Clean Room Classification

Clean Room Classification aspects of cleanroom requirements in aseptic Process in

manufacturing of sterile dosage form.

Classification of clean room:

Clean rooms in pharma industry are arias which are specially designed to reduce and eliminate,
particulate and microbial contamination in environment in aria used for manufacturing of pharmaceutical
dosage forms, especially parenteral dosage forms which are required to be injected directly into body
and blood stream are required to be free from viable microbial contamination as well as comply with the
requirement of particulate matter present in addition to requirement of absence of pyrogen. Controlling
particle count in air supplied to manufacturing aria is achieved by filtering air entering inside through high
efficiency particulate filters which are capable of removing 99.997 % of particles from air in rage of 0.3
micron and more. There are many techniques combined together to achieve required level of cleanness,
like more is the volume of air filtered per unit time (per hour) more is the cleanness of clean room. ULFA
filters are used to remove lower size particles.

Particulate matter are carriers of microorganisms hence reducing particle count directly reduce microbial
contamination.

Requirements of or classifications of clean rooms in pharma manufacturing unit where sterile dosage
forms are being manufactured are laid down by various agencies worldwide of which following are
important one.

Guidelines used worldwide to classify clean rooms in pharma industry.

1) US Federal standards in good manufacturing practices guidelines in 21 CFR Part 210 211.
2) British Standards for clean room classification. BS 5295
3) European Union standards as described in Guide for good manufacturing practices for medicinal
products manufactured for sale in Europe.
4) ISO 14644-1 Standards.

We have given below in details major requirements of each guidelines, and their advantages.

1. United states Federal Standards 209E.

Clean room are classified as class 10 to 10000000, read below in detail.
These guidelines also classify clean room aria as 1) Controlled aria clean room 2) Critical aria.
Controlled area is used for compounding, and manufacturing process where in final product is made
sterile in later stage, by terminal or filtration sterilisation( Class 1000, 10000). While critical aria is an aria
where in products is exposed to external environment hence this environment must prevent
contamination of product, (Class 100), Microbial count requirements as per US Federal regulation.
Critical area: Not more that 0.1 colony forming units per cubic foot.
Controlled area: Not more that 2.5 colony forming units per cubic foot.

2. European Community Guidelines.

Clean rooms are classified as
1) Class A --- At rest Particle count NMT 3500 particles of 0.5micron and above in cubic meter, at
Operation 3500 particles of 0.5micron and above in cubic meter ( Equivalent to Class 100 , Laminar
airflow bench)
2) Class B equivalent to Class 1000
3) Class C equivalent to Class 10000
4) Class D. equivalent to Class 100000

EU guidelines require that clean room meets standards in operation and as well as at rest.
It also require that particle count measurements should be done with in one foot distance of working
point.

Regulatory requirements of a clean room in pharmaceutical

High Efficiency Particulate Air filter, A velocity of 0.45 meters/second (90 feet per minute) has generally
been established, with a range of plus or minus 20 percent around the set point. Higher velocities may
be appropriate in operations generating high levels of particulates.

Two clean room or clean areas are of particular importance to sterile drug product quality: the critical
area and the supporting clean areas associated with it.

Clean Room Classification :-

Critical Area – Class 100 (ISO 5) and minimum Class 10,000 (ISO 7) , can be Class 1,000 (ISO 6)

1.All clean room classifications are based on data measured in the vicinity of exposed materials/articles
during periods of activity.

2.ISO 14644-1 designations provide uniform particle concentration values for clean rooms in multiple
industries. An ISO 5 particle concentration is equal to Class 100 and approximately equals EU Grade A.

3.Values represent recommended levels of environmental quality. You may find it appropriate to
establish alternate microbiological action levels due to the nature of the operation or method of analysis.

4. The additional use of settling plates is optional.

5. Samples from Class 100 (ISO 5) clean room environments should normally yield no microbiological
contaminants.

A. Critical Area – Class 100 (ISO 5) clean aria classification

Environmental conditions that must be designed to maintain pharmaceutical product sterility (CFR
211.42(c)(10)). Activities conducted in such areas include manipulations (e.g., aseptic connections,
sterile ingredient additions) of sterile materials prior to and during filling and closing operations. This
area is critical because an exposed product is vulnerable to contamination and will not be subsequently
sterilized in its immediate container. To maintain product sterility, it is essential that the environment in
which aseptic operations (e.g., equipment setup, filling) are conducted be controlled and maintained at
an appropriate quality. One aspect of environmental quality is the particle content of the air. Particles are
significant because they can enter a product as an extraneous contaminant, and can also contaminate it
biologically by acting as a vehicle for microorganisms Appropriately designed air handling systems
minimize particle content of a critical area. Air in the immediate proximity of exposed sterilized
containers/closures and filling/closing operations would be of appropriate particle quality when it has a
per-cubic-meter particle count of no more than 3520 in a size range of 0.5 µm and larger when counted
at representative locations normally not more than 1 foot away from the work site, within the airflow, and
during filling/closing operations. This level of air cleanliness is also known as Class 100 (ISO 5) clean
room.

