A short presentation on containments which is used for potent drug manufacturing facilities and labs also these are used in biotechnology industries. Very useful for BSL facility and Oncology/Hormones/Esteroids manufacturing and research.

2. OUTLINE OF CONTAINMENTS (RABS & ISOLATORS
An Isolator is a fully sealed unit, The handling of drug
substances and the filling of vials take place within a closed
system that completely isolates the operator and the
surrounding environment from the drug product.
RABS is a type of barrier system for aseptic processing of
pharmaceutical products that reduces or eliminates
interventions into the critical zone providing:
Rigid wall enclosure (for physical separation of aseptic
processing operations from operators) with interlocked doors;
Unidirectional air flow systems (to reach a class A environment
to the critical area);
use of glove ports to access all areas of the enclosure during
operations;
High level of asepsis (through unidirectional air flow with High
Efficiency Filters) and / or monitoring of the internal particle
contamination (through particles counting equipment).

3. DESCRIPTION OF ISOLATORS
 Manufacturing facility consists of three things mainly:
 Pharmaceutical material
 Personnel
 Environment surrounding
Isolator is the medium which isolate the pharmaceutical
material from personnel and surrounding environment.
 Types of Isolator-
 Negative Isolator-
Negative pressure maintained inside the isolator against
surroundings.
 Positive Isolator-
Positive pressure maintained inside the isolator against the
surroundings.

4. NEGATIVE ISOLATOR
2. Recirculation1. 100% Exhaust
 Two type of isolators designed as per the air circulation mention below:

5. POSITIVE ISOLATOR
Positive Isolators

6. SIGNIFICANCE OF ISOLATORS
 Product Quality- Prevent cross-contamination /contamination and provided minimum human
intervention.
 Operator protection- Separate surroundings from the impact of product.
 Operational hygiene- Fully closed & leak proof system provided high degree of assurity and
the operation inside isolator done through the leak proof glove assemblies.
 Environment safety- Leak proof system so it protect the surroundings from the product
contamination.
 Operational improvement- Personnel as well as product safety on the priority.
 Personnel Health- Health of personnel working in the area it may not on risks.
 ISO 5 (Grade A) environment maintained inside the isolator to support the aseptic
environment for the product, Even in grade D uncontrolled environment.

7. CHANGES IN PHARMACEUTICAL INDUSTRY
 Pharma industries tend to increase their production of certain
therapeutic drugs whose activity is higher (HPAPI’s) due to
compete the market demands.
 Isolators & RABS are not only for powders but also aerosols
potentially generated during liquid filling.
 Production of smaller quantity of product due to their activity and
demand.
 Safety of personnel who work on the product.
 Protection to environment from these potent drug entities.
 Product transfer to the Isolator and RABS without affecting sterility
and quality of product.
 Technique need to be transferred from nuclear industry.

8. CATEGORIZATION OF RABS
RABS provide protection by delivering an aerodynamic barrier over a
critical process zone and bio-decontamination is usually manual.
RABS when installed in ISO 7 environments already have a good
class of cleanroom air therefore the RABS can provide ISO 5 in the
critical zone.
Because RABS are being used more and more in aseptic processing
they have developed to an extent where there are various types of
RABS including:
 Passive RABS-
Uses the cleanrooms ventilation system as the source of clean air,
because it has no own air recirculation, filtering, or conditioning.
Air flows downward from the ceiling and returns to the surrounding
room through openings under the doors.
The air from the room returns through air ducts to the room air-
handling unit

9. CATEGORIZATION OF RABS
 Active RABS- Provides clean air via HEPA
filters by forced ventilation from an on board
fan.
Always contains a HEPA filter with laminar air
flow.
Air-conditioning expenditure is limited because
fresh air is drawn from the clean room.
 Passive Closed RABS- Utilize the clean
room's ventilation system and ceiling supply
filter to maintain the ISO 5.
Return through pre filter to room’s air
conditioning system
Used for less toxic and dusty product
applications.

10. CATEGORIZATION OF RABS
 Closed Active RABS- Air when it passes over the critical
zone it is filtered before it spills into the cleanroom or is
recirculated back through the supply filters of the RABS.
Can be operated with positive or negative pressure with
good sealing.
Closed RABS is similar to an isolator for such
application.
Used with toxic and dusty product applications.
Note-
1. Rapid Transfer ports (RTP) are used in the Isolators &
Closed RABS for material transfer to maintain the sterility of
material.
2. Gloves/glove ports used in the separation wall of
ISOLATOR & RABS for control of manual interventions.

