Aseptic Room Validation

BY

Dr. K. Prabhakar Reddy, Associate Professor,

St Peter’s Institute of Pharmaceutical Sciences, Vidyanagar, Hanamkonda.

1Pharmaceutical Equipment and Validation

CONTENTS

1. INTRODUCTION

2. THE DESIGN OF ASEPTIC AREA

3. FACILTY DESIGN

4. SOURCES OF CONTAMINATION

5. MAINTENANCE OF ASEPTIC ROOM

6. STANDARDIZATION OF ASEPTIC AREA

7. ASEPTIC AREA VALIDATION

8. CONCLUSION

9. REFERENCES.


Asepsis is the practice to reduce or eliminate contaminants (such as bacteria, viruses, fungi, and parasites) from entering the operative field in surgery or medicine to prevent infection.

Ideally, a field is "sterile" — free of contaminants — a situation that is difficult to attain. However, the goal is elimination of infection, not sterility.

Aseptic Processing is the processing of drug components ( drug product, containers, excipients, etc.) in a manner that precludes microbiological contamination of the final sealed product.

INTRODUCTION


Contd…..

The parenteral preparations differ from all other drug dosage forms because of the unique requirements imposed.

They must be exceptionally pure and free from physical chemical and biological contaminations.

These products are require to be sterile. These requirements place a heavy

responsibility on pharmaceutical industry and pharmacist .


Ideal methods of manufacturing of these sterile products fall into two categories

1. Terminal Sterilization Process

2. Aseptic Processing Operation


Comparison of terminal sterilization and aseptic processing

Terminal sterilization Product containers are filled

and sealed under high-quality environmental conditions designed to minimize contamination, but not to guarantee sterility.

Product in its final container is subject to a sterilization process such as heat or irradiation.

Aseptic processing Drug product, container, and

closure are subject to sterilization separately, and then brought together.

Because there is no process to sterilize the product in its final container, it is critical that containers be filled and sealed in an extremely high –quality environment.


TERMINAL STERILIZATION

Sterile Drug

Product !

Sterilization Process must be compatible with all components !7Pharmaceutical Equipment and Validation

Aseptic Processing

Drug Product

Sterilization Process

Container

Closure

Excipient




Sterile Closure

Sterile Excipient

Aseptic Processi

ng

Sterile Drug

Product

SterileContainer

Sterile Final

Product

Can use multiple sterilization processes each optimized for the individual component


THE DESIGN OF ASEPTIC AREA


THE DESIGN OF ASEPTIC AREA FOR THE MANUFACTURE OF PARENTERALS

The design should possess following characteristics:

The aseptic room must be designed to prevent contamination of parenteral during processing.

It must be cleanable and sanitizable, with a minimum of particles shedding and crevices or other sites where dirt can accumulate.


The structural surfaces are of ceiling, walls, and floors, and works surfaces and storage surfaces should be smooth and resistant to cleaning and sanitizing.

Work surfaces should be stainless-steel and epoxy coated structural surfaces are preferred.

Equipments and instruments also should meet these general requirements. Equipments with non-cleanable, particle-shedding mortars, gears, preferably in stainless steel enclosures.


The design of aseptic room includes Site, size, windows, door, surfacing material ( floors, walls & ceiling and bench tops), services, furniture

SITE As far as possible from rooms to

which non pharmaceutical staff have access.

It must be adjacent to support areas so that an efficient flow of components may be achieved.

Be away from stairs, lift shafts and corridors.

Access to the room should be through one or more rooms with washing and changing facilities.

THE DESIGN OF ASEPTIC ROOM


SIZE

Size of the room is the maximum number of people using at any one time.

Provide adequate space for work load anticipated.

Generally large size is preferred but the capital and maintenance cost of the equipment for controlling the microbial content, temperature, humidity of atmosphere are reduced by keeping the room small.


WINDOWS Should be made of non-shedding

materials preferably with double panel and should flush with walls.

Large windows of clear glass are most preferable but they must not open. DOORS

Should be air lock with double doors about one meter apart. This prevent a sudden inrush of air when the door is opened.

They are two types of doors 1. Sliding Doors 2. Swinging

Doors


Surface Materials:

The floors, walls, and bench tops of an asepsis room must be easily cleaned, smooth, impervious, resistant to chemicals, non shedding, non flaking, non cracking, and unbroken

1. FLOORS Flooring should be unbroken and provided with a cove both at the junction between the wall and floors as well as the wall and ceiling.The most suitable materials are terrazzo, linoleum and plastics.

