media fill protocol luisa stoppa manufacture of sterile medicines – advanced workshop for sfda gmp...

Media Fill Protocol

Luisa Stoppa

Manufacture of sterile medicines – Advanced workshop for SFDA GMP inspectors

Nanjing, the People’s Republic of China – 16-20 November 2009

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Manufacture of sterile medicines – Advanced workshop for SFDA GMP inspectorsNanjing, the People’s Republic of China, 16-20 November 2009

Objectives of the hands-onObjectives of the hands-on

To evaluate the aseptic process simulation (media fill or broth fill)

To know the key points in designing a media fill protocol to meet regulatory expectations

To learn how failures have to be evaluated and which consequences they have

To discuss in a team

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aseptic process:

Objectives of the hands-onObjectives of the hands-on

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Principles of media fillPrinciples of media fill

Why the validation of aseptic process is required by pharmaceutical regulations?

A “sterile product” is defined as “free of viable organisms”

As it is not practical examine every unit for confirmation of sterility, all efforts are made to minimise the risk of contamination (finishing, HVAC, pressure differentials, cleaning procedure, monitoring programme)

Despite such measures, contamination is an ever-present danger because aseptic processing is a process being operated in a controlled –but not sterile- environment and sample numbers are too small; so that only gross contamination is likely to be detected

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Pharmaceutical regulations:- Regulations: FDA “guidance for industry, sterile drug products produced by aseptic processing – cGMP” - - EU GMP Part I annex 1 – - PIC/S PI 007-2 “recommendations on the validation of aseptic process”

Although media fills must duplicate aseptic manufacturing conditions, it is not possible for them to be conducted in exactly the same way as the manufacture of a production batch of a pharmaceutical product

What medium? How many units? How long?

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In aseptic processing, the greatest risk comes from the personnel working in the clean room: the operators have to participate in media fills

Which qualifications to the operators need and when can operators be considered qualified?

Environmental monitoring activities are required during aseptic filling operations

Are there additional monitoring activities necessary or not?

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It is usual to include the “worst case” conditions that can occur in production runs

Which kind of interventions have to be considered ?

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washing machine

depirogenating tunnelfilling machine

stoppering machinecapping machine

Hands-on: aseptic liquid filling lineHands-on: aseptic liquid filling line

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Hands-on: Media Fill Protocol Hands-on: Media Fill Protocol

Key elements to be taken into account include:

• Number and frequency of runs• Medium culture (to replace the product)• Number of units filled • Container (vial) size• Fill volume• Line speed (or filling speed)• Duration of fill• Operators shifts• Monitoring activities• Interventions –both routine and non-routine-• Incubation method• Acceptance criteria

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Hands-on: Media Fill Protocol Hands-on: Media Fill Protocol

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Key element: number and frequency of runs

In start up simulation at least three consecutive separate successful runs should be performed (it is recommended they are performed in different days).

For on-going simulation, a routine semi-annual qualification is recommended (one run)

Extraordinary media fill should be performed after all changes to a product or line changes evaluated as a potential danger for the aseptic process.

Media fill Protocol Media fill Protocol

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Key element: medium culture (1/3)

The medium needs to support the growth of a wide variety of microorganisms, including aerobic bacteria, yeasts and moulds (non-selective medium).

Guidance notes the use of Soybean Casein Digest Medium (SCD) also known as tryptone soya broth (TSB).

If the product is being filled in anaerobic conditions, usually in a nitrogen environment, an anaerobic medium is used: fluid thioglycollate medium (FTM).


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The media have to support the growth of microorganisms (growth promotion test). The organisms to be tested are stated by pharmacopoeia (USA, EU, Japan). Moreover the use of one or two of the most common environmental isolates is recommended. Generally at the end of incubation period, some vials (taken from the beginning, at half and at the end of the process) are inoculated with < 100 CFU and incubated for 3 days (bacteria) and 5 days (yeast and mould).


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About prion contamination from component of animal origin found within media, it is important that medium supplier can provide the certification for confirming materials are sourced from “BSE-free” countries.

