PROCESS SIMULATION TEST

IN INJECTABLE
VALIDATION

(MEDIA FILL)

DR. AJMAL NASIR

Director Technical

BF Biosciences ltd
LAHORE

INJECTABLES
Injections must be sterile and as per definition shall be Free
from Organisms,
Sterility is One of the most examined areas of pharmaceutical
and medical device manufacturing .
The favored method to assure sterility is Terminal Sterilization
( Autoclaving or Irradiation etc).
Materials sensitive to heat can not be terminally sterilized.

Hence ASEPTIC PROCESS and ASEPTIC FILLING .

ASEPTIC PROCESSING?
The Production Of Sterile Drug Products By Bringing
Together The Product, Container & Closure sterilized
through different Sterilization Methods Separately, And
Assembled In An Extremely High Quality Environment
By Skilled Personnel Using The Right Equipment And
Techniques.

ASEPTIC FILLING
Aseptic filling is an Aseptic Process
that requires the close coordination
and complex interaction between :
Personnel,
Sterilized Product,
The Fill/Finish Equipment System,
Cleanroom / Support Facilities,
Sterilized Filling Components.

HISTORICAL BACKGROUND
SEPTEMBER 2004:

After 17 years FDA replaced older guidelines In September

2004, with an updated document,

Guidance for Industry - Sterile Drug Products

Produced by Aseptic Processing - Current Good
Manufacturing Practices.
Europe made some changes to the :
'EC Guide to Good Manufacturing Practice, Annex
I, Manufacture of Sterile Medicinal
Products
These documents initiated a thought process at various
forums.
More focus on sterility Assurance

Aseptic Process Simulations faced significant

changes as compared to other parts .

ASEPTIC FILLING

Aseptic processing is a process being operated in a controlled

but NOT STERILE ENVIRONMENT .

Probability of non-sterility cannot be calculated.

Involves a great deal of process control, with sensitive handling

of components until they are sealed within their final
containers.

All efforts are made to minimize the risk of contamination.

The industry works to a recognized, accepted contamination
levels,
Probability of viable contamination is recognized and calculated

To Ensure The Sterility Of Products

Purporting To Be Sterile, Sterilization,
Aseptic Filling And Closing Operations Must
Be Adequately Validated ( 211.113).
The goal of even the most effective
sterilization processes can be defeated if the
sterilized elements of a product (the drug
formulation, the container, and the closure)
are brought together under conditions that
contaminate any of those elements.

STERILITY ASSURANCE CHALLENGE

Contamination Is An Ever-Present Threat .
Materials , surfaces and inefficiencies in air filtration may
carry organisms.
Potential viable contamination comes from operators
running the process.
Sterility testing Sample numbers are too small to detect
low level contamination.
Only GROSS contamination is likely to be detected.
Sterility Assurance need another Tool to Guarantee .
Process Simulations (Media Fills) supported by
environmental monitoring and other related processes
demonstrates control of the process to the industry
standard for allowable contamination levels.

STERILITY TEST VERSUS MEDIA FILL

Sterility Test

Media Fill

Guidance
Documents

FDA Guidance for Industry

Code of Federal
Regulations
Multiple Pharmacopeia

FDA Guidance for Industry

EU Guide to Good
Manufacturing Practice
Revision to Annex 1

Sample size/Lot

Maximum 20

All

Media

Soybean-Casein Digest
Medium (TSB)
Fluid Thioglycollate Medium
(FTM)

Tryptic Soya Broth(SoybeanCasein Digest Medium (TSB))

(FTM in case anaerobes
detected in sterility Failure)

Incubation
conditions

TSB 14 days @ 20-25C

FTM 14 days @ 30-35C

TSB 7 days @ 20-25C

TSB 7 days @ 30-35C

Growth promotion

Yes

Yes

Test method

Destructive:
Sample contents transferred

Non-destructive:
Integral container

Sensitivity

False positives
Detects high level sterility
failure

No false positives
Detects single vial failure

PROCESS SIMULATION

Commonly Known as MEDIA FILL.

Carried out to demonstrate that the process is capable
and robust enough to produce products with highest
degree of Sterility Assurance.

Media fills utilize culture media in place of product to

evaluate contamination levels and validate Aseptic
Processing .
Media Fill Program Provides Evaluation Of Multiple
Systems.
The media fill should be designed to mimic, as closely as
possible to the aseptic processes used in practice.

Process Simulation is ONE element in the

validation of an aseptic process.

