The document provides a detailed overview of media fill and process simulation in the context of manufacturing injectable products, emphasizing the significance of aseptic processing and regulatory requirements. It describes various types of injectable devices, the manufacturing and packing processes, and establishes the purpose of media fill testing, which is to validate the sterility of these processes. Additionally, it outlines specific regulatory guidelines and expectations for media fill testing to ensure contamination-free production and compliance with industry standards.

1. Media Fill/Process simulation.
A Comprehensive Overview
Mohan B.

2. Index
• Importance of Injectable and safety.
• FDA waring & Recall.
• Different type of injectable product /Devise.
• Overview of manufacturing process.
• What is media fill testing & its Importance.
• Regulatory requirement for media fill testing and expectations.
• Grade A,B,C,D.
• Steps of aseptic process (Media+API).
• Aseptic media fill.
• Study design.
• Frequency and Run (as per FDA).
• Size of Runs and Line speed.
• Interpretation and test result.

3. NEWS:-

5. DIFFERENT TYPE OF INJECTABLE DEVICES
• Syringes, Auto-Injector, Pen Injectors, Infusion Pumps, Pre-Filled Syringes, Vaccine Pens etc.
…

6. Injectable devises

7. Overview of manufacturing and packing process:-
 Dispensing:---
 Dispensing of various types of Loads:-
 Manufacturing loads;- (Media Fill Load)
*API (Active Pharmaceutical Ingredient) (Pass Box)
 Excipients Load:-
 Packing materials Load (Vials,
Ampoules, bottles, Cartridges, Rubber,
stopper, dosing vessel filter, Tubes, pipes etc.)
 Prefill syringe load
 Accessories Loads :- Nuts Bolts, Rods.
 Sterilization Loads : Beaker,
Aluminium foil, agar media etc.

8. •Preparation: Items to be sterilized are placed in the autoclave chamber. They
must be cleaned and properly arranged to allow steam penetration.
•Sterilization Cycle: The autoclave heats water to produce steam, which is
then pressurized within the chamber. Common conditions are 121°C (250°F) at
15 psi for 15-30 minutes, but parameters can vary based on the
material and load.(Alternatives sterilizations may be used (sanitizations/decontaminations )

9. Overview of manufacturing and packing process:-
 Preparation of Raw Materials & API
(Compounding vessel, stirrer, jacket, valves, connector)
 Water For Injection. (Preparation)
 Quality control sample for Bioburden test and BET test
(Bacterial Endotoxin test)
 QC Approval-
 Filtration vassal (sterile filtration with 0.2 micron)
 IPQA Sampling (In process sampling, IP Spec test)
(Description assay degradation product, BET test)
 Connector – isolator
 Interventions*

10.  Assemble isolator - Load inputs : glass vials, PFS, ampules etc.
 vaporized Hydrogen peroxide used for sterilization of isolator
 Having a container dosing vessels (taps)(white tubes show in image)
 Having a tunnels :- Depyrogention tunnels only for glass vials –washing zone --
preheat Zone- mid temperature- zone approx.- then Depyrogention zone(high
heat)-Colling Zone.
 Tunnel glass vials –isolator
 Convers belt –Filling –stoppering (for Liquid)
 Nitrogen purging* (Not in Media)
 Inspection-AQL-Labeling –Inspection-QC

11. What is Media Fill Testing?
 Media fill testing is a procedure used to validate the sterility of the manufacturing process for
injectable products. It involves filling containers with a nutrient-rich medium instead of the
actual drug product to simulate and detect potential contamination.
 Purpose:- The primary goal of media fill testing is to ensure that the aseptic processing
environment and procedures are capable of producing sterile products, thereby safeguarding
patient health.
 Importance:
 Sterility Assurance: Ensures that the final injectable product remains free of harmful
microorganisms.
 Regulatory Compliance: Meets industry standards and regulatory requirements to maintain
product safety and efficacy.

12.  Process Validation: Validates the effectiveness of the aseptic processing and handling
procedures.
 Aseptic Processing:-A manufacturing process designed to keep products sterile by
preventing contamination from microorganisms during production.
 Nutrient-Rich Medium (Media):-A liquid used in media fills that supports microbial growth,
allowing detection of any contamination that might occur during the filling process.
(growth promotion test data required from Micro before this process.)
 Regulatory Guidelines: Standards and requirements set by agencies like the FDA (U.S.
Food and Drug Administration)
 EMA (European Medicines Agency) to ensure the quality and safety of pharmaceutical
products.
 Simulation:-The act of mimicking actual conditions or processes in a controlled
manner to test for potential issues without using the actual product.
 Pass/Fail Criteria:-Standards used to determine whether a media fill test meets the
required sterility assurance, with a "pass" indicating no contamination and a "fail"
indicating potential issues.

