Air velocities for containment systems and unidirectional

(To verify unidirectional flow protection systems to be measured.
flow or containment In accordance with ISO 14644-3 Annex B4
conditions)

Filter leakage tests Filter penetration tests to be carried out by a competent

(To verify filter integrity) person to demonstrate filter media, filter seal and
filter frame integrity. Only required on HEPA filters. In
accordance with ISO 14644-3 Annex B6

Containment leakage Demonstrate that contaminant is maintained within a room

(To verify absence of by means of:
cross-contamination) • airflow direction smoke tests
• room air pressures.
In accordance with ISO 14644-3 Annex B4

Recovery Test to establish time that a cleanroom takes to recover

(To verify clean-up time) from a contaminated condition to the specified cleanroom
condition. Should not take more than 15 min.
In accordance with ISO 14644-3 Annex B13*

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 Test parameter Test procedure
Airflow visualization Tests to demonstrate air flows:
(To verify required airflow • from clean to dirty areas
patterns) • do not cause cross-contamination
• uniformly from unidirectional airflow units
Demonstrated by actual or video-taped smoke tests.
In accordance with ISO 14644-3 Annex B7

8.2.15 Requalification should also be done when any change, which could
affect system performance, takes place.
8.2.16 Clean-up or recovery times normally relate to the time it takes to “clean
up” the room from one condition to another, e.g. the relationship between “at-
rest” and “operational” conditions in the clean area may be used as the criteria
for clean-up tests. Therefore, the clean-up time can be expressed as the time
taken to change from an “operational” condition to an “at rest” condition.
8.2.17 If energy-saving procedures such as reducing the airflow during
non-production hours are used, precautionary measures should be in place
to ensure that the systems are not operated outside the defined relevant
environmental conditions.
These precautionary measures should be based on a risk assessment to
ensure that there is no negative impact on the quality of the product.
8.2.18 Documents that should be included in the qualification manuals
should include system airflow schematics, room pressure cascade drawings,
zone concept drawings, air-handling system allocation drawings, particle
count mapping drawings, etc.

8.3 Maintenance
8.3.1 There should be a planned preventive maintenance programme,
procedures and records for the HVAC system. Records should be kept.
8.3.2 Operating and maintenance (O&M) manuals, schematic drawings,
protocols and reports should be maintained as reference documents for any
future changes and upgrades to the system. These documents should be
kept up to date, containing any system revisions made.
8.3.3 Maintenance personnel should receive appropriate training.
8.3.4 HEPA filters should be changed either by a specialist or a trained
person, and then followed by installed filter leakage testing.
8.3.5 Any maintenance activity should be assessed critically to determine
any impact on product quality including possible contamination.

258
 8.3.6 Maintenance activities should normally be scheduled to take place
outside production hours, and any system stoppage should be assessed with
a view to the possible need for requalification of an area as a result of an
interruption of the service.

9. Premises
9.1 As the efficient operation of the air-handling system and cleanliness
levels attained are reliant on the correct building layout and building
finishes, the following items should be considered:
• adequate airlocks, such as personnel airlocks (PAL) and/or material
airlocks (MAL), change rooms and passages should be provided to
protect passage between different cleanliness conditions . These should
have supply and extract air systems as appropriate;
• areas such as airlocks, change rooms and passages, should be designed so
that the required pressure cascades can be achieved;
• detailed diagrams depicting pressure cascades, air flow directions
and flow routes for personnel and materials should be prepared and
maintained;
• where possible, personnel and materials should not move from a higher
cleanliness zone to a lower cleanliness zone and back to a higher
cleanliness zone; (if moving from a lower cleanliness zone to a higher
cleanliness zone, changing /decontamination procedures should be
followed); and
• the final stage of the changing room should, in the “at rest” state, be the
same GMP classification grade as the area into which it leads.

References
1. Good manufacturing practices for pharmaceutical products: main principles.
In: WHO Expert Committee on Specifications for Pharmaceutical Preparations
Thirty-seventh report. Geneva, World Health Organization, 2003 (WHO
Technical Report Series, No. 908), Annex 4. http://whqlibdoc.who.int/trs/WHO_
TRS_908_eng.pdf; Quality assurance of pharmaceuticals. A compendium
of guidelines and related materials. Volume 2, Second updated edition.
Good manufacturing practices and inspection. Geneva, World Health
Organization, 2007; and Quality assurance of pharmaceuticals. A compendium
of guidelines and related materials. Geneva, World Health Organization, 2010
(CD-ROM).
2. Expert Committee on Specifications for Pharmaceutical Preparations.
Fortieth report. Geneva, World Health Organization, 2005 (WHO Technical
Report Series, No. 937). http://whqlibdoc.who.int/trs/WHO_TRS_937_eng.pdf.

259
Further reading
Quality assurance of pharmaceuticals. A compendium of guidelines and
related materials, Volume 1. Geneva, World Health Organization, 1997.
Quality Assurance of Pharmaceuticals. A compendium of guidelines and
related materials, Volume 2, Second updated edition. Good manufacturing
practices and inspection. Geneva, World Health Organization, 2007. http://
www.who.int/medicines/areas/quality_safety/quality_assurance/production/
en/index.html; and Quality Assurance of Pharmaceuticals. A compendium of
guidelines and related materials. Geneva, World Health Organization, 2010
(CD-ROM).
World Health Organization. Supplements and updates available at:
www.who.int/medicines.
ASHRAE handbook 1999. HVAC Applications, SI edition. Atlanta, GA,
ASHRAE, 2007. http://www.ashrae.org/technology/page/548.
ASHRAE handbook 2000. HVAC Systems and Equipment. Atlanta, GA,
ASHRAE, 2008. http://www.ashrae.org/technology/page/548.
Daly BB. Woods practical guide to fan engineering. Colchester, Woods
of Colchester Ltd. Third impression, June 1985. Cambridge, Cambridge
University Press. www.flaktwoods.com.
European Commission. The rules governing medicinal products in the
European Community, Volume IV. Good manufacturing practice for medicinal
products. European Commission, Brussels, 2005. http://www.cen.eu/cenorm/
sectors/sectors/healthcare/index.asp.
ISPE Baseline® pharmaceutical engineering guides, Volume 2. Oral solid
dosage forms, Second Edition / November 2009, International Society for
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ISPE Baseline® pharmaceutical engineering guides for new and renovated
facilities, Volume 5. Commissioning and qualification, 1st ed. Tampa, Fl,
International Society for Pharmaceutical Engineering, 2001.
http://www.ispe.org/.
International Cleanroom Standards, ISO 14644. Geneva, International
Organization for Standardization. http://www.iso.org/iso/standards_
development.htm.
Luwa. Introduction to high efficiency filtration. Bulletin 50.10.10, Sheet 020.
Pharmaceutical Inspectorate Convention/Pharmaceutical Inspection
Co-operation Scheme. Guide to Good Manufacturing Practice for Medicinal
Products. PH 1/97 (Rev. 3), 15 January 2002.
PIC/s GMP Guide (PE 009) http://www.picscheme.org/publication.php?id=4
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World Health Organization. Draft working document QAS/10.376: “Guidelines
on quality risk management", 2010 (in preparation).

