Process Simulation (Media Fill) Test

Company Name:: Version No.
Humanwell Pharmaceutical Ethiopia PLC 1
Document No: Title: Page No.

SOP/SL/013 Process Simulation (Media Fill) Test Page 1 of 15
Effective Date Revision Date
Process Simulation (Media Fill) Test
Prepared by： X X
Department Name Date
Approve by： Y Y
Authorized by： Z Z

1. Purpose
This procedure describes methods and procedures for the conduct of process simulation tests
of sterile products, including media fill procedures, media selection, fill volume, incubation,
time and temperature, inspection of filled volume, documentation, interpretation of results,
and possible corrective action required where its main purposes are to
o Demonstrate control over the aseptic operations
o Qualify aseptic processing personnel
o Qualify aseptic techniques
o Comply with cGMP requirements
2. Scope
This procedure is applicable to - sterile products dosage forms in Humanwell

Pharmaceutical Ethiopia PLC
3. Validity
This SOP is valid only until next revision date and if it bears control seal.
4. Responsibility
It is the responsibility of all concerned departmental managers to follow the procedure. The
quality assurance (QA) manager is responsible for SOP compliance.
5. Material and Equipment
Media to be used
 Soybean casein digest broth shall be used for normal media runs, but thioglycolate
broth shall be used for the detection of anaerobic organisms, especially when a filtered
nitrogen purge is used to ensure anaerobic conditions. Prepare the media according to

manufacturing direction. Adjust pH; the medium is sterilized by 0.2 . Sterile

filtration technique is used.
6. Procedure
6.1. Manufacturing methods/process simulation
The manufacturing methods/process simulation include the following steps (see
individual MFM [manufacturing formula and method] for media fill run):
 Filling equipment preparation
 Media preparation
 Sterilization
 Sterile filtration
 Washing of commodity
depyrogenation
 Vial stoppering, unloading, and scaling
 Filling
 Media addition
The filling operation of nonlyophilized product shall be completed in 12 hours (two

shifts) as follows:
Table 1 Nonlyophilized Products

Runs Filling 12 Bre
Hours ak
Run I Shift 1/day 1 Shift 2/day 3–4
1
Run II Shift 1/day 2 Shift 2/day 3–4
2
Run III Shift 1/day 3 Shift 2/day 3–4
3
6.2. Verification of medium sterility

 Perform sterility test for the bulk media after filtration and bioburden count before
filteration.
 Aseptically, QA inspector shall take samples (each 100 ml of media) at the beginning
each 2-hour period and near the end of the process.
6.3. Filling operations

o The containers and closures shall be cleaned and sterilized per current SOPs.
o The filling machine is operated at the predetermined and validated fill rate for
the container size utilized.
o During filling, all major and minor interventions will be performed as defined
in approved manufacturing direction.
o Power shutdown is to be simulated by stopping the laminar flow unit and the
main air handling unit (AHU) for 30 seconds.
o The containers are sealed and the medium-filled units are collected in
sequentially numbered trays or boxes (notified to the filling time).
o The filled units should be briefly inverted and swirled after filling to assure
closure contact with the medium.
o Increase the size of filling crew to more than the number necessary to fill the
batch (maximum of three people).
o All routine activities that take place on the filling line should be a part of the
test, i.e., weight adjustments, replacement of containers, addition of
components, change of filling needle, installation of machine parts,
installation of sterile filter, etc.
6.4. Powders
Selection of placebo powder. The chosen material must be easily sterilizable,

dispersible, or dissolvable in the chosen medium. The principal sterile placebo
materials (irradiated in a final container) are irradiated lactose, mannitol,
polyethylene glycol 6000, and sodium chloride. The material should pass the
solubility testing at the desired concentration with suitable amount and time of
agitation
6.4.1. Compounding operations


A quantity of an appropriate sterilized placebo powder is blended with sterile

excipients prior to filling (if needed), in a manner similar to the production
process being simulated.
The medium is passed through the run as though it were an actual product batch,
and all routine procedures used in manufacture of a batch are performed.
Once the medium has been processed, it is held for a period of time at least equal
to that for aseptically produced materials.
Any of aseptic manipulations performed during and at the end of the hold period
should be simulated hold times and product recalculation.
6.4.2. Filling operations

 The containers, and closures are cleaned and sterilized using SOPs.
 The filling machine is operated at the predetermined fill rate for the container
size utilized, as well as at the fastest speed (maximization) and the slowest
speed (handling difficulty).
 The containers are sealed and the medium-filled units are collected in
sequentially numbered trays or boxes (notified to the filling time).
 All routine activities that take place on the filling line should be a part of the
test, i.e., weight adjustments, replenishment of containers, addition of
components, change of filling agar, installation of machine parts, etc.
 Increase the size of filling crew to more than the number necessary to fill the
batch.
6.4.3. Powder reconstitution
Before incubation of the vials, powder showed reconstituted with adapted media
(TSB or thioglycolate broth) using aseptic technique under laminar flow. The

reconstitution volume is according to the volume described in original formula. A

strict environmental monitoring should be followed through this step.
6.4.3.1. Media-filled vial/ampoule analysis