11. SAFETY & HEALTH
 Health Hazardous Band-

12.  OEB Level 1-
It is the lowermost OEB level, where the toxicity of the material will not able to hazard the operator.
 OEB Level 2-
In this OEB level there is a slight risk from the toxicity of the material. Pragmatic steps should be taken to
prevent uncontrolled operator exposure.
For example, in addition to previous containment methods, drums should be opened within a down flow /
crossflow booth or with dedicated local exhaust ventilation. Alternatively the drums can be fitted with cone
valves for enabling dust-tight transfer.
‘OEL’ & CONTAINMENT DESIGN

13. ‘OEL’ & CONTAINMENT DESIGN
 OEB Level 3-
At this OEB level, the material is mildly hazardous, hence measures should be taken to
prevent operator exposure. Flexible isolators and glove bags are frequently used to provide
a simple barrier, whilst continuous liner systems can also provide a suitable method of
contained discharge. Material transfer is commonly performed using Split Butterfly Valves.
 OEB Level 4-
At this OEB level, the material is hazardous, hence significant measures should be taken to
prevent operator exposure. There should be a physical barrier between the operator and the
material. Isolators and Restricted-Access Barrier Systems (RABS) are common, whereby
material can be openly handled inside the dedicated enclosure, with the operator
manipulating the equipment and material externally through glove ports.
 OEB Level 5-
At this OEB level, the material is highly hazardous, hence significant measures should be
taken to prevent operator exposure. There should be a permanent physical barrier between
the operator and the material. Closed material handling and transfer should be undertaken
inside Isolators and Restricted-Access Barrier Systems (RABS) via glove ports. Even inside
the isolator, material should not be openly handled instead utilising RTPs and split valves for
contained handling.

14. PRODUCT RELATED CONSIDERATION
 The most common value used in the pharmaceutical industry to define a product-specific exposure
limit, which an operator is allowed to be exposed to, is the Occupational Exposure Limit (OEL).
 A pharmaceutical working group calculates each OEL as soon as a new product is defined and
categorized.
 No Observable Effect Level (NOEL) is basis of OEL calculation. NOEL value is determined by testing
the new active pharmaceutical ingredient (API) on individuals.
 The daily dosage in mg of active/(kg bodyweight x day) is increased day by day until the first
individual shows a reaction (lead effect).
 This NOEL is then multiplied by the average bodyweight of a human being to calculate the acceptable
exposure amount for an operator. Considering that the operator will mainly absorb the airborne
product by breathing, the previously calculated value is subsequently divided by the volume that
human breathes per day.

15. PRODUCT RELATED CONSIDERATION
 OEL can be calulated as follows-
OEL = NOEL x Body Weight / ( V x SF1 x SF 2 )
NOEL: No observable effect level
V: Volume of air breath by human per day (8-10 m3/8-hour
workday)
SF1 and SF2: Safety factors
 This OEL calculation now gives a value for the amount of airborne product
particles in the working environment that an operator/personnel can be exposed
to on a daily basis, for their entire life, without any risk to their health.

16. EQUIPMENT RELATED CONSIDERATION
 The real exposure levels of equipment can’t be calculated, but they can be determined by
measurement. This is done using special air sampling methods; the amount of collected airborne
particles is then determined by analysis.
 This amount is divided by the volume of air that passes through the air sampler during sampling,
which gives a value in µg/m3.
 This value only represents an average exposure level for the specific sampling time, a time
weighted average (TWA).
 Generally, the pharmaceutical industry works with two different TWAs:
1. The Short Term Time Weighted Average (STTWA), based on a sampling time of 15 minutes;
2. The Long Term Time Weighted Average (LTTWA), based on a sampling time of 8 hours.
• Until recently, no guide existed that described how to take these measurements.
Note- A guide, initiated by GEA and created by an international working group, is now published by the ISPE and is
known as SMEPAC (Standardised Measurement of Equipment Particulate Airborne Concentration). This guide now
defines the required test processes and parameters, and remains as close as possible to actual operating conditions.
Data obtained by SMEPAC-based tests provide the Short Term Time Weighted Average (STTWA) for a 100% pure active
material.