It is recommended that measurements to confirm air cleanliness in critical areas be taken at sites where
there is most potential risk to the exposed sterilized product, containers, and closures. The particle
counting probe should be placed in an orientation demonstrated to obtain a meaningful sample. Regular
monitoring should be performed during each production shift. It is recommended conducting nonviable
particle monitoring with a remote counting system. These systems are capable of collecting more
comprehensive data and are generally less invasive than portable particle counters.

As provided for in the regulations, separate or defined clean areas of operation in an aseptic processing
facility should be appropriately controlled to attain different degrees of air quality depending on the
nature of the operation. Design of a given area involves satisfying microbiological and particle criteria as
defined by the equipment, components, and products exposed, as well as the operational activities
conducted in the area.

Clean area control parameters should be supported by microbiological and particle data obtained during
qualification studies. Initial clean room qualification includes, in part, an assessment of air quality under
as-built, static conditions. It is important for area qualification and classification to place most emphasis
on data generated under dynamic conditions (i.e., with personnel present, equipment in place, and
operations ongoing). An adequate aseptic processing facility monitoring program also will assess
conformance with specified clean area classifications under dynamic conditions on a routine basis. The
following table summarizes clean area air classifications and recommended action levels of
microbiological quality

B. Supporting Clean Areas

Supporting clean areas can have various classifications and functions. Many support areas function as
zones in which nonsterile components, formulated products, in-process materials, equipment, and
container/closures are prepared, held, or transferred. These environments are soundly designed when
they minimize the level of particle contaminants in the final product and control the microbiological
content (bioburden) of articles and components that are subsequently sterilized.

The nature of the activities conducted in a supporting clean area determines its classification, clean
room classification. FDA recommends that the area immediately adjacent to the aseptic processing line
meet, at a minimum, Class 10,000 (ISO 7) standards (see Table 1) under dynamic conditions.
Pharmaceutical Manufacturers can also classify this area as Class 1,000 (ISO 6) or maintain the entire
aseptic filling room at Class 100 (ISO 5). An area classified at a Class 100,000 (ISO 8) air cleanliness
level is appropriate for less critical activities (e.g., equipment cleaning).

C. Clean Area Separation

An essential part of contamination prevention is the adequate separation of areas of operation. To

maintain air quality, it is important to achieve a proper airflow from areas of higher cleanliness to
adjacent less clean areas. It is vital for rooms of higher air cleanliness to have a substantial positive
pressure differential relative to adjacent clean rooms of lower air cleanliness. For example, a positive
pressure differential of at least 10-15 Pascals (Pa)(Equal to 0.04-0.06 inches of water gauge.) should be
maintained between adjacent rooms of differing classification (with doors closed). When doors are open,
outward airflow should be sufficient to minimize ingress of contamination, and it is critical that the time a
door can remain open to be strictly controlled . In some cases, the aseptic processing room and
adjacent clean rooms have the same classification. Maintaining a pressure differential (with doors
closed) between the aseptic processing room and these adjacent rooms can provide beneficial
separation. for example a clean room of class 100 which is a aseptic filling aria, laminar air flow bench
have higher pressure than its adjustment clean room of class 1000, so that contaminated air do not get
in to critical area, In any facility designed with an unclassified room adjacent to the aseptic processing
room, a substantial over pressure (e.g., at least 12.5 Pa) from the aseptic processing room should be
maintained at all times to prevent contamination. If this pressure differential drops below the minimum
limit, it is important that the environmental quality of the aseptic processing room be restored and
confirmed. The Agency recommends that pressure differentials between clean rooms be monitored
continuously throughout each shift and frequently recorded. All alarms should be documented and
deviations from established limits should be investigated. Air change rate is another important clean
room design parameter. For Class 100,000 (ISO 8) supporting rooms, airflow sufficient to achieve at
least 20 air changes per hour is typically acceptable. Significantly higher air change rates are normally
needed for Clean room class of a Class 10,000 and Class 100 areas. Higher the air changes filtration of
room is more and eventually the final set operational stage quality of air after required air changes is of
good quality. Clean room of classification of class 100 needs about 450 to 500 air changes.

A suitable facility monitoring system will rapidly detect atypical changes that can compromise the
facility’s environment. An effective system facilitates restoration of operating conditions to established,
qualified levels before reaching action levels. For example, pressure differential specifications should
enable prompt detection (i.e., alarms) of an emerging low pressure problem to preclude ingress of
unclassified air into a classified room.

What is a Clean room , its classification of clean room.

Clean Area- It is an area in a pharmaceutical manufacturing company which is supposed to meet a

cleanness standard of a defined particle and microbiological count and intern the cleanliness and clean
room in pharma are classified based on particle count, the cleanness is achieved by circulation of filtered
air through HEPA filter of efficiency 99.99%

Clean room- In pharmaceutical manufacturing room is designed and controlled and maintained to attain
a highest level of clean environment so as to prevent microbial (bacterial and viral ) and particulate
matter contamination of drug products being handled in the clean room. Typically clean rooms are
assigned a class depending up on its degree cleanness or its of capability to provide cleanness
reproducible and are required to meet an appropriate standard of air cleanliness classification. Clean
rooms have great importance in sterile dosage form or injectable dosage form manufacturing.

