Media Fill validation Protocol for Liquid Injection (Ampoule)
Index
| Sr. No. | Contents | Page No. |
| 1. | Approvals | |
| 2. | Overview | |
| 3. | Responsibility | |
| 4. | Pre Requisite Checks | |
| 5. | Filling and Stoppering Operations | |
| 6. | Methodology Of Validation | |
| 7. | Precautions | |
| 8. | Revalidation | |
| 9. | Frequency | |
| 10. | Final Report | |
| 11. | Report Approval | |
| 12. | Appendix |
1.0 Objective
The objective of this protocol is to ensure that the aseptic filling process of the Ampoules on the Ampoule Filling and Sealing Machine is under control and confirms to acceptability of the aseptic filling procedure in protecting the product from adventitious microbial contamination.
2.0 Scope
This protocol is applicable for the Ampoule Filling and Sealing machine with Equipment ID No.: ——- supplied by M/s ———– installed in the Ampoule Filling and Sealing Room of the Aseptic Liquid Injection Filling Facility.
3.0 Responsibility
The validation group comprising of representatives from each of the following departments shall be responsible for the overall compliance of this protocol:
Department Responsibility
| Department | Responsibility |
|
Validation |
i. Preparation of the validation protocol.
ii. Execution of the protocol after approval from QA. iii. Scheduling of validation. iv. Conducting of validation runs. v. Compilation of the data and review. vi. Preparation of Final Report and recommendation thereafter (if required). vii. Carry out investigation in case the media fills trials fails. viii. Schedule for revalidation. |
|
Maintenance
|
i. Review of the protocol. ii. Ensuring all utility related control during run. iii. Carrying out the repairs or modification (if required) prior to validation runs.
|
|
Quality Control |
i. Review of the protocol. ii. Preparation and qualification of the media. iii. Carrying out the microbial monitoring (Settle plate, Air sampling, Swab Testing and Personnel Monitoring) in critical areas. iv. Inoculation and Incubation of empty Ampoules and stoppers to confirm its sterility. v. Incubation of the medium used to confirm its sterility.
|
|
Quality Control |
i. Carry out Growth Promotion Test for the media (GPT). ii. Incubation of the filled Ampoules for 14 days. iii. Observation of the incubated filled Ampoules after 7 and 14 days. iv. Detecting the organism in the Ampoules showing microbial growth (if any). v. Carry out post media fill GPT.
|
|
Production |
i. Review of the protocol. ii. Filtration of the media. iii. Providing samples wherever necessary. iv. Carrying out the aseptic filling process. v. Carrying out Optical Inspection for Ampoules after filling and Stoppering. vi. Cleaning and sanitization of the area and equipments before and after media fill. vii. Destruction of the media filled Ampoules after authorization from QA. |
|
Quality Assurance
|
i. Approval of the Record. ii. Failure investigation and corrective actions (if any). |
4.0 Pre- requisite Checks
4.1 Prior to taking up the validation of aseptic filling, it should be ensured that all the following areas, equipments, systems and components have been qualified, validated and approved.
Areas
| Room Name | Room Number |
| Ampoule Filling and Sealing Room | ——- |
| Cooling Zone | ——- |
| Ampoule Washing and Sterilization | ——– |
Equipments
| Equipment Name | Equipment ID |
| Ampoule Filling and Sealing machine | ——– |
| Ampoule Washing Machine | ——— |
| Tunnel | ——- |
| Optical Inspection Board | ——– |
| BOD Incubator (22±2°C) | ——– |
| Bacteriological Incubator (32±2°C) | ——— |
Systems
| System Name | System ID |
| WFI System | ——- |
| Pure Steam Generation System | ——– |
Components
| Items Name | Type | Manufacturer/ Supplier | Sizes | Quantity
(For each trial) |
| Glass Ampoules | USP Type -1 | Ampoules | 1 ml | 10000 |
| 2 ml | 10000 | |||
| 3 ml | 10000 |
4.2 The following reference SOP’s / STP’s shall be used during the media fill runs. Attach the copies of SOP’s / STP’s along with Annexure No. I.