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© World Health Organization
WHO Technical Report Series, No. 961, 2011

Annex 6
WHO good manufacturing practices for sterile
pharmaceutical products

Introduction
Following implementation of these WHO good manufacturing practices (GMP)
guidelines (1) within the context of the WHO Prequalification of Medicines
Programme, clarifying, editorial modifications have been proposed. These
changes were adopted for maintenance purposes. In order to ease reading the
full guideline has been reproduced again as an Annex to the current report of
the WHO Expert Committee on Specifications for Pharmaceutical Preparations.

WHO good manufacturing practices

for sterile pharmaceutical products
1. General considerations

2. Quality control

3. Sanitation

4. Manufacture of sterile preparations

5. Sterilization

6. Terminal sterilization

7. Aseptic processing and sterilization by filtration

8. Isolator technology

9. Blow/fill/seal technology

10. Personnel

11. Premises

12. Equipment

13. Finishing of sterile products

References

Further reading

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1. General considerations
1.1 The production of sterile preparations should be carried out in clean
areas, entry to which should be through airlocks for personnel and/or for
equipment and materials. Clean areas should be maintained to an appropriate
standard of cleanliness and supplied with air that has passed through filters
of the required efficiency.
1.2 The various operations of component preparation (such as those
involving containers and closures), product preparation, filling and
sterilization should be carried out in separate areas within the clean area.
These areas are classified into four grades (see section 4).
1.3 Manufacturing operations are divided here into two categories:
— first, those where the product is terminally sterilized; and
— second, those which are conducted aseptically at some or all stages.

2. Quality control
2.1 The sterility test applied to the finished product should only be
regarded as the last in a series of control measures by which sterility is
assured. The test should be validated for the product(s) concerned.
2.2 Samples taken for sterility testing should be representative of the whole
of the batch but should, in particular, include samples taken from parts of
the batch considered to be most at risk of contamination, for example:
• for products that have been filled aseptically, samples should include
containers filled at the beginning and end of the batch and after any
significant interruption of work;
• for products that have been heat sterilized in their final containers,
consideration should be given to taking samples from that part of the
load that is potentially the coolest.
2.3 The sterility of the finished product is assured by validation of
the sterilization cycle in the case of terminally sterilized products, and
by “media simulation” or “media fill” runs for aseptically processed
products. Batch-processing records and, in the case of aseptic processing,
environmental quality records, should be examined in conjunction with
the results of the sterility tests. The sterility test procedure should be
validated for a given product. Pharmacopoeial methods should be used for
the validation and performance of the sterility test. In those cases where
parametric release has been authorized in place of sterility testing special
attention should be paid to the validation and the monitoring of the entire
manufacturing process.

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 2.4 For injectable products the water for injection and the intermediate,
if appropriate, and finished products should be monitored for endotoxins,
using an established pharmacopoeial method that has been validated for
each type of product. For large-volume infusion solutions, such monitoring
of water or intermediates should always be done, in addition to any tests
required by an approved monograph for the finished product. When a sample
fails a test, the cause of the failure should be investigated and necessary
action should be taken. Alternative methods to those in the pharmacopoeias
may be used if they are validated, justified and authorized.
2.5 The use of rapid microbiological methods to replace the traditional
microbiological methods, and to obtain earlier results on the microbiological
quality of, for example, water, the environment or bioburden, could be
considered if appropriately validated and if a comparative assessment of the
proposed rapid method is performed against the pharmacopoeial method.

3. Sanitation
3.1 The sanitation of clean areas is particularly important. They should
be cleaned frequently and thoroughly in accordance with an approved
written programme. Where disinfectants are used, more than one type
should be employed. Monitoring should be regularly undertaken to detect
contamination or the presence of an organism against which the cleaning
procedure is ineffective. Interactions between different cleaning materials
should be validated. Appropriate cleaning validation should be carried out
to ensure disinfectant residuals can be detected and are removed by the
cleaning process.
3.2 Disinfectants and detergents should be monitored for microbial
contamination; dilutions should be kept in previously cleaned containers
and should only be stored for defined periods unless sterilized. Disinfectants
and detergents used in Grade A and B areas should be sterile before use.
3.3 A disinfectant programme should also include a sporicidal agent since
many common disinfectants are ineffective against spores. The effectiveness
of cleaning and disinfectant procedures should be demonstrated.
3.4 Fumigation of clean areas may be useful for reducing microbial
contamination in inaccessible places.

4. Manufacture of sterile preparations

4.1 Clean areas for the manufacture of sterile products are classified
according to the required characteristics of the environment. Each
manufacturing operation requires an appropriate level of environmental

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 cleanliness in the operational state to minimize the risks of particulate or
microbial contamination of the product or materials being handled.
4.2 Detailed information on methods for determining the microbiological
and particulate cleanliness of air, surfaces, etc., is not given in these
guidelines.
ISO 14644-1 (2) should be used for classification of cleanliness according
to concentration of airborne particles (determination of number of sample
locations, calculation of sample size and evaluation of classification from
the data obtained). Table 1 should also be used to define the levels to be
used as the basis for monitoring clean areas for airborne particles.
4.3 For the manufacture of sterile pharmaceutical preparations, four
grades of clean areas are distinguished as follows:
• Grade A: The local zone for high-risk operations, e.g. filling and making
aseptic connections. Normally such conditions are achieved by using a
unidirectional airflow workstation. Unidirectional airflow systems should
provide a homogeneous air speed of 0.36–0.54 m/s (guidance value) at a
defined test position 15–30 cm below the terminal filter or air distributor
system. The velocity at working level should not be less than 0.36 m/s.
The uniformity and effectiveness of the unidirectional airflow should be
demonstrated by undertaking airflow visualization tests.
• Grade B: In aseptic preparation and filling, this is the background
environment for the Grade A zone.
• Grades C and D: Clean areas for carrying out less critical stages in the
manufacture of sterile products or carrying out activities during which
the product is not directly exposed (i.e. aseptic connection with aseptic
connectors and operations in a closed system).
A unidirectional airflow and lower velocities may be used in closed isolators
and glove boxes.
4.4 In order to reach the B, C and D air grades the number of air changes
should be appropriate for the size of the room and the equipment and
personnel present in it.
4.5 High-efficiency particulate air (HEPA) filters should be subjected
to an installed filter leakage test in accordance with ISO 14644-3 (3) at a
recommended interval of every 6 months, but not exceeding 12 months. The
purpose of performing regular leak tests is to ensure the filter media, filter
frame and filter seal are free from leaks. The aerosol selected for HEPA leak
testing should not support microbial growth and should be composed of a
sufficient number or mass of particles. HEPA filter patching is allowed at
the filter manufacturer and in situ operation provided that the patch sizes
and procedures follow the recommendations of ISO 1822-4 (4).

