Pharmaceutical Technology (PHR 402)

Cleanroom Technology

What is a cleanroom?
According to International Organization for Standardization (ISO) standard 14644-1, a cleanroom
is defined as:

Importance of Cleanrooms
• People, production machinery and the building structure generate contamination.
• A cleanroom controls this dispersion and allows manufacturing to be carried out in a clean
environment, thereby, protecting the products from being adversely affected by contaminants
• Cleanrooms are an important part of manufacturing processes, scientific research and quality
control. Provide the technological solution for contamination control.
– They are designed to protect products, personnel, and equipment from being adversely affected
by contaminants, through creating clean and controlled areas.
– Cleanrooms are also used in other sectors such as….

Contamination & Cross-contamination

Contamination
Definition?

Cross-contamination
According to WHO definition, cross-contamination is defined as …..

Sources of Contamination
Possible sources of contamination are:
• Atmosphere

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 • Operator/Personnel
• Raw materials
• Equipment

• Atmosphere is heavily contaminated with particles and microorganisms.

a) Contaminants in outside air → Originate from soil and carry soil organisms including-
1. Bacteria spores (Bacillus spp, Clostrium spp)
2. Mould spores (Penicillium, Mucour, Aspergilius)
3. Yeasts and Micrococci
b) Contaminants in indoor air → Originate from human body and clothing’s including-
1. Bacteria spores on human skin (Staphylococcus spp, Streptococcus spp)
2. These will also occur in droplets expelled out into the air from respiratory tract by
talking, coughing, sneezing etc.

• Operator (most risky factor)

The skin, hair, clothing and droplets of moisture from breathing and coughing of the operator are potent
sources of particulate and microbial contamination. Organisms found on the skin and transmitted on the
skin particles are –
• Staphylococcus
• Diphtheroids
• Malassezia yeasts (Lipophillic yeasts)

• Raw Materials
1. Drugs which are obtained from natural sources, example:
• Plant source – saprophytic bacteria, yeast, moulds;
• Animal source –pathogenic bacteria / spores.
2. Packaging materials and closures contaminate specially the parenteral solutions.
3. Pigments: Salmonella
4. Starches: Coliforms
5. Gums: Actinomyces
6. Water – prime source of particulate contamination
• Equipment
Particles and droplets from atmosphere may sediment on to the internal and external surface of equipment.

Sources of Cross-Contamination
1. Poorly designed, operated or maintained air handling systems and dust extraction systems
2. Inadequate procedures for personnel
3. Insufficiently cleaned equipment

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Why is there a need for cleanrooms?

What is a clean area?

Cleanroom Classification Systems

Cleanrooms are classified based on the quantity and size of particles per cubic meters of air inside a
controlled environment.
Cleanrooms can be classified according to
• US Federal Standard 209E ( FS 209E )
• International Organization for Standardization 14644-1 (ISO 14644-1 )
• EU cGMP

Each cleanroom class is denoted by a maximum concentration of particles per cubic meter of air.

ISO 1 is the “cleanest” class and ISO 9 is the “dirtiest” class. Even if it’s classified as the “dirtiest” class, the ISO 9 clean room
environment is cleaner than a regular room.

All pharmaceutical facilities belong to one or other class of cleanroom.

Equation for the IS0 classification?

Types of Cleanrooms
Cleanrooms can be categorized by their methods of ventilation (way supply air is distributed/airflow
direction) and occupancy states.
1. Method of ventilation/Direction of airflow
(a) Unidirectional/Laminar flow Cleanrooms

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 - Horizontal air flow
- Vertical air flow
(b) Non-unidirectional flow cleanrooms/Turbulently ventilated cleanrooms
(c) Combination systems (Mixed airflow cleanrooms)

2. Occupancy states
(a) as built
(b) at rest
(c) in operation

Methods of Ventilation
Turbulently ventilated cleanroom

Turbulently ventilated cleanroom receives clean filtered air (filtered by high efficiency air filters) through
air diffusers in the ceiling. This air mixes with the room air and removes airborne contamination through
air extracts at the bottom of the walls.
Airflows with varying velocity and in nonparallel direction. The air change rate are normally equal to, or greater
than, 20 per hour