Terrazzo (Cement + Crushed Marble): Most suitable materials for flooring.It is spread in plastic form on the site or is available as tiles In both cases expansion joints of ebonite (black) or PVC (various colors) are essential to prevent cracking.


Linoleum Available as a sheet and tile

forms in many colors. It can be surface sealed to

reduce treading in of dust and marking rubber heels

Plastics The non- slip or matt-finish

grades of PVC are suitable for asepsis rooms.

Obtained as sheets or tiles& the joints can be welded.


2. walls and ceilings: The possible surfaces are Tiles,

hard gloss paint on smooth plaster, plastic laminated board.

Walls shall be suitable for flushing some special requirements like electric sockets, gas points, sterilizers

Ceilings shall be suitable for light fittings, air-grills flushing; not hanging from the ceiling.

3. Bench tops: The most popular surfaces for

asepsis work are stainless steel, plastic laminates.


SERVICES: The location of these services and their

servicing or repairing shall not pose any threat to the integrity of the facility.

An asepsis laboratory will require many, if not all, of the following

Ventilation, Electricity, Gas, Compressed Air or Vacuum, Nitrogen, Water, A Method of Dealing with Waste.

ELECTRICITY: It is used for lighting and sometimes for

a hot plate, UV lamp, aerosol producer or vacuum pump.

Switches and sockets should be flush fitting and have finger plates of plastic because metals are less resistant to the fluids used for cleaning and disinfection.

Most of the controls can be outside the room.

Light Fixtures:


GAS: Used for Bunsen burner For easy operating / usage wall fitted service outlets and

separate benches are arranged, so that the operator need not to getup to use them.

Gas cock should not place under the bench top because the tubing comes up from the front and there is a risk that if the burner is near to edge, therefore it will be pulled off by the elasticity of the tubing or by accidentally catching the latter with arm.

COMPRESSED AIR AND VACUUM: Used for clarification and bacterial filtration. For these services rotary pumps can provide separately. They are best housed outside the lab because pumps

are noisy.


NITROGEN: Used for replacement of air by an inert gas. A cylinder can be kept near to the vacuum pump and

connected via a reducing valve.

WATER: It is difficult to justify a water supply in the laboratory

itself. Hand washing facilities are undesirable, because of the

risk of splashing organisms from the hands into the air and onto working surfaces or equipment.

If water system is necessary, elbow operated taps, foot operated taps, knee operated taps are arranged along tiled walls.

In all above types there is no need to use the cleaned hands for turning of the water.


Waste disposal: Foot operated waste bin is popular (but it need partly

sliding from the chair and also the mouth is very small i.e., can’t receive large pieces of paper).

The better method is to fit the bin at the side of knee space having a metal ring that can be swiveled under or out from the bench.

FURNITURE: Shall be smooth, washable and made of stainless steel

and any other appropriate materials other than wood.


BENCHES: The conventional benches may be replaced by tables, wall

mounted work shelves are preferred to remove the dust. These tables and work shelves are easily over looked the

dust under surface of top.

SEATS:

Must be adjustable and comfortable Adjustable chairs ensures that the users face is well above

the front opening of the screen, therefore the breath is kept away from the materials underneath.

The fabric must be washable. Comfort is best assured by chairs rather than stools and by

upholstered rather than wood or metal, seats and backs.


TROLLEYS: Trolleys with removable trays (preferably of

stainless steel) are better than tables benches or cupboards for holding spare equipment.

DOOR MAT: It is necessary, because the outdoor shoes are

heavily contaminated with dust born organisms. It is better useful if the mats are in the air lock and

immersed in a detergent disinfectant solution. These mats are available in two types 1) Honeycomb structure like mats and 2) Disposable adhesive faced pad.


Screens: Usually aseptic technique is carried out

under screen 1) Shake type and 2) fume cupboard type The shake type consisted of wooden case

with sloping front of glass but the most common fabrication material is Perspex.

The fume cupboard type used to fit the tall operators like burettes.


