RedLotus Pharmtech Private Limited

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RedLotus, "Pharma Technical Services Company" brings in years of
collective experience in the areas of a drug substance as well as a
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technology transfer, manufacturing, quality assurance & control,
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 Process Simulation Test (Media Fill)
Model Report

TABLE OF CONTENT

1.0 EXECUTIVE SUMMARY................................................................................................................................................. 3

2.0 BACKGROUND............................................................................................................................................................... 4
3.0 PURPOSE....................................................................................................................................................................... 6
4.0 OBJECTIVE.................................................................................................................................................................... 7
5.0 SCOPE............................................................................................................................................................................ 7
6.0 INTRODUCTION............................................................................................................................................................. 7
7.1 OVERVIEW OF ASEPTIC MANUFACTURING PROCESS STEPS...............................................................................7
7.2 VIAL WASHING................................................................................................................................................................................................................7
7.3 VIAL STERILIZATION AND DEPYROGENATION..........................................................................................................................................................9
7.4 STERILE DRY POWDER FILLING & RUBBER STOPPERING................................................................................................................................10
7.5 VIAL SEALING...............................................................................................................................................................................................................12
7.6 VIAL VISUAL INSPECTION..........................................................................................................................................................................................13
8.1 PROCESS SIMULATION TEST PROCEDURE............................................................................................................ 13
8.2 OVERVIEW.....................................................................................................................................................................................................................13
9.1 PROCESS SIMULATION – OBSERVATIONS, FINDINGS & RESULTS......................................................................14
9.2 GENERAL CONSIDERATIONS.....................................................................................................................................................................................14
9.3 CONTAINER CLOSURE SYSTEM...............................................................................................................................................................................17
9.4 WORST CASE SITUATION..........................................................................................................................................................................................17
9.5 PROCESS SIMULATION BATCH SIZE.......................................................................................................................................................................18
9.6 MEDIA SELECTION.......................................................................................................................................................................................................18
9.7 COMPOUNDING............................................................................................................................................................................................................19
9.8 COMPONENT, MACHINE PARTS & ACCESSORIES STERILIZATION..................................................................................................................20
9.9 FILLING...........................................................................................................................................................................................................................20
9.9.1 FILLING SPEED.........................................................................................................................................................................................................20
9.9.2 FILLING DURATION..................................................................................................................................................................................................21
9.9.3 FILLING WEIGHT AND VOLUME..............................................................................................................................................................................21
9.9.4 INERT GASSING.......................................................................................................................................................................................................22
9.9 INTERVENTIONS...........................................................................................................................................................................................................22
9.10 VIALS RECONCILIATION...........................................................................................................................................................................................24
9.11 ENVIRONMENTAL MONITORING.............................................................................................................................................................................25
9.12 PERSONNEL MONITORING......................................................................................................................................................................................28
9.13 POST FILL VISUAL INSPECTION.............................................................................................................................................................................29
9.14 VIAL LABELING..........................................................................................................................................................................................................29
9.15 INCUBATION................................................................................................................................................................................................................29
9.15.1 ORIENTATION OF MEDIA FILLED UNITS DURING THE INCUBATION.............................................................................................................29
9.15.2 INCUBATION CONDITIONS...................................................................................................................................................................................29
9.16 POST INCUBATION....................................................................................................................................................................................................30
9.16.1 INSPECTION............................................................................................................................................................................................................30
9.16.2 STASIS TEST (POST INCUBATION MEDIA GROWTH PROMOTION TEST)............................................................................33
9.17 IN-COMING MATERIAL AND IN-PROCESS SAMPLING.........................................................................................................................................34
9.18 ASEPTIC PRACTICES................................................................................................................................................................................................38
10.0 DISCUSSION, ANALYSIS AND INTERPRETATION.................................................................................................. 40
11.0 CONCLUSION............................................................................................................................................................ 44
12.0 RECOMMENDATIONS............................................................................................................................................... 44

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1.0 EXECUTIVE SUMMARY

In recent USFDA inspection, subsequent Q&A’s (on the response/ corrective actions) and third-party
(RedLotus Pharmtech Pvt. Ltd.) expert reviews, gaps/ deficiencies were identified in the media fill
program. These deficiencies were primarily related to; [a] interventions (mismatch with respect to type,
frequency with routine product batch manufacturing and inadequately explained), [b] media fill not
accurate simulation of the routine product batch manufacturing process, and [c] ambiguity in
reconciliation of media filled units.
In view of the above observations/ gaps/ deficiencies, a thorough and critical review of the media fill
program, was done in the period of Dec. 2016 to Feb. 2017. This opportunity was also utilized to
compare/ verify the existing media fill program and all associated documentation with respect to the
current requirements specified in: (a) USFDA guidance document, “Guidance for Industry Sterile Drug
Products Produced by Aseptic Processing — Current Good Manufacturing Practice”, Sep. 2004, (b) PDA
Technical Report No. 22, “Process Simulation for Aseptically filled Products, 2011, (c) PICS
recommendation on the validation of aseptic process, PI 007-6/ 1 Jan. 2011”, (d) WHO technical report
series 961, 2011, (e) EU GMP guideline for manufacture of sterile medicinal products, vol. 4, Annex – 1,
(f) experience and knowledge on this subject, (d) current industry trends.
Subsequent to making all changes/ improvements and trainings as mentioned above two media fill
batch(s) MF/01 and & MF/02 were manufactured on 31st Mar. 2017 and 01st Apr. 2017 respectively.
Before manufacturing of these media fill batches, a series of trainings by in-house experts as well as
RedLotus Pharmtech Pvt. Ltd. were conducted between Dec. 2016 to Feb. 2017 on aseptic area
practices as well as related topics like data integrity – media fill as specific reference.
Media fill batches have been manufactured as per the (Revised) SOP # QA/MF/010-R09 to accurately
simulate the routine product batch manufacturing process, including the interventions by actually
replacing the product with sterile placebo powder (Mannitol) followed by filling sterile, liquid nutrient
media (SCDM). Thus, filled and sealed vials were incubated at 20°C – 25°C for 168 hrs. (7 days) in
inverted condition followed by incubation at 30°C – 35°C for 168 hrs. (7 days) in upright condition.
Before inspection, vials were subject to 100% visual inspection. All good vials along with the cosmetic
rejects (integral rejects) like particulate contamination and cosmetic defects were also incubated with
good vials. The rejects that are non-integral were rejected. All reject units removed automatically either
by the machine or manually, that may have fallen off the line during transportation (between the vial
filling and before the final product inspection step) were considered non-integral, thus not considered for
incubation and disposed exactly in the same manner as the routine production vials.
The process simulation test result and manufacturing process details recorded in a specifically designed
(Revised) batch production record. Other supporting documents, such as the environmental monitoring
reports, sterilization reports etc. are enclosed with the corresponding batch manufacturing records. A
high- level summary is presented in the following table.
Summary of media fill batch(s) MF/01 and & MF/02
Inspection Defects Contaminated
Batch # Fill Date Inspected Defect-11 Defect-22 Incubated
units
MF/01 31-03-2017 15,486 0 227 15,486 03
MF/02 01-04-2017 15,137 0 244 15,137 0

1 Defects affecting the container closure integrity like – cracked vials, missing stoppers/ missing seals
2 Defects not affecting the container closure integrity like – cosmetic defects (scratch, moulding defects etc.) and visual particulate matter
3 One cracked vial was found damaged during the inspection of the vials on 7 th day of incubation. As this vial, can be representative of drug product released

to the market and that, no growth/ turbidity/ contamination was noted in this vial, it was not excluded and allowed to remain for the rest of the incubation
period. An investigation is completed to find out the root cause and identify appropriate CAPA for the cracked vial – refer incident/ investigation report no.
IIR/2017/021