DEFINITION
The "Media Fill", or "Broth Fill", technique, is one
in which a liquid microbiological nutrient growth
medium is prepared and filled in a simulation of a
normal manufacturing operation. The nutrient
medium processed and handled in a manner which
simulates the "normal" manufacturing process as
closely as possible with the same exposure to
possible contamination (from operators,
environment, equipment and surfaces) as would
occur during routine manufacture. The sealed
containers of medium thus produced are then
incubated under prescribed conditions and
examined for evidence of microbial growth, and
thus of an indication of the level of contaminated
units produced. Health Canada

PROCESS SIMULATION TEST

Test shall be designed to accurately represent the
aseptic process.
Following Parameters shall be addressed in Test
Design:
STUDY DESIGN
MEDIA FILL DOCUMENT
FREQUENCY
MEDIA FILL RUN SIZE & SPEED
ENVIRONMENTAL CONTROL
MEDIA QUALITY
INCUBATION & FILLED UNITS ANALYSIS
RESULTS INTERPRETATION

PRE - REQUISITES
Validated manufacturing process.
Validated equipment.
Validated sterilization cycles.
Trained and qualified Personnel.
All materials suitable for sterile manufacturing /
processing.

STUDY DESIGN
Shall cover all possible contamination Risks.
Shall closely simulate process.
Worst case scenarios incorporated .
Shall contain:

Longest possible runs

Normal and non routine interventions.
Lyophilization (if applicable).
Assembling of equipment ( start up and during processing)
Number of workers / activities
Aseptic additions ( container , closure Charging, transfers)
Gown changes, shift Changes,
Aseptic sampling
Line speed
Weight checks
Container closure systems

MEDIA FILL DOCUMENT

A written approved Batch Record.
Same vigilance as In Routine Run.
Clear Definition of Rationales.
A separate Document approval for each Run.
Detailed instructions for the test .
Complete list of planned interventions

LIST OF PERMITTED INTERVENTIONS

DURINGFILLING
LIST
PERMITTED
INTERVENTIONS
LIST OF
OF PERMITTED
INTERVENTIONS
DURINGFILLING
DURINGFILLING
Adjustment of weight in the dosing wheel.
Adjustment of Stopper holding spring.
Transfer of liquid from container to hopper.
Transfer of stoppers from bag to hopper.
Picking un stoppered vials from the out feed back to conveyor for
stoppering.
Adjustment of separator.
Picking up of fallen vials from turn table.
Cleaning of machine with vacuum cleaner.
Adjustment of stoppering channel height.
Adjustment of stoppering pressure rollers.

LIST OF PERMITTED INTERVENTIONS

DURINGFILLING
LIST
OF PERMITTED

INTERVENTIONS DURINGFILLING
Replacing of a piston from the wheel.
Adjustment of turn table over load sensor.
Lifting and closing of the LAF cabinet door.
Checking the weight in balance.

Adjustment of dosing air.

Cleaning of vacuum pot.

Pushing stopper in the channel by forceps.

Power interruption (samples should be marked)

Number of personnel in sterile filling area (8 persons).

Multiple dosing of the liquid.
Running the machine at a slower speed than actual production
run.
Covering the working shift usually 12 hours and shift Change
over
Entry of maintenance person for repairing of M/C

FREQUENCY
Initially at least 3 consecutive, separate, successful, runs on
each line.
Routine semi Annual Qualification for each Line .
All authorized personnel for aseptic processing including
technicians and maintenance personnel, should participate in a
media fill at least once a year.
Participation should be consistent with the nature of each
operators duties during routine production.
Each Change shall be controlled through written Change
control.
Changes and events with potential to effect sterility should be
assessed through additional Media Fill.

FREQUENCY (Contd.)
CAUSES OF FOR REVALIDATION OF THE
SYSTEM:
Facility and equipment modifications,
Line configuration changes,
Significant changes in personnel,
Anomalies in environmental testing results,
Container closure system changes,
Extended shutdowns,
End product sterility testing showing contaminated
products.

MEDIA FILL RUN SIZE & SPEED

Run size should be sufficient to provide a high probability of detecting a low
incidence of microbial contamination.
A generally acceptable starting point for run size is in the range of 5,000 to
10,000 units.
For operations with production sizes under 5,000, the number of media
filled units should at least equal the maximum batch size made on the
processing line.

The media fill program should adequately address the range of line
speeds employed during production.
Each media fill run should evaluate a single line speed, and the speed
chosen should be justified.
For example, use of high line speed is often most appropriate in the
evaluation of manufacturing processes characterized by frequent
interventions or a significant degree of manual manipulation.

