VAL-190 Guideline for the Validation of Aseptic Processing

1. Purpose

The purpose of this document is to provide a set of guidelines to be used when determining a suitable approach for the qualification of aseptic processing validations at the manufacturing facility involving in aseptically filled medicinal products.

2. Scope

These guidelines provide suggestions and considerations for the design and preparation of aseptic media fill validations protocols and reports. They are intended to be used as guidelines only, alternative approaches for validation may be appropriate and can be used provided that the principals of validation as outlined in the company Validation Master Plans are followed.

Out of scope are closed aseptic processing systems where no aseptic connections are made post sterilization and temperature and pressure are monitored during Steam In Place (SIP) sterilization.

3. Safety

All safety requirements for relevant production areas must be followed at all times.

4. References

PICS 2009 Guide to Good Manufacturing Practice for Medicinal Products Annex 1, Current BP <A266> and USP <1211> Monographs.

5. Introduction

The sterility assurance for aseptic processing during final formulation and filling cannot be measured by the sterility test alone and therefore it is critical to routinely challenge the actual aseptic process under “worst case” conditions. Aseptic processing is challenged using microbiological growth media under simulated conditions.

The media fill is a simulation of the entire aseptic formulation and filling process, which substitutes a microbiological growth medium for a sterile product. The media fill also provides a way to evaluate changes made to aseptic processing operations which may affect the sterility assurance of the final product and the performance of aseptic filling personnel under operational conditions. 

6. Pre-requisites for Aseptic Process Validation (Media Fills)

Before aseptic assembly or processing, different parts of the final product are generally subjected to different sterilization processes, such as dry heat, moist heat, and sterile filtration.

Each stage of the aseptic filling operation requires validation and control as a pre-requisite to aseptic processing. Each also introduces the possibility of error that might ultimately lead to the distribution of contaminated product. Any manual or mechanical manipulation of the sterilized drug, components, containers, and closures prior to or during aseptic assembly poses a risk of contamination and thus necessitates careful control.

GMP facility ensures that a combination of full equipment qualification, component and equipment protection during handling and storage and rigorous personnel training are essential to the safe manufacture of aseptically manufactured products.

Pre-requisites for Process Simulations

– Critical areas (Grade B /Class 10,000 and Grade A / Class 100) are qualified and HVAC HEPA filters certified

– Environmental monitoring procedures are qualified

– Environmental monitoring media is approved for use

– Validated equipment and component sterilization steps are completed

– Validated sterile filtration of bulk products is completed

– Media used in simulations is qualified

– Staff participating in the simulation are qualified in aseptic gowning

– Staff entering the filling areas are trained in aseptic technique – includes filling operators, maintenance staff, QC staff and environmental monitoring staff.

Occupation States

Occupation states list below defines the condition in which an aseptic environment is operating for the purpose of testing and classification. This document sets out the requirements for “in-operation” testing or classification only.

– The “at rest” state means that there are no activities being conducted and the area is un-manned (no operating personnel is present). Monitoring at this state is required to establish the effectiveness of the sanitization and the integrity of the HAVAC and supporting systems.

– The “in operation” state means that the activities being conducted are routine and associated with the processes of product manufacture. It is used to establish the microbial quality of air that the product is exposed to during routine manufacturing activities.

– Examples of operations to be carried out during the “in operation” state are defined in Table 1

Table 1 – “In operation”

7. Frequency and Requirements for Challenges

7.1 New Production Lines

Prior to release of a facility, filling line or process line for commercial use acceptable results from a minimum of three consecutive successful media fill trials should be achieved to demonstrate the reproducibility of the process.

7.2 Changes to Existing Production Lines

There should be change control evaluation, including microbiological/sterility assurance evaluation before any change is introduced. If the change could impact the sterility assurance of the process line then media fill trials should be performed. (Refer to QMS-125 “Change Management System”).

The number of trials required as part of re-qualification due to changes is at the discretion of the Quality Assurance Manager and the Validation Manager, and depends on the significance of the change(s). This should be determined as part of change control.