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A thorough review and critical analysis of all observations and study findings confirms that, in the
manufacturing of the media fill batch(s) MF/01 and & MF/02:
 The steps preceding filling and sealing have been designed and executed in manner to
incorporate all conditions, product manipulations, and interventions that could impact on the
sterility of the product.
 All preparation and sterilization procedure followed were same as those being followed in
manufacturing of a routine production batch
 The vials preparation, filling, stoppering and sealing machine and aseptic manufacturing area used
in the media fill batches were used in exactly the same manner as normally done in the routine
manufacturing batch
 The studies incorporated all product manipulations, additions, and procedures involving
exposure of product contact surfaces to the environment.
Based on a thorough review, analysis and discussions, it is assessed/ judged that the process simulation
study results derived from the media fill batch numbers MF/01 and MF/02 manufacturing are reliable,
accurate simulation of the routine product batch manufacturing process, and meeting the acceptance
criteria as defined in the SOP # QA/MF/010-R09. Thus, demonstrating adequate controls in place to
protect the product during manufacturing. The aseptic manufacturing process is considered validated
and qualified if no contamination of a media filled vial is observed.
Based on the overall assessment/ judgement, it is concluded that: (a) the aseptic manufacturing
process/ techniques/ technology, (b) manufacturing and containment controls, (c) environmental
conditions and controls, (d) personnel involved/ engaged, (e) applicable corresponding SOP’s and
procedures available and/ or being followed at Model Life Sciences Limited, Nagpur, Unit VI
demonstrates the on-going capability of producing sterile drug product by aseptic processing.
However, a thorough review and critical analysis has resulted in identification of few improvement areas,
which have been described in the “Recommendations” section.
2.1 BACKGROUND
To ensure the sterility of products purporting to be sterile, sterilization, aseptic filling and closing
operations must be adequately validated. 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.
To demonstrate the process of bringing together all these sterilized elements to produce a sterile drug
product at Model Life Sciences (MLS), Unit VI, Nagpur, an aseptic processing operation is validated
using a sterile placebo powder (Mannitol) and a sterile microbiological growth medium (SCDM) in
substitution of the product. This process simulation, is also known as a media fill, and includes exposing
the sterile placebo powder to the product contact surfaces of equipment, container closure systems,
critical environments, and process manipulations to closely simulate the same exposure that the product
itself will undergo. The sealed containers filled with the sterile placebo powder and a growth medium are
then incubated to detect microbial contamination. Results are then interpreted to assess the potential for
a unit of drug product to become contaminated during actual operations (e.g., start-up, sterile ingredient
additions, aseptic connections, filling, and over sealing).
In recent USFDA inspection, subsequent Q&A’s with USFDA (on the response/ corrective actions) and
third-party (RedLotus Pharmtech Pvt. Ltd.) expert reviews, gaps/ deficiencies were identified in the
media fill program. Summary of the observations/ gaps/ deficiencies is presented below, please refer –
Attachment 1 for details.
 Number of interventions did not reflect what takes place during aseptic filling (USFDA)
 Process interventions performed do not represent the actual manufacturing process (USFDA)

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 Routine interventions identified in the validation protocol were not carried out in a representative
manner (USFDA)
 Response was inadequate in that it does not provide details, as how MLS intend to improve the
procedure for future MF (USFDA)
 Difference noted in process simulation and routine MF batches – MF not close simulation,
particularly with respect to Shifts, machine speed & interventions (RedLotus Pharmtech Pvt. Ltd.)
 MF vials reconciliation – confusing & complex (RedLotus Pharmtech Pvt. Ltd.)
In view of the above observations/ gaps/ deficiencies, a thorough and critical review of the media fill
program, was done in the period of Dec. 2016 to Feb. 2017. Summary of specific steps taken/
improvements done are as follows:
 “Aseptic Process Intervention – Identification & Risk Assessment” study initiated with approved
protocol # PRO/MF/010.
 Before execution of the protocol # PRO/MF/010, training was given by RedLotus Pharmtech Pvt.
Ltd. to each team member participating in the review, assessment and reporting.
 Trend review of aseptic process interventions observed and reported in all product batches
manufactured in the year 2016 performed. The list of interventions defined in SOP No. MG/MF/050-
R05 was considered as a basis for the trend analysis of each intervention. This exercise helped in
measuring frequency of occurrence for each intervention. The maximum number of occurrences for
each intervention was taken as a benchmark for finalizing the number of times each intervention to
be simulated in the next media fill batches.
 Brainstorming done on each intervention identified/ reported in year 2016 for its identification,
categorization, based on a structured risk assessment.
 A final intervention list was prepared based on team brainstorming on pre-defined questions and a
decision tree, that helped in identification of each intervention with respect to its nature as well as
potential risk to sterility assurance.
 The above reviews and results were used to finalize the list of interventions and their frequency to
be simulated in the next media fill batches. This List is now a part of a new SOP # MG/IN/071-R02,
“Aseptic Process Intervention – Lifecycle Management” and the old list of interventions (SOP #
QA/MF/010) has now become obsolete.
This opportunity was also utilized to compare/ verify the existing media fill program and all associated
documentation with respect to the current requirements specified in: (a) USFDA guidance document,
“Guidance for Industry Sterile Drug Products Produced by Aseptic Processing — Current Good
Manufacturing Practice”, Sep. 2004, (b) PDA Technical Report No. 22, “Process Simulation for
Aseptically filled Products, 2011, (c) PICS recommendation on the validation of aseptic process, PI 007-
6/ 1 Jan. 2011”, (d) WHO technical report series 961, 2011, (e) EU GMP guideline for manufacture of
sterile medicinal products, vol. 4, Annex – 1, (f) experience and knowledge on this subject, (d) current
industry trends. This review resulted in making improvements in the following documents/ systems/
processes:
 Revision of media fill SOP # QA/MF/010 – included and/ or improved instructions/ section for: (a)
providing an overview and process description, (b) planning of the media fill based on review of
past results/ trends – this is in line with the USFDA current thinking on the process validation, i.e.,
continuous process verification, (c) vis-à-vis comparison of routine product batch and a media fill
batch manufacturing process, (d) media fill batch size definitions clarified depending upon the
normal working hours and line speed, (e) process rejects – definition, identification and handling, (f)
media fill vials, post-incubation inspection procedure – need of training, (g) decision tree –
interpretation of the media fill results, and (h) media fill report preparation.

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 Revision of media fill batch manufacturing record – revised to improve/ include instructions/ section
for: (a) summary of the batch on top of the batch manufacturing record, (b) comparable with the
product batch manufacturing record, (c) vials reconciliation
Additionally, series of trainings by in-house experts as well as RedLotus Pharmtech Pvt. Ltd. were
conducted between Dec. 2016 to Feb. 2017 on aseptic area practices as well as related topics like data
integrity – media fill as specific reference.
It is very well understood that, the successful completion of a process simulation test(s) cannot be
considered as a sole-criteria for validation of aseptic manufacturing processing in same sense that a
performance qualification effort involving biological challenge and temperature measurement can
support a steam sterilization process. Aseptic processing relies heavily on personnel intervention
practices, equipment features, facility design/ control and procedures that in combination serve to
exclude micro- organisms from sterile components and products. These elements of aseptic processing
cannot be as rigorously controlled as a sterilization process, resulting in a lingering risk of contamination.
Thus, the aseptic process simulation is only a demonstration of the capability with respect to the sterility
assurance of the process to produce sterile products aseptically at the time of its execution using the
defined process, materials, facility, equipment and personnel.
There are multiple programs followed at following programs that are followed as a holistic approach to
ensure sterility of the products. This incorporates many systems as indicated below, to control and
assure sterility of the products:
 Product, equipment and component sterilization
 Personnel training and certification of cleanroom gowning, practices and aseptic technique
 Equipment and facility sanitization program, including the isolators
 Environmental systems, microbial levels, differential pressures, air flow pattern, velocity,
temperature & humidity
 Personnel, material and equipment flow
 Standard operating procedures
 An underlying quality system approach to process control
These systems are routinely monitored for verification of their continued acceptable performance, by
which sterility assurance of the manufactured product is established. Hence, all of the related sanitization
and sterilization process(s) are (re)validated independently, such as sterilization/ Depyrogenation of the
product, container, closure, and all product contact and indirect product contact surfaces (e.g.,
component hoppers).
Subsequent to making all changes/ improvements and trainings as mentioned above two media fill
batch(s) MF/01 and & MF/02 were manufactured on 31st Mar. 2017 and 01st Apr. 2017 respectively.
This study is an effort to evaluate the sterility assurance of the sterile products manufacturing process at
Model Life Sciences, Nagpur, Unit VI with respect to its capability to produce a sterile product by bringing
together sterilized elements of a product (the drug formulation, the container, and the closure) under
conditions that can potentially contaminate any of these elements.
3.1 PURPOSE
 Report results of the media fill study/ batch(s) manufactured subsequent to making changes /
improvements as explained in the above section 1.
 Identification of the gaps/ deficiencies and recommendations (if any) for the continuous improvement.