| SOP/ STP Title | SOP/ STP Number |
| Operation and Cleaning of Ampoule Filling and Sealing Machine. | |
| Cleaning and Disinfection of Aseptic Filling Facility | |
| Entry and Exit Procedure into Aseptic Filling Facility | |
| Area Monitoring in Aseptic Filling Facility |
|
| Preparation of culture suspension for media suitability test |
|
| Operation and Cleaning of Ampoule Washing Machine | |
| Operation and Cleaning of Dry Heat Sterilizer | |
| Filter Integrity Test |
|
| Test For Sterility |
|
| Procedure for Optical Inspection of Filled Ampoules and Pre Filled Ampoules | |
| Procedure for destruction of waste materials |
5.0 Filling, Stoppering and Sealing Operation
Ensure that the nitrogen gas valve are open and connected to the Ampoule filling and sealing machine. The pressure is within the acceptable range. Switch on the Machine. All the sterilized accessories like needle with silicon tubing’s is connected to their respective positions. Filtered media in a sterile stainless steel pressure vessel shall be brought under Laminar Air Flow Unit and connected to the peristaltic pump.All the activities shall be carried out aseptically.
Ensure that the washing and sterilization activity has been started at least two hours before starting ampoule filling activity. Wait until the sterilized Ampoules are reached to the table of ampoule filling machine.
Start the peristaltic pump and adjust the fill volume. The filled Ampoules are targeted to deliver target fill volume of 1ml ( for 1 ml amp.), 2 ml( for 2 ml amp.) and 3 ml( for 3 ml amp.) By using weight basis method all Ampoules filled volume is verified. Adjust the fill volume accordingly through PLC of machine. Once the volume is set, check for the quality of sealing of the Ampoules.
Start the filling and sealing activity. Check for the volume and sealing quality intermittently and collect ampoules for incubation at the interval of 15 minutes from each filling head ( Rest of the ampoules are to be stored in the temperature 20-25°C for 7 days and further at 30-35°C for 7 days in a controlled room).Collect the filled and sealed Ampoules in tray and pass the filled tray through pass box to the ampoule collection area (Ampoules inspection area).
6.0 Methodology Of Validation
6.1 Process Flow
6.2 Preparation and Qualification of media
6.3 Sterilization of Accessories
6.4 Sterility of the Components
6.5 Process Simulation
6.6 Planned Interventions During Media Fill Run
6.7 Monitoring During The Validation Runs
6.8 Inspection of The Filled Ampoules
6.9 Incubation Control
6.10 Post Incubation Growth Promotion Test
6.11 Interpretation Of Results
6.12 Acceptance Criteria
6.13 Failure Investigation
6.14 Corrective Actions
6.2 Preparation and Qualification of Media
6.2.1 Selection of the Media
6.2.1.1 Soyabean Casein Digest Media (SCDM) shall be used for media fill runs.
6.2.1.2 Justification for using sterile SCDM to simulate the filling process:
1. Low selectivity, clarity, media concentration and filterability.
2. Solubility of SCDM in water.
3. Flow ability of SCDM through the filling needle.
4. Ability to support growth of a wide range of microorganisms.
5. It does not inhibit the growth of microorganisms at the selected concentration.
6.2.2 Preparation of Media
6.2.2.1 Ready made media after checking the appearance and the expiry date shall be used.
| Ampoules Size | Size of runs | Quantity of Medium Required
For each run
|
| 1 ml | 10000 (+ 200) | 4.700 lts |
| 2 ml | 10000 (+ 200) | 9.400 lts |
| 3 ml | 10000 (+200) | 14.100 lts |
(+) additional numbers to be filled
6.2.2.2 The percentage of the media shall be 3 %.
6.2.2.3 Weigh and dissolve the medium in about half of the required quantity of WFI and after dissolving make up the volume with same water.
6.2.2.4 Stir the media for 10 minutes.
6.2.2.5 Check and record the pH. SCDM medium with pH of 7.3 0.2 shall be used.
6.2.2.6 Make adjustment in the pH with 0.1N NaOH or 0.1N HCl solutions if required.
6.2.2.7 Use clean dried glass containers for media preparation.
6.2.2.8 Record the observations in the ‘Media Preparation Record’. The format for Media Preparation Record is attached as Annexure No. II.
6.2.3 Filtration of the Media
6.2.3.1 The media solution shall be filtered through a pre sterilized 0.22µ membrane filter into a previously sterilized SS pressure vessel under Laminar Air Flow Unit.