13. Regulatory requirements for media fill testing
 U.S. Food and Drug Administration (FDA)
 Guidelines:-21 CFR Part 211: This regulation outlines (cGMP), This regulation including the
need for aseptic processing validation.
 Guidance for Industry – Sterile Drug Products Produced by Aseptic Processing
 Key Points: Media fills should be conducted under conditions that simulate the worst-case
scenario to challenge the sterility of the process.
 The media used should be capable of supporting microbial growth and should be
evaluated for effectiveness.
 European Medicines Agency (EMA) (Guidelines)
 Annex 1 to the EU GMP Guidelines – Manufacture of Sterile Medicinal Products: This
annex covers requirements for aseptic processing and media fill testing. It provides
detailed guidance on the design and execution of media fills, including the need for
appropriate media, environmental monitoring, and evaluation criteria.
 Key Points: The test should be repeated regularly and whenever there is a significant
change in the process or equipment.

14.  International Council for Harmonization (ICH) Guidelines: ICH Q7 – Good Manufacturing
Practice Guide for Active Pharmaceutical Ingredients: This guideline emphasizes the need
for process validation, including media fills, to ensure the sterility of active pharmaceutical
ingredients and finished products.
 Guidelines' Chapter <71> – Sterility Tests: Provides general requirements for sterility testing
of sterile products, which includes media fill testing as a method for validating aseptic
processing.
 USP Chapter <1116> – Microbiological Control and Monitoring of Aseptic Processing
Environments: Offers guidance on the control and monitoring of environments used for
aseptic processing, including media fill requirements.
 Key Points: Specifies the need for rigorous testing procedures and regular review of media
fill results to ensure ongoing compliance with sterility standards.

15. GENERAL REGULATORY EXPECTATIONS:
• Validation: Media fills must be validated under conditions that simulate actual production
processes to challenge the sterility of the manufacturing environment.
• Documentation: Detailed records of media fill tests, including preparation, execution, and
results, must be maintained for regulatory review. (Audit points TBD.)
• Frequency: Media fills should be conducted regularly and whenever significant changes occur in
the manufacturing process or equipment.(Container change, Machine parts change, stopper
etc.)
• Worst-Case Conditions: The testing should consider worst-case scenarios, including the
maximum potential for contamination, to ensure that the process can handle such situations
without compromising sterility.
• These requirements ensure that injectable products are manufactured under conditions that
maintain their sterility and meet the safety and efficacy standards set by regulatory agencies.

16. Comparison of the different grades, focusing on key differences such as cleanliness
requirements, particulate contamination limits, and typical uses:
Grade Particulate Contamination
Limits
Air Quality Typical Uses Environmental
Control
Grade A
Very stringent; minimal allowable
particles. For example, in a 0.5 µm
particle size, the limit might be
3 particles/m³.
Highest level of air cleanliness
with stringent monitoring
requirements. Continuous or
frequent monitoring.
Direct product contact
areas, sterile zones, critical
operations (e.g., aseptic
filling).
Strict control measures;
continuous monitoring;
frequent cleaning and
maintenance.
Grade B
Less stringent than Grade A but
still controlled. For example, in a
0.5 µm particle size, the limit might
be 10 particles/m³.
High air quality; less stringent
than Grade A but still requires
rigorous monitoring.
Support areas for Grade A
operations, such as
preparation areas or areas
close to Grade A zones.
High control measures;
regular monitoring;
cleaning protocols in place.
Grade C
More lenient than Grade A and B.
For example, in a 0.5 µm particle
size, the limit might be
100 particles/m³.
Moderate air quality; less
frequent monitoring compared to
Grade A and B.
Areas for less critical
operations, such as
equipment preparation,
and storage of materials.
Controlled environment;
periodic monitoring;
cleaning protocols.
Grade D
Least stringent; higher allowable
particle counts. For example, in a
0.5 µm particle size, the limit might
be 350 particles/m³.
Basic air quality standards;
monitoring less frequent.
Non-critical areas, such as
personnel facilities,
storage, and support
activities.
Basic control measures; less
frequent monitoring;
standard cleaning
procedures.

17. • Steps of aseptic process:
Dispensing
Manufacturing
(Compounding)
(Media)
Sterilization
(Flirtations using
0.22 µ Filter)
Not in Media
Washing and Depyrogention of
vials In Case PFS
sanitizations/decontaminations
of PFS tubes
pre-filled syringes (PFS)
Aseptic Filling
(Isolator/RABS*/LAF)
Sealing and
Collection
Visual Inspection
and packing
(Incubation for
media)

18. Aseptic Media Fill:-
 What is aseptic media Fill simulation or Aseptic media Fill Validation or Process Simulation ?
 It is close simulation of routine aseptic manufacturing operations incorporating worst case
conditions that provide a challenge to routine manufacturing operation.
 Simulate :- Pretend I To mimic. (a real process or situation)
 Purpose of Media Fill:-
 Ensure that qualified process and personnel can aseptically produce drug product that
consistently and reliably meets the required quality
 Assess the capability of facility, personnel, equipment, procedures and process controls to
aseptically produce a sterile drug product
 Assess the robustness of the overall process when challenged with worst-case microbial
contamination risk
 Comply with current GMP requirements