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This document and comprehensive review of the media fill data will become basis and a framework for
“continuous process verification – sterility assurance”. This is a requirement as per the USFDA Guidance
for Industry, “Process Validation: General Principles and Practices, Jan. 2011”
4.0 OBJECTIVE
To evaluate the capability of aseptic manufacturing process followed at Model Life Sciences, Unit VI,
Nagpur in achieving high degree of sterility assurance, based on review of the executed process
simulations tests/ media fill batch(s) MF/01 and & MF/02 manufactured on 31 st Mar. 2017 and 01st Apr.
2017 respectively.
5.0 SCOPE
The scope of this study is limited to sterility assurance evaluation based on the review of media fill study/
batch(s) MF/01 and & MF/02 manufactured on 31st Mar. 2017 and 01st Apr. 2017 respectively.
Review of the other programs that contribute to the finished product sterility assurance as explained in
the section 1, is not included in the scope of this review study. Theses reviews are performed individually
as a separate subject.
6.1 INTRODUCTION
The Unit VI, Dry Powder Injectable Manufacturing facility is operated in two shifts; the third shift is
utilized for cleaning, sanitization and all preparatory work. Hence, the interventions observed and/ or
expected in two shifts, including during the shift changeover are being considered in this assessment
exercise.
The process of dry power injection manufacturing at Model Life Sciences is divided in following steps:
 Inspection and loading of empty glass vials
 Washing of the empty glass vials in a closed automatic vial washing machine located in an ISO 7
(Class 10,000) environment
 Online Depyrogenation/ Sterilization of washed glass vials using a tunnel sterilizer with in-built
HEPA filters to maintain ISO 5 (Class 100) environment inside the tunnel
 Aseptic filling and stoppering of sterile dry powder in glass vials under laminar airflow unit (ISO 5 or
Class 100) environment. The powder filling machine is located in ISO 6 (Class 1000) environment
 Pre-and post-purging of sterile filtered nitrogen or any other gas according to the process need is
purged to flush empty vial and/ or fill the headspace
 Filled and stoppered vials exit into an ISO 7 (Class 10,000) vial sealing room, where the vials are
sealed using Aluminum overseals under ISO 5 (Class 100) Laminar Airflow Unit
 Inspection of filled and sealed vials for cosmetic defects as well as gross contaminants using semi-
automatic, online visual inspection machine
 Labeling and packaging of the inspected vials
7.1 OVERVIEW OF ASEPTIC MANUFACTURING PROCESS STEPS
7.2 Vial Washing
Ambica high-speed linear vial washing machine is used for washing of the vials. Vials are washed
according to a set and validated sequence at different washing stations using sterile filtered compressed
air, re-circulated water, Purified Water, and final rinse with Water for Injection. The vials washing
machine has a maximum capacity to wash 300 vials per minute. All parts coming in contact with washing
zone/ jet are constructed of stainless steel 316. The vial washing machine is controlled by a PLC, which
contains validated program(s) for the washing of different size glass vials. High-pressure spray
jet/nozzles enter into the vials and ensure the perfect wash and entire washing process can be observed

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from a transparent glass window.

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Figure 2: Vial Washing Machine (PIVWM-01)

The vials are washed from both sides i.e., inside and outside of the vial by using high pressure
jet/nozzle, containing media defined for that specific washing station, i.e., filtered, (re)circulated purified
water, purified water, compressed air and water for injection. A typical washing method at a washing
station is shown in the following figure 3.
Figure 3: A Typical Vial Washing Station

The vials from the in-feed gets inverted and positioned into a pocket, which then moves to the sequence
of washing stations. At each washing station, the vial stops, the nozzles manifold raises up and nozzles
enter into the mouth of each vial. A solenoid valve in the water and compressed air lines is activated and
a jet of air/water is introduced inside and outside of the vials. A standard washing cycle sequence is
explained in following table 1:

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Table 1: Vial Washing Sequence

Vial Surface
Station
Internal External
1 Sterile filtered compressed air --
2 Filtered re-circulated water Filtered re-circulated water
3 Filtered re-circulated water --
4 Purified water --
5 Purified water Purified water
6 Water for Injection --
7 Water for Injection Water for Injection
8 Sterile filtered compressed air --

Following figure 4 explains the complete washing sequence schematic diagram.

Figure 4: A schematic diagram of vial washing machine operation

7.2 Vial sterilization and Depyrogenation:

Out-feed of the vial machine is integrated with the Macofar T800, vials sterilizing tunnel. Macofar T800,
tunnel is manufactured by Macofar, Italy and has been designed to sterilize and depyrogenate glass
vials of various sizes (10 ml, 15 ml, 20 ml, 50 mL and 100 ml) by means of a continuous process using
HEPA filtered, dry and hot unidirectional air-flow. The air temperature within the sterilizing zone of the
tunnel sterilizer is maintained in a rage of 320°C to 360°C. An ISO 5 (Class 100) environment according
to ISO 14644-1 requirements is maintained inside the entire length of the tunnel.
Washed glass vials are transported into the sterilizing tunnel by a conveyor belt constructed of stainless-
steel wire mesh with lateral guides that avoids the vial from sliding along the tunnel walls. The tunnel is
constructed of stainless steel AISI 316 and AISI 304 steel panels. The conveyor speed can be adjusted
according to the vial size and configuration. Maximum conveyor speed of 200 mm per minute can be
achieved.

Figure 5: Macofar T800 Sterilizing Tunnel (PISTT-01)

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Salient features of the Macofar T800, sterilizing tunnel are as follows: The tunnel has 3 zones, i.e. (a) in-
feed chamber – vials are preheated in this zone, (b) heating chamber – vials are heated enough to reach
set Depyrogenation temperature in this zone, and (c) cooling chamber – dehydrogenated vials are
cooled gradually to reach to the room temperature at tunnel exit.
Figure 6: Schematic diagram of Macofar T800 Tunnel showing different zones

The sealing of the HEPA filters is achieved by using ceramic fiber gaskets. The area around the frames
of the HEPA filters is connected to the air inlet of the corresponding fan to create a required pressure
gradient across the HEPA filter, this helps avoiding accidental infiltration of non-sterile air through the
gaskets.
Adjustable flaps within the hot chambers ensures uniformity of air velocity and laminarity of the inlet air
filtered through the HEPA filters, this helps ensuring uniform heat distribution across the heating
chamber.
All process control parameters are continuously monitored, automatically controlled and printed.
7.3 Sterile Dry Powder Filling & Rubber Stoppering:
The out-feed of the Macofar T800, sterilizing tunnel is integrated with the Macofar Micro 18 automatic
vial filling and rubber stoppering machine. The Macofar Micro 18, machine fills sterile dry powder into
various size (10 ml, 15 ml, 20 ml, 50 mL and 100 ml) glass vials using vacuum and pressure system.
Figure 7: Macofar Micro 18, Vial Filling and Bunging Machine (PIVFB-01)

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The sterilized and dehydrogenated vials are received from an integrated tunnel out-feed onto the vial
filling and bunging machine in-feed turntable. This in-feed turntable is housed in an openable, plexiglass
cabinet, with Laminar Air-flow. HEPA filter unit on the ceiling to maintain ISO 5 (Class 100) environment.
Vials from the in-feed turntable are guided through the online weighing, empty vial gas flushing, powder
filling, filled vial online weighing, post-fill headspace gas flushing, and rubber stoppering station by a
system of star wheels to ensure vial remains firmly upright and to allow the correct identification of each
vial during the automatic online weight check process. The vials are distributed in two identical tracks on
both sides and conveyed by star wheels under the powder-dosing disc.
Figure 8: Different Stations and Steps in the Vial Filling and Bunging Process

The Macofar Micro 18 machine has in-built statistical weight control system for on-line fill weight
measurement and control. This system ensures the high standard of quality control over the fill weight of
vial during vial filling operation. Dosing ports are automatically adjusted during the operation in
accordance with feedback from the online weighing stations. The dosing ports can also be manually
adjusted from the control panel without opening of the machine cabinet.
Figure 9: Online Fill Weight Measurement & Control System