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Unidirectional flow cleanroom

High efficiency filters are installed across a whole ceiling (or wall in some systems) and these supply air.
This air sweeps across the room in a unidirectional way (parallel airstreams) with a uniform velocity of
around 0.4 m/s (80 ft/min) and exits through the floor, thus removing the airborne contamination from the
room. No diffusers are used here.
This system uses much more air than the turbulently ventilated cleanroom (higher air change rate) but,
because of the directed air movement, it minimizes the spread of contamination about the room and sweeps
it out through the floor and thus gives superior cleanliness.
Unidirectional flow rooms are of two types, namely horizontal or vertical flow. In the horizontal system,
the airflow is from wall to wall and in the vertical system it is from ceiling to floor.
Unidirectional airflow is used in cleanrooms when low airborne concentrations of particles or
microorganisms is required.

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Mixed airflow cleanroom
A mixed flow room is one where the ventilation of the background room area is non unidirectional but has
unidirectional flow cabinets, workstations, or isolators within the room.

Occupancy States
As built

The condition where the installation is complete with all services connected and functioning, but with no production
equipment, materials or personnel present.

At rest

The condition where the installation is complete with equipment installed and operating in a manner agreed upon, but
with no personnel present.

Operational

The condition where the installation is complete and equipment is functioning in the specified manner, with the
specified number of personnel present and working in the manner agreed upon (installation complete, equipment
present, personnel present).

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Cleanroom Design & Operation
1. Preparation Area
- This room is used for the purpose of preparing components, i.e. to open the outer wrappings and to sanitize the
materials.

- All components should be taken in or out of the aseptic area through hatches or pass-box.

- The air quality in this area should be maintained at a minimum level of Class 100,000.

- Sinks and other related facilities should be made available in this room.

2. Mixing Area
- This room is used for mixing medicaments

- Sinks and other related facilities should be made available in this room.

3. Changing Area
- This room is used for personnel to change into clean apparel before entering the aseptic area.

- The air quality in this area should be maintained at a minimum level of Class 100,000.

- This room should be designed as an air lock with 3 distinct areas. The 3 areas should be clearly designated
either with a cross-over bench or a line on the floor. The areas can be classified as black, grey and white
area.

- Some changing area may also contain air shower. An air shower is designed to remove particles from
clothing and hence reduce dispersion in a cleanroom.

4. Aseptic Area:
- Filling medicaments into final containers using aseptic technique.

- Air quality in this area should be maintained at a minimum of Class 100.

- Minimum area of the room is 50 sq.ft. depending on the size of the laminar airflow cabinet and the number of
personnel.

- This room should have the highest pressure compared to the outside (or other rooms) and there should be a net
flow of air from this room to other areas.

- This room should be kept dry and clean.

- Sinks or pipes should not be located within this area.

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Cleanroom Construction
A cleanroom requires a higher standard of construction than many other conventional buildings.

• A cleanroom should be built with an airtight structure.

• The internal surface finish should be smooth, easy-to-clean and not harbor dirt.

• The internal surface finish should be sufficiently tough to resist chipping or powdering when impacted or
abraded.

• In some cleanrooms, electro-dissipative construction materials will be required.

• In some cleanrooms, construction materials that give a minimum of ‘outgassing’ will be required.

Cleanroom Air Pressure

To ensure that contaminated air does not come into the room from dirtier adjacent areas, the cleanroom must be
maintained at a positive air pressure.

In a cleanroom with positive air pressure (pressure inside cleanroom higher than outside), air always moves from the
cleanroom to less-clean adjacent areas.

Differential pressures of 10 Pa between two cleanrooms, and 15 Pa between a cleanroom and an unclassified
area, are usually maintained.

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The Three Components of Cleanroom Technology

Figure: Various parts of cleanroom technology and their interconnections

HVAC System
Maintenance of cleanroom air purity and the necessary requirements for temperature and humidity adjustment is
controlled using an HVAC system.

An HVAC system is Heating, Ventilation and Air Conditioning system. That is used for conditioning of air in a
big area where a usual air conditioner cannot be used. It contains a boiler, chiller cooling tower and air handling
unit.

