A
Seminar
On
Validation of Integrated line by Media Fill
Test
INDEX
 Introduction
• What is media fill test ?
• Principle of media fill test
• Protocol
 Validation
• Objectives
• Scope
• Responsibilities
• Pre-requisites
• Equipment/ system description
• Study design
• Procedure
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13 April 2015
Media Fill Test
What is Media Fill Test ?
Aseptic media fill test is used to quantify the aseptic technique of
compounding personnel or processes and to insure that the
processes used are able to produce a sterile product without
microbial contamination
During this test microbiological growth medium such as Soybean
Casein Digest Medium (SCDM) is substituted for the actual drug
product to simulate admixture compounding
The final container is then incubated and checked for turbidity
which indicate the microbial contamination
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13 April 2015
Principles 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)
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13 April 2015
Principles of Media Fill
Despite of 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
So the sterility of the product is major requirement, But the
sterility test of the whole batch is not possible to check
whole batch because it is destructive method.
It is better to validate the integrated line by media fill test
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13 April 2015
Media Fill Protocol
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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
13 April 2015
Validation of Integrated line by Media
Fill test
 OBJECTIVE
The Objective of validation protocol is to establish
documented evidence that the process employed for aseptic
processing of Parenterals liquid/Ophthalmic solution will produce
the desired results consistently, within the specified acceptance
limits, when performed as per the latest Standard Operating
Procedures.
 SCOPE
The Validation protocol describes the procedure for the total
Process Simulation (Media Fill) for integrated line.
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RESPONSIBILITIES
Sr. no .
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Responsibility
Name of the
department
1
Preparation of Protocol
QC
2
Provision of qualified personnel to assist in the
protocol preparation and execution
QC, QA, Production
and Maintenance
3
Verification of Protocol
QC and Production
4
Approval of protocol
QA
5
Final determination of System Acceptability
QA
6
Review and assembling of data into a final report
QA
13 April 2015
PRE-REQUISITES
Approved Soybean casein digest broth
Environmental Monitoring of manufacturing areas by Plate
Exposure, Air sampling and surface monitoring procedures and
its SOP’s.
Qualified and validated manufacturing equipments, system
facility (i.e. HVAC, water, compressed gases) CIP and SIP
procedures.
Trained operating personnel’s.
Approved BMR for media fill trial.
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EQUIPMENT / SYSTEM
DESCRIPTION:
 Location :Manufacturing Area integrated line ( Mixing room
and filling room)
 Equipments :Mixing Tank, Holding Tank, Filtration housings,
connected product line and FFS machines
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IDENTIFICATION OF CRITICAL CONTROL
MONITORING PARAMETER
Check and ensure that•
•
•
•
•
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The equipment and system facility is validated.
The HVAC system, compressed air, CIP and SIP procedures are
qualified.
All operations, cleaning/sanitization procedures are established and
operating personnel are trained.
Media used for Process Simulation is passed for GPT
The WFI used for preparation of batch is complied to USP/IP
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STUDY DESIGN
1. Worst Case Consideration
2. Frequency, Duration, Number of runs & Fill Volume
Environmental Consideration
4. Media
5. Incubation and examination of filled units
6. Interpretation of Test Result:
3.
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STUDY DESIGN
1. Worst Case Consideration
a.
b.
c.
d.
e.
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Allow maximum number of personnel in the aseptic processing
area , including the maintenances and house keeping personnel
Increased the time period to start the filling operation
Increase the Duration of the media fill trial than that required
for routine manufacturing operation.
Simulating Process / Power breakdown during the process
simulation test
Shift changes and breaks
13 April 2015
STUDY DESIGN
2. Frequency, Duration, Number of runs & Fill Volume
Must be performed on semi-annual basis for each aseptic process
and additional media fill trials should be performed in case of any
change in procedure, practices or equipment configuration .
b. Filled units in Media Fill run should be 10,000 units or more.
Fill minimum 3000 units in each production shift.
c. The duration of Media Fill run must cover all the three
operational shifts in each run turn by turn including worst cases
as stated in steps
d. Fill volume for Media Fill run for SVP is 10 ml.
a.