6.2.3.2 The integrity of the sterilized filter should be confirmed immediately after use as per SOP No. . The result of the integrity test shall be entered in the record
6.2.4 Media Sterility Test
100 ml of the sterilized media shall be collected in a sterile container (in duplicates) and incubated at 20-25°C for 7 days and further at 30-35°C for 7 days to show that there is no contamination in the media after sterilization. Record the observations in the ‘Media Sterility Test Record’. The format for Media Sterility Test Record is attached as Annexure No. III.
6.2.5 Growth Promotion Test Of The Filtered Media
6.2.5.1 100 ml of the sterilized media shall be collected in sterile container (3 no) for Growth Promotion Test. The Growth Promotion Test shall be carried out with Aspergillus niger (ATCC 1344 ), Bacillus subtilis (MTCC 441 ) and Candida albicans (MTCC 227 ).
6.2.5.2 The Growth Promotion Test should demonstrate that the medium supports recovery and low number of microorganisms ie. 10-100 cfu / unit or less.
6.2.5.3 Prepare culture suspensions as per SOP No. . and select suitable dilution and volume that gives 10-100 cfu / unit or less.
6.2.5.4 Incubate the above inoculated containers at specified incubation conditions as mentioned in Table1 below.
6.2.5.5 Observe the medium daily for growth.
6.2.5.6 The test medium is valid, if it shows adequate growth within specified incubation period.
6.2.5.7 The medium is considered invalid if it shows inadequate growth response.
6.2.5.8 Record the observations in the ‘Growth Promotion Test Record’. The format for Growth Promotion Test Record is attached as Annexure No
Table: 1
|
Test media |
Test Organisms |
Incubation | |
| Temperature / Days | Conditions | ||
|
Soyabean Casein Digest Medium (SCDM) |
Aspergillus niger (ATCC 1344) | 22.5 ± 2.5 °C for 5 days. |
Aerobic |
| Bacillus subtilis (MTCC 441 ) | 22.5 ± 2.5 °C for 3 days. |
Aerobic |
|
| Candida albicans (MTCC 227 ) | 22.5 ± 2.5 °C for 5 days. |
Aerobic |
|
6.3 Sterilization of Accessories
6.3.1 Sterilization of Filling & Sealing machine parts (Filling needle, Silicone tube), SS forceps, etc shall be done before commencing Media filling.
6.3.2 The sterilization of accessories shall be carried out as per SOP No.
6.4 Sterility of the Components
6.4.1 Ampoules shall be collected separately in sterile containers just before the start of the filling operations to confirm the sterility of the components.
6.4.2 The test shall be carried out as per SOP No. MIC/SOP/OO2. Result of the test shall be entered in the Data record.
6.5 Process Overview
6.5.1
6.5.1.1 The filling and sealing machine shall be operated as per the SOP No
6.5.1.2 Filtered media in the SS vessel shall be brought under the Laminar Air Flow Unit of the filling and sealing machine.
6.5.1.3 The fill volume of the vial to be set for targeted fill volume to deliver 1 ml ( for 1 ml amp. ) 2 ml ( for 2 ml amp.) and 3 ml (for 3 ml amp.) from each syringe.
6.5.1.4 The filling operation is started and the Ampoules are filled with media solution.
6.5.1.5 The filled Ampoules are then sealed on line.
6.5.1.6 Check the quality of sealing on line.
6.5.1.7 The filled and sealed Ampoules are transferred through pass box outside the aseptic filling area.
6.5.1.8 Collect the Ampoules in SS trays.
6.5.1.9 Each tray shall be labelled with Run Number / Session / Serial Number / Number of Ampoules Filled, Date and Sign.
6.5.2 Number of Media Fill Runs & Filling Quantity
6.5.2.1 Three media fill runs shall be conducted in three different days.
6.5.2.2 Three run shall be carried out with Soyabean Casein Digest Medium.
6.5.2.3 Size of Runs
| Ampoules Size | Minimum Ampoules to be filled |
| 1 ml | 10000 |
| 2 ml | 10000 |
| 3 ml | 10000 |
6.5.2.4 Each run shall be covering two shifts and considering all possible interventions.
6.5.3 Filling Speed
6.5.3.1 Filling speed and their records shall be maintained during all the three media fill runs.
6.5.4 Filling Session
Table 3
|
Session |
Number of Total Ampoules To Be Filled | |||
|
1 ml |
2 ml |
3 ml |
||
| Session A (Morning- Around 11.00 hrs) | 3500 | 3500 | 3500 | |
| Session B (Evening- Around 17.00 hrs) | 3500 | 3500
|
3500
|
|
| Session C (Night- Around 23.00 hrs) | 3500 | 3500 | 3500 | |
6.5.5 Number of People Involved
6.5.5.1 Persons (Supervisors and Operators) from different departments like Production, Quality Assurance, Quality Control, Mantainance shall be present – by rotation in trials to cover all the persons. Minimum number of persons permitted at a time during operations shall not be less than 5.
