Aseptic Media Fill or Process simulation.

A media fill, also known as an aseptic process simulation, is a critical test used in pharmaceutical manufacturing to ensure that the aseptic production process is capable of producing sterile products consistently. In this test, a nutrient-rich microbiological growth medium (instead of the actual product) is filled into containers under aseptic conditions. This medium serves as a substitute for the product and provides a means to detect any microbial contamination that might occur during the filling and packaging process. The idea is to mimic the actual manufacturing process as closely as possible, including personnel interventions, equipment operation, and environmental conditions.

Overview
A media fill, or aseptic process simulation, is a comprehensive test used in pharmaceutical and biotechnology manufacturing to validate the aseptic production process. It is designed to ensure that the entire production line can consistently produce sterile products without contamination. During this test, a sterile growth medium, usually a nutrient broth that promotes microbial growth, is used in place of the actual product. This medium is filled into containers (e.g., vials, ampoules, syringes) under normal production conditions. The test aims to mimic the routine manufacturing process as closely as possible, including all aseptic manipulations, environmental conditions, and personnel interventions, to simulate both standard and worst-case scenarios.
The media fill process includes several key steps: preparation of the growth medium, aseptic filling under controlled conditions, and sealing of the containers. Once filled, the containers are incubated for a specified period, typically 14 days, to detect any microbial growth. If contamination occurs, it indicates a breach in the aseptic process, prompting a thorough investigation to identify and rectify the source of contamination. Successful media fills, characterized by no microbial growth in any container, demonstrate that the aseptic process is capable of producing sterile products reliably. This test is a regulatory requirement and a critical component of quality assurance in aseptic manufacturing, ensuring patient safety by confirming that the product meets sterility 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:-

4. Different type of Injectable Devices S yringes, Auto-Injector, Pen Injectors, Infusion Pumps, Pre-Filled Syringes, Vaccine Pens etc.…

5. Injectable devises

6. 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.

7. 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 )

8. 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*

9. 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

10. 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.

11. 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.

12. 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.

13. 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.

14. 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.

15. 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.

16. 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)

17. 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

18. 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.

19. 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.)

20. 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

21. 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 #

22. 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.

23. 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

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

25. Thank you……..!