“MICROBIOLOGICAL CONTROL AND MONITORING OF ASEPTIC PROCESSING ENVIRONMENTS & PRODUCT” Microbiologically controlled environments are used for a variety of purposes within the healthcare industry. This general information chapter provides information and recommendations for environments where the risk of microbial contamination is controlled through aseptic processing. Products manufactured in such environments include pharmaceutical sterile products, bulk sterile drug substances, sterile intermediates, excipients and in certain cases, medical devices. Aseptic processing environments are far more critical in terms of patient risk than controlled environments used for other manufacturing operations-for example, equipment and component preparation, limited bio-burden control of non-sterile products, and processing of terminally sterilized products. The type of aseptic processing is differentiated by the presence or absence of human operators. An advanced aseptic process is one in which direct intervention with open product containers or exposed product contact surfaces by operators wearing conventional clean-room garments is not required and never permitted. Clean rooms have a very special place and purpose in aseptic areas such as sterility lab. The establishment, maintenance, and control of the microbiological quality of the clean room and the controlled environments require engineering and science based approach. Aseptic area maintained by when microorganisms entry are prevented by providing high quality of air in the room, using HEPA (High Efficiency Particulate Air) filters that provide Laminar Air Flow.  Surface monitoring of Walls and floor and equipments lying in class 10000 or Grade C shall be done by this method.  Area of 100cm2 shall be selected for taking swabs.  Swabs shall either be pre- sterilized or non sterile swabs can be sterilized at about 121 oC for 20 min or as per validation cycle.  Sterilized swabs shall be placed in 10mL of sterile 0.9% saline solution.  Rub the swab gently over the surface which is to be monitored.  Keep them back in the tubes containing 0.9% saline solution. Each tube shall be marked with the location and date of sampling.  Take 1mL of saline suspension containing swab sample in, pre sterilized Petri dish.  Pour molten Soyabean casein digest agar (SCDA) media, in duplicate. Mark the plates indicating location and date of sampling.  Let the plates solidify. Incubate them first at 30 oC to 35 oC temperature for 48 hours and then at 20 oC to 25 oC temperature for next 72 hours.  Observations shall be recorded first after 48hrs and then after 120hrs.  Multiply the count observed in each plate with 10, so as to get the actual count of that area.  All the environmental parameters shall be verified and established at the time of installation and thereafter monitored at periodic intervals. The recommended frequencies of periodic monitoring shall be as follows: -Particulate Monitoring in Air – cont..

1. “MICROBIOLOGICAL CONTROLAND
MONITORING OF ASEPTIC PROCESSING
ENVIRONMENTS & PRODUCT”
Presented by- ALOK YADAV
M.Sc. Microbiology
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ALOK YADAV

2. Microbiology Laboratory Equipments
ALOK YADAV 2

3. ALOK YADAV 3
Microbiology is a crucial part of pharmaceutical quality assurance and is used to prevent and
control microbial contamination during the manufacturing, storage, and distribution of
pharmaceuticals.
What is the Microbiology ?
What Microbiologist do?
*Microbiologists in the pharmaceutical industry work to ensure the safety and efficacy of
pharmaceutical products.
*They perform rigorous testing protocols to assess the sterility, stability, and overall
microbiological quality of pharmaceutical formulations.

4. ALOK YADAV 4
*Microbiologists in the pharmaceutical industry are members of the quality department.
*They ensure the quality of raw materials before they are processed, monitor the
microbiological quality of the environment and water, and validate the test methods used
in testing finished products.
*They also test finished products from a microbiological perspective.