Standards used in classification of Clean rooms are as follows

1.US FED STD 209E Clean room Standards. 2.Clean room standards ISO 14644-1
 Here is the comparison chart for above two standards.

3.Clean room standards.BS 5295

Requirements for designing clean room .

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FUMIGATION OF ASEPTIC AREA STERILITY TESTING ROOM
SOP for fumigation of Aseptic area sterlity testing room.
OBJECTIVE : To maintain the microbial load to a minimum of sterility testing room by fumigation by use
of formalin and potassium permagnate powder

UNIT OF RESPONSIBILITY :- QA Executive and trained QA Assistants

IMPORTANT POINTS /NOTES /PRECAUTIONS :-

1 Handle the formalin solution carefully as it is corrosive in nature

2 Use hand gloves while handling the formalin

MATERIAL :-

1 Formalin solution 37 % ( 50 to 75 ml )'

2 Potassium paermagnate powder

3 Seprtating flask 125 ml capacity

4 Petridishes and Tripod stand

PROCEDURES :

1 Before entering into the airlock remove the footwear outside the room

2 Ensure that the door closes completely on releasing when you enter the airlock

3 Remove the normal clothing and put it on the hanger

4 Take one set of sterile uniform Wear sterile uniform as per the instructions given below

a) Put on the headgear

b) Put on the bushshirt and then pyjama

c) Headgear to be tucked into the bushshirt and the in turn to be tucked in the pyjama

d) Put on the footwear and tie it properly

e) Take the pair of sterile gloves of proper size and wear them properly

5 Rinse the gloved hands with 70 % Isopropyl alcohol

6 Open the door with care while entering the sterility testing room

7 Before the start of fumigation ensure that the Air handling unit is shut off to prevent

the fumes of formalin to enter the other areas through Air handling unit
8 Fill the seperator with suffficient quantity of 50 ml of formalin ( 1 : 1 )

9 Take around 25 gm of potassium permagnate powder in dry pertridish without lid

Keep the seperator filled with formalin on a tripod stand and the pertridish with potassium

below it at the centre of the area which is to be fumigated

10 Loosened the stopcock so that formalin will fall drop by drop on the potassium permagnate

powder

11 Immediately come out of the room, lock the door

12 While coming out remove the sterile uniform in the reverse order, that is fisrt gloves,

second footwear, third pyjama, and forth bushshirt and lastly the headmask

13 Put your normal clothing kept on the hanger Before opening the door of the airlock corfirm

that door of sterility room is closed

14 These uniform gloves are to be washed and resterilised before using for next operation

15 The fumigation record is recorded in the ataachment 1

PERSONNEL TO BE REPORTED IN CASE OF NON COMPLIANCE :- Department head

FREQUENCY OF OPERATION :- Weekly

NO OF ATTACHMENT :-

RETENTION OF RECORDS :- 5 Years

SOP CLEANING ASEPTIC AREA STERILITY TESTING ROOM

SOP for cleaning sterility testing room
OBJECTIVE :- To clean the sterility testing room by removing the extraneous material and to disinfect
the area making it condusive to carry out the operation

UNIT OF RESPONSIBILITY :- Trained QA personal

IMPORTANT POINT /NOTE /PRECAUTIONS :-

1 Handling of disinfectant should be done carefully

REQUIREMENT :-
1 25 % v/v dettol solution
2 25 % v/v lysol solution

3 01 % v/v benzalkonium solution ( BKC)

4 70 % v/v Isopropyl alcohol (IPA )

5 Lint free duster

6 Sterile gown and gloves

PROCEDURE :-

1 Prepare the solution of disinfectant of different concentration

2 Wear the sterile gown as mentioned in SOP for technique to follow in sterility testing room and
enter the room

3 Take about 25 litres of 25 % dettol solution and wipe then walls , the ceiling and then the floor
of the room with sterilised lint free duster, then wipe the walls ceiling and the floor with either
of the solutions mentioned :-

a) 25 % v/v lysol ( 25 ml of the lysol in 100 ml of DM water )

b) 01 % v/v bezalkonium chloride solution ( 1 m of BKC IN 1 L OF DM water)

4 Clean the table surface and the glasspanes and the laminar air flow bench with 70 % IPA
solution

5 After cleaning is over, spray the entire area with 70 % IPA solution

6 Come out of the sterility testing room and gown as per SOP for technique to follow in sterility
testing room

7 Record the cleaning details in the attachment 1

OTHER RELEVANT DETAILS :-

At the weekends fumigation is done with formalin and potassium permagnate powder
PERSONNEL TO BE REPORTED IN CASE OF NON COMPLIANCE :- Department head
FREQUENCY OF OPERATION :- The cleaning is done 3 times a week with alternate disinfactant

NO OF ATTACHMENT :-

RETENTION PERIOD OF RECORDS :- 5 years

Following are some of articles which will be useful for you in further understanding of aspects of sterile
dosage form manufacturing