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 Clean room and clean-air device classification
4.6 Clean rooms and clean-air devices should be classified in accordance
with ISO 14644 (2–3, 5–7).
4.6.1 Classification should be clearly differentiated from operational
process environmental monitoring. The maximum permitted airborne
particle concentration for each grade is given in Table 1.

Table 1
Maximum permitted airborne particle concentrat
Maximum permitted number of particles per m3 greater
than or equal to the tabulated size

At resta In operationb
Grade 0.5 μm 5.0 μm 0.5 μm 5.0 μm
A 3 520 20 3 520 20
B 3 520 29 352 000 2 900
C 352 000 2 900 3 520 000 29 000
D 3 520 000 29 000 Not defined Not defined
a
The “at rest” state is the condition where the installation is complete with equipment installed and
operating in a manner agreed upon by the customer and supplier, but with no personnel present.
b
The “in operation” state is the condition where the installation is functioning in the defined operating
mode and the specified number of personnel is present. The areas and their associated environmental
control systems should be designed to achieve both the “at rest” and “in operation” states.

4.6.2 For classification purposes in Grade A zones, a minimum sample

volume of 1 m³ should be taken per sample location. Referring to Table 1, for
Grade A the airborne particle classification is ISO 4.8 dictated by the limit for
particles ≥ 5.0 μm. For Grade B (at rest) the airborne particle classification is
ISO 5 for both particle sizes considered. For Grade C (at rest and in operation)
the airborne particle classification is ISO 7 and ISO 8, respectively. For Grade
D (at rest) the airborne particle classification is ISO 8. For classification
purposes ISO 14644-1 (2) methodology defines both the minimum number
of sample locations and the sample size based on the class limit of the largest
particle size considered and the method of evaluation of the data collected.
The sample volume should be determined according to ISO 14644-1 (2)
clause B.4.2. However, for lower grades (Grade C in operation and Grade
D at rest) the sample volume per location should be at least 2 litres and the
sample time per location should be not less than 1 minute.
4.6.3 Portable particle counters with a short length of sample tubing should
be used for classification purposes to avoid the loss of particles ≥ 5.0 μm.
Isokinetic sample heads should be used in unidirectional airflow systems.
4.6.4 “In operation” classification may be demonstrated during normal
operations, simulated operations or during media fills as worst-case simulation

265
 is required for this. ISO 14644-2 (6) provides information on testing to
demonstrate continued compliance with the assigned cleanliness classification.

Clean room and clean-air device monitoring

4.7 Clean rooms and clean-air devices should be routinely monitored
while in operation and the monitoring locations based on a formal risk
analysis study and the results obtained during the classification of rooms
and/or clean-air devices.
4.7.1 For Grade A zones, particle monitoring should be undertaken for
the full duration of critical processing, including equipment assembly,
except where justified by contaminants in the process that would damage
the particle counter or present a hazard, for example, live organisms and
radiological hazards. In such cases monitoring during routine equipment
set-up operations should be undertaken before exposure to the risk.
Monitoring during simulated operations should also be performed. The
Grade A zone should be monitored at a frequency and sample size such
that all interventions, transient events and any system deterioration would
be captured and alarms triggered if alert limits are exceeded. It is accepted
that it may not always be possible to demonstrate low levels of ≥ 5.0 μm
particles at the point of fill when filling is in progress, due to the generation
of particles or droplets from the product itself.
4.7.2 It is recommended that a similar system be used for Grade B zones,
although the sample frequency may be decreased. The importance of the
particle monitoring system should be determined by the effectiveness of
the segregation between the adjacent Grade A and B zones. The Grade B
zone should be monitored at a frequency and with a sample size such that
changes in levels of contamination and any deterioration of the system
would be captured and alarms triggered if alert limits are exceeded.
4.7.3 Airborne particle monitoring systems may consist of independent
particle counters; a network of sequentially accessed sampling points
connected by manifold to a single particle counter; or multiple small
particle counters located near monitoring points and networked to a data
acquisition system. Combinations of systems can also be used. The system
selected should be appropriate for the particle size considered.
Where remote sampling systems are used, the length of tubing and the radii of
any bends in the tubing should be considered in the context of particle losses
in the tubing. The selection of the monitoring system should take account of
any risk presented by the materials used in the manufacturing operation, for
example, those involving live organisms or radiopharmaceuticals.
4.7.4 The sizes of samples taken for monitoring purposes using automated
systems will usually be a function of the sampling rate of the system used.

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 It is not necessary for the sample volume to be the same as that used for
formal classification of clean rooms and clean-air devices.
4.7.5 The airborne particle conditions given in Table 1 for the “at rest” state
should be achieved in the absence of the operating personnel after a short
“clean-up” or “recovery” period of about 15–20 minutes (guidance value),
after completion of the operations. The particulate conditions given in Table
1 for Grade A “in operation” should be maintained in the zone immediately
surrounding the product whenever the product or open container is exposed
to the environment. The “clean-up” or “recovery” test should demonstrate
a change in particle concentration by a factor of 100 within the prescribed
time (ISO 14644-3 clause B.12) (3).
4.7.6 In order to demonstrate control of the cleanliness of the various
clean areas during operation, they should be monitored for airborne particles
and microbial contamination. In addition to “at rest” and “in operation”
classification, airborne particles should be monitored periodically “in
operation” at critical locations. The sampling plan need not be the same as
that used for classification. Locations and sample sizes should be determined
based on an assessment of the process and contamination risk.
4.7.7 The monitoring of Grade C and D areas in operation should be
performed in accordance with the principles of quality risk management. The
requirements and alert/action limits will depend on the nature of the operations
carried out, but the recommended “clean-up period” should be attained.
4.7.8 Other characteristics such as temperature and relative humidity
depend on the product and nature of the operations carried out. These
parameters should not interfere with the defined cleanliness standard.
4.7.9 Examples of operations to be carried out in the various grades are
given in Table 2 (see also sections 4.12–4.20).
Table 2
Examples of operations to be carried out in the various grades
Grade Examples of operations for terminally sterilized products
(see sections 4.12–4.15 )
A Filling of products when unusually at risk
C Preparation of solutions when unusually at risk. Filling of products
D Preparation of solutions and components for subsequent filling

Grade Examples of operations for aseptic preparations

(see sections 4.16–4.20)
A Aseptic preparation and filling
C Preparation of solutions to be filtered
D Handling of components after washing