HVAC controls parameters such as Temperature, Relative Humidity, Differential Pressures, and Air Flow for a
cleanroom.

This system is used to control the temperature of a designated space and it also controls the contamination of
airborne particles by regulating the movement of air.

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HVAC Parameters

Parameter Recommended Limit

Temperature 20±5°C

Humidity 50 ±5 % in all manufacturing areas and 20% for effervescent products

Air Class As per the required International standards; i.e. Federal standards, ISO standards; British standards etc.

Pressure Gradient 10 Pa between two cleanrooms, and 15 Pa between a cleanroom and an unclassified area

Air Quality Dust and Odor free

Sound Level Within 20 db

Typical System for Supplying Clean Air

1. Intake of fresh air
2. Prefiltration
3. Temperature adjustment
4. Humidification
5. Final filtration

What is Air Handling Unit (AHU)?

Final filtration is achieved by using a “High Efficiency Particulate Air” (HEPA) filter. HEPA filter has
an efficiency of 99.97% in removing small airborne particles of size 0.3 µm and greater.
▪ High Efficiency Particulate Air Filter, or HEPA filter has an efficiency of 99.97% in removing small
airborne particles of size 0.3 µm and greater.

▪ More efficient filters than HEPA filters are known as Ultra Low Penetration Air (ULPA) filters. An ULPA
filter have an efficiency greater than 99.999% against 0.1-0.2 µm particles.

It is generally accepted that

• For cleanrooms of IS0 Class 6 (Class 1000) and poorer quality, HEPA filters are used

• For IS0 Class 5 (Class 100), HEPA filters are used

• For IS0 Class 4 (Class 10) or lower, ULPA filters should be used

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Most HEPA filters are made up of interwoven mesh of glass fibers.

• There are four mechanism of filtration

1. Sieving (Straining/Screening)
2. Impaction (Impingement)
3. Interception
4. Diffusion

Straining
If the pores of the filter medium through which the air is flowing are smaller than the particles, which is to be
removed then the particles will be retained.

Impingement
• As air passes the filter fiber, the air flow pattern is disturbed as shown in the following figure.

• Suspended particles with in-sufficient momentum may not follow the air path, but impact (impinge) on the
filter fibre and are retained due to the attractive forces between the particle and the fibre.

Interception
If a particle strikes a fibre as it passes it, i.e. tangentially, it will be captured and retained due to the attractive forces
between the particle and the fibre.

Diffusion
Particles collide with other particles in the air and with particles already retained in the filter. This bombardment
produces random motion of the particles. As the particles touch the fibers on its way or previously captured
particles, they are retained.

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Testing Of Clean and Aseptic Rooms
a) Commissioning Tests
• Final filter installation test: This is done to demonstrate that the filter is not damaged and that the filter
mounting frame does not leak at the gasket flange or the connection to the ducting.

• Induction leak test: This test demonstrates that particles cannot enter the room from leaks in construction
joints or by back-streaming from openings.

• Filter efficiency test: Aerosol photometers and the generation and detection of DOP(Dioctyl pthalate)
smoke or sodium chloride crystals are usually used for these tests.

• Particulate contamination control test: This is used to demonstrate that the number and size distribution
of particles in the clean room air do not exceed the levels specified for the particular class of room.

Particle counter is used for this type of test.

- counts no. of particles in the air of a cleanroom

- measures size of particles in the air of a cleanroom

• Air pressure test: This test determines the differential pressure between the clean area and adjacent areas.
This is usually measured using a sensitive manometer. 10 Pa between two cleanrooms, and 15 Pa between
a cleanroom and an unclassified area

• Temperature and humidity tests: Measurements are usually made with a sling psychrometer.

- Temperature: 20±5ºC unless there is any specific recommendation for any special operation.

- It is recommended to maintain RH within 50±5 % in all manufacturing areas, unless there is any specific
recommendation for any special operation. For example, for Effervescent product manufacturing it is
recommended to maintain the relative humidity around 20%.

• Air flow tests: Air flow (volume of air entering the room and air velocity) in the cleanroom can be
measured with a pitot tube and anemometer. Air change rates can be demonstrated by tracer gas decay
rate.