Facility Design


Facilities

Establishing and Maintaining an aseptic environment

Use clean-rooms of various classes to establish an aseptic area

Clean rooms use combinations of filtration, air exchange, and positive pressure to maintain “clean” environment

Lower quality clean areas should not be placed next to high quality areas


Facilities: General Clean room Design

HEPA / ULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors, walls, and ceilings constructed of smooth hard

surfaces that can be easily cleaned Limited equipment, fixtures and personnel Layout of equipment to optimize comfort and movement of

operators


Facilities: HEPA Filters

High Efficiency Particulate Air

Minimum particle collection efficiency: 99.97% for 0.3µm diameter particles.

Disposable

Filter made of pleated borosilicate glass


Facility Design: Clean Area Classification

FS209 Cleanroom classification

ISO 14644-1 Cleanroom classification ≥0.5um particles/m3

Microbiological Active Air Action Levels

(cfu/m3)

Microbiological Settling Plates Action Levels (diam.

90mm; cfu/4hours)100,000 8 3,520,000 100 5010,000 7 352,000 10 51000 6 35,200 7 3100 5 3,520 1 1


Class 10,000 clean room

http://www.americancleanrooms.com/am/photogallery_08.html

Class 100 clean room

Facilities: Clean room Classification


Facilities: Clean room Classification

FS209 Cleanroom

classification

ISO 14644-1 Cleanroom

classification≥0.5um

particles/m3

Viable Microbes(cfu/m3)

Ave Airflow Velocity

(fpm)Air

changes/hr

100,000 8 3,520,000 100 5-10 5-48

10,000 7 352,000 10 10-15 60-90

1000 6 35,200 7 25-40 150-240

100 5 3,520 1 40-80 240-480


Facilities: Pressure Differentials

Used to maintain airflow in the direction of higher cleanliness to adjacent less clean areas

A minimum of 10-15 Pascals should be maintained between the aseptic area and an adjacent rooms with differing clean room classifications (doors open)


http://news.thomasnet.com/images/large/451/451402.jpg

Facilities: Air Lock

Permits the passage of objects and people into a clean room.

Consists of two airtight doors in series which do not open simultaneously.

Spray down materials with 70% IPA before placing in the airlock


Fiber-shedding materials such as cardboard and paper Cardboard packaging must be removed and items placed into

non-cardboard containers.

Wood (i.e. wooden pallets) Undesignated charts

Facilities: Material NOT permitted in a Clean room


Facilities: Cleaning

Water should be changed FREQUENTLY

1. Vacuum all accessible surfaces

3. Mop floors using a lint free polyester mops attached to stainless steel handles

2. Wipe surfaces with a cleaning solution


Isolators

The use of isolators prevents direct contact with product

However, the use of isolators can lead to relaxation of aseptic practices!


SOURCES OF CONTAMINATION

1. The Atmosphere

2. The Breath

3. The Hands

4. Clothing

5. The Hair

6. The Working Surface

7. Equipment


MAINTENANCE OF ASEPTIC ROOM

To maintaining the clean and sanitized conditions of aseptic areas requires diligence (care) and dedication of expertly trained custodians.

The design of the facilities to be cleanable and sanitizable, a carefully planned schedule should be developed depending on location and its relation to the most critical class 100 areas.


Contd…

Tools used should be non- lining designed for clean room use, held captive to the area and preferably sterilized.

The maintenance of aseptic room depends on cleaning methods, personnel, garments, and environmental control.


CLEANINIG METHODS:

They are necessary to remove the dirt.

The cleaning methods includes; dry- cleaning, wet

cleaning, and sanitization / disinfection.

Dry cleaning:

Vacuum cleaners are more prefer, because they

prevent tiny particles from passing through the fabric.

Mops impregnated with dust retaining oils also used

but they need regular cleaning and reimpregnated.


Contd…. Brooms are unsuitable because they disperse

large amount of dust into the air.

If the floor is semi porous surfaces (such as

unpolished linoleum) occasionally treatment

with spindle oil.

Wet cleaning:

Floors must be mopping with hot water

containing detergent

Walls and furniture must be cleaned with clean

cloth with slight dampness.


Disinfection / sanitization:

This is necessary to possess good hygienic conditions in aseptic area

The employees must be trained for this purpose.

The sanitization is produced by using different concentrations of sanitizing agents like alcohols, aldehydes, cationic surfactants, chlorhexidine, dyes, phenols, oxidizing agents and halogens.

Mostly liquid disinfectants are preferable because their reliable activity against inherent environmental microorganisms.

They should be recognized as supplements to good house keeping.

Industrially “three bucket” system used to sanitized the aseptic room


The UV light rays of 237.5nm radiated by germicidal lamps are an effective surface disinfectant.

