Protocol for validation of Aseptic Media filling Process

1.1 Objective:

To evaluate and know the requirements to design a media fill protocol in compliance with regulatory standards.

1.2 Principle

Every product may be contaminated despite the controlled conditions maintained but in aseptic process they may be in less number.

Pharmaceutical regulations should be followed which are given by FDA as “Guidance for industry, sterile drug products produced by Aseptic Processing – cGMP” and also by EU GMP Part annexure I. PIC/S PI 007-2 also gives “Recommendations on validation of aseptic process”

Even the media fills maintain the aseptic conditions but it cannot be replicated to the fullest.

The personnel during the aseptic processing tend to cause risk by not maintaining hygienic conditions.

Monitoring environment is the crucial activity during the process

Worst case should be identified which may occur during the media filling process

1.3 Media fill Protocol – Key Factors

1.3.1 Frequency and number of runs

Initially at simulation there should be at least three separate successful runs should be done on different days.

When the simulation is ongoing then semiannual qualification should be done on routine basis.

 When product or line changes occur then extraordinary media fill should be done otherwise it may be danger to aseptic process.

1.3.2 Medium culture

The medium should be able to support the growth of variety of microorganisms like bacteria, yeasts, moulds.

When using soybean casein digest medium / tryptone soya broth guidelines should be followed

When filling the product in anaerobic conditions or nitrogen environment then anaerobic medium like fluid thioglycollate is used.

The microorganisms used for the growth promotion test should be in accordance with pharmacopoeia.

At the end of incubation period some of the vials are inoculated with less than 100 cfu and incubated for 5 days for yeast& mould and 3 days for bacteria.

If the medium is of animal origin then the supplier should take certification as ‘BSE free’ from the countries where they are sourced.

Alternatively vegetable origin medium like TSB can be used

SDC powder may be subjected to mycoplasma contamination which can’t be removed by sterile filtration.

1.3.3 Units to be filled

The number of units to be filled should be in compliance with the real batch size and sometimes less number of units can be filled when the number is enough to reflect the maximum number of interventions & impact of potential operator fatigue.

But according to the regulations given the number of units to be filled should be in accordance with the batch production size.

1.3.4 Container size

Larger containers have huge opening which allows for more microbial contamination. On the other hand small containers present difficulty in handling and also are likely to break and jam in the equipment.

Initial qualification should include two runs using larger container and smaller container for third container.

These containers should be included for validation

Containers should be clear rather than amber containers which allow for visually detecting the microorganisms

1.3.5 Fill volume

The volume to be filled should be sufficient so that it allows for the visually detection of any growth of microbes and also the container and the closure seal the surfaces

Small containers shouldn’t be filled up to the brim which may prevent sufficient air to be available to the headspace in the container and this in turn may cause growth of aerobic organism

1.3.6 Filling speed

During the production the line speed should be addressed. The speed chosen should be justified.

For manufacturing process which are characterized by manual manipulation requires high filling speed and manufacturing process which are characterized by exposure of sterile components in aseptic area require low filling speed.

In routine evaluation the speeds may vary and the initial validation of filling may be performed at low speed and two at high speed.

1.3.7 Fill duration

The media fills are to be for long periods typically at 3-4 hrs and must reflect the potential operator fatigue.

Ideally the fill should use more units then in the product being simulated.

Some empty or water filled – blank units are used to maintain operating conditions during large batches or campaigns. This technique can be used to validate processes which run for many days.

1.3.8 Operator Shifts

Every operator who performs aseptic processes participate in the media filling

Set up and line operators should not part of more than one process simulation in a year. The same applies to the environment samplers & line mechanics.

In case of multiple shifts, both second & third shifts should be mentioned in the media fill programme

In manual operations every line operator should participate in all initial validation runs and should run at least one re validation

1.3.9 Environmental Monitoring

Microbial conditions should be as stated by the regulatory authorities and pharmacopeia

Sampling of air should be by either active or passive methods and should be done during process execution

The personnel should be monitored for their hygienic conditions

Monitoring of particles and microorganisms should be done during media filling process by using the same methods in force

1.3.10 Routine and non-routine interventions

Interventions should indicate what happens in production run. All interventions should be documented.

Routine interventions identify the first containers which indicate the problem with aseptic assembly.

Non routine interventions indicate the interventions which occur randomly like sensor failure, weight and rail adjustments, jam stoppers, operator break, glove changes and environmental monitoring

1.3.11 Incubation methods

The filled units should be examined before the incubation and the defects which hinder the container closure or any non-integral units should be rejected & documented.

Incubation should be performed at 20-30^oC for 14 days and at 20-25^oC for 7 days and for another 7 days at 30-35^oC. Later it is moved to the filled containers at 20-25^oC.

The units are incubated in inverted position during the first half of incubation period and turned upside down for remaining period.

1.3.12 Acceptance criteria

Even the aim is for zero growth but still the contamination should be less than 0, 10% with confidence level of 95% is accepted

PDA and FDA agree the aim for zero contaminated units excluding the size of the run

Any failure should be investigated thoroughly

The following table indicates maximum permitted number of contaminated units per various media filling run sizes which indicate the acceptable level as given above

Media filling testing is part of quality assurance and a robust media fill programme is necessary to validate the process

Validation of aseptic media filling process, acceptance criteria, incubation methods, environmental monitoring, fill duration, filling speed, fill volume