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 4.8 To control the microbiological cleanliness of Grades A–D in
operation the clean areas should be monitored. Where aseptic operations
are performed, monitoring should be frequent using methods such as
settle plates, volumetric air and surface sampling (e.g. swabs and contact
plates). Sampling methods used in operation should not interfere with zone
protection. Results from monitoring should be considered when reviewing
batch documentation for finished product release. Surfaces and personnel
should be monitored after critical operations. Additional microbiological
monitoring is also required outside production operations, e.g. after
validation of systems, cleaning and sanitization.
4.9 Levels of detection of microbial contamination should be established
for the purpose of setting alert and action limits and for monitoring the
trends in environmental cleanliness in the facility. Limits expressed in
colony-forming units (CFU) for the microbiological monitoring of clean
areas in operation are given in Table 3. The sampling methods and numerical
values included in the table are not intended to represent specifications, but
are for information only.
Table 3
Recommended limits for microbial contaminationa
Grade Air sample Settle plates Contact plates Glove print
(CFU/m3) (diameter 90 mm) (diameter 55 mm) (5 fingers)
(CFU/4 hours)b (CFU/plate) (CFU/glove)
A <1 <1 <1 <1
B 10 5 5 5
C 100 50 25 –
D 200 100 50 –
CFU, colony-forming units.
a
These are average values.
b
Individual settle plates may be exposed for less than 4 hours.

4.10 Appropriate alert and action limits should be set for the results of
particulate and microbiological monitoring. If the action limits are exceeded
or a trend is identified in the alert limits, investigation should be initiated
and the appropriate corrective actions should be taken, as prescribed in the
operating procedures.
4.11 The area grades as specified in sections 4.12 to 4.20 should be
selected by the manufacturer on the basis of the nature of the process
operations being performed and validation runs (e.g. aseptic media fills or
others types of process simulations) are used to establish processing hold
times and a maximum fill duration. The determination of an appropriate
process area environment and a time limit should be based on the microbial
contamination (bioburden) found.

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Terminally sterilized products
4.12 Components and most products should be prepared in at least a
Grade D environment to ensure low microbial bioburden and particulate
counts prior to filtration and sterilization. Where the product is at unusual
risk of microbial contamination (e.g. because it actively supports microbial
growth, must be held for a long period before sterilization, or is necessarily
processed mainly in open vessels), the preparation should generally be done
in a Grade C environment.
4.13 The filling of products for terminal sterilization should generally be
done in at least a Grade C environment.
4.14 Where the product is at unusual risk of contamination from the
environment (e.g. because the filling operation is slow, the containers are wide-
necked or are necessarily exposed for more than a few seconds before sealing),
the filling should be done in a Grade A zone with at least a Grade C background.
4.15 The preparation and filling of ointments, creams, suspensions and
emulsions should generally be done in a Grade C environment before
terminal sterilization.

Aseptic preparation
4.16 Components after washing should be handled in at least a Grade D
environment. The handling of sterile starting materials and components,
unless subjected to sterilization or filtration through a microorganism-
retaining filter later in the process, should be undertaken in a Grade A
environment with a Grade B background.
4.17 The preparation of solutions which are to be sterile-filtered during
the process should be undertaken in a Grade C environment (unless a closed
system is used, in which case a Class D environment may be justifiable). If
not sterile-filtered (therefore an aseptic manipulation) the preparation of
materials and products should be undertaken in a Grade A environment
with a Grade B background.
4.18 The handling and filling of aseptically prepared products, as well as
the handling of exposed sterile equipment, should be undertaken in a Grade
A environment with a Grade B background.
4.19 The transfer of partially closed containers, as used in freeze-drying,
before stoppering is completed, should be undertaken either in a Grade A
environment with a Grade B background or in sealed transfer trays in a
Grade B environment.
4.20 The preparation and filling of sterile ointments, creams, suspensions
and emulsions should be undertaken in a Grade A environment with a Grade
B background when the product is exposed and is not subsequently filtered.

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 Processing
4.21 Precautions to minimize contamination should be taken during all
processing stages, including the stages before sterilization.
4.22 In general, preparations containing live microorganisms should not be
made, nor should containers be filled in areas used for the processing of other
pharmaceutical products. However, if the manufacturer can demonstrate
and validate effective containment and decontamination of the live
microorganisms, the use of multiproduct facilities may be justifiable. Vaccines
consisting of dead organisms or of bacterial extracts may be dispensed into
containers in the same premises as other sterile pharmaceutical products,
provided that the inactivation procedure has been properly validated.
When multiproduct facilities are used to manufacture sterile preparations
containing live microorganisms and other sterile pharmaceutical products, the
manufacturer should demonstrate and validate the effective decontamination
of the live microorganisms, in addition to precautions taken to minimize
contamination.
4.23 Validation of aseptic processing should include a process simulation
test using a nutrient medium (media fill). Selection of the nutrient medium
should be made based on dosage form of the product and selectivity, clarity,
concentration and suitability for sterilization of the nutrient medium.
4.24 The process simulation test should imitate as closely as possible the
routine aseptic manufacturing steps except where the activity may lead to
any potential microbial contamination.
4.25 Process simulation tests should be performed as part of validation by
running three consecutive satisfactory simulation tests. These tests should
be repeated at defined intervals and after any significant modification to
the heating, ventilation and air-conditioning (HVAC) system, equipment or
process. Process simulation tests should incorporate activities and interventions
known to occur during normal production as well as the worst-case situation.
The process simulation tests should be representative of each shift and shift
changeover to address any time-related and operational features.
4.26 The number of containers used for media fills should be sufficient to
enable a valid evaluation. For small batches the number of containers for
media fills should at least equal the size of the product batch. The target
should be zero growth and the following should apply:
• when filling fewer than 5000 units, no contaminated units should be
detected.
• when filling 5000–10 000 units:
— one contaminated unit should result in an investigation, including
consideration of a repeat media fill;

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 — two contaminated units are considered cause for revalidation
following investigation;
• when filling more than 10 000 units:
— one contaminated unit should result in an investigation;
— two contaminated units are considered cause for revalidation
following investigation.
4.27 For any run size, intermittent incidents of microbial contamination
may be indicative of low-level contamination that should be investigated.
Investigation of gross failures should include the potential impact on the
sterility assurance of batches manufactured since the last successful media fill.
4.28 Care should be taken to ensure that any validation does not
compromise the processes.
4.29 Water sources, water-treatment equipment and treated water should
be monitored regularly for chemicals, biological contamination and
contamination with endotoxins to ensure that the water complies with the
specifications appropriate to its use. Records should be maintained of the
results of the monitoring and of any action taken (8).
4.30 Activities in clean areas, especially when aseptic operations are in
progress, should be kept to a minimum and the movement of personnel should
be controlled and methodical, so as to avoid excessive shedding of particles and
organisms due to over-vigorous activity. As far as possible, personnel should
be excluded from Grade A zones. The ambient temperature and humidity
should not be uncomfortably high because of the nature of the garments worn
and to reduce the risk of contamination liberated from the personnel.
4.31 The presence of containers and materials liable to generate fibres
should be minimized in clean areas and avoided completely when aseptic
work is in progress.
4.32 Components, bulk-product containers and equipment should be
handled after the final cleaning process in such a way as to ensure that they
are not recontaminated. The stage of processing of components as well as
the bulk-product containers and equipment should be properly identified.
4.33 The interval between the washing and drying and the sterilization of
components, bulk-product containers and equipment, as well as between
sterilization and use, should be as short as possible and subject to a time-
limit appropriate to the validated storage conditions.
4.34 The time between the start of the preparation of a solution and its
sterilization or filtration through a bacteria-retaining filter should be as short
as possible. A maximum permissible time should be set for each product that
takes into account its composition and the prescribed method of storage.