Media-filled vials will be incubated in an upright position at two
temperatures for a minimum of 14 days. The vials/ampoules will be
inspected for growth at two separate times, once at a minimum of 7 days
of incubation at 22.5±2.5oC and again at the end of the 14 days of
incubation at 32.5 ± 2.5oC. Record of these inspections will be
documented in QC notebook. Any vials found positive for growth will be
investigated as detailed in the acceptance criteria section of the SOP.
6.4.3.2. Growth promotion of media-filled vials
 Growth promotion of the media-filled vials will be performed on
representative vials of each media-fill run after the completion of
the 14-day incubation period per instructions given in standard test
method (provide number). The media shall be challenged after
being subjected to the same condition of the other media fill units.
 Growth promotion ability of the medium in final filled containers
must be demonstrated using ten randomly selected containers for
each challenge organism (more than 100 microorganisms per test
container).
 The units used for growth testing must be subject to the same
processing steps (e.g., cleaning, depyrogenation, sterilization,
filtration, filling, lyophilization, reconstitution) up to the point at
which they are placed into incubation.
 In case of growth-promotion controls, growth must be observed in
at least 95% of the test containers for all the challenge
microorganisms.

 If no growth is observed in all of the ten challenged containers, a

second-stage test must be conducted to rule out the laboratory. In
the second-stage test, both containers must support growth.
6.4.3.3. Negative control
 Media from the bulk media container is incubated at 32.5 ± 2.5°C
for 14 days as negative control for liquid media fill to check its
sterility.
 500 containers filled only with the reconstitution medium serve as
negative control for powder fill run.No growth should be
demonstrated in negative controls to indicate the acceptability of
media fill. If growth is observed, the discrepancy should be
investigated and documented.
6.4.3.4. Incubation and inspection of media-filled units
o Leaking or damaged media-fill evaluation units shall be removed,
and a record made of such removal, following processing and prior
to incubation of the media.
o Media-filled evaluation units shall be incubated for a minimum of
14 days.
o Incubation temperatures shall be appropriate for the specific
growth requirements of microorganisms that are anticipated in the
aseptic filling area. Note: Environmental monitoring data can assist
in identifying the optimum incubation temperatures. Frequently
used temperature ranges for incubation are 22.5 o±2.5oC or 32.5 ±
2.5oC. Media-filled units shall be stored or manipulated to allow
contact of the media with all product contact surfaces in the unit.

o After completion of the incubation period, the media-filled

containers shall be visually inspected for the presence of microbial
growth.
o Microorganisms present in contaminated units shall be identified to
help determine the likely source of the contamination.
Note: Media-filled units should be chronologically identified within the

batch to help identify the time at which a contaminated unit was filled.
6.5. Acceptance Criteria
A minimum of 4800 units will be filled and incubated. For multiple shift fills, at
least 3000 vials/ampoules are to be filled per shift. Table 3 shows the acceptance
criteria for initial performance qualification of an aseptic processing line. Table 4
shows the acceptance criteria for requalification of an aseptic processing line.
Table 5 shows alert and action levels when a 0.1% contamination rate is attained
for large numbers of media-filled units, i.e., when one elects to media-fill more
than 3000 units.
7. Abbreviations
 SOP - Standard Operating Procedure
8. Related Documents /Applicable document/
Related Title
Documents./
SOP/SL/013 /F01 Media Fill Request Form
9. Revision History

Version
Revision Description
No.
1 New

Table 3 Initial Qualification Alert and Action Levels

No. Batch Size Minimum Media Fill Alert Media Fill Action
(No. Units) No. Level Level
Media Fill and Required
Action
4800–7500 Three fills One Two contaminated
contaminated units in a single run
unit in any run: or one each in two
investigate runs: investigate
cause, conduct cause; repeat initial
one additional qualification media
run (repeat fills
performance
qualification if
the additional
run fails)
7501–10,000 Three fills One Two contaminated
cause; conduct cause; repeat initial
one additional qualification media
run (repeat fills
performance
qualification if
the additional
run fails)
10,001+ Three fills One Two contaminated
cause; conduct cause; repeat initial
one addi-tional qualification media
run (repeat fills
performance
qualification if
the additional
run fails)
Notes:Any recovered microorganisms shall be investigated related to the reason for them and their
possible origin.
A sufficient number of units should be media filled to permit most interventions and worst-
case conditions that may occur during production conditions.