An alternative approach would be to use medium TSB derived from vegetable materials.

SDC powder is generally subjected to mycoplasmi contamination that are not removed by sterile filtration; in this case previous sterilised powder (with gamma rays) can be used.


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Key element: number of units filled

Number of units filled should reflect the real batch size; it is allowed to fill a lowest number of units provided that the number of units filled is sufficient to reflect the effect of potential operator fatigue and adequately represents the maximum number of interventions.

Some regulations suggest the number of units to be filled in consideration of batch production size.


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Key element: container size (1/2)

The extremes of size containers should be considered.

The largest container (often filled at the lowest speed because of its large fill volume) often has the large opening , so the potential for microbial entry from the environment should be the greatest for that size.

The smallest container (often filled at the highest speed for its lower fill volume) represents the greatest handling difficulty; the smaller container are more fragile and less stable and be more subjected to breakage and jamming in the equipment.


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Key element: container size (2/2)

In the initial qualification two runs might be performed using the largest container and the third run using the smallest container

In routine evaluation of the line, any container should be included in the validation program

Clear containers should be used as a substitute for amber containers to allow visual detection of microbial growth


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Key element: fill volume

The volume of media filled into the containers need not the routine fill volume.

It should be sufficient to contact the container-closure seal surfaces (when the unit is inverted and swirled) and sufficient to allow visual detection of microbial growth post incubation.

Smaller containers should not be over-filled as sufficient air must be available in the container headspace to support the growth of aerobic organisms (generally 25% of volume is not filled).


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Key element: line (or filling) speed

The media fill should address the range of line speeds employed during production. Sometimes more than one line speed should be evaluated.

The speed chosen for each batch during simulation should be justified.

Use of high line speed is justified for manufacturing processes characterized by frequent interventions or a significant degree of manual manipulation.

Use of low speed is justified for manufacturing processes characterized by prolonged exposure of sterile components in the aseptic area.

In the initial validation of a filling line, one run might be performed at the lowest speed and two at the highest speed.

In routine evaluation of the line, the speeds would be alternated


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Key element: duration of fill

Even if the most accurate model would simulate the actual production run, other models cab be justified.

In general media fills should be long enough to include all of the required interventions and stoppage and should reflect the potential operator fatigue: a typical media fill might be at least 3-4 hours long. Ideally a media fill should use more units than are in the product being simulated (for all batches up to 5000 units). For very large batches or long campaigns, some blank units (either empty or water filled) are used to maintain operating conditions during the simulation: this technique can be used to validate processes that may run for several days in order to validate the full length of the longest approved campaign


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Key element: operators shifts

Each operator performing aseptic processes are requested to participate in media fill.

Set-up and line operators should be part of not less than one process simulation per year. Operators such as line mechanics and environmental samplers should be managed in a similar manner.

A maximum number of personnel present in the aseptic processing room should be established.

When a firm operates on multiple shifts, the second and third shift should be included in the media fill programme.

In case of manual operations (filling), each line operator should participate into all three initial validation runs and at least one run in re-validation (every six months)


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Key element: environmental monitoring activities

There are regulatory and pharmacopoeia references that states the microbial conditions.

Air sampling using either active and passive sampling methods should be performed during the execution of the process. Surface sampling is best performed at the end of aseptic process. Also personnel should be monitored

Microbiological monitoring (air, surfaces, personnel) and particle monitoring should be performed during media fill employing the same procedure in force

Sometimes the number of sampling locations might be increased respect the routine procedure


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Key element: interventions –both routine and non-routine-

The interventions should simulate what occurs in a production run; media fill records should document all interventions performed and the number of units removed.

Routine interventions: aseptic line set-up in which sterilised parts are removed from protective materials and installed is a potential danger; it is common to identify the first containers filled as they may be more indicative of potential problem with aseptic assembly.

Other routine interventions: stoppers bowl feeding, remove fallen vials, remove jam stoppers, operators breaks, gloves change, environmental monitoring.