17. INTERPRETATION OF ‘OEL’ AND ‘TWA’ VALUES
 We now know that the employer must ensure, using suitable equipment, that the real
equipment exposure level is lower than the product-specific exposure Limit.
 It must be shown that the Long Term Time Weighted Average (LTTWA) exposure derived from
the equipment is lower than the Occupational Exposure Limit (OEL).
LTTWA < OEL
 However, For certain applications, such as charging or discharging, equipment-related
exposure only occurs during the docking, transfer and undocking process, which is generally
considered to take less than 15 minutes.
 So, to calculate now a Long Term Time Weighted Average (LTTWA) based on a known Short
Term Time Weighted Average (STTWA) for that equipment, we simply divide the STTWA by
32 (8 h = 32 x 15 minutes) and multiply by the actual number of cycles.
LTTWA = (STTWA/32) x number of cycles
 In addition, more pharmaceutical companies are also considering the dilution factor of the
handled product. As the equipment exposure data given by SMEPAC is based on a test with a
pure API, diluted material will only release an amount of active material that correlates with
the dilution factor (assuming the active is released at the same rate).

18. INTERPRETATION OF ‘OEL’ AND ‘TWA’ VALUES
 That means that the real LTTWA for a diluted material can be calculated as follows:
LTTWA = (STTWA/32) x number of cycles x dilution factor
 This calculated LTTWA is now lower or equal to the product-specific OEL.
 For example-
An operator works for a complete shift with a discharge station. The product is Ethinyl
Estradiol, which has an OEL of 0.035 µg/m3, and there are eight cycles (makes and
breaks) per shift. At this stage of the process, the material is already diluted to a
concentration of 5% active. The required STTWA for a make and break connection
for this process step is
STTWA req. = OEL x 32/number of cycles x dilution factor
STTWA req. = 0.035 (µg/m3) x 32/8 x 0.05
STTWA req. = 2.8 µg/m3

19. FACTOR ENFLUENCING CONTROL DESIGN/SELECTION

20. VALIDATION APROACH
Qualification of Isolators and RABS follow below mention sequence:
1. Design Qualification (DQ) defines the functional and operational specifications
of an equipment.
2. Installation Qualification (IQ) ensures that an equipment is received as
designed and specified. It documents the installation in the selected user
environment.
3. Operation Qualification (OQ) demonstrates that an equipment will function
according to its operational specification in the selected environment.
4. Performance Qualification (PQ) demonstrates that an equipment consistently
performs according to a specification appropriate to its routine use.
5. Maintenance Qualification (MQ) describes and documents any maintenance
required on the autoclave or equipment.

21. EQUIPMENTS FOR VALIDATION
 Validation equipment's used in the performance qualification and requalification of
2. Aerosol
Generator
7. Dry Ice
4. Hygrometer3. Photometer1. Particle Counter
5. Anemometer 6. Flow Hood 8. Gloves Integrity Tester

22. VALIDATION APROACHES
 Performance Qualification-
 Air Velocity Test
 Filter Integrity Test
 Airflow Pattern Test
 Particle counter
 Active Air Sampling
 Glove Integrity Test
 Chamber Leak Test (Isolator/CRABS)
 Humidity & Temperature of Chamber
(Isolator/CRABS)
 Differential Pressure Test
9. Active Air Samplers
9. Differential Pressure Measuring
Devices

23. VALIDATION APROACHES
 Air velocity test of the HEPA filter installed inside the Isolator and RABS shall be
verified during the PQ and periodically as per the validation plan.
 Equipment's Used for the Velocity Tests are Anemometer and Flow Hood.
 Anemometer is used for measuring the average air velocity at the face supply or
return air grilles. The air velocity is displayed digitally and the reading is held as long as
the button remains depressed.
Velocity range 0.25 to 30 m/sec or 50 to 6000 ft/min. Accuracy ±1% of reading ± 1 digit
 Flow Hood System is used to measures air flow and may compensate for density
and backpressure effects, allowing direct air flow readings in cubic feet per minute (cfm)
and liter per second (L/s) corrected for local air density. The measurement range is 25-
2500 cfm supply and 25-1500 cfm exhaust.
Accuracy is ±3% of reading ±7 cfm from 100 to 2000 cfm.
Air velocity of HEPA filter is approx. 0.45 m/s or 90 FPM.

24. VALIDATION APROACHES
 Air Changes Per Hour- Air changes is how many times air enters & exit
from the room through the help of HVAC system in one hour.
 ACPH calculation:
Average velocity (V) = (V1+V2+V3+V4+V5) / 5
If V is in m/s then it shall be changed in to FPM.
Average Air Velocity (FPM) = V x 3.28 x 60
Total Air Volume Per Minute (CFM) = FPM x Area of Filter (L x W)
ACPH = (CFM x 60) / Room Volume (L x W x H)
 Note- FPM = Feet Per Minute
CFM = Cubic Feet Per Minute

25. VALIDATION APROACHES
 Filter Integrity Test- It is also called as DOP test or PAO test, DOP
means Dispersible Oil Particulate Teste.
 Used chemical is PAO (Poly Alfa Olefin).
 Particle measured upstream up to 100% and downstream 0.01%
concentration through the Photometer.