Whenever a significant change is introduced into an existing aseptic process, the change should be validated.  Examples of significant changes include:

– Changes to handling procedures for components or equipment

– Changes to line set up practices

– Variations or additions to containers and closures processed on the line

– Extensions of line runs beyond standard (validated) hours of operations

The following changes would normally require three trials to be conducted:

– Changes to validated parameters

– Changes to aseptic techniques

– Changes to room operation

At least one successful media fill trial should be completed prospectively before any manufacturing is recommenced after changes to existing production lines.  Where more than one media fill trial is required, subsequent trials may be conducted concurrently with manufacture.  No batches are to be released until the results of all media fills have been reported and the report reviewed and approved by Quality Assurance.

7.3 Routine Re-validation of Aseptic Filling Lines

Routine revalidation media fills involve concurrent validation. If any unit from a single run is positive then Quality Assurance intervention, investigation and corrective action must be implemented immediately.  Refer to Section 13 for acceptance criteria and responses.

Each aseptic filling line shall be subjected to a media fill challenge according to GMP guides. For maximum time interval for each filling line, refer to Table 2. It is permissible to bracket container/closure systems for each line, however the worst case combinations should be considered eg. bottle with the widest neck/largest fill (longest exposure) in combination with the smallest container/closure system (hardest to manipulate).

For the filling lines two simulation studies (media fills) are required in order to evaluate the sterility confidence for each manufacturing process used in the filling room.

One of the simulation studies will be performed every 6 months to simulate routine aseptic manufacturing process for products manufactured for the EU market. In this simulation study the filling operator will use sterile forceps to pick up the pillow packs from the hopper as described in production procedures.

The second simulation study will be performed every 12 months to simulate routine aseptic manufacturing process for products manufactured for the non-EU and non US markets. In this simulation study the filling operator will use sterile gloved hand to pick up the pillow packs from the hopper and load on the carousel.

Table 2 – Media Fill Frequency

7.4 Facility Shutdowns and Recommissioning

In the event of a filling line shutdown and recommissioning at least one media fill should be conducted prospectively (before commencement of aseptic filling).  The results of at least the 7day inspection should be known before aseptic filling of product commences, though approval to fill on risk can be proceeded through the Deviation Procedure.

For major changes to the dispensing facility three consecutive media fills should be conducted to completely requalify the line.  (Two of these may be conducted concurrently with manufacture). Any product manufactured before the results of the entire media fill series are known should be quarantined.

7.5 Routine Clean Room Classification

Room classifications are performed to demonstrate that the quality of air within an environment routinely meets the grade required to carry out critical operations. The process of classification involves the sampling of non-viable particulate material to define the size and quantity of particles within the environment. The quality of air within the room is directly related to the composition of non-viable particles and defined as either a room grading under GMP’s or an ISO class under ISO 14644:2.

Routine “in operation” classification for Grade A and B zones within aseptic filling areas are performed on a 6 monthly basis. During classification the room should be in an “in operation” state as defined in section 6.3 and can be performed during normal operations, media fills or under simulated operations.

For each room the highest classification to achieve is the requirement state in the GMP code for PIC’S products. This provides the maximum permitted airborne particle concentration for 0.5 and 5.0µm particle for each grade as given in the following table:

Table 3 – Non-viable Particles Classification (PIC’S)

For the classification purposes, a minimum sample volume of 1m³ should be taken per sample location. The minimum number of sample locations has been defined according to EN/ISO 14644-1 for classification purposes and is summarized in Table 4 and in Diagram 1.

Particulate monitoring in the Grade A area should be carried out near the crimping station in filling rooms to demonstrate that the hydraulic crimping does not present a particulate risk to the product.

Diagram 1 – Non-viable particulate monitoring positions for Filling Rooms

8. Production Records

Validation will maintain a schedule of all required media fill operations and submit the required schedule on an annual basis to the production scheduling department.

The scheduling department will schedule the required trial to be conducted and production will request documentation from the documentation department.

The execution of the trial is performed through the production record. The production record should give detailed instructions on how to perform the simulation. All interventions and stoppages must be recorded as the type of intervention, time it occurred and duration.