271
 4.35 Any gas that is used to purge a solution or blanket a product should
be passed through a sterilizing filter.
4.36 The bioburden should be monitored before sterilization. There should
be working limits on contamination immediately before sterilization, which
are related to the efficiency of the method to be used. Bioburden assay
should be performed on each batch for both aseptically filled products and
terminally sterilized products. Where overkill sterilization parameters are
set for terminally sterilized products, bioburden might be monitored only
at suitable scheduled intervals. For parametric release systems, bioburden
assay should be performed on each batch and considered as an in-process
test. Where appropriate, the level of endotoxins should be monitored. All
solutions, in particular large-volume infusion fluids, should be passed through
a microorganism-retaining filter, if possible sited immediately before filling.
4.37 Components, bulk-product containers, equipment, and any other
articles required in a clean area where aseptic work is in progress, should
be sterilized and wherever possible passed into the area through double-
ended sterilizers sealed into the wall. Other procedures that prevent the
introduction of contamination may be acceptable in some circumstances.
4.38 The efficacy of any new processing procedure should be validated
and the validation should be repeated at regular intervals thereafter or when
any significant change is made in the process or equipment.

5. Sterilization
5.1 Whenever possible products intended to be sterile should be terminally
sterilized by heat in their final container. Where it is not possible to carry
out terminal sterilization by heating due to the instability of a formulation
or incompatibility of a pack type (necessary to the administration of the
product, e.g. plastic eye-dropper bottles), a decision should be taken to use
an alternative method of terminal sterilization following filtration and/or
aseptic processing.
5.2 Sterilization can be achieved by the use of moist or dry heat, by
irradiation with ionizing radiation (noting that ultraviolet irradiation is
not normally an acceptable method of sterilization), by ethylene oxide (or
other suitable gaseous sterilizing agents), or by filtration with subsequent
aseptic filling of sterile final containers. Each method has its advantages
and disadvantages. Where possible and practicable, heat sterilization is the
method of choice. In any case the sterilization process must be in accordance
with the marketing and manufacturing authorizations.
5.3 The microbial contamination of starting materials should be minimal
and their bioburden should be monitored before sterilization. Specifications

272
 should include requirements for microbiological quality when the need for
this has been indicated by monitoring.
5.4 All sterilization processes should be validated. Particular attention
should be paid when the adopted sterilization method is not in accordance
with pharmacopoeial standards or other national standards, or when it is
used for a preparation that is not a simple aqueous or oily solution, for
example, colloidal suspensions.
5.5 Before any sterilization process is adopted, its suitability for the
product and its efficacy in achieving the desired sterilizing conditions in
all parts of each type of load to be processed should be demonstrated by
physical measurements and by biological indicators, where appropriate.
The validity of the process should be verified at scheduled intervals, at least
annually, and whenever significant modifications have been made to the
equipment. Records should be kept of the results.
5.6 For effective sterilization the whole of the material should be subjected
to the required treatment and the process should be designed to ensure that
this is achieved.
5.7 Biological indicators should be considered only as an additional
method of monitoring the sterilization process. They should be stored and
used according to the manufacturer’s instructions, and their quality checked
by positive controls. If they are used, strict precautions should be taken to
avoid any transfer of microbial contamination from them.
5.8 There should be a clear means of differentiating products that have
not been sterilized from those which have. Each basket, tray, or other carrier
of products or components should be clearly labelled with the name of the
material, its batch number and an indication of whether or not it has been
sterilized. Indicators such as autoclave tape may be used where appropriate to
indicate whether or not a batch (or sub-batch) has passed through a sterilization
process, but they do not give a reliable indication that the batch is in fact sterile.
5.9 Validated loading patterns should be established for all sterilization
processes.
5.10 Sterilization records should be available for each sterilization run.
They should be approved as part of the batch-release procedure.

6. Terminal sterilization
Sterilization by heat
6.1 Each heat-sterilization cycle should be recorded by means of
appropriate equipment of suitable accuracy and precision, e.g. on a time/
temperature chart with a suitably large scale. The temperature should

273
 be recorded by a probe situated at the coolest part of the load or loaded
chamber, this point having been determined during the validation; the
temperature should preferably be checked against a second independent
temperature probe located at the same position. Sterilization records should
be available for each sterilization run and should be approved as part of the
batch release procedure. Chemical or biological indicators may also be used
but should not take the place of physical controls.
6.2 Sufficient time should be allowed for the whole of the load to reach
the required temperature before measurement of the sterilizing time
is started. This time should be determined for each type of load to be
processed.
6.3 After the high-temperature phase of a heat sterilization cycle,
precautions should be taken against contamination of a sterilized load
during cooling. Any cooling fluid or gas in contact with the product should
be sterilized.
Sterilization by moist heat
6.4 Both temperature and pressure should be used to monitor the process.
Control instrumentation should normally be independent of monitoring
instrumentation and recording charts. Where automated control and
monitoring systems are used for these applications they should be validated
to ensure that critical process requirements are met. System and cycle
faults should be registered by the system and observed by the operator.
The reading of the independent temperature indicator should be routinely
checked against the reading on the chart recorder during the sterilization
period. For sterilizers fitted with a drain at the bottom of the chamber, it
may also be necessary to record the temperature at this position throughout
the sterilization period. There should be regular leak tests on the chamber
when a vacuum phase is part of the cycle.
6.5 The items to be sterilized, other than products in sealed containers,
should be wrapped in a material that allows the removal of air and the
penetration of steam but prevents recontamination after sterilization.
Specially designed autoclavable stainless steel containers, that allow steam
to enter and air to leave, can also be used. All parts of the load should be
in contact with water or saturated steam at the required temperature for the
required time.
6.6 Care should be taken to ensure that the steam used for sterilization
is of suitable quality (chemical, microbiological and endotoxin analysis of
condensate and physical examination of steam (such as dryness, superheat,
and non-condensable gases) and does not contain additives at a level that
could cause contamination of the product or equipment. Steam used for
sterilization should be tested regularly.