Different sanitizing shall be used in rotation with sufficient frequency.

Dilution of sanitizing agents with fresh distilled water (above 700C) used.

Eg; 70% alcohol or 60-70%of isopropyl alcohol used as hand sprays.

Formaldehyde or any other equally effective fumigant is recommended for the fumigation of aseptic areas.

All these diluted disinfectants shall bear the label clearly like “use before” etc.

Some germicides are recommended for floors, surfaces are synthetic phenols, quaternary ammonium compounds

and iodophors.


PERSONNEL

Must be neat, orderly and reliable.

They should be in good health and free from

dermatological conditions.

If they show symptoms of a head cold, allergies or

similar illness are not allowed into aseptic area until

recovery.

Aseptic area operators should be given thorough,

formal training in the principles of aseptic processing.

Personnel should have sufficient knowledge and

skills in aseptic techniques to be employed.


CLOTHING: The garments used in asepsis lab includes,

gowns and trousers, head dress, masks, gloves, goggles & foot wear.

The design of all these to facilitates the contaminants discharged from the body of the operator.


ENVIRONMENTAL CONTROL

The facility should be centered on the control of air borne and surface contamination both viable and nonviable.

Current aseptically processed parenteral drugs should provided a level of sterility assurance of 1in 3000units. This level is still potentially demanding standard.

Therefore to achieve this level of sterility assurance the adequate environmental control program to be designed.

Environmental control should be provided by, HVAC (Heating Ventilation and Air Conditioning system) Temperature and humidity control Air filtration (High Efficiency Particulate Air) Air handling system (Central-Air Conditioning) Air motion (laminar air flow) Pressure control system (pressure gradients)


The design of suitable installation of above systems provide bacteriologically clean air.

HVAC systems: It should be designed to provide control over the

air borne viable and non viable particles To prevent air borne contaminants from entering

the clean or aseptic environment ,all air supplied for the environment must be filtered.

The level and type if filtration needed depends on level of cleanliness required.


Temperature and humidity control: Temperature control is required to offer primarily a

comfortable working environment for operators. Generally 68-74 0F(19-23 0C) is acceptable range. Lower temperature are preferred in poorly ventilating

manufacturing environment. Certain areas of manufacturing facility (like those ware

autoclaves and dry heat sterilization tunnels and ovens) are located ,should provide higher heat loads to a system.

Temperature control in asepsis room by the ventilation system is important. This is advantage of heating because the elimination of radiator which are dust collectors and distributors.


Humidity control: This is very important in asepsis lab because the

asepsis room may become unpleasantly dry if several Bunsen's are burning or the heating unit is operating .

Generally 45-55%RH is preferred for asepsis. Normal humidity levels can be easily achieved

with air conditioning systems. Air dryers can be used to maintain lower than

normal humidity levels. Controlled spaces requiring humidity control

should be built with vapor proof barrier materials to ensure the minimum of water migrations.


Air filtration It is important for air borne contamination control with

using various types of air filters.

Bacteriologic ally clean air can be produced by removing the microorganisms by mechanical filtration or electrostatic precipitations or by destroying them with heat , U.V light or chemicals.

Filtration mechanisms:

Inertial impaction, Direct impaction, Diffusion ,Impingement, Electrostatic forces

Types of filters: Three types,

Coarse filters, High efficiency filters and Absolute filters.

High efficiency filters:

When a higher standard is desired ,these are used.

Eg: HEPA


HEPA filter design:

It is a disposal, extended media, dry type filter in a rigid frame .

Construction of HEPA Frame: with Board, Plastic, Metallic

material. Filter medium: glass micro fiber. Separators: Kraft paper, Al-alloy

and plastic. Adhesives: Poly urethane cones

and silicate adhesive Gasket: cell neoprene foam or

Teflon

Most of the micro glass filter media on the market have fire- retardant and water proof properties.


Principle: Brownian diffusion for small particles. Inertial effect for intermediate particles. Sieving effect for large particles.

Functioning : Recirculation of air or dilution of

contaminants. To provide enough capacity to ensure an

adequate level of cleanliness by recirculating the air contained with in the environment


Air Handling Systems (Central-Air-Condition)

It can be described as the core of the heating, ventilation, & central air condition system.

Central systems used for control areas normally employ an air-to-air or air-to-water cooling systems.

The filter configuration in the air handling system shall be suitable designed to achieve the following grades of air.