19. HOW?
 Use of sterile microbial growth supportive medium Compounding / Manufacturing.
 Sterile filtration. (Use of 0.22 micron filter)
 Aseptic packing into sterile containers.
 Entire sterile manufacturing process should be simulated to assure the sterility of the aseptically
filled drug product.
 Media Fill includes all aseptic activities performed during dosage form compounding.
 Why microbial growth supportive medium?
 As the microbial growth supportive medium promotes the growth of microorganisms , which
create the worst case scenario for aseptic processing.
 To prove that aseptic process is consistency producing sterile units even when microbial growth
supportive medium is used instead of product.
 Example of media used: Soyabean casein digest (SCDM)*/ tryptic soy broth (TSB) for aerobic
process Fluid Thioglycolate Medium (FTM) for anaerobic process
Anaerobic media simulations are performed for oxygen sensitive products.

20. Study Design of Media Fill:-
 Factor associate with the longest permitted run on the processing lines that can pose
contaminations risk (e.g. Operative fatigue)
 Representative number, type and complexity of normal interventions that occurred with each run
as well as nonroutine intervention and events (e.g. maintenance, stoppage, equipment's and
adjustment)*
 Lyophilization when applicable (Liquid unstable product convert to Dry powder, Video)
 Aseptic assembly of equipment's (e.g. at start up, During processing)
 Number of personnel and there activities. (operators 2-3- same activity)
 Representative number of aseptic addition (e.g. charging container and closure as well as sterile
ingredients) or transfer
 Shift changes, breaks and gown changes. (as when applicable)
 Type of aseptic equipment's disconnection /connections.
 Aseptic sample collections
 Line speed and configurations (Media-exhibit-commercial)
 Weight checks
 Container closure system (e.g. size type compatibility with equipment.)

21. Frequency and Number of Runs
 At least three consecutive separate successful runs shall be performed during initial line
qualifications.
 Subsequently, routine semi-annual qualification conducted for each processing line will
evaluate the state of control of control aseptic process
 Any changes and event that have the potential to affect the ability to the aseptic process to
exclude contaminations from, the sterilized product should be assed through additional
media fills.
 For example Facility and equipment's modification lines configuration changes, significant
changes in personnel , container closure system changes, extended shutdown or end
product sterility testing showing contaminated products may be cause for revalidation of the
systems
 Durations of Runs
 The duration of the media fill run should be determined by the time it takes to incorporated
manipulation and intervention , as well as appropriate considerations of the duration of the
actual aseptic processing operation.
 Interventions that commonly occur should be routinely simulated, while those occurring rarely
can be simulated periodically

22. Size of Runs & Line speed
 The simulation run sizes should be adequate to mimic commercial production conditions and
accurately assess the potential for commercial batch contamination.

#For large commercial batches the number of units can be Statistically determined to
represent the manufacturing process under normal conditions.
 Line speed : The media fill program should adequately address
the range of line speeds employed during production.
 Example:- smallest container at Highest speed
or Largest container at lowest speed
Number of units filled
Production batch description Product on Batch Size
( # Filled units)
Minimum APS* batch size
( Filled units)
Small scale Less than 5000 units Less than 5000 units
Mid –scale 5000-10000 Units 5000-10000 Units
Large –Scale More than 10000 Units More than 10000 Units #

23. Incubation and Examination of Media-Filled Units
 Incubation temperature should be suitable for recovery of bioburden and environmental
isolates and should at no time be outside the range of 20-35°C (NLT 14 days.)
 If two temperatures are used for the incubation of the media filled units, the units should
be incubated for at least 7 days at each temperature (starting with the lower
temperature). (For example:-7 days at 20 -25 °C & then next 7 days at 30 - 35 °C.)
 Each media-filled unit should be visually examined for microbiological contamination by
qualified personnel.
 All integral units should proceed to incubation.
 Units found to have defects not related to integrity (e.g., cosmetic defect) should be
incubated; units that lack integrity should be rejected.

24. INTERPRETATION, OF TEST RESULTS
When filling less than 5000 units no contaminated units should be detected
–1 contaminated unit is consider cause for revalidation following and investigations
When filling from than 5000 to 10000 units no contaminated units should be
detected
–1 contaminated unit should result in investigation and Repeat media fill.
- 2 contaminated units are consider cause for revalidation following investigation.
When filling more than 10000 units.
-1 contaminated unit should result in an investigation
2 contaminated unit are considered cause for revalidation following investigation

25. References:-
1. FDA Guidance for Industry-“Sterile Drug Products Produced by Aseptic Processing –
Current Good Manufacturing Practices.” September 2004.
2. Guidance for Industry :-for the Submission Documentation for Sterilization Process
Validation in Drug Products
3. Guidance : Media Fills for Validation of Aseptic Preparations for Positron Emission
Tomography (PET) Drugs
4. Aannexure 6 : WHO good manufacturing practices for sterile pharmaceutical products
5. Annex 1 EU:- Manufacture of Sterile Medicinal Products
6. PDA Technical Report No. 22 –“Process Simulation for Aseptically Filled Products.” 2011.

26. Thank you……..!