Each production record will be identified by a unique validation identification number that will be issued from Validation department.

The following essential information must also be recorded in the batch and or media fill records:

– Identification and description of the line, room and process to be simulated

– Type of container/closure combination used, including Lot numbers and all sterilization run numbers

– Line conditions – speed and settings

– Number of containers to be filled, fill volume and actual number filled – the containers should be sequentially numbered (at least in tray order)

– List of the proposed and actual interventions (planned and unplanned), holds and stoppages during set up, operation and close down (this may best be achieved by a line log.)

– List of all personnel participating – including new and support personnel, and activities (e.g. aseptic set ups) or responsibilities

– Media used – batch number, records of preparation and sterilization and quality control

– Environmental monitoring planned and actually performed including all records of results

– Incubator identification, incubation conditions, time for incubation, inspection dates and identity of inspectors

– Agreement between Quality Assurance and Production management on the worst case events.

Reconciliation requirements of process simulation units should be equivalent to the requirement for production. A range between 98-102% reconciliation/accountability of all filled units should be the target. Any excursion must be investigated and documented, however a variance is not an automatic invalidation of a process simulation test.

9. Duration of the Run and Number of Containers to be Filled

9.1 Duration

The duration of the run should simulate the expected time for routine manufacture and should include all production shifts. If the normal dispensing occurs over more than one shift, then the media fill must also occur over the subsequent shift.

The duration of the process simulation should be dictated by the time needed to fill the required number of units (i.e. 5001 units) under worst-case conditions (e.g., lowest or highest filling speed). The process simulation should be of sufficient duration to ensure that the necessary number of units and interventions (planned and unplanned) are included. 

9.2 Size of runs

The number of units in the media fill should be large enough to yield a high probability of detecting low incidences of contamination. 

A minimum of 5001 containers, should be included in any one simulation using Filling Machine FM01. (See Section 13.2 – Acceptance Criteria and Responses).

A minimum of 5001 containers, should be included in any one simulation using Filling Machine FM02. (See Section 13.3 – Acceptance Criteria and Responses).

9.3 Filling

At the beginning of each fill day, flush filling line by filling media into pillow packs until minimum of 20 litres of media is filled. These packs should be placed in a separate container and must be incubated with the main packs in the same conditions and will be included in the final count of units filled during this process simulation run.

During the filling operation, any unit that lack integrity should be rejected and reason for rejection clearly documented (e.g. pack number, reason for being discarded).

10. Simulation Conditions

The design of initial Media Fill must consider all line interventions and include worst case to demonstrate that the manufacturing process is capable of excluding contamination from the sterilized product. The validation may for example include slower line speed, routine interventions, personnel breaks, simulated equipment adjustment, maximum number of processing personnel in attendance, cleaning operations etc.

All planned interventions which would occur during normal filling must be simulated for every fill.

The simulated worst case conditions should be described in the media fill production records.

10.1 Aseptic Processing Operators and Support Personnel

All personnel who are authorized to enter the aseptic processing room during manufacturing, including fill operators, new fill operators under training and maintenance personnel, should participate in a media fill at least once a year. Participation should be consistent with the nature of each operator’s duties during routine production. Names of all personnel in attendance should be clearly documented. Maximum two (2) personnel are allowed in each fill room during in-operation conditions. Names of the personnel permitted (qualified) to enter the filling rooms are displayed on Form-840 “List of Permitted Personnel to Enter Filling Clean Rooms” located on the filling suite doors.

10.2 Procedures

Wherever possible, use materials, components and closures that have been stored in the area for the maximum allowed time.  Alternatively, provide a reference to the validation of the maximum storage times.

Fill the smallest unit at the fastest speed (handling difficulty).

Fill the largest fill at the slowest operating speed (maximum exposure).

Program the run over the maximum nominated holding and dispensing times allowed. If necessary, suspend filling during the run to ensure the equipment and filter systems are exposed for the maximum time.