Grade At Rest In Operation

The maximum number of permitted particles per cubic meter equal to or above

0.5µm 5µm 0.5µm 5µm

A 3520 29 3500 29

B 35200 293 352000 2930

C 352000 2930 3520000 29300

D 3520000 29300Not

Defined Not Defined


The minimum air changes for grade B and grade C areas shall not be less than 20air changes for hour in a room with good air flow pattern and appropriate HEPA filters.

For grade A, LAF work stations the air flow rate shall be 0.3m/sec ± 20%(for vertical flows) and 0.45m/s±20%(for horizontal flows).


Air Motion (Laminar air Flow):The required environmental control of aseptic area has been made

possible by the use of LAF originally through a HEPA filter . It bathes the total space with very clean air , sweeping away

contaminants. Velocity: 90 fpm ± 20%. It produce clean air in a vertical, horizontal or curvilinear fashion. LAF clean benches should supply class 100 air. The velocity meter – to check the air flow rate. Plastic curtains are installed to maintain unidirectional air flow i.e.,

the area out side the curtains can be maintain at a slightly lower level of cleanliness than that inside.


Today it is accepted –to meet class 100 standards.

LAF will do an excellent job of maintaining the sterility of an article bathed in the air flow.

For most sterility testing operations- horizontal LAF are more preferred than vertical.

ENTRY PROCEDURES: Entry point is changing room. Changing room should not be used

for the entry of anything except personnel.

The change room is most conveniently arranged to provide separate three areas – black, grey, and white areas for successive stages in the entry process.

The black room used to remove and store outer clothing and Jewelers and to change the shoes.


The grey room is used for a preliminary hand wash using detergent based disinfectants such as Chlorhexidine solution.

White room: separated from grey room by a low threshold-bench.& Used for clean room clothing may be worn.

Change room doors shall not be opened simultaneously.

For Communication An appropriate inter locking system

and a visual and or audible warning system may be installed to prevent the opening of the door more than one time.

Intercom telephones or speak-phones shall be used.


By Simulation method, calculation method. Mostly used method is Simulation method.

Simulation Method/ Media fill method: It is desirable to be able to quantify the risk of

contamination in any aseptic fill or transfer procedure.

In this method to simulate the aseptic transfer by substituting a microbial growth medium for the product.

It involves preparation and sterilization of sterile triptycase soya broth. and filling this broth in to the sterile container under conditions similarly.

Incubate at 20 0C - 25 0C and 30-35 0C at least 14 days.

Standardization of Asepsis


If growth occurs contaminants has entered the container during process.

To pass the test at 95% confidence not more than 0.1%of the challenged units.

This test is a very stringent evaluation of efficiency of an aseptic filling process.

Advanced aseptic processing It is designed to isolate aseptic operations from personnel. Ex: Isolation (barrier) technology.

In this technology operations are performed within windowed, sealed walls with operators working through gloves ports.

The sealed enclosures are pre -sterilized, usually with para-acetic acid, hydrogen peroxide vapour or steam.


Sterile supplies are introduced from Sterilizable movable modules through uniquely engineered transfer ports or directly from attached sterilizer, including autoclaves and hot air sterilizing tunnels.

Results have been very promising ,giving expectation of significantly enhanced control of the aseptic processing environment.

This is used to increase frequency in the processing of sterile products.


Advantages

To maintain human intervention Increase dramatically the assurance of

sterility Inexpensive Allowed for aseptic processing with out the

construction of large processing areas, sterile suites or gowning areas.


ASEPTIC AREA VALIDATION


PURPOSE To provide the guideline for classification of

areas to prevent mix–ups or cross contamination

RESPONSIBILITY It is the responsibility of all departmental

managers to follow the procedure. The quality assurance manager is responsible

for SOP compliance.


PROCEDURE TestingThe controlled areas will be subjected to

the following set of performance tests: Air flow velocity HEPA filter/leak (DOP) Airborne particle count Air flow patterns Recovery Airborne microbial Surface bio burden


The instruments used for these tests should

be calibrated and included with the report.

The tests will be performed at rest, dynamic, and stress conditions.


Air Flow Velocity Test

Equipment Air flow velocity: electronic micro

anemometer or vane-type anemometer.

Procedure Divide the work zone entrance plane

into a grid of equal areas. Individual areas should not exceed 0.37 m2 (4 ft2).

Support the anemometer sensor probe with a suitable stand.