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The sterile powder container lid is replaced with sterile butterfly valve under laminar airflow unit in the
cooling zone, which is then secured and transferred to the vial filling machine, where it is connected
aseptically to the main powder hopper in-feed chute. The powder is fed to the intermediate hopper and
the intermediate hopper, which is equipped with an agitator. Dosing chambers are located at the dosing
disk that is just below the intermediate powder hopper. Dosing wheel consists of 24 radial chambers in
two rows, each consisting 12 dosing chambers. Each dosing port contains a dosing piston, which can be
adjusted according to required fill weight.
Rubber stoppering unit consists of a vibrating bowl for correct orientation of the rubber stoppers, double
feeding chute and rubber stoppering heads for holding the rubber stoppers using vacuum. Vacuum is
released automatically when the rubber stopper is perfectly aligned with the vial mouth. Further the
rotary head presses the stopper further to ensure complete stoppering of the vials. Two pneumatic
deviators, positioned after the flat transfer star wheels, channel the correctly packaged vials onto the
belts towered the downstream machine. Non-conforming vials i.e., incorrect dose, empty vial, un-
stopper, improperly positioned stopper are automatically detected and diverted into two rejection
channels. The complete vial filling and stoppering machine including the in-feed turntable, connecting
conveyors and out-feed turntable have laminar air-flow units and an openable plexiglass & transparent
curtains enclosure to ensure maintenance of ISO 5 (Class 100) environment and air flow laminarity.
7.4 Vial Sealing
The out-feed of the vial filling and bunging machine leads to the vial sealing machine through a small
mouse hole in a rigid wall between the vial filling and sealing rooms. The vial sealing room is ISO 7
(Class 10,000) grade hygiene zone with ISO 5 (Class 100) laminar air-flow unit mounted above the vial
sealing machine. The Macofar CSP 18 vial sealing machine has a capability to seal various size glass
vials with aluminum, flip-off or tear-off overseals. Pre-sanitized Aluminum overseals are charged into the
sealing machine vibratory bowl, where the overseals are properly oriented and guided to the feeding
chute. While the vials are transferred from the timing screw to the star wheel, the caps are pulled off
from the vibrating chute by the moving vials.
Figure 10: Vial Sealing Machine (PIVSM-01)

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Each container entering the machine is clamped between a stationary base plate and a rotating head
cap is then positioned on the container, and then tightly and securely fastened as head rotates. Vial
sealing process is shown in the following figure 11.
Figure 11: Vial Sealing Process Steps

The Macofar CSP 18, vial sealing machine is highly instrumented, operated through a PLC and has
capability of adjusting the capping pressure automatically.
7.5 Vial Visual Inspection:
Vial sealing machine out-feed is integrated with an online, semi-automatic vial visual inspection machine.
Trained and qualified operators use this machine for the inspection of sealed vials for such as cosmetic
defects (Molding defects & sealing defects), breakage & cracks, fill weight variation, foreign particles,
dent on seal, dirty vials, and missing aluminum seals as per SOP No. PI-SG-014
This machine is suitable for four operators, two operators on the left side and other two on the right side.
The out-feed of the vial sealing machine leads to the vial visual inspection machine through a small
mouse hole in a rigid wall between the vial sealing and vial visual inspection machine. Sealed vials are
received on an in-feed turntable of the vial visual inspection machine from where the vials are guided to
the vial visual inspection machine. Vials are oriented in about 3000 angle on rotating nylon rollers to help
operator inspect complete vial surface. Glass mirrors provided on strategic sites to help the operator to
also inspect top of the vials as well. Each operator has a segregated inspection area, which is
adequately illuminated
8.1 PROCESS SIMULATION TEST PROCEDURE
8.2 Overview
Media fill has been performed as per the SOP # QA/MF/010-R09 to simulate aseptic manufacturing
process including the most probable and common interventions by actually replacing the product with
sterile placebo powder (Mannitol) followed by filling sterile, liquid nutrient media (Soybean Casein Digest
Medium – SCDM). Once filled with media, the vials are incubated at 20°C – 25°C for 168 hrs. (7 days) in
inverted condition followed by incubation at 30°C – 35°C for 168 hrs. (7 days) in upright condition.
Each filled, stoppered and sealed vial that exit the filling line are subject to the 100% visual inspection.
All good vials along with the rejects that are not related to integrity i.e., particulate contamination and
cosmetic defects are also incubated. The rejects that are non-integral, such as cracks are rejected.
All reject units removed automatically either by the machine, as explained in above section Error!
Bookmark not defined. or that may have fallen off the line during transportation (between the vial filling
and before the final product inspection step) are considered non-integral in nature, thus not considered
for incubation and disposed exactly in the same manner as the routine production vials.
The aseptic manufacturing process is considered validated and qualified if no contamination of a media
filled vial is found. SOP # QA/MF/010-R09 unambiguously defines the acceptance criteria and necessary
instructions for investigations and actions required in case the process simulation test fails.

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The process simulation test result and manufacturing process details are recorded in a specifically
designed batch production record. Other supporting documents, such as the environmental monitoring
reports, sterilization reports etc. are enclosed with the corresponding batch manufacturing records.
A high-level summary of media fill study/ batch(s) MF/01 and MF/02 manufactured on 31 st Mar. 2017 and
01st Apr. 2017 respectively is presented in the following
Table 1.
Table 1: Summary of media fill batch(s) MF/01 and & MF/02
Filled & Sealed Visual Inspection Defects Vials
Vials Transfer Transferred Contaminated
Batch # Fill Date
to Visual Defect-14 Defect-25 for units
Inspection Incubation
MF/01 31-03-2017 15,486 0 227 15,486 06
MF/02 01-04-2017 15,137 0 244 15,137 0

9.1 PROCESS SIMULATION – OBSERVATIONS, FINDINGS & RESULTS

Each process simulation run was performed to cover all normal processing operations, including the line
set-up, possible interventions (routine as well as non-routine) that are realistically encountered during
the product filling operations.
The aspects specifically covered, performed and/ or simulated during the manufacturing of media fill
batches to establish comparison with the normal aseptic manufacturing process are summarized in the
following sub-sections.
9.2 General Considerations
Process simulation test covered all normal processing steps right from sterile material issuance and
transfer into the sterile area to the vial labeling steps. A typical aseptic manufacturing process flow
diagram is presented in the following Figure 1.

4 Defects affecting the container closure integrity like – cracked vials, missing stoppers/ missing seals
5 Defects not affecting the container closure integrity like – cosmetic defects (scratch, moulding defects etc.) and visual particulate matter
6 One cracked vial was found damaged during the inspection of the vials on 7 th day of incubation. As this vial, can be representative of drug product released

to the market and that, no growth/ turbidity/ contamination was noted in this vial, it was not excluded and allowed to remain for the rest of the incubation
period. An investigation is completed to find out the root cause and identify appropriate CAPA for the cracked vial – refer incident/ investigation report no.
IIR/2017/021

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Figure 1: A typical manufacturing process flow diagram

API Rubber Stopper Glass Vials Seals

Approved Rubber Stoppers Approved Aluminum Seals

Approved API sealed canisters dispensed By Warehouse Approved Vials Dispensed by Dispensed by Warehouse
as such by warehouse Warehouse personnel
personnel personnel

Transfer the intact canister to Transfer to Preparation and De-Cartoning and Disinfection
IRM store sterilization room Transfer to Decartoning Room of bags

Loading of RFS/Non RFS Rubber

Surface Disinfection and loading Stoppers by opening non-sterile Inspected Decartoned Vials transfer Load in dynamic pass box a
to Vial Washing Room
in to dynamic pass box door hold for 30 minutes

Hold for 30 minutes and Washing/Sterilization & Drying in Unload and transfer to
unloading in sampling room autoclave cum Bung Processor Vial Washing Sealing Room

Load into the Hopper

Disinfect and transfer to filling Unload sterilized rubber stoppers Depyrogenation of vials
(EN ISO 14644-1/
room via dynamic pass box by opening sterile door in Tunnel Sterilizer
Grade A/Class 100)

Load to the hopper

Transfer to Filling Room via Vials Collected onto the Turn Table
(EN ISO 14644-1/
dynamic pass box in Filling Room
Grade A/Class 100)

Load to the Hopper

(EN ISO 14644-1/ Vials fed to Filling Machine
Grade A/Class 100)

Powder Filling &

Compressed gas purging
EN ISO 14644-1/
Grade A/Class 100)

Bunging of filled vials

Sealing Under LAF

Visual Inspection

Vial Labeling

Final Packing

F.G. Quarantine

In order to demonstrate that, the media fill process simulates the manufacturing process a vis-à-vis
comparison of both process(s) is shown in the following table 3.