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Sterilization by dry heat
6.7 Sterilization by dry heat may be suitable for non-aqueous liquids or
dry-powder products.
The process used should include air circulation within the chamber and the
maintenance of a positive pressure to prevent the entry of non-sterile air. If
air is supplied it should be passed through a microorganism-retaining filter
(e.g. a HEPA filter). Where sterilization by dry heat is also intended to remove
pyrogens, challenge tests using endotoxins are required as part of the validation.

Sterilization by radiation
6.8 Sterilization by radiation is used mainly for heat-sensitive materials
and products. Many pharmaceutical products and some packaging materials
are radiation-sensitive, so this method is permissible only when the absence
of deleterious effects on the product has been confirmed experimentally.
Ultraviolet irradiation is not an acceptable method for terminal sterilization.
6.9 If sterilization by radiation is done by an outside contractor, the
manufacturer is responsible for ensuring that the requirements of section
6.8 are met and that the sterilization process is validated.
6.10 During the sterilization procedure the radiation dose should be
measured. The dosimeters used for this purpose should be independent of the
dose rate and should provide a quantitative measurement of the dose received
by the product itself. Dosimeters should be inserted in the load in sufficient
number and close enough together to ensure that there is always a dosimeter
in the chamber. Where plastic dosimeters are used they should be used within
the time-limit of their calibration. Dosimeter absorbance should be read
shortly after exposure to radiation. Radiation-sensitive colour discs may be
used to differentiate between packages that have been subjected to irradiation
and those that have not; they are not indicators of successful sterilization. The
information obtained should constitute part of the batch record.
6.11 Validation procedures should ensure that consideration is given to
the effects of variations in the density of the packages.
6.12 Material-handling procedures should prevent any mix-up of
irradiated and non-irradiated materials. Each package should carry a
radiation-sensitive indicator to show whether or not it has been subjected to
radiation treatment.
6.13 The total radiation dose should be administered within a predetermined
period.

Sterilization by gases and fumigants

6.14 Sterilization by gases and fumigants should only be used for finished
products where there is no suitable alternative.

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 6.15 Various gases and fumigants may be used for sterilization (e.g. ethylene
oxide and hydrogen peroxide vapour). Ethylene oxide should be used only
when no other method is practicable. During process validation it should
be shown that the gas has no damaging effect on the product and that the
conditions and time allowed for degassing are such as to reduce any residual
gas and reaction products to defined acceptable limits for the type of product or
material concerned. These limits should be incorporated in the specifications.
6.16 Direct contact between gas and microorganisms is essential;
precautions should, therefore, be taken to avoid the presence of organisms
likely to be enclosed in materials such as crystals or dried protein. The nature
and quantity of packaging materials can significantly affect the proces.
6.17 Before exposure to the gas, materials should be brought into
equilibrium with the humidity and temperature required by the process.
This requirement should be balanced against the need to minimize the
waiting time before sterilization.
6.18 Each sterilization cycle should be monitored with suitable biological
indicators, using the appropriate number of test pieces distributed throughout
the load. The information thus obtained should form part of the batch record.
6.19 Biological indicators should be stored and used according to the
manufacturer’s instructions and their performance checked by positive
controls.
6.20 For each sterilization cycle, records should be made of the time taken
to complete the cycle, of the pressure, temperature and humidity within
the chamber during the process and of the gas concentration. The pressure
and temperature should be recorded on a chart throughout the cycle. The
records should form part of the batch record.
6.21 After sterilization, the load should be stored in a controlled manner
in ventilated conditions to allow concentrations of residual gas and reaction
products to fall to their prescribed levels. This process should be validated.

7. Aseptic processing and sterilization by filtration

7.1 The objective of aseptic processing is to maintain the sterility of a
product that is assembled from components, each of which has been
sterilized by one of the above methods (see sections 5 and 6).
7.2 The operating conditions should be such as to prevent microbial
contamination.
7.3 In order to maintain the sterility of the components and the product
during aseptic processing, careful attention needs to be given to:

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— the environment;
— personnel;
— critical surfaces;
— container/closure sterilization and transfer procedures;
— the maximum holding period of the product before filling into the final
container; and
— the sterilizing filter.
7.4 Certain solutions and liquids that cannot be sterilized in the final
container can be filtered through a sterile filter of nominal pore size
0.22 micron (or less), or with at least equivalent microorganism-retaining
properties, into a previously sterilized container. Such filters can remove
bacteria and moulds, but not all viruses or mycoplasmas. Consideration
should be given to complementing the filtration process with some degree
of heat treatment. Filtration alone is not considered sufficient when
sterilization in the final container is possible. Of the methods currently
available, steam sterilization is preferred.
7.5 Owing to the potential additional risks of the filtration method
as compared with other sterilization processes, a double-filter layer or
second filtration through a further sterilized microorganism-retaining filter
immediately prior to filling may be advisable. The final sterile filtration should
be carried out as close as possible to the filling point.
7.6 The fibre-shedding characteristics of filters should be minimal
(virtually zero). Asbestos-containing filters should not be used under any
circumstances.
7.7 The integrity of the sterilized filter should be verified before use and
should be confirmed immediately after use by an appropriate method such
as a bubble point, diffusive flow or pressure hold test. The time taken to
filter a known volume of bulk solution and the pressure difference to be used
across the filter should be determined during validation and any significant
differences from these during routine manufacturing should be noted
and investigated. Results of these checks should be included in the batch
record. The integrity of critical gas and air vent filters should be confirmed
after use. The integrity of other filters should be confirmed at appropriate
intervals. Consideration should be given to increased monitoring of filter
integrity in processes that involve harsh conditions, e.g. the circulation of
high-temperature air.
7.8 The same filter should not be used for more than one working day
unless such use has been validated.
7.9 The filter should not affect the product either by removing ingredients
from it or by releasing substances into it.

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8. Isolator technology
8.1 The use of isolator technology to minimize human interventions in
processing areas may result in a significant decrease in the risk of microbial
contamination of aseptically manufactured products from the environment.
There are many possible designs of isolators and transfer devices. The
isolator and the background environment should be designed so that the
required air quality for each zone can be realized. Isolators are constructed
of various materials more or less prone to puncture and leakage. Transfer
devices may vary from single-door to double-door designs to fully-sealed
systems incorporating sterilization mechanisms.
8.2 The transfer of materials into and out of the unit is one of the greatest
potential sources of contamination. In general the area inside the isolator
is the local zone for high-risk manipulations, although it is recognized that
unidirectional airflow may not exist in the working zone of all isolators and
transfer devices.
8.3 The air classification required for the background environment
depends on the design of the isolator and its application. It should be
controlled, and for aseptic processing it should be at least Grade D.
8.4 Isolators should be introduced only after appropriate validation.
Validation should take into account all critical factors of isolator technology,
for example, the quality of the air inside and outside (background) the
isolator, sanitization of the isolator, the transfer process and isolator integrity.
8.5 Monitoring should be done routinely and should include frequent leak
testing of the isolator and the glove/sleeve system.