Introduce “worst case” manipulations and interventions per the Media Fill SOP and Batch Record Form. Document the planned manipulations and interventions and record the actual activities.

Fill volume should be approximately 50% of the container capacity and all packs and containers should be inverted ensuring that there is sufficient wetting of all internal surfaces of the container / closure. 

Note: Ensure that this does not significantly increase the rate of filling and hence minimize the container exposure time.

10.3 Equipment – Maintenance and Breakdown Simulation

Simulate adjustments to equipment expected to be conducted during routine operations.

Adjust equipment, conveyor tracks, stopper and bottle tracks etc.

Simulate clean ups, stoppages and line clearances.

Simulate line intervention by gowning qualified maintenance staff.

10.4 Environmental Monitoring

Routine environmental monitoring will be conducted as per normal aseptic filling requirements. This will involve both viable and non-viable monitoring through out the entire filling process.

Full screen microbiological environmental monitoring and physical monitoring should be conducted during the media fill, both in the Grade A and B areas. Monitoring shall be conducted during set up (e.g. continuous settle plates are exposed) as well as during operation until the completion of the fill.  Post fill monitoring of product contact surfaces is required.

Non-viable particle monitoring shall be performed throughout filling operations with the Grade A filling zone.

All personnel involved in the media fill should be monitored for bioburden (prints during processing and gowns on exiting the room for each session).

Particle monitoring of both viable and non-viable particles will be carried out as defined in environment monitoring procedures.

11. Media

The most common media used is Tryptone Soy Broth for aerobic process simulations.

The media must be capable of supporting growth – this must be demonstrated by growth promotion testing using either EP or USP methods. The units of media used for growth promotion testing must be subjected to the same processing conditions as the trial units.

Growth promotion samples should be selected randomly from the filling line ensuring that samples are taken from the start, middle and end of the filling run and upon completion of 14 day incubation.

The media should be formulated and sterilized by using the same method use in production. All routine procedures used in the manufacture should be performed e.g.. sampling, filter integrity testing etc.

Sterilized filter hold times before filling should mimic the maximum bulk product hold time allowed.

Any spillage of media in the clean rooms or critical space must be cleaned up and disinfected as soon as practical to prevent any residual media supporting microbial growth.

12. Inspection and Incubation of Filled Containers

The containers should not be completely filled with medium in order to provide sufficient oxygen for the growth of obligate aerobes. Similarly, containers should not be overlaid with inert gases even though the product may be (unless trials to detect strict anaerobes are being conducted).

Containers should be subjected to a routine inspection process and examined for physical damage before incubation.

Any units that will be discarded in routine production runs should be discarded.

The reason for discarding units should be clearly documented. A written justification for excluding vials from the trial batch must be prepared as part of the Validation Report.  Rejected units must be checked and verified by the Microbiology Department and reconciled.

Prior to incubation, the containers with the microbiological growth medium should be inverted or otherwise manipulated to ensure that all internal surfaces, including the internal surface of the closure, are thoroughly wetted by the medium. All filled units should be sufficiently manipulated to assure the contact of all internal surfaces by the growth media prior to incubation. Such manipulations should be documented. This inversion of test units should be sufficient to capture any microorganisms present on the upper inner surfaces of the container/closure system.

The incubation temperature chosen should be based upon its ability to recover organisms normally found in the environment or in the product bioburden. A two-temperature incubation schedule should be used. The first stage is to incubate at 20-25°C for a minimum of 7 days followed immediately by incubation at a higher temperature range at 30-35°C for a total minimum incubation time of 14 days. However if the circumstances do not allow, higher temperature range can be used as first stage followed immediately by a lower temperature range. Thirty (30) units must be inspected for signs of microbial contamination prior to incubating at the secondary temperature range.

Data should be available to show the suitability of the selected incubation temperature to support the growth of standard USP ATTC strains and of typical environmental and pre-sterilization bioburden isolates.

The selected temperature should be monitored continuously during the incubation period.

100% visual inspection to be performed by QC personnel after 14 days of incubation. Each unit should be physically inspected for signs of physical or microbial contamination with gentle swirling.