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Table 3: Vis-à-vis Comparison of Normal Vial Fill Process with Media Fill Process
Manufacturing Operation / Step Routine Production Process Simulation
Manufacturing Process
Dispensing (Intact API containers issued by the warehouse)  7
Media Solution Preparation & Sterilization  8
Vial Filling – Powder  
Vial Filling – Liquid Media  5
Gassing # *
Rubber Stoppering  
Aluminum Cap Over-sealing  
Vial Visual Inspection  
Vial Labeling  9
Vials Incubation for 14 days (22.5 ± 2.5°C for 7 days followed by 32.5 ±
 
2.5°C for seven days)
Vials Inspection (for contamination on 7th day of incubation)  
Vials Inspection (for contamination on 14th day of incubation)  
Manufacturing Support Process(s)
Container External Surface Sanitization/ Decontamination & Transfer to
 
the Cooling Zone
Process & Filters Integrity Testing &Sterilization  
Media Solution Vessel Vent Filters Integrity Testing (Pre & Post Use)  2
Filling Machine Parts Sterilization &Transport to Machine  
Interior Vials Washing & Depyrogenation  
Rubber Stoppers Sterilization& Transport to Machine  
Aluminum Overseal Sanitization  
Media Solution Vessels Cleaning & Sterilization  
In-Process Controls
Nutrient Media Solution Sampling for Sterility &GPT  
Post Incubation Media Fertility Test (GPT Test)  
Environmental Monitoring  
Fill Weight Check  

In the manufacturing of the media fill batch(s) each media fill batch MF/01 and & MF/02 all preparation
and sterilization procedure followed were same as those being followed in manufacturing of a routine
production batch(s). The vials preparation, filling, stoppering and sealing machine and aseptic
manufacturing area used in the media fill batches were used in exactly the same manner as normally
done in the routine manufacturing batch(s).

7 Placebo powder Sterile Mannitol is issued

8 Sterile nutrient media solution is filled subsequent to the placebo powder fill. It is not part of routine batch process
# May / may not be required, depending upon the manufacturing process

* Simulated by substituting the inert gas with sterile filtered compressed air

9 Simulation of the labeling process only, labels not applied on the vials

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Comparison of the manufacturing process followed for a routine product and media fill batch
manufacturing indicates; that the media fill manufacturing process closely simulates, sterile drug product
manufacturing process.
9.2 Container Closure System
Following table 4 presents vis-à-vis comparison of container/closure used in the normal product
production and media fill batch(s) MF/01 AND MF/02.
Table 4: Vis-à-vis Comparison of Container Closure System
Presentation
B. No Rubber A.R. No. Equipment Train Used
Vial
Stopper

10 ml Tubular USP Vial Washer → Sterilizing Tunnel

MF/01 PM/16/2520
Type – I → Vial Filling & Stoppering
20 mm GBBR
Machine → Vial Sealing Machine
(RFS)
15 ml Tubular USP → Semi- automatic vial visual
MF/02 PM/10/1932 inspection machine
Type – I

The container and closure system are selected for the process simulation test according to the criteria
defined in SOP # QA/MF/010-R09, “Process Simulation by Media Fill Study” and it is same as being
used in the manufacturing of routine product batches, thus accurately simulates sterile drug product
manufacturing process.
9.3 Worst Case Situation
During the manufacturing of media fill batch(s) No’s MF/01 & MF/02 no worst-case situations with
respect to the sterilized components hold time were planned for simulation. However, comparison of the
hold times of the sterilized components before use in the media fill batch No’s MF/01 & MF/02 with the
established/ validated hold times (study/ SOP reference - REP/MIS/005/00 & REP/MIS/006/00) is
presented in the following
Table 2: Comparison of sterilized component hold times
Batch No. Holding Time

Hold Time MF/01 MF/01 Std. Hold Hold Time (min)

Reason /
Time10
From To From To Purpose MF/01 MF/01 Remark
(NMT)

16:18 01:22 13:32 01:10 Awaiting

Sterile area use in the
30-03- 01-04- 31-03- 02-04- 72 Hrs. 34:04 Hrs. 35:38 Hrs. --
garments vial filling
2017 2017 2017 2017 machine

21:17 01:22 00:17 01:10 Awaiting

Rubber
use in the
stoppers in 30-03- 01-04- 01-04- 02-04- 72 Hrs. 28:05 Hrs. 24:53 Hrs. --
vial filling
RFS bags 2017 2017 2017 2017 machine

03:29 01:22 05:51 01:10 Awaiting

Machine use in the
31-03- 01-04- 01-04- 02-04- 72 Hrs. 21:53 Hrs. 19:19 Hrs. --
Parts vial filling
2017 2017 2017 2017 machine

10 Established as per the report no. RPT/HT/SG/01/00 & RPT/HT/SI/01/00.

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Batch No. Holding Time

Hold Time MF/01 MF/01 Std. Hold Hold Time (min)

Reason /
Time10
From To From To Purpose MF/01 MF/01 Remark
(NMT)

06:06 01:22 08:19 01:10 Awaiting

Miscellaneou use in the
31-03- 01-04- 01-04- 02-04- 72 Hrs. 25:16 Hrs. 16:51 Hrs. --
s Media vial filling
2017 2017 2017 2017 machine

23:30 01:22 21:30 01:10 Awaiting

Liquid Media
30-03- 01-04- 31-03- 02-04- use in the 72 Hrs. 49:52 Hrs. 51:40 Hrs. --
Load
2017 2017 2017 2017 vial filling

Comparison of the hold times of sterilized components and equipment before their use in the
manufacturing of media fill batch No’s MF/01 & MF/02 with the established/ validated hold time
indicates; that the hold times of the sterilized components were observed within the established hold
time limits as per the “Hold Time Study” report no. REP/MIS/005/00 & REP/MIS/006/00 and are
comparable with the routine product manufacturing batches.
9.4 Process Simulation Batch Size
Batch size of the media fill batch No’s. MF/01 & MF/02 were selected in accordance with the with the
media fill batch size criteria defined in the SOP # QA/MF/010-R09. The SOP, instructs to use the
rationale presented in the following Table 3 for deciding the media fill batch size.
Table 3: Media fill batch size selection rationale
Criteria If the production batch size is; Then, the media fill batch should be;
1 < 5,000 > actual product batch size
2 > 5,000 15,000

Comparison of a routine product and media fill batch size is presented in the following Table 4.
Table 4: Comparison of routine product and media fill batch sizes
Target Batch Size (No. of Vials)
Media Fill Batch Remark
Container Closure
Routine Product (Range) (Plan/ Incubated) (Ref. Table 3)
MF/01 MF/02
10mL
Tubular, 16,500
20mm, ---
USP Type (15,486)
I, Glass Grey, Meets batch size
Bromo 5,034 to 83,333vials criteria 2
15mL
Butyl,
Tubular, 16,500
RFS ---
USP Type (15,127)
I, Glass

Comparison of the routine product and media fill batch sizes indicates; that the batch sizes of the media
fill batch No’s. MF/01 & MF/01 vials meet the batch size criteria of SOP # QA/MF/010-R09, which is in
line with the USFDA guidance for industry, “sterile drug products produced by aseptic processing –
cGMP”, Sep. 2004 and PDA technical report no. 22, revised 2011.
9.5 Media Selection
The details of microbial growth medium used in each media fill batch(s) batches are presented in the
following Table 5.