Air Flow Velocity Test (Contd…)

Measure the air flow velocity at each test position.

Allow at least five seconds for each measurement and record the average reading during that period.

Acceptance criteria: The average air flow velocity, should be within ±5% of the value specified for the clean room


HEPA Filter/Leak Test (DOP)

Equipment

DOP aerosol. DOP aerosol generator.

Aerosol photometer.


Method Introduce DOP aerosol upstream of the filter

through a test port and search for leaks downstream with an aerosol photometer.

Acceptance criteria HEPA filters 99.99% : challenge aerosol

penetration is lower or equal to 0.01% of the upstream concentration

HEPA filters 99.97% : challenge aerosol penetration is lower or equal to 0.03% of the upstream concentration

HEPA filters 95%: challenge aerosol penetration is lower or equal to 5% of the upstream concentration.


Repairs No repair is authorized without the

acceptance of the contractor.

If the contractor agrees to repair the

filter, the medium to repair the filter should be agreed upon.

Each repair must be properly documented on the worksheet.


In any case, the maximum surface to be repaired is less than 5% of the visible surface of the filter and any dimension of any repair is maximum 4 cm.

Other criteria for repair can be agreed upon with the contractor.

FREQUENCY

Initial validation: once Revalidation: every six months for class 100

areas; every year for class10,000 and 100,000 areas respectively.


Airborne Particle Count Test

Equipment CI-500 laser particulate counter with printer

Method

1. Using the particle analyzer, count particles greater than or equal to 0.5 µm in diameter

2. If the particle count in the 0.5 µm range is less than 50 per cubic foot of air, four additional counts at this location are taken to place these particle counts within a 50% confidence interval.

Next repeat steps 1 and 2 with operational personnel present The minimum volume of air to be sampled can be read from Table A

The minimum number of sampling points can be read from the Table B


Table ATesting for particles Air volume required≥ 0.1 µm 0.1 cft or 2.8 l≥ 0.3 µm 0.1 cft or 2.8 l≥ 0.5 µm 0.2 cft or 5.6 l≥ 5 µm 0.3 cft or 8.4 lTable BSquare feet Square meter Sampling points100 9.2 4200 18.4 8400 36.8 161,000 92 402,000 184 804,000 368 16010,000 920 400


Federal Standard 209 E class limits in particles per cubic foot of size equal to or greater than particle size shown

Measured Particles size (µm)Class 0.1 0.2 0.3 0.5 5.0 1 35 7.5 3 1 NA 10 350 75 30 10 NA 100 NA 750 300 100 NA 1,000 NA NA NA 1,000 7 10,000 NA NA NA 10,000 70 100,000 NA NA NA 100,000 700


Acceptance criteria At any of the designated critical locations the

particulate count shall not exceed 100 particles 0.5 µm in diameter and larger per cubic foot of air.

The same test should be repeated at ancillary environments.

Ancillary environments shall not exceed a particle count of 100,000 particles 0.5 µm in diameter and larger per cubic foot of air .

Frequency Initial validation: once Revalidation: annually


Air Cleanliness Level: Definition of ClassesMaximum number of particles0.5 µm 5 µmClass Per cf Per cm Per cf Per cm .

100 100 3.5 — —10,000 10,000 350 65 2.3100,000 100,000 3500 700 25

Cf-cubic footCm- cubic meter


Guidelines for Cleanliness Levels Required during Manufacturing of a Parenteral Drug

Operation Class Cleanliness level

(particles 0.5 µm and larger)

Warehousing — —

Preparation 100,000 `Not more than 100,000

Filtration 100,000 Not more than 100,000

Filling area 100,000 or better Not more than 100,000

Filling line (point of use) 100 Not more than 100


Air Flow Patterns Test Equipment White visible or yellow smoke generator,

anemometer, 35-mm camera or videotape recorder

Method If the system operates according to

the specified operating parameter, begin to generate white visible smoke at the critical locations..

Generate white smoke inside and over each component.


Smoke should flow through these critical areas. If the air returns (back-flows) due to turbulence, the system cannot be accepted and must be rebalanced or adjusted.

Frequency Initial validation: once Revalidation: annually


RECOVERY TESTEquipment Visual smoke generator, particles counter, and hot

wire anemometerMethod With smoke generation output tube located at a pre

designated location, generate smoke for 1 to 2 min and shut off.