Table 4 Periodic Requalification — Media Fill Acceptance Criteria

Production No. Minimum Media-Fill Media-Fill Action Level
Batch Size Med No. Units Alert Level
(No. Units) ia per Media
Fills Fill
500 3 Max. batch
One Two contaminated units in
sizea contaminated a single run: cease
unit in any requalification; investigate
run: cease cause; repeat initial
requalification qualification per Table 3
; investigate
cause; repeat
periodic
requalification
500–2999 1 Max.a As above See Table 3 for maximum
batch size Repeat the action level values
media-fill run
3000 1 3,000 Repeat the See Table 5 for maximum
media-fill run action level values
if the alert
values in
Table 5 are
exceeded
a
Fewer than 3,000 units would be filled in any one run, so the 95% confidence limit table cannot be directly
applied; however, on an empirical accumulative basis, no positive units in each of the individual media-fill
runs would suggest a low contamination level. See Table 5.
Table 5 Alert and Action Levels for Large Numbers of Media-Filled Units
No. Units Alert Levela Action Levelb
in Single (No. Contaminated Units (No. Contaminated Units
Media-Fill Test in a Single Media-Fill Run) in a Single Media-Fill Run)
3,000 Not applicable 1
4,750 1 2
6,300 1 3
7,760 1 4
9,160 1 5
10,520 2 6
11,850 2 7
13,150 3 8
14,440 3 9
15,710 4 10
16,970 4 11
a
This alert level is based on selection of a 0.05% contamination rate.

b
Contamination rate of >0.1% at a 95% confidence level.
9.1. Media-Fill Runs Exceeding Action Levels

9.1.1. Investigation
When media-fill action levels are exceeded, an investigation shall be conducted and
documented regarding the cause. If action levels are exceeded, there shall be a prompt
review of all appropriate records relating to aseptic production between the current media
fill and the last successful one.
The investigation should include, but not be limited to, consideration of the following:
 Microbial environmental monitoring data

 Particulate monitoring data
 Personnel monitoring data (finger impressions, etc.)
 Sterilization cycles for media, commodities, and equipment
 HEPA filter evaluation (airborne particulate levels, smoke-challenge testing,
velocity measurements, etc.)
 Room airflow patterns and pressures
 Operator technique and training
 Unusual events that occurred during the media fill
 Storage conditions of sterile commodities
 Identification of contaminants as a clue to the source of the contamination
 Housekeeping procedures and training
 Calibration of sterilization equipment
 Pre- and postfilter integrity test data and/or filter housing assembly
 Product and/or process defects and/or limitation of inspectional processes
 Documented disqualification of samples for obvious reasons prior to final reading
9.1.2. Corrective actions


 Media-fill tests that exceed action levels in Table 3 shall require action.
 Decisions on whether or not to take action against product being held and/or
distributed shall be based upon an evaluation of all the information available and
shall be documented.
 All products produced on a line following the media fill shall be quarantined until a
successful resolution of the media fill has occurred.
 The materials and equipment used for manufacturing this batch will be documented
in the batch record.
9.1.3. Failure investigation and corrective actions
A contaminated container should be examined carefully for any breach in the container
system. All positives (from integral containers) should be identified to at least genus and
to species whenever possible. The identification of contaminant should be compared to
the database of the organisms recently identified. The biochemical profile of the
contaminant can then be compared to that of microorganisms obtained from the sterility
tests and bioburden and environmental monitoring programs, in order to help identify the
potential sources of the contaminant.
 If media-fill contaminant is same as sterility test contaminant: increase media-fill vial

quantities and routine filling environmental monitoring to identify the source of
contamination. Review environmental data obtained during line set up.
 If media-fill contaminant is same as media fills environmental contaminant: increase
routine environmental monitoring to determine whether the contamination potential
exists during routine filling operation.
 If media-fill contaminant is same as routine environmental contaminant: increase
media fill environmental monitoring (in the same location) to confirm the contaminant
source.

 If sterility test contaminant is same as media fill environmental contaminant: increase

routine environmental monitoring (in the same location) and number of media-fill
vials to conform.
 If sterility test contaminant is same as routine environmental contaminant: the sterility
test is voided. Investigate sterility test procedures and room sanitation/sterilization
methods to eliminate cause.
 If media-fill environmental contaminant is same as routine environmental
contaminant: increase the number of media-fill vials in media fill to determine the
product risk potential. Review monitoring technique for possible problem. Review
personnel practices, gowning, sanitation, and sterilization.
 If media-fill environmental contaminant is same as sterility test contaminant and if
routine environmental contaminant is same as sterility test contaminant: check
environmental monitoring methods and techniques closely for problems. Review
personnel practices, gowning, sanitation, and sterilization.
 If the failure repeated represent a potential for product concern: a corrective action
system should be activated. This system should contain provision for the following:
o Critical systems (HVAC, compressed air/gas, water, steam) should be
reviewed for documented changes.
o Calibration records should be checked.
o All HEPA filters in the filling area should be inspected and rectified, if
warranted.
o Training records for all individuals (production, maintenance, cleaning)
involved in the fill should be reviewed to assure proper training was
provided.
o The root cause is assignable; corrective action needs to be taken and
documented.

o If three consecutive runs over action levels occur, a media-fill failure

investigation and corrective action report must be issued.
o For template microbiology laboratory reports refer to Attachment No. 1800.10(A).
10. RECORDS
 Media Fill Request Form
 Media Fill Reporting Form
11. RELATED DOCUMENTS
None
12. Training: Training will be given for QA, Microbiology and Production personnel

Process Simulation (Media Fill) Test

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Company Name:: Version No.

Humanwell Pharmaceutical Ethiopia PLC 1

Document No: Title: Page No.

Effective Date Revision Date