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Table 5: Microbial growth medium used in the media fills

Specification Supplier Lot No’s.
Pre-sterilized, Soybean Casein Digest Medium, 0000971247
HIMEDIA make. AR # RM/20/0161

Pre-sterilized Soybean Casein Digest Medium (SCDM) is selected due to the reason(s); (a) it is
essentially the same formulation as the medium that is used for routine environmental monitoring
(Soybean Casein Digest Agar), (b) it is a non-selective media and (c) demonstrates to promote the
growth of wide range of microorganisms including low levels of the challenged organisms.
In order to ensure the media used for the process simulation test (media fill) promotes growth of the
organisms, growth promotion test of the media is performed at three levels, as explained below:
 Each incoming media lot on receipt is accepted only if it meets the acceptance criteria for growth
promotion test by inoculating with a <100 CFU challenge. If the growth promotion testing fails, the
incoming media lot is not accepted. This ensures, the use of right media for the process simulation
test (media fill). Results of the incoming GPT for media lot # 0000971247, AR # RM/20/0161
indicates compliance with the test acceptance criteria, refer test report # GPT/RM/17/253
 Sample of bulk media solution manufactured for process simulation test (media filling) is collected
post sterilization from the dosing needle to demonstrate the growth test by inoculating with a <100
CFU challenge. This helps in establishing the fact that, the media solution retains its growth
promotion property when it undergoes through a manufacturing process, as well as sterility of the
media dosing system is maintained. Results of the GPT for media sample collected post
sterilization from the dosing needle indicates compliance with the test acceptance criteria, refer test
report # GPT/BK/17/0163, GPT/BK/17/0176, GPT/BK/17/0186 & GPT/BK/17/0187.
 Similarly, vials of the successful media fill batch post 14-days incubation are also tested for the
growth promotion test by inoculating with a <100 CFU challenge. This helps establishing a fact that,
the media filled into the vial retained its growth promotion properties even past its transition through
the range of incubation temperatures. Results of the GPT for media vials collected post incubation
indicates compliance with the test acceptance criteria, refer test report # MB/17/0223 &
MB//17/0228
 Organisms selected for the media growth promotion test represent USP indicator organisms, as
well as the most commonly observed environmental isolates in the aseptic processing area.
Review of the Table 5 indicates appropriate media was used in the media fill batch No’s. MF/01 & MF/01
and that, it retained its growth promotion property throughout the stages of manufacturing as well as
incubation.
9.6 Compounding
Media solution (3%) is prepared by dissolving the dehydrated media in Water for Injection at room
temperature and mixed for a nominal 30 minutes according to the SOP # QA/MF/056-R02. Before
sterilizing the media, in-process sample was withdrawn to check media solution pH. Actual compounding
conditions and in-process test result ranges (minimum and maximum values) observed are presented in
the following Table 6.
Table 6: Compounding Conditions and In-process Test Results
Media Solution Manufacturing Details
Conditions / Results
MF/01 MF/02
Quantity of media used (Kg) 3.465 5.445
Quantity of WFI Used (L) 115.5 181.5
pH (Limit: 7.2 + 0.2) 7.36 7.37
GPT result as per SOP # QA/QC/124-R07 Pass Pass

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9.7 Component, Machine Parts & Accessories Sterilization

The component, machine part & rubber stopper is sterilized by steam sterilization in a manner to the
production process being simulated. All the load is sterilized with defined load pattern with defined load
number as per SOP No. # QA/MF/025-R03.
pre-washed and pre-siliconized rubber stoppers are received in ready for sterilization (RFS), Tyvek
bags. The rubber stoppers are sterilized using steam sterilizer (SS-01), transferred and stored under
LAF in cooling zone until used in the manufacturing.
Over-seals are transferred into the vial sealing room through dynamic pass-box. The bags containing
over- seals are subject to hold under UV light for 30 minutes in the dynamic pass-box before they are
used for sealing.
Similarly, the filling machine parts/ accessories (such as, dosing disc, star-wheel, powder hoppers,
interconnecting tubes etc.) are washed, dried, wrapped in lint-free bags and sterilized in a steam
sterilizer (SS-01) using a validated sterilization cycle. The sterilized machine parts/ components are
transferred and stored under LAF in cooling zone until used in the manufacturing.
Comparison of sterilization cycles followed for the sterilization of components and machine parts in
routine product batch manufacturing and media fill batch No’s. MF/01 & MF/01 is presented in the
following Table 7 .
Table 7: Comparison of routine product and media fill batch sizes
Cycle Name
Load/ Component Routine Product Media Fill Batch
Batch MF/01 MF/02
Sterile area garments HPHV HPHV HPHV
Rubber stoppers in RFS bags HPHV HPHV HPHV
Machine parts HPHV HPHV HPHV
Miscellaneous HPHV HPHV HPHV
Liquid Media --- STANDARD STANDARD
Review of the sterilization records indicate using same sterilization cycle(s) that are being used for
sterilization of machine parts and components in the routine product batch manufacturing.
9.8 Filling
9.8.1 Filling Speed
The filling and over-sealing machine were operated as per the same procedure and simulated speed
range normally followed for filling the routine production batches. As the liquid fill unit is used in the
media fill batches only, the vial filling speed lowers significantly as compared to the machine speed in
routine product batch manufacturing and also that the routine product batch sizes are significantly
higher, a concept of alternate filling between media filled and empty/ powder filled vials has been
followed. The empty vials were filled at maximum speed to experience interventions that are usually
experienced due to running the filling machine at its maximum speed, immediately on correcting the
intervention vials were filled with media with an intent to evaluate the impact due to the intervention and
its correction. Such vials were appropriately identified and included into the subsequent inspection as
well as incubation. This practice is in-line with the SOP # QA/MF/010-R09, USFDA guidance for
industry, “sterile drug products produced by aseptic processing – cGMP”, Sep. 2004 and PDA technical
report no. 22, revised 2011.
However, the planned number of vials (about 15,000) have been filled to represent the manufacturing
process under normal conditions. Comparison of filling speed in the routine product batch and media fill
batch No’s. MF/01 & MF/01 is presented in the following

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Table 8: Vial filling speed comparison in routine product batch and media fill batch(s)
Vial Filling Machine Speed (VPM)
Routine Product Media Fill Batch (Average/ speed simulation)
(Average/ range) MF/01 MF/02
60
(simulated @ 160 for 10 min, media
filled vials subsequent to the speed ---
simulation trial collected in tray no’s
140 15 and 16)
(160 to 240) 60
(simulated @ 200 & 240 for 15 min
--- each, media filled vials subsequent
to the speed simulation trial collected
in tray no’s 15 & 42 respectively)

The sealed and media filled vials are collected in HDPE trays. Each vial is then subject to on-line visual
inspection visual inspection.
Review of the vial filling machine speeds indicate close simulation of the normal operating machine
speeds that is used for the filling of routine product batch(s). The vial filling machine speed range
simulated were adequate to mimic commercial production conditions, accurately assess the potential for
commercial batch contamination and simulate activities that are representative of the manufacturing
process. Simulation of;
(a) high line speed helped in evaluating a significant degree of manual manipulation due to the
frequent interventions, and (b) slow line speed helped in evaluating prolonged exposure of the
sterile drug product and containers/closures in the aseptic processing area.
9.8.2 Filling Duration
The overall length of the vial filling duration simulated in media fill batches was similar to the filling
duration for routine production batches. Comparison of vial filling duration for the routine production
batches and the media fill batch(s) is presented in the following Table 9.
Table 9: Vial filling duration comparison in routine product batch and media fill batch(s)
Vial Filling Duration – including set-up and breaks (Hrs.)
MF/01 Media Fill Batch MF/02
Routine Product

16 17:23 ---
(2 shifts) --- 16:51

Review of the vial filling duration indicate appropriate simulation of the vial filling duration and that the
routine product batch filling duration and the media fill batch duration are comparable. The duration of
the media fill runs was adequate to incorporate manipulations and interventions, as well as
appropriate consideration of the duration of the actual aseptic processing operation. All
interventions that commonly occur in a routine product batch manufacturing were simulated in the
entire duration of filling each media fill batch.
9.8.3 Filling weight and volume
Media fill volume filled during the media fill batch(s) manufacturing is set in accordance with the media fill
SOP # QA/MF/010-R09, (fill volume should not be less than 70% as per the vial size). However, the
volume of media filled in batches is adequate to facilitate contact with the entire internal surface area,
and to allow the visual inspection to easily detect any microbial growth / contamination. Following
Table 10 presents the sterile placebo powder (Mannitol) and sterile, liquid media fill volume and
comparison with the suggested values.
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Table 10: Powder & media fill volumes

Review of the sterile placebo powder (Mannitol) and sterile, liquid media fill weight and volume
respectively indicate actually filled weight and volume is comparable with the values suggested in the
SOP # QA/MF/010-R09 and closely simulates the sterile powder filling step/ process.
9.8.4 Inert Gassing
While the use of sterile filtered inert/ a specified gas like Nitrogen/ Carbon Di-oxide respectively is used
for the flushing of empty vials and purging the powder from the dosing wheel in the manufacturing of
each product, post fill flushing of the vial headspace with Nitrogen or any other gas is not required in the
manufacturing process of few products. As, the use and presence of inert gas in the manufacturing
process and in the vial, could adversely impact the microbiological growth promotion the inert gas is
replaced with sterile filtered compressed air in the manufacturing of media fill batches for a close
process simulation. Following Table 11 presents the sterile placebo powder (Mannitol) and sterile, liquid
media fill volume and comparison with the suggested values.
Table 11: Comparison of Inert Gassing Parameters