Record the particle count. If it is not 100 per cubic foot or less, repeat the test with the wait interval increased in increments of 0.5 min until counts are less than 100 per cubic foot.

Acceptance criteria The recovery time should be not more than 2 min.Frequency Initial validation: once Revalidation: annually


Airborne Microbial TestEquipment Solid surface impactor with a rotating

collection surface or staged plates (Anderson-Slate)

Method Aseptically prepared collection plates are

placed in the sampling apparatus. Sampling time should be 20 min at every

location. After the sampling is complete, remove the

collection plates, cover, and identify them After incubation, the number of colonies on

each plate is counted.


Acceptance criteria The following table provides the Federal Standard

209 E for cleanliness level:

Air Cleanliness Guidelines in Colony-Forming Units (cfu) in Controlled Environments (Using a Slit-to-Agar Sampler or Equivalent)

class cfu per cm of air cfu per cf of air

100 Less than 3 Less than 0.1 10,000 Less than 20 Less than 0.5 100,000 Less than 100 Less than 2.5.


Suggested Frequency of Sampling on the Basis of Criticality of Controlled Environment

Sampling Area Frequency of Sampling

Class 100 or better room designations Each operating shift

Supporting areas immediately adjacent To Class 100 (e.g., class 10,000) Each operating shift Other support areas (class 100,000) Twice/week

Potential product/container contact areas Twice/week

Other support areas to aseptic processing areas but non product contact (class 100,000 or lower) Once/week


Surface Bioburden Test

Equipment Cotton swabs or RODAC plate (nutrient

agar culture medium)

Method Take the swab stick from the tube and

gently swab 25 cm2 of area (walls, floor, equipment, etc.) and place back in tube containing 5 mL sterile buffer and test or incubate per official procedure.


Acceptance criteria The maximum number of colonies per square

foot should not exceed the limits in following Tables

Surface Cleanliness Guidelines of Equipment and Facilities in cfu Controlled Environment

Class cfu per Plate 100 3 10,000 5

Contact plate areas vary from 24 to 30 cm2. When swabbing is used in sampling, the area covered should be greater than or equal to 24 cm2 but no larger than 30 cm2.


Surface Cleanliness Guidelines in Controlled Environments of Operating Personnel Gear in cfu

class cfu per plate*

U.S. Customary Gloves Clothing

100 3 5

10,000 10 20

Frequency Initial validation: once Revalidate: annually


CONCLUSION

Sterile products ,being very critical and sensitive

in nature, a very high degree of precautions,

preventions and preparations are needed.

Dampness, dirt and darkness are to be avoided

to ensure aseptic conditions in all areas.

There shall be strict compliance in the prescribed

standards especially in the matter of supply of

water ,air, active material and in the maintenance

of hygienic environment.


The aim of aseptic technique is to prevent the

access of microorganisms during the

preparation and testing of pharmaceutical

products.

Therefore, the aseptic area requires

construction features designed for maximum

microbial and particulate control.

Finally conclusion should be drawn based on the results of above tests and documentation.


REFERENCES

1 Leon Lachman, Herbert A. Lieberman, J0seph L.Kanig.

The theory and Practice of Industrial Pharmacy, pp. 618-

638.

2. Remington: the science and Practice of Pharmacy Vol.II,

20th . Edition, pp. 770-820, 2020-2030.

3. Leon Lachman and Lieberman: Pharmaceutical Dosage

Forms- Parenteral Medications Vol. I, 2nd Edition, Revised

and Expanded; pp. 325-342.

4. James Swarbrick and James C. Boylan, Marcel Decker

Inc., Encyclopedia of Pharmaceutical Technology, Vol. I,

pp. 351-368.

5. Michael E Alton: 623-631. Pharmaceutics: the science of

Dosage Form Design, pp. 638-641.


6. Cooper and Gunn’s, Dispensing for Pharmacy Students,

Edited by S.J Cater, 12th Edition, pp. 623-631.

7. United States Pharmacopoeia / National Formulary,

2002 Edition, pp. 2206-2211.

8. CGMPs for pharmaceutical Preparations.

9. Validation Standard Operating Procedures-Second Edn-Syed Imtiaz Haider-pp -151, 357

10. US FDA, General Principles of Validation, Center for Drug Evaluation and Research (CDER), Rockville, MD (1987).

11. www.germfree.com

12. www.gonka.com.

13. www.pharmatech.com


Thank you


Aseptic Room Validation

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