5. MICROBIOLOGICAL CONTROL AND MONITORING OF
ASEPTIC PROCESSING ENVIRONMENTS
*Microbiologically controlled environments are used for a variety of purposes within the
healthcare industry.
*Risk of microbial contamination is controlled through aseptic processing.
*Aseptic processing facilities can control microbial contamination by regulating air
supply and equipment, and processing and packaging products without further
contamination.
*Equipment can be sterilized with heat and chemicals.
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Heat sterilization
*Autoclave
A pressure cooker that uses high-pressure, saturated steam to sterilize medical devices.
The items are placed in a chamber and the steam is maintained at around 121°C for at least
30 minutes.
This method is effective, fast, safe, and affordable.
*Dry heat
Uses a heating cabinet or conveyor tunnel to expose devices to high temperatures for a
long time (176–232°C for around 60 minutes).
This method doesn't require steam or water, so there's no risk of corrosion, and the devices
remain dry.
Dry heat is less efficient than wet-heat sterilization, so it may require higher temperatures
or longer times.

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Autoclave
Dry heat Oven

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Chemical sterilization
*Ethylene oxide
A flammable, colorless gas with a sweet smell that's often used to sterilize medical
equipment.
*Formaldehyde
A highly reactive gas that kills microorganisms by modifying proteins. However, it's a
mutagen and carcinogen.
*Ozone
Mainly used to decontaminate medical devices and the air.
*Nitrogen dioxide
Used in the medical device, biotechnology, and pharmaceutical industries to sterilize
temperature-sensitive materials.

9. ALOK YADAV 9
*Aseptic processing facilities have clean
rooms with filtered air and positive
pressure to prevent contamination.
*Aseptic area maintained by when
microorganisms entry are prevented by
providing high quality of air in the room,
using HEPA (High Efficiency Particulate
Air) filters that provide Laminar Air Flow.
*This type of air filter can theoretically
remove at least 99.97% of dust, pollen,
mold, bacteria, and any airborne particles
with a size of 0.3 microns (µm).

10. ALOK YADAV 10
All the environmental parameters shall be verified and established at the time of
installation and thereafter monitored at periodic intervals. The recommended frequencies
of periodic monitoring shall be as follows:
• Particulate Monitoring in Air – Half Yearly
• HEPA Filter Integrity Testing (Smoke Test) – Yearly
• Air Change Rates – Half Yearly
• Air Pressure Differentials – Daily.
• Temperature and Humidity – Daily
• Microbiological Monitoring by Settle Plates and/or Swabs in aseptic areas– Daily, and at
decreased frequency in other areas.
NOTE: -The above frequencies of monitoring shall be changed as per the requirements and load in
individual cases.

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AIR BORNE VIABLE PARTICLE MONITORING
Environmental microbiological monitoring should reflect the facility used (Air Lock) and
include a combination of air and surface sampling methods appropriate to the facility,
such as:
*Passive air sampling by Plates Exposure Methods,
*Surface monitoring by RODAC (Replicate Organism Detection and Count) plate or by swabs,
*Personnel Monitoring (Gloves etc.) by RODAC plate.

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Surface Monitoring by Swab Method
Surface monitoring of Walls and floor and equipments
lying in Grade C shall be done by this method.
*Area of 100cm2 shall be selected for taking swabs.
*Swabs shall either be pre- sterilized or non sterile swabs can be sterilized at about 121oC
for 20 min or as per validation cycle.
*Sterilized swabs shall be placed in 10mL of sterile 0.9% saline solution.
*Rub the swab gently over the surface which is to be monitored.
*Keep them back in the tubes containing 0.9% saline solution.

13. ALOK YADAV 13
*Each tube shall be marked with the location and date
of sampling.
*Take 1mL of saline suspension containing swab
sample in, pre sterilized Petri dish.
*Pour molten Soyabean casein digest agar (SCDA)
media, in duplicate. Mark the plates indicating location
and date of sampling.
*Let the plates solidify. Incubate them first at 30 oC to 35 oC temperature for 48 hours and
then at 20 oC to 25 oC temperature for next 72 hours.
*Observations shall be recorded first after 48hrs and then after 120hrs.
*Multiply the count observed in each plate with 10, so as to get the actual count of that
area.

14. ALOK YADAV 14
THANK YOU