Inspectors should be qualified to inspect media fill containers and supervised by QC personnel. Record of personnel eye sight checks is documented in the Form-845 “Eye Test Record”.

All positive units (units showing evidence of microbial growth) should be subjected to inspection for physical damage to the container/seal and the organism(s) identified to species level.

13. Validation Report of Aseptic Processing

A final report will be produced to summaries the data from the production record and environmental monitoring samples. The following should be documented in the final report:

– Number of units filled and number inspected

– Number of units incubated

– Number of units discarded prior to incubation with reasons for rejection

– Reconciliation

– Number of positive units

– Identification of the contaminant to genus and species, when attainable

– Media growth promotion results

– Environmental monitoring results

Based upon the information in the records a final conclusion regarding the acceptability of the manufacturing process, personnel and facility should be made. This report must be submitted to the Production Manager, Validation Manager and the Manager of Quality Operations for review and approval.  

14. Acceptance Criteria and Responses

14.1 General Requirements

The target is zero positives. Any positive unit indicates a potential sterility assurance problem, regardless of run size. All positive units should be identified and should result in a thorough, documented investigation. If the positive units are indicative of an unacceptable practice (e.g., an incorrect or inappropriate type of intervention) it should be corrected.

Initial qualification requires a minimum of 3 consecutive successful process simulation tests with no positives on any run.  If there are any contaminated units in a single run, investigate the cause; carry out corrective action as appropriate and then repeat the qualification process. 

14.2 PIC’S – Acceptance Criteria

Target:       

– NIL positives in 5000 units

Limits:

– When filling < 5000 units, no contaminated units should be detected.

– One (1) positive results in an investigation, halt to production (of European product) and revalidation;

– When filling 5000 to 10 000 units:

– One (1) contaminated unit should result in an investigation, including   consideration of a repeat media fill;

– Two (2) contaminated units are considered cause for revalidation, following             investigation and halt to production (of European product).

– When filling more than 10 000 units:

– One (1) contaminated unit should result in an investigation;

– Two (2) contaminated units are considered cause for revalidation, following investigation and halt to production (of European product);

The following table lists the number of positive containers allowed based on the number of units incubated:

14.3 Other – Acceptance Criteria

Target:     NIL positives in 5000 units

Limits:      When filling 5000 units

– 1 – 5 positive units results in an investigation (alert).

– Six (6) positive units results in an investigation, halt to production and remediation/ revalidation (failure).

The following table lists the number of positive containers allowed based on the number of units incubated:

The target should be zero growth, but production need not be halted if a contamination rate of less than 0.1% is observed.

14.4 Notification of a Media Fill Out of Specification (OOS)

In the event of media fill positive units exceeding pre defined acceptance criteria the Validation Engineer should immediately notify the QA Manager and provide the following details:

– Affected filling room

– Date of last successful media fill validation (PIC/s and Other)

– Number of units incubated, number of positive units and which acceptance criteria has not been met (PIC/s and / or Other)

In the event of Other acceptance criteria being met but PIC/s acceptance criteria not being met the QA Manager or delegate will place on HOLD all products, both released and under inspection, manufactured for Europe in the affected time frame and still under the control of GMP site.

In the event of both Other and PIC/s acceptance criteria not being met the QA Manager or delegate will place on HOLD all products manufactured in the affected time frame.

Products will remain on hold until an assessment of the impact of the positive units is completed in accordance with the QMS-035 “Deviation Report System”.

15. Cancelling of Process Simulation Runs

Media Fills may be cancelled in the following circumstances:

– Insufficient number of containers filled. However, in this situation, it is necessary to proceed through to the end of the Media Fill.  Another Media Fill must be scheduled to ensure at least 5000 units are filled.

– Failure of the physical conditions in the aseptic processing area (e.g., power outage)

Cancellation must occur before incubation, unless in the case that growth promotion testing has failed – (provided there are no positive units in the process simulation).

Deviations that occur during a Media Fill should be handled in accordance with QMS-035 “Deviation Report System”.

REFERENCE Documents