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STUDY DESIGN
3. Environmental conditions
a.
Cleaning of Area must be done by using routine cleaning agent
and disinfectant solution, as per latest SOP
b.
Microbiological Environmental Monitoring should be carried out
by •
•
•
•
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Settle plate
Air sampling
Swab test and
personnel monitoring as per the latest SOP.
13 April 2015
STUDY DESIGN
4.
Media:
a.
Soybean Casein Digest Medium, manufactured by Hi Media
Laboratories should be used for Media fill trial
b.
The media must be passed the test for GPT as per SOP No.
APL/QC/SOP/185 to promote the growth of gram-negative
and gram-positive bacteria and yeast and molds
c.
For anaerobic microbs Fluid Thioglycollate Medium (FTM)
is used
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STUDY DESIGN
5.
Incubation and examination of filled units:
Incubate all media filled units in normal position after leak test at
of 20 to 250C for 7 days. Incubation temperature should be
maintained within 22.5 ± 2.50C .
b. After completion of 7 days Incubation at 20 to 250C, invert the
units and incubate them at 30-350C for next 7 days. Incubation
temperature should be maintained within 32.5±2.50C .
c. Each media filled unit should be examined by trained
Microbiologist after 3rd day, 7th day, 10th day and 14th day.
d. All suspect units identified during the observation should be
brought to the immediate attention of the QC Microbiologist.
a.
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STUDY DESIGN
6. Interpretation of Test Result:
Any contaminated unit should be considered objectionable and
investigated. The microorganism should be identified to species
level.
b. The investigation should survey the possible causes of
contamination.
c. When filled units up to 10000, one contaminated unit should
result in an investigation, including consideration of a repeat test.
a.
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VALIDATION PROCEDURE
Main steps for theValidation of the integrated line by
media fill test

1.
2.
3.
4.
5.
6.
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
Cleaning of the line
Dispensing of Soybean Casein Digest Medium for 150 L batch
size
Batch Preparation 150 L
Filling And Sealing
Incubation and Examination of Media Filled Units
Interpretation of Results
13 April 2015
VALIDATION PROCEDURE
1. Cleaning of the SVP line
Carry out cleaning of SVP mixing tank and holding tank along with
product line and bottle pack machine as per respective SOP for CIP.
At the end of cleaning, collect last rinses sample from sampling point and
send to QC department with written information for testing of previous
product traces.
After getting approval report from QC, affix status label on the tank
“READY FOR STERILIZATION”.
Immediately carry out the sterilization of SVP holding tank along with
final filter and product line of bottle pack machine as per respective SOP.
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VALIDATION PROCEDURE
2. Dispensing of Soybean Casein Digest Medium for 150 L
batch size
Enter to dispensing room as per SOP for entry exit procedure to
dispensing area.
Check for the clearance of the area from any unwanted materials.
Check for the cleanliness of the area, LAF, weighing pan as per
checklist. Put “ON” the reverse LAF unit 15 minutes before
dispensing of material.
Check the availability of clean containers, pressure differentials, and
temperature & humidity should be not more than 250C and 45 to
60% RH respectively.
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VALIDATION PROCEDURE
Calibrate the balance as per SOP of Balance Calibration.
Take the Approved Soybean Casein Digest Medium in predispensing room, place on SS pallet and check the label of container
for correctness and Approval of material.
Transfer the material to Dispensing room, place the empty clean
container on the balance and record the tare weight. Press “ZERO”
of the balance and weigh the required quantity of material, note the
weighed material and then remove the container from balance and
press Zero.
Close the dispensed material, affix the weighing tag and
transfer the material in dispensed material storage room.
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VALIDATION PROCEDURE
After dispensing, put “OFF” the balance and LAF. Clean the surrounding
area, balance and spray with 70% IPA solution.
Reseal the original container and shift to their original place.
3. Batch Preparation 150 L:
Ensure that the area and product line is clean and free from the
traces of previous product.
Recheck gross weight of Soybean Casein Digest Medium (SCDM)
to be used for manufacturing and ensure that they match as per
entries made in the BMR weighing sheet.