26. VALIDATION APROACHES
 Airflow pattern test shall be used for the verification
of the direction of the airflow. Two type of air flow patterns
are describe below.
 Unidirectional Airflow Pattern- In this type of flow
pattern, Air shall move with uniform velocity and in a single
direction with parallel air streams within a close or confined
area. It is applied in Class 100 and lower area.
 Turbulent Airflow Pattern- In this type of flow patterns,
air shall not move in unidirectional way within the confined
area due to multiple pass direction, varying velocity and
non parallel flow pattern. It is mainly applicable in Class
10000 and others sometime it is used in the CRABS and
Isolators during the VHP cycle.
 Smoke generator and Dry Ice used for the verification of
the flow pattern.

27. VALIDATION APROACHES
 Particle count test shall be used for the verification of
the particle size which are present under the LAF within the
ISO standard limit.
 ISO Standard For clean area
 Particle counter sensor should be kept at the working height
under LAF within the defined sampling point.

28. VALIDATION APROACHES
 Air Sampling (Microbial)- Two type of air sampling defined for microbial
detection.
 Active Air Sampling- A microbial air sampler is used to force air into, or onto its
collection medium (e.g., Petri Dish with nutrient agar based test media) over a specified
period of time. The collected culture can then be incubated and analysed (ie., count
bacterial and/or fungal, colony forming units (CFU), and identify if required).
 Passive Air Sampling- Settle plates (Petri dishes) are opened and exposed to the air
for specified periods of time to determine what microbiological particles may be present
in the environment, as they may settle out of the ambient air, and onto the media
surface of the Petri Dish. These plates are then incubated and analysed.
Active air sampling done for the detection of microbes in the controlled area during working
time.
 Two types of media used for the air sampling:
 Soybean-Casein Digest Medium for bacteria detection and
 Malt Extract Agar Medium for fungi detection
Both are applicable at high risk compounding area.

29. VALIDATION APROACHES
 Glove Breach Test- The airflow through one open glove port shall be
measured by placing an anemometer at the centre of the glove port. The
velocity shall be agreed between customer and supplier (guidance value: 0.5
m/s).
 Glove Integrity Test- Glove integrity tester machine is used for the glove
integrity test. Pressurise the Glove at 1000 Pa with compressed air and leave it
for few minute to stabilize. After stabilization maintain the 1000 Pa and hold it
for approx. 10 minute. A drop in this pressure will indicate a leak through the
glove fabric or securing arrangement.
If the glove/gauntlet is sound, then the reading shown on the manometer will
remain static within 2 Pa to 10 Pa.
If the glove/gauntlet is damaged, then the reading shown on the manometer will
fall (i.e. 500 Pa, 495 Pa, 490 Pa). This trend will be distinct and progressive. The
rate of change will be proportional to the level of damage to the integrity of the
glove.

30. VALIDATION APROACHES
 Chamber Leak Test- Chamber leak test is applicable for the rigid closed
systems (CRABS and Isolators).
 Formula for Leak test mention below-
Leakage = Starting Pressure – End Pressure X 60 min. X 100%
End Pressure Test Time mins.
 Differential Pressure Verification- In this test the differential pressure
between the containment and environment measured with a calibrated
standard Magnahelic pressure gauge or sensor.
 Pressure of containment is more than the surroundings in Positive Isolators
and pressure of containment is less than the surrounding environment, but all
the parameters of containments should meet the ISO standard.

31. VALIDATION APROACHES
 Humidity & Temperature Control- Containments shall legible to
control the humidity and temperature of their own area, if they have separate
air handling unit.
 Working Temperature is about 22±2°C
 Relative Humidity in between 30 to 60%, it will be changed as per the
product requirements.
• Other tests which are perform during the qualification of containments are-
 Power Failure Test
 Area Recovery Test
 Sound Level Test
 Light Intensity Test

32. ROUTINE MONITORING & SAFETY
 Periodic tests: Must be subject to a schedule of periodic tests at various
levels and intervals.
 Daily,
 Weekly,
 Quarterly
 Yearly
 The yearly test schedule is essentially a revalidation schedule. It provides for
performance requalification (PRQ) tests to confirm that data collected during
performance qualification remain valid.
 Safety Measures:
 Follow the area gowning procedure
 Wear all personnel protective equipment for safety
 If any casualty call to maintenance engineer.

33. REFERENCES
ISO 14644- 1
ISO 14644- 3
ISO 14644- 7
ISO 10648- 2
PIC/S PI 014-3
Extract technology Containments
GEA technology Containments