9. Blow/fill/seal technology
9.1 Blow/fill/seal units are purpose-built machines in which, in one
continuous operation, containers are formed from a thermoplastic granulate,
filled and then sealed, all by the one automatic machine. Blow/fill/seal
equipment used for aseptic production which is fitted with an effective Grade
A air shower may be installed in at least a Grade C environment, provided
that Grade A or B clothing is used. The environment should comply with
the viable and non-viable limits at rest and the viable limit only when in
operation. Blow/fill/seal equipment used for the production of products which
are terminally sterilized should be installed in at least a Grade D environment.
9.2 Because of this special technology, particular attention should be paid
to at least the following:
— equipment design and qualification;

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 — validation and reproducibility of cleaning-in-place and sterilization-in-
place;
— background clean room environment in which the equipment is located;
— operator training and clothing; and
— interventions in the critical zone of the equipment including any aseptic
assembly prior to the commencement of filling.

10. Personnel
10.1 Only the minimum number of personnel required should be present
in clean areas; this is particularly important during aseptic processes. As
far as possible, inspections and controls should be conducted from outside
such areas.
10.2 All personnel (including those concerned with cleaning and
maintenance) employed in such areas should receive initial and regular
training in disciplines relevant to the correct manufacture of sterile products,
including hygiene and the basic elements of microbiology. When outside
staff who have not received such training (e.g. building or maintenance
contractors) need to be brought in, particular care should be taken over their
instruction and supervision.
10.3 Staff who have been engaged in the processing of animal-tissue
materials or of cultures of microorganisms other than those used in the
current manufacturing process should not enter sterile-product areas
unless rigorous and clearly defined decontamination procedures have been
followed.
10.4 High standards of personal hygiene and cleanliness are essential
and personnel involved in the manufacture of sterile preparations should
be instructed to report any conditions that may cause the shedding of
abnormal numbers or types of contaminants; periodic health checks for
such conditions are desirable. The action to be taken in respect of personnel
who might be introducing undue microbial hazards should be decided by a
designated competent person.
10.5 Changing and washing should follow a written procedure designed
to minimize the contamination of clean-area clothing or the carry-through
of contaminants to clean areas. The clothing and its quality should be
appropriate for the process and the grade of the working area. It should be
worn in such a way as to protect the product from contamination.
10.6 Outdoor clothing should not be brought into changing rooms leading
to Grade B and C rooms. For every worker in a Grade A/B area, clean sterile
(sterilized or adequately sanitized) protective garments should be provided
at each work session. Gloves should be regularly disinfected during

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 operations. Masks and gloves should be changed at least every working
session. Operators working in Grade A and B areas should wear sanitized
goggles.
10.7 Wrist-watches, cosmetics and jewellery should not be worn in clean
areas.
10.8 The clothing required for each grade is as follows:
• Grade D. The hair and, where relevant, beard and moustache should be
covered. Protective clothing and appropriate shoes or overshoes should be
worn. Appropriate measures should be taken to avoid any contamination
from outside the clean area.
• Grade C. The hair and, where relevant, beard and moustache should be
covered. A one-piece jumpsuit, gathered at the wrists and with a high
neck, and appropriate shoes or overshoes should be worn. The clothing
should shed virtually no fibres or particulate matter.
• Grades A and B. Entry of personnel into Grade A areas should be
minimized. Headgear should totally enclose the hair and, where relevant,
beard and moustache. A one-piece jumpsuit, gathered at the wrists and
with a high neck, should be worn. The headgear should be tucked into
the neck of the suit. A facemask should be worn to prevent the shedding
of droplets. Sterilized, non-powdered gloves of appropriate material and
sterilized or disinfected footwear should be worn. Trouser-bottoms should
be tucked inside the footwear and garment sleeves into the gloves. The
protective clothing should shed virtually no fibres or particulate matter
and should retain particles shed by the body.
10.9 Clothing used in clean areas should be laundered or cleaned in such
a way that it does not gather additional particulate contaminants that can
later be shed. Separate laundry facilities for such clothing are desirable. If
fibres are damaged by inappropriate cleaning or sterilization, there may be
an increased risk of shedding particles. Washing and sterilization operations
should follow standard operating procedures.

11. Premises
11.1 All premises should as far as possible be designed to avoid the
unnecessary entry of supervisory or control personnel. Grade A and B
areas should be designed so that all operations can be observed from
outside.
11.2 In clean areas all exposed surfaces should be smooth, impervious
and unbroken to minimize the shedding or accumulation of particles or
microorganisms and to permit the repeated application of cleaning agents
and disinfectants, where used.

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11.3 To reduce the accumulation of dust and to facilitate cleaning, there
should be no uncleanable recesses and a minimum of projecting ledges,
shelves, cupboards and equipment. Doors should be carefully designed
to avoid uncleanable recesses; sliding doors may be undesirable for this
reason. Swing doors should open to the high-pressure side and be provided
with self-closers. Exceptions are permitted based on egress and site
environmental, health and safety containment requirements.
11.4 False ceilings should be sealed to prevent contamination from the
void space above them.
11.5 Pipes and ducts and other utilities should be installed so that they
do not create recesses, unsealed openings and surfaces that are difficult
to clean. Sanitary pipes and fittings should be used and threaded pipe
connections should be avoided.
11.6 Sinks and drains should be avoided wherever possible and should
be excluded from Grade A and B areas where aseptic operations are carried
out. Where installed they should be designed, located and maintained so as
to minimize the risks of microbial contamination; they should be fitted with
effective, easily cleanable traps and with air breaks to prevent backflow. Any
floor channels should be open and easily cleanable and be connected to drains
outside the area in a manner that prevents the ingress of microbial contaminants.
11.7 Changing rooms should be designed as airlocks and used to provide
physical separation of the different stages of changing and so minimize
microbial and particulate contamination of protective clothing. They should
be flushed effectively with filtered air. The final stage of the changing room
should, in the at-rest state, be the same grade as the area into which it leads.
The use of separate changing rooms for entering and leaving clean areas is
sometimes desirable. In general hand-washing facilities should be provided
only in the first stage of the changing rooms.
There should not be a change of more than one grade between airlocks or
passages and changing rooms, i.e. a Grade D passage can lead to a Grade C
airlock, which leads to a Grade B changing room, which leads to a Grade B
clean room. Changing rooms should be of a sufficient size to allow for ease of
changing. Changing rooms should be equipped with mirrors so that personnel
can confirm the correct fit of garments before leaving the changing room.
11.8 Airlock doors should not be opened simultaneously. An interlocking
system and a visual and/or audible warning system should be operated to
prevent the opening of more than one door at a time.
11.9 A filtered air supply should be used to maintain a positive pressure
and an airflow relative to surrounding areas of a lower grade under all
operational conditions; it should flush the area effectively. Adjacent rooms