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VALIDATION PROCEDURE
Check the status board affixed on the tank “READY FOR USE”, also
verify the records and ensure that the bottom outlet valve of the
mixing tank is closed.
Send the entry point sample of WFI from the user point to QC
department for testing along with BMR.
On approval of WFI sample from QC department, affix a status
board on the Mixing tank “UNDER MANUFCTURING” with
Product name and B. No.
Collect approx 50 L water for injection at 80 to 850C in a
manufacturing tank fitted with stirrer.
Start the stirrer and add SCDM through the mainhole of the tank.
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VALIDATION PROCEDURE
Continue stirrer for complete dissolution of ingredients.
Stop the stirrer.
Make up the volume to the 150 L with water for injection.
Start the stirring for complete dissolution of SCDM and
homogeneous bulk solution (generally required 10 minutes).
Collect sample of bulk solution in a sterile sampling bottle and
send it to QC for testing of color clarity, pH and bioburden
along with bulk intimation slip.
After getting clearance of bulk analysis from Quality Control,
start the filtration from mixing tank to Holding tank of line with
the help of pump.
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VALIDATION PROCEDURE
After getting clearance of bulk analysis from Quality Control,
start the filtration from mixing tank to Holding tank of line with
the help of pump.
Perform the bubble point test of the final filter after holding tank
as per SOP of Bubble point test.
4. Filling And Sealing:
Start the filtration from holding tank to FFS machine using
pump.
Drain one buffer tank approx 1.3 liters of bulk solution from
filling nozzle to eliminate any possibility of dilution of bulk by
condensates in product line of the machine post SIP.
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VALIDATION PROCEDURE
Check online cartridge filter integrity test as per its respective
SOP.
Start Machine line and discard initial 15 shots.
Collect first cassette of vials from next shot and send the sample
with written information to QC for testing.
Arrange the out coming cassettes of vials sequentially in vacuum
chamber tray and verify the results of testing from QC
department.
Now start the filling and sealing continuously as per SOP for
Filling and sealing.
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VALIDATION PROCEDURE
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Collect the filled and sealed containers coming out of the filling
area in plastic crates.
During filling operation keep the filled ampoules separately for
each breakdown, shift change, power breakdown, stoppage etc
and assign lot number.
Arrange the cassettes of vials lot wise in SS trays vertically in
vacuum leak testing chamber tray and carry out the leak testing
at 650 – 720 mm Hg for 30 minutes. Do not use the leak vials
for further media fill study.
After leak test, transfer the goods vials in the clean plastic crates
horizontally in cassette from one above the other, lot wise
separately
13 April 2015
VALIDATION PROCEDURE
5. Incubation and examination of filled units:
Incubate all media filled units in normal position after leak
test at of 20 to 250C for 7 days. Incubation temperature should
be maintained within 22.5 ± 2.50C .
b. After completion of 7 days Incubation at 20 to 250C, invert
the units and incubate them at 30-350C for next 7 days.
Incubation temperature should be maintained within
32.5±2.50C .
c. Each media filled unit should be examined by trained
Microbiologist after 3rd day, 7th day, 10th day and 14th day.
d. All suspect units identified during the observation should be
brought to the immediate attention of the QC Microbiologist.
a.
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VALIDATION PROCEDURE
6.
Interpretation of Results:
When filling fewer than 5,000 units, no contaminated units
should be detected.
When filling 5,000 to 10,000 units :
 One contaminated unit should result in an investigation, including
consideration of a repeat media fill
 Two contaminated units are considered cause for revalidation,
following investigation.
When filling more than 10,000 units :
 One contaminated unit should result in an investigation;
 Two contaminated units are considered cause for revalidation,
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following investigation.
13 April 2015
REFERENCES
 Syed Imtiaz Haider, “ Validation Standard Operating
Procedures ” 314-321
 R. A. Nash and A. H. Wachter “Pharmaceutical Process
validation”; Third edition
 Agalloco James, Carleton J. Fredric
“Validation of
Pharmaceutical Processes”;Third edition
 Pharmaguideline.blogspot.com
 www.milipore.com
 www.nsdl.niscair.res.in/bitstream
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