Review of the filling process as shown in the above table 11 indicates that, the use of inert gas in routine
product manufacturing process was replaced with the sterile filtered compressed air without changing/
modifying the normal control parameters, thus closely simulating a routine product manufacturing
process.
9.9 Interventions
The interventions that may normally occur during a routine product batch manufacturing process have
been simulated as per their nature. The interventions are basically categorized in 2 types as follows:
 Routine/ Normal (Inherent Intervention)
 Non-routine (Corrective Intervention)
The interventions that usually occur during the course of aseptic processing in the manufacturing
process, and that must be simulated in a media fill batch manufacturing are defined and listed in the
SOP # MG/IN/071-R02, “Intervention – Life Cycle Management”.
Section 5.5 of the SOP # QA/MF/010-R09, “Process Simulation by Media Fill Study” provides
instructions for following a media fill execution plan, which also includes the list of aseptic interventions to
be simulated in the corresponding media fill batch. The list of aseptic interventions and frequency to be
simulated is determined based on the intervention trends review of the interventions recorded in
routine production

11
As specified in the SOP # QA-SG-073-R09 (section 5.5.7.3)
12
As specified in the SOP # QA-SG-073-R09 (section 5.5.7.4)

Depending upon the product and/ or process

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batch(s) manufactured since the last media fill batch(s). This requirement is defined in the section 5.3 of
the SOP # MG/IN/071-R02, “Intervention – Life Cycle Management”.
Vial Type
Sterile Mannitol (g) Nominal media fill volume (mL)
As explained in 0 above, SOP # MG/IN/071-R02, “Intervention – Life Cycle Management” is an outcome
Size (mL) Neck (mm)
of a detailed exercise including
Target11 risk
MF/01 assessment
MF/02and trend
Targetanalysis
12 of the interventions
MF/01 MF/02observed in all
routine
10 product 20batch(s) manufactured
0.525 0.52575 in the year
--- 2016 and
7 has been made
6.860 to 7.140 effective in---March 2017. As
media
15 fill batch20numbers0.825
MF/01 & ---
MF/02 are0.82534
being manufactured
11 first---after implementation of this SOP,
11.01 to 11.04
it was decided to simulate all identified interventions in these media fill batches.
However, in the subsequent media fill batch(s) type and frequency of the interventions will be simulated
based on the intervention trends review of the interventions recorded in routine production batch(s)
manufactured since the last media fill batch(s).
Details of the interventions occurred/ simulated in media fill batch No’s. MF/01 & MF/02 are presented in
the following Table 12.
Table 12: Interventions
Defined Frequency Occurrence/ simulation in MF
Intervention (Ref. Media Fill Batch #
Details
No. Execution Plan for MF/01
& MF/02) MF/01 MF/02 Remark
Routine/ Normal (Inherent Intervention)
R-01 Machine Parts Assembly 1 1 1 ---
Rubber stoppers charging into the rubber stopper
R-02 20 20 20 ---
infeed bowl Batch Details
R-03 Powder Charging into the Powder Hopper Routine Production 10 10 Fill Batches
Media 10 ---
R-04 Conditions / Parameters
Fill weight adjustment (On PLC) Min Max 8 Min 8 8 Max ---
Removal of empty fallen/ broken/0.5 cracked vial 0.5 0.5 0.5
R-05 anywhere between the infeed turntable and  12 12 12 ---
Sterile filtered Nitrogen Sterile filtered Compressed Air Limit (0.2 to
the infeed star-wheel
Inert Gassing
(0.2 to 1.2 bar) 1.2 bar)
R-06 Movement of swing conveyor 18 18 18 ---
Removal of powder spilled on conveyor or outfeed
R-07 7 7 7 ---
turntable line using vacuum cleaner
To perform in process sampling (for verification of
R-08 14 14 14 ---
online check-weigher)
R-09 Environmental Monitoring by the Microbiologist 5 5 5 ---
Non- routine (Corrective Intervention)
NR-01 Compressed gas dosing pressure adjustment. 8 8 8 ---
NR-02 Adjustment of line speed 6 6 6 ---
NR-03 Adjustment of dosing wheel height 10 10 10 ---
Removals of tilted/ fallen, filled/unfilled vials from
NR-04 12 12 22
the out-feed turntable
Rubber stoppers chute adjustment during the
NR-05 7 7 31
process
Adjustment of rubber stopper stoppering heads
NR-06 6 6 6 ---
during the process
NR-07 Removal of jammed stopper in stopper chute/ bowl 12 12 12 ---
Removal of vials under the dosing disc with sterile
NR-08 4 4 4 ---
forceps
NR-09 Power failure for 5 minutes 2 2 2 ---
Cleaning of choked powder dosing piston from the
NR-10 2 2 2 ---
dosing disc
Breakdown of upstream and/ or downstream
NR-11 4 4 4 ---
machine for about 30 minutes
NR-12 Clutch adjustment for the star wheel 2 2 2 ---

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Interventions presented in the above Table 12 are representative of all the activities and interventions
representative of each shift, and shift changeover. While interventions and/or stoppages are normally
recorded in the batch record, the manner of documenting these occurrences in the media fill bath
manufacturing record is slightly different. Each intervention is documented with associated time, duration
and of the tray number in which the vial(s) filled immediately after the intervention event were collected.
As explained in section 9.8.1, the vial filling speed is comparatively low in the manufacturing of media fill batch(s)
due the use of liquid fill unit, which is used in the media fill batches only. A concept of alternate filling between
media filled and empty/ powder filled vials has been followed to simulate/ experience interventions, that usually
occur due to running the filling machine at its maximum speed. The empty vials were filled at maximum speed to
experience the normal interventions, immediately on correcting the intervention vials were filled with media with
an intent to evaluate the impact due to the intervention and its correction. Such vials were appropriately identified
and included into the subsequent inspection as well as incubation.
Review of the interventions as shown in the above Table 12 indicates, simulation of all interventions that normally
and routinely have been identified in a routine product batch manufacturing process, thus closely simulating a
routine product manufacturing process.
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.
9.10 Vials Reconciliation
The total number of vials manufactured, rejected at different stages of manufacturing, vials incubated
and inspected post-incubation has been recorded and reconciled in the corresponding media fill batch
manufacturing records. A summary is presented in the following Table 13.
Table 13: Vials Reconciliation
Post Incubation Visual
Filling, Stoppering & Sealing (Vials) Visual Inspection (Vials)
Theoretical Inspection (Vials)
Filling
Batch Batch Size To Reject- Reject- To
Date Filling Sealing
(Vials) Inspection 114 215 Incubation Inspected Contaminated
Rejects13 Rejects
(A) (B) (C) (D) = (A – B)
31-03-
MF/01 16,500 146 42 15,481 0 227 15,481 15,481 016
2017
01-04-
MF/02 16,500 260 82 15,137 0 244 15,137 15,137 0
2017

Review of the vial reconciliation indicates, the vials reconciliation for the media fill vial batch no’s. MF/01
& MF/02is done in a manner it is done for the routine drug product batches. The only exception is, the
integral rejects of vial visual inspection step, such as visible particulate matter and other cosmetic
defects are also accounted and incubated along with the good vials. This is in accordance with: (a)
USFDA guidance document, “Guidance for Industry Sterile Drug Products Produced by Aseptic
Processing — Current Good Manufacturing Practice”, Sep. 2004, (b) PDA Technical Report No. 22,
“Process Simulation