• Noise level tests: British Standard stipulates a maximum level of 65 dB

• Lighting test: The quality of the general illumination within the area and also at the work bench is
measured using a portable photoelectric photometer.

Microbiological tests

- Settle plate method

- Contact plate and swabbing method
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b) Monitoring tests:
• Air pressure, temperature, and humidity measurements should be recorded continuously.
• Particulate contamination should be determined daily for aseptic rooms and weekly for clean rooms
• Tests for air flow velocity should be carried out at 3-monthly intervals.
• Tests for filter efficiency should be conducted yearly. Tests should also be repeated after repairs or
maintenance
• Settle plates, agar contact plates and swabs should be carried out during each work shift to monitor
environmental air for microbes.

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Microbiological Tests
1. Settle plate method

• A method for environmental monitoring of microbial count.

• Petridishes containing nutrient agar medium are exposed to cleanroom environment for several hours (apx.
4 hr), thus allowing microbe-carrying particles to deposit onto them.

• The petridishes are incubated for a specific period of time (e.g. 2-3 days) and the colony forming units are
counted.

2. Contact surface method

Contact plates containing agar are used when the cleanroom surface to be sampled is relatively flat. The agar is
rolled over the cleanroom surface to be sampled. Microorganisms will stick to the agar and when the dish is
incubated for a suitable time and temperature, the micro-organisms will grow into colonies that can be counted.

3. Swabbing method

To swab uneven surfaces, a commonly used method is the application of a bud swab made from material such as
cotton. A sterile swab is randomly rubbed over the cleanroom surface to be sampled and then rubbed over an agar
plate. The plate is then incubated and the microbial count determined.

Contamination Control

To be able to control contamination in a cleanroom it is first of all necessary to assess the various risks.

A number of risk management systems exist for contamination control in cleanrooms. They are-

• Fault Tree Analysis (FTA)

• Failure Mode and Effect Analysis (FMEA)
• Hazard Analysis and Critical Control Point (HACCP)

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HACCP System
HACCP has an eight-step approach:

1. Identify the sources of contamination in the cleanroom, e.g. room air, surfaces, people, equipment

2. Assess whether, or not, these sources are hazards that need to be controlled.

3. Identify methods that can be used to control these hazards e.g. HEPA filters, correct airflow movement,
disinfecting surfaces, proper gowning by personnel, sterilizing machines and equipment

4. Determine valid sampling methods to monitor either the hazards, or their control methods, or both e.g.
HEPA filter integrity tests, air supply velocity, air change rates, room pressure differentials, particle counts

5. Establish corrective actions, where appropriate, when particle and microbial levels are exceeded.

6. Verify that the contamination control system is working effectively by reviewing the product rejection rate
and sampling results and, where appropriate, modifying control methods.

7. Establish and maintain appropriate documentation.

8. Train the staff.

Principles of Cleanroom Testing

Re-testing to Demonstrate Compliance

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Risk factors for assessing hazards
A risk assessment is required to answer the question ‘how great a hazard are cleanroom walls’.

A set of variables known as risk factors should be determined:

– the amount of contamination on, or in, the source that is available for transfer (risk factor A)
– the ease by which the contamination is dispersed or transferred (risk factor B)
– the proximity of the source to the critical point where the product is exposed (risk factor C)
– how easily the contamination can pass through the control method (risk factor D)

Each of these factors (A to D) should be assessed and given a score of 0 to 2.

A risk rating can therefore be obtained for each contamination source and this rating can be used to determine the
importance of each source and whether it is a hazard to the product.

The risk rating can be assigned a ‘low’, ‘medium’ or ‘high’ category.

– A risk rating of less than 4 can be considered ‘low’

– A risk rating between 4 and 12 can be considered ‘medium’
– A risk rating higher than 12 can be considered ‘high’.

Significance?
Problems?
What is the difference between a sterile product and an aseptic product?

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Explain Table 3.4

Critical area and controlled area?

Typical Materials used in cleanroom?

Reference:
Cleanroom Technology: Fundamentals of Design, Testing and Operation by W. Whyte

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