6.5.6 Garments
6.5.6.1 All the gowns used during the media fill runs shall be sterilized at least one day prior to the media fill runs.
6.6 Planned Interventions During Media Fill Run
Following interventions shall be covered in the entire media fill runs:
6.6.1 Failure of Power Supply for two minute.
6.6.2 Change of Operator.
6.6.3 Break down of the filling machine.
6.6.4 Removing Ampoules for fill volume check.
6.6.5 Changing of hand gloves.
6.6.6 Others
Record all the observations in the ‘Planned Interventions Record’. The format for ‘ Planned Interventions Record’ is attached as Annexure No.
6.7 Monitoring During The Validation Runs
6.7.1 Personnel Monitoring
6.7.1.1 Ensure that personnel entering the area have followed the gowning and entry procedures as per SOP No.
6.7.1.2 Personnel monitoring shall be carried out as per SOP No.
6.7.1.3 During personnel monitoring if any organisms are observed, the same organisms shall be isolated and preserved in the Quality Control (Microbiology) Laboratory for Identification.
6.7.2 Microbial Air Monitoring
6.7.2.1 Microbial Air Monitoring of the area shall be carried out as per SOP No. ——-. The results of the microbial air monitoring shall be entered in the data recording sheet.
6.7.2.2 During area monitoring if any organisms are observed, the same organisms shall be isolated and preserved in the Quality Control (Microbiology) Laboratory for Identification.
6.7.3 Non Viable Particulate Count
6.7.3.1 Non Viable Particulate Count Monitoring of the area shall be carried out. The results of the Non Viable Particulate Count Monitoring shall be entered in the record sheet.
6.7.4 Environmental Conditions Monitoring
6.7.4.1 Monitor the environmental conditions for temperature, differential pressure and relative humidity.
6.7.4.2 Record the observations in the ‘Environmental Conditions Record’. The format for ‘Environmental Conditions Record’ is attached as Annexure No.
6.8 Inspection of the Filled Ampoules
6.8.1 Inspection of the filled and sealed Ampoules shall be done by qualified personnel as per SOP No. —–
6.8.2 Rejection analysis shall be done and reason of rejection to be identified and recorded.
6.8.3 Record all the observations in the ‘Inspection of Filled Ampoules Record’. The format for ‘Inspection of Filled Ampoules Record’ is attached as Annexure No. VII.
6.8.4 100 % inspection to be carried out prior to incubation.
6.9 Incubation Control
6.9.1 Incubate all the filled Ampoules except the ones having sealing defects, develops cracks during the filling and rejected during optical inspections.
6.9.2 The number of filled Ampoules kept for incubation should not be less than 360 Ampoules for all the pack size.
6.9.3 Invert and swirl to ensure that all the internal surface are in contact with media.
6.9.4 Incubate the filled good Ampoules for 7 days at 20-25°C.
6.9.5 After 7 days, inspect the Ampoules for contamination. If all the Ampoules are clear invert and swirl to ensure that all the internal surface are in contact with media.
6.9.6 Incubate the Ampoules at 30-35°C for further 7 days.
6.9.7 Visually inspect the Ampoules for growth of microorganism against black and white background and record in validation report.
6.10 Post Incubation Growth Promotion Test Of The Medium
6.10.1 Post Incubation Growth Promotion Test of the media used during simulation studies shall be carried out on completion of the incubation period for 14 days to demonstrate the ability of the media to sustain growth.
6.10.2 100 ml of the filtered media shall be collected in sterile container (3 no) for Growth Promotion Test. The Growth Promotion Test shall be carried out with Aspergillus niger (ATCC 1344), Bacillus subtilis (MTCC 441) and Candida albicans (MTCC 227).