Non routine interventions (occur randomly): glass breakage, change / reset of filling needles, interventions on weight adjustments, sensor failure, rail adjustments.


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Key element: incubation methods

Any filled units should be inspected prior to incubation; any defects that compromise the container closure or non-integral units are rejected and documented.

Divergence in industry practice: incubation is performed for 14 days at 20-35°C (+/- 2,5°C): it is performed for 7 days at 20-25°C and further 7 days at 30-35°C; it is performed for 7 days at 30-35°C and then move the filled containers to 20-25°C

The lack of agreement suggest that the selection of incubation conditions employed.

Units are incubated in an inverted position for the first half of the incubation period and then returned to an upright position for the remainder.


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Key element: acceptance criteria

The target should be zero growth but a contamination rate less than 0,10% with 95% confidence level is acceptable (approx. 1 contaminated unit in 5000 filled units).

FDA and PDA agree that the target should be zero contaminated units regardless of size of run

It is important to note that “invalidation” of a media fill run should be a rare occurrence

Each failure should be investigated


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Key element: acceptance criteria


Filled units per run

Contaminated units permitted (action level)

30000

47501

63002

77503

91504

105105

118406

131507

144308

157109

1696010

The table indicates the maximum permitted number of contaminated units per various Media Fill “run sizes” to indicate a 0,10% contamination limit with a 95% confidence level

Health Canada

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ContaminationContamination

The root cause of a failure (contamination), or at least the most probable one, must be identified

It is important to be able to isolate and identify (to species level) the microorganisms

An appropriate corrective action / preventive action plan must be implemented

The impact of the failure on product lots already released (if any) must be evaluated

After the corrective actions have been implemented, a new media fill study is performed to confirm their efficacy

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Filling chronology (time or units)


Contamination rate increasing during filling

This could indicate a contamination originated in the liquid media path (i.e. wrong aseptic connection to the media tank, contamination in the recirculation loop). It is important to keep under aseptic conditions the media bulk tank at the end of filling, while waiting the media fill results, for verifying this hypothesis

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Contamination rate decreasing during filling

This could be indicative of a contamination occurred during the line set-up (stopper bowl / guides, dosing pumps / needles), partially or totally “washed out” during filling.

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Contamination spike during filling

A spike (some contaminated units in a short time interval could be linked to a not correctly performed critical intervention during filling. For this reason it is crucial to have an accurate time traceability of the filled units and the interventions performed (videotaping the filling operations can be helpful)

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Contamination spike followed by an increase

Similar to the previous example, but in this case the wrong intervention could have affected the liquid path/loop (i.e. change of media tank, replacement of a pump / needle)

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Isolated events (few contaminated units per run)

Unfortunately, this occurrence is the most common and also the most difficult to be investigated. If repeated on multiple runs, it could indicate that this profile is representative of the “background noise” of the process, so requiring radical actions (i.e. change of garments/dressing code, disinfectants, air flow patterns)

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ConclusionConclusion

Even if all media fills are negative in a Company, it does not necessarily indicate that no products were ever possibly contaminated because the frequency requirement of media fill testing is so minimal that it is not statistically significant

A robust media fill programme is a necessary step to validate process

Media fill testing is just one part of a necessary overall quality assurance program (HVAC qualification, cleaning procedure, sanitization, personnel training, …)

36 |Workshop on WHO prequalification requirements for reproductive health medicines, Jakarta, October 2009

[email protected]

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Reference documentsReference documents

WHO GMP guidelines – Technical Report series n. 937

EU GMP guidelines, Part I annex 1 & 15

ICH Q7A or EU GMP Part II chapter 13

FDA Guidance for Industry: sterile drug products produced by aseptic processing cGMP

PIC/S Recommendations PI 007-5

USP <797> ‘media fill testing’ / <71> ‘growth promotion test’

EP 2.6.1 ‘sterility’

media fill protocol luisa stoppa manufacture of sterile medicines – advanced workshop for sfda gmp...

Documents

sfda gmp inspectors

peoples republic of

sterile product

principles of media

sterile environment

media fills

validation of aseptic

eu gmp