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 of different grades should have a pressure differential of approximately
10–15 Pascals (guidance value). Particular attention should be paid to the
protection of the zone of greatest risk, i.e. the immediate environment to
which the product and the cleaned components in contact with it are exposed.
The recommendations regarding air supplies and pressure differentials may
need to be modified where it becomes necessary to contain certain materials,
e.g. pathogenic, highly toxic, radioactive or live viral or bacterial materials
or products. The decontamination of the facilities and the treatment of air
leaving a clean area may be necessary for some operations.
11.10 It should be demonstrated that airflow patterns do not present a
contamination risk; for example, care should be taken to ensure that
particles from a particle-generating person, operation or machine are not
conveyed to a zone of higher product risk.
11.11 A warning system should be operated to indicate failure in the air
supply. Indicators of pressure differentials should be fitted between areas
where this difference is important, and the pressure differentials should be
regularly recorded and failure alarmed.
11.12 Consideration should be given to restricting unnecessary access to
critical filling areas, e.g. Grade A filling zones, by means of a physical barrier.

12. Equipment
12.1 A conveyor belt should not pass through a partition between a Grade
A or B clean area and a processing area of lower air cleanliness, unless the
belt itself is continuously sterilized (e.g. in a sterilizing tunnel).
12.2 Whenever possible, equipment used for processing sterile products
should be chosen so that it can be effectively sterilized by steam or dry heat
or other methods.
12.3 As far as possible, equipment fittings and services should be designed
and installed so that operations, maintenance and repairs can be carried out
outside the clean area. Equipment that has to be taken apart for maintenance
should be re-sterilized after complete reassembly, wherever possible.
12.4 When equipment maintenance is carried out within a clean area,
clean instruments and tools should be used and the area should be cleaned
and disinfected again, where appropriate, before processing recommences,
if the required standards of cleanliness and/or asepsis have not been
maintained during the maintenance work.
12.5 All equipment such as sterilizers, air-handling and filtration systems,
air vent and gas filters, water treatment, generation, storage and distribution

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 systems should be subject to validation and planned maintenance; their
return to use should be approved.
12.6 Water-treatment plants and distribution systems should be designed,
constructed and maintained so as to ensure a reliable source of water of an
appropriate quality. They should not be operated beyond their designed
capacity. Consideration should be given to including a testing programme in the
maintenance of a water system. Water for injection should be produced, stored
and distributed in a manner which prevents the growth of microorganisms, e.g.
by constant circulation at a temperature above 70 °C or not more than 4 °C (8).

13. Finishing of sterile products

13.1 Containers should be closed by appropriately validated methods.
Containers closed by fusion, e.g. glass or plastic ampoules, should be
subject to 100% integrity testing. Samples of other containers should be
checked for integrity according to appropriate procedures.
13.2 The container closure system for aseptically filled vials is not
fully integral until the aluminum cap has been crimped into place on the
stoppered vial. Crimping of the cap should, therefore, be performed as soon
as possible after stopper insertion.
13.3 As the equipment used to crimp vial caps can generate large quantities
of non-viable particulates, the equipment should be located at a separate
station equipped with adequate air extraction.
13.4 Vial capping can be undertaken as an aseptic process using sterilized
caps or as a clean process outside the aseptic core. Where this latter approach
is adopted, vials should be protected by Grade A conditions up to the point
of leaving the aseptic processing area, and thereafter stoppered vials should
be protected with a Grade A air supply until the cap has been crimped.
13.5 Vials with missing or displaced stoppers should be rejected prior
to capping. Where human intervention is required at the capping station,
appropriate technology should be used to prevent direct contact with the
vials and to minimize microbial contamination.
13.6 Restricted access barriers and isolators may be beneficial in assuring
the required conditions and minimizing direct human interventions into the
capping operation.
13.7 Containers sealed under vacuum should be tested for maintenance of
that vacuum after an appropriate, predetermined period.
13.8 Filled containers of parenteral products should be inspected individually
for extraneous contamination or other defects. When inspection is carried

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 out visually this should be done under suitable and controlled conditions of
illumination and background. Operators doing the inspection should pass regular
eyesight checks, using personal corrective lenses (e.g. spectacles or contact
lenses) as required, and be allowed frequent breaks from inspection. Where
other methods of inspection are used, the process should be validated and the
performance of the equipment checked at intervals. Results should be recorded.

References
1. Good manufacturing practices for sterile pharmaceutical products. In: WHO
Expert Committee on Specifications for Pharmaceutical Preparations. Thirty-sixth
report. Geneva, World Health Organization, 2002 (WHO Technical Report Series,
No. 902), Annex 6; and in Quality assurance of pharmaceuticals. A compendium
of guidelines and related materials. Vol. 2. 2nd updated ed. Good manufacturing
practices and inspection. Geneva, World Health Organization, 2007; and in
Quality Assurance of Pharmaceuticals. A compendium of guidelines and related
materials. Geneva, World Health Organization, 2010 (CD-ROM).
2. ISO 14644-1. Clean rooms and associated controlled environments. Part 1:
Classification of airborne particles. Geneva, International Organization for
Standardization.
3. ISO 14644-3. Clean rooms and associated controlled environments. Part 3:
Test methods. Geneva, International Organization for Standardization.
4. ISO 1822-4. High efficiency air filters (HEPA and ULPA). Determining leakage
of filter elements (scan method).
5. ISO 14644-4. Clean rooms and associated controlled environments. Part 4:
Design, construction and start-up. Geneva, International Organization for
Standardization.
6. ISO 14644-2. Clean rooms and associated controlled environments. Part 2:
Monitoring for continued compliance with ISO 14644-1. Geneva, International
Organization for Standardization.
7. ISO 14644-5 Clean rooms and associated controlled environments. Part 5:
Cleanroom operations. Geneva, International Organization for Standardization.
8. Good manufacturing practices for pharmaceutical products: water for
pharmaceutical use. In: WHO Expert Committee on Specifications for
Pharmaceutical Preparations. Thirty-ninth report. Geneva, World Health
Organization, 2005 (WHO Technical Report Series, No. 929), Annex 3; and
in Quality assurance of pharmaceuticals. A compendium of guidelines and
related materials. Vol. 2. 2nd updated ed. Good manufacturing practices and
inspection. Geneva, World Health Organization, 2007.

Further reading
FDA Guidance for Industry. Sterile drug products produced by aseptic
processin g — cGMP. US Food and Drug Administration, 2004.
Guidance for industry. Sterile drug products produced by aseptic
processing. Japan, 2005.
Manufacture of sterile medicinal products. In: The rules governing medicinal
products in the European Union Vol. 4. EU guidelines to good manufacturing
practice medicinal products for human and veterinary use. Annex 1, Brussels, 2008.

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