6.10.4 Growth Promotion Test of the above medias shall be carried out as per steps under section 6.2.5 above.
6.10.5 The environment isolates can also be used for the Growth promotion test, if any found.
6.10.6 If the Post Incubation Growth Promotion Test results fail to show the ability of the media to sustain growth; the media fill studies shall be repeated until all four consecutive runs result complies.
6.11 Interpretation Of Results
6.11.1 Observe the growth/turbidity in all the media filled Ampoules, after 24 hrs, 3rd, 7th, 8th, 12th & 14th days of incubation.
6.11.2 If contamination occurs, check the integrity of the sealing quality.
6.11.3 Record the observations in the ‘Media Fill Test Report’. The format for ‘Media Fill Test Report’ is attached as Annexure No.
6.12 Acceptance Criteria
| Ampoules Size | Number of Units Filled | Alert Level (number of contaminated units in a single media fill run) | Action Level (number of contaminated units in a single media fill run) |
| 1 ml | 10000 | 1 | 2 |
| 2 ml | 10000 | 1 | 2 |
| 3 ml | 10000 | 1 | 2 |
6.12.1 The target is Zero positives.
6.12.2 Any positives unit indicates a potential sterility assurance problem.
6.12.3 If the level of contamination is found to be 2 Ampoules per 10000 Ampoules, then the cause of contamination should be identified and should result in a through documented investigation.
6.12.5 If the number of contaminated units exceeds 2 per 10000 Ampoules, then the entire media fill studies shall be considered as invalid.
6.12.6 Cease the media fill runs, investigate the reasons of failure, rectify and then repeat the trials.
6.12.7 The aseptic filling process shall be considered validated if all the 3 successful consecutive runs are completed.
6.13 Failure Investigation
6.14.1 Incase of failure, the media fill studies shall be considered as invalid and comprehensive investigations shall be carried out.
6.14.2 The investigation shall be carried out by the Quality Assurance Department along with the other concerned departments surveying all possible causes of the contamination and find out the reasons of failure.
6.14.3 The contaminant shall be identified up to species level (if possible).
6.14.4 The microbial monitoring data shall be reviewed and the environmental isolate (if any) found during personnel or area monitoring shall also be identified up to species level (if possible) to check for the similar type.
6.14.5 The cause of contamination shall be identified and recorded. The recording shall be carried out in the ‘Failure Investigation Report’. The format for ‘Failure Investigation Report’ is attached as Annexure No. IX.
6.14 Corrective Actions
6.14.1 The media fill runs shall be repeated again until all the three consecutive runs comply.
6.14.2. The recording for corrective actions shall be carried out in the ‘Corrective Actions Report’. The format for ‘Corrective Actions Report’ is attached as Annexure No. X.
7.0 Precautions
7.1 Check the absence of media in the filling area after completion of media fill run.
7.2 Destroy all used Ampoules and media used during media fill studies as per SOP No. PI-012.
7.3 Clean the area after media fill run and wait at least for 5 days, till the environmental monitoring results meets the acceptance criteria.
8.0 Revalidation
8.1.1 Revalidation shall be performed after any significant modification to the equipment, area, process, number of shifts and change in operators.
9.0 Frequency
9.1 Once in a 6 month.
10.0 Final Report
10.1 Results and Evaluation
Results shall be documented in the Test Report provided as Annexure to the Protocol and all Test Data Sheet mentioned in the protocol. Based on the observations recorded in the qualification tests, evaluation of the results shall be carried out.
All the results meeting the acceptance criteria establish that the aseptic filling process for Ampoules filling and Stoppering machine qualifies for all relevant parameters and can be used for carrying out the Filling and Stoppering process for Ampoules.
10.2 Summary
On the basis of the results and evaluation, a summary report shall be prepared. The summary report shall draw a conclusion to state the successful qualification of the aseptic filling process for Ampoules filling and Stoppering operation in a consistent manner (or) re qualification (if required in case of failure). The summary report shall be attached with the protocol along with the test reports.
11.0 Report Approval
The summary report shall prepared by Validation, and reviewed by Production, Quality Control and Engineering. The Quality Assurance shall do the final approval of the report.
12.0 Appendix
12.1 Abbreviations
GPT – Growth Promotion Test
WFI – Water For Injection
USP – United States Pharmacopoeia
IPA – Iso Propyl Alcohol
SCDM – Soyabean Casein Digest Medium
ATCC – American Type Culture Collection
CFU – Colony Forming Units
LAF – Laminar Air Flow Unit