Use of slow line speed is generally appropriate for evaluating

manufacturing processes with prolonged exposure of the sterile
drug product and containers/closures in the aseptic area.

ENVIRONMENTAL CONTROL
Media fills should be adequately representative of Actual
Process Conditions
Stressful conditions Permitted as per SOPs shall be
simulated.(e.g., maximum number of personnel present
and elevated activity level), .
Challenges shall be supportive to the study design.
Stressful conditions do not include artificially created
environmental extremes, such as reconfiguration of HVAC
systems to operate at worst-case limits .

MEDIA QUALITY
Generally soybean casein digest medium, is used.
Anaerobic growth media (e.g., fluid thioglycollate medium)
should be considered in special circumstances.
Media must Promote promote growth of gram-positive, gramnegative bacteria, yeast and mold (e.g., USP indicator
organisms).
Environmental monitoring and sterility test isolates can be
substituted (as appropriate) or added to the growth promotion
challenge.
Growth promotion units should be inoculated with a <100 CFU
challenge.
In case of failure of Growth Promotion test the origin of any
contamination found during the simulation should nonetheless
be investigated and the media fill promptly repeated.
Each unit should be filled with enough quantity and type of
microbial growth medium to contact the inner container closure
surfaces (when the unit is inverted or thoroughly swirled) and
permit visual detection of microbial growth.

INCUBATION & FILLED UNITS ANALYSIS

Incubation temperature should be at no time be
outside the range of 20-35oC.
Incubation temperature should be maintained
within +2.5oC of the target temperature.
Incubation time should not be less than 14 days.
If two temperatures are used for the incubation
of the media filled units, the units should be
incubated for at least 7 days at each temperature
(starting with the lower temperature).
TSB 7 days @ 20-25C
TSB 7 days @ 30-35C

INCUBATION & FILLED UNITS ANALYSIS

Filled units to be inspected by Trained staff only
under supervision of Qualified Microbiologist.
Each Positive unit must be brought in notice of
QC Microbiologist
Only integral units shall be incubated.
All intervention units must be recorded in the
Media fill document separately .

RESULTS & INTERPRETATION

When filling fewer than 5000 units, no contaminated
units should be detected.
One (1) contaminated unit is considered cause for
revalidation, following an investigation.

When filling from 5,000 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.

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.
For any run size, intermittent incidents of microbial contamination in
media filled runs can be indicative of a persistent low-level
contamination problem and that should be investigated.

VARIATIONS FOR
DIFFERENT DOSAGE
FORMS

STERILE POWDERS

Two possible procedures:

Fill the chosen inert powder into the containers (e.g.
ampoules/vials) which are already filled with sterile liquid
medium.
Fill the inert powder first, and then add the sterile liquid
medium.
Commonly sterile PEG 8000 is used as powder .
In both these variations, a powder fill is simulated, but an
additional, non-routine step (i.e. the filling of the liquid
growth medium) is involved.

SUSPENSION PRODUCTS:
Simulate the entire normal process as closely
as possible,
Use sterile inert powder in place of the
normal powder product.
Micronize etc. (if part of the normal process)
and form suspension.
Use sterile liquid growth medium in place of
the normal liquid phase of the suspension
product.
Fill as normal and incubate.

FREEZE-DRIED PRODUCT:
Simulate the entire normal process i.e.
preparation of bulk solution,
filling of solution,
loading of freeze-dryer,
running of freeze-drying cycle,
sealing/closing of containers,
inspection.
Actual freeze-drying of the medium solution is
not practicable, but exposure, holding times in
the freeze dryer should be as normal.
.

SEMI-SOLID PRODUCTS

(e.g. sterile ointments and creams):

Simulate the normal process cycle as

closely as possible,
Fill a sterile liquid growth medium made
to similar consistency as the normal
product by the addition of agar
(approximately 4 g. per liter) or
carboxymethylcellulose

DOs AND DON'Ts

Validate Maximum number of workers in the area.
Run line at slower speed to give maximum exposure to
open containers.
Do planned interventions
Validate all sizes of containers specially with large mouth
size.
Loading of containers and stoppers shall be an essential
part of study.
Don not use inert gases to in the media fill run. Instead
use filtered air.

IT PAYS IN THE LONG RUN

MORE CONFIDENCE
REGULATORY APPROVALS
NO REJECTION.

THANKS