13 These are the vials rejected either automatically by the vial filling machine and/ or fallen during transportation, over sealing. The vials that are automatically
rejected at the vial filling stage are non-integral process rejects, due to not fulfilling the conditions defined in the vial filling machine controls, such as no
stopper, no over seal, low/ high fill etc. These vials are on-line rejects.
14 Defects affecting the container closure integrity like – cracked vials, missing stoppers/ missing seals. Such vials are excluded from the incubation.
15 Defects not affecting the container closure integrity like – cosmetic defects (scratch, moulding defects etc.) and visual particulate matter. Such vials are
included for the incubation.
16 One cracked vial was found during the inspection of the vials on 7th day of incubation. As this vial, can be a representative of drug product released to
the market and that, no growth/ turbidity/ contamination was noted in this vial, it was not excluded and allowed to remain for the rest of the incubation
period. An investigation is completed to find out the root cause and identify appropriate CAPA for the cracked vial – refer incident/ investigation report
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no. IIR/2017/021
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for Aseptically filled Products, 2011, (c) our experience and knowledge on this subject, and (d) current
industry trend.
Review of the above data, also indicates that vial quantities for the individual media fill batches are
accurately reconciled and no discrepancies observed. This fact is confirmed by reconciliation of the
overall sum total numbers, as well and that no contaminated unit was found in the batch no’s. MF/01
and MF/02 after a 14-day incubation period.
9.11 Environmental Monitoring
Environmental monitoring including airborne aerobic microbial monitoring using settle plates, active air
monitoring, swabs/ contacts (at the end of processing) was performed during the media fill batches to
simulate the environmental monitoring process followed during the routine aseptic manufacturing
process.
Environmental monitoring is performed and results reported as per the corresponding SOP. The original
reports are available with the corresponding batch records, however summary of the environmental
monitoring results by settle plate, active air & surface monitoring for the vial filling room, vial sealing and
other production area used during the manufacturing of media fill batches are presented in the following
Table 14, Table 15 and Table 16.
Table 14: Environmental monitoring results of settle plate (90 mm, per 4 hours)
Results (dia. 90mm; CFU/ 4 hours)
Settle plate study and monitoring details
MF/01 MF/02
Limit (dia. 90mm;
Room / Area Sample CFU/ 4 hours) No. of No. of
Grade Min Max Min Max
Details17 Location ID Plates Plates
Alert Action
SP01 A -- 1 4 0 0 4 0 0
SP02 A -- 1 5 0 0 5 0 0
SP03 A -- 1 5 0 0 5 0 0
SP04 A -- 1 5 0 0 5 0 0
Vial filling room SP05 A -- 1 4 0 0 4 0 0
SP06 B 2 3 4 0 0 4 0 0
SP07 B 2 3 4 0 0 4 0 0
SP34 A -- 1 4 0 0 4 0 0
SP08 B 2 3 1 0 0 1 0 0
SP09 B 2 3 1 0 0 1 0 0
Sterile buffer SP10 B 2 3 1 0 0 1 0 0
SP11 C/A/B 2 3 1 0 1 1 0 0
SP12 B/A/B 2 3 1 0 0 1 0 0
Sterile passage SP13 B 2 3 1 0 0 1 0 0
SP14 A -- 1 1 0 0 1 0 0
SP15 A -- 1 1 0 0 1 0 0
RM sampling room
SP16 B 2 3 1 0 0 1 0 0
SP17 UC/A/B 2 3 1 0 1 1 0 0
Change room-1 SP19 C 4 5 1 0 24 1 0 26
Change room -2 SP20 C 4 5 1 0 1 1 0 2
SP21 B 2 3 1 0 0 1 0 1
Change room-3
SP22 B/A/B 2 3 1 0 0 1 0 0
Entry buffer SP23 B 2 3 1 0 0 1 0 0
Exit buffer SP24 C 4 5 1 0 2 1 0 0
Return change room SP25 D 35 50 1 0 21 1 0 25

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17
Refer Annexure – 1 for the aseptic processing area layout.

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Results (dia. 90mm; CFU/ 4 hours)

Settle plate study and monitoring details
MF/01 MF/02
Limit (dia. 90mm;
Room / Area Sample CFU/ 4 hours) No. of No. of
Grade Min Max Min Max
Details17 Location ID Plates Plates
Alert Action
Mobile LAF (Cooling SP32 A -- 1 1 0 0 1 0 0
Zone) SP33 A -- 1 1 0 0 1 0 0
SP26 C 4 5 1 0 1 1 0 0
SP27 A -- 1 1 0 0 1 0 0
SP28 UC/A/C 4 5 1 0 2 1 0 1
Vial sealing room
SP29 C 4 5 1 0 0 1 0 0
SP30 C 4 5 1 0 0 1 0 0
SP31 D 35 50 1 0 20 1 0 16
SP32 A/C 4 5 1 0 1 1 0 0
Garment Washing
SP33 C 35 50 1 0 10 1 0 12
SP34 A/C 4 5 1 0 2 1 0 0
Equipment Washing SP35 A/C 4 5 1 0 0 1 0 0
& sterilization Room SP36 C 35 50 1 0 7 1 0 11
SP37 A/C 4 5 1 0 1 1 0 0
Vial Washing Room SP38 C 35 50 1 0 13 1 0 15
De-Cartoning SP39 D 70 100 1 0 39 1 0 40

Table 15: Environmental monitoring results of Active Air Sampling

MF/01 Results (CFU/ M3) MF/02

Active Air Sampling study and monitoring details

Limit (CFU/
Room / Area Sample M3) No. of No. of
Grade Min Max Min Max
Details Location ID Samples Samples
Alert Alert
VA01 A -- 1 1 0 0 1 0 0
VA02 A -- 1 1 0 0 1 0 0
VA03 A -- 1 1 0 0 1 0 0
VA04 A -- 1 1 0 0 1 0 0
Vial filling room VA05 A -- 1 1 0 0 1 0 0
VA06 B 2 3 1 0 1 1 0 1
VA07 B 2 3 1 0 0 1 0 0
VA34 A -- 1 1 0 0 1 0 0
VA08 B 2 3 1 0 0 1 0 0
VA09 B 2 3 1 0 2 1 0 0
Sterile buffer VA10 B 2 3 1 0 2 1 0 0
VA11 C/A/B 2 3 1 0 0 1 0 0
VA12 B/A/B 2 3 1 0 0 1 0 0
Sterile passage VA13 B 2 3 1 0 0 1 0 0
VA14 A -- 1 1 0 0 1 0 0
VA15 A -- 1 1 0 0 1 0 0
RM sampling room
VA16 B 2 3 1 0 0 1 0 1
VA17 UC/A/B 2 3 1 0 0 1 0 0
Change room-1 VA19 C 4 5 1 0 31 1 0 30
Change room -2 VA20 C 4 5 1 0 3 1 0 2
VA21 B 2 3 1 0 0 1 0 0
Change room-3
VA22 B/A/B 2 3 1 0 0 1 0 1
Entry buffer VA23 B 2 3 1 0 0 1 0 0
Exit buffer VA24 C 4 5 1 0 4 1 0 3

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MF/01 Results (CFU/ M3) MF/02

Active Air Sampling study and monitoring details

Limit (CFU/
Room / Area Sample M3) No. of No. of
Grade Min Max Min Max
Details Location ID Samples Samples
Alert Alert
Return change
VA25 D 35 50 1 0 26 1 0 26
room
Mobile LAF VA32 A -- 1 1 0 0 1 0 0
(Cooling Zone) VA33 A -- 1 1 0 0 1 0 0
VA26 C 4 5 1 0 3 1 0 3
VA27 A -- 1 1 0 0 1 0 0
VA28 UC/A/C 4 5 1 0 4 1 0 0
Vial sealing room VA29 C 4 5 1 0 0 1 0 4
VA30 C 4 5 1 0 2 1 0 0
VA31 D 35 50 1 0 40 1 0 36
VA32 A/C 4 5 1 0 2 1 0 0
Garment Washing
VA33 C 35 50 1 0 14 1 0 12
VA34 A/C 4 5 1 0 4 1 0 3
Equipment Washing VA35 A/C 4 5 1 0 1 1 0 0
& sterilization Room VA36 C 35 50 1 0 19 1 0 15
VA37 A/C 4 5 1 0 1 1 0 4
Vial Washing Room VA38 C 35 50 1 0 18 1 0 19
De- Cartoning VA39 D 70 100 1 0 69 1 0 62

Table 16: Environmental monitoring results of surface monitoring

MF/01 Results MF/02

Surface swab sampling study date/and monitoring details

Room / Area Sample Alert Action No. of No. of

Grade Min Max Min Max
Details Location ID Level Levels Samples Samples
SS01 A -- 1 1 0 0 1 0 0
SS02 A -- 1 1 0 0 1 0 0
SS03 A -- 1 1 0 0 1 0 0
Vial filling room
SS04 A -- 1 1 0 0 1 0 0
SS05 A -- 1 1 0 0 1 0 0
SM20 A -- 1 1 0 0 1 0 0
SS06 B 2 3 1 0 0 1 0 0
Sterile buffer SS07 C/A/B 2 3 1 0 0 1 0 0
SS08 B/A/B 2 3 1 0 0 1 0 0
SS09 A -- 1 1 0 0 1 0 0
RM sampling SS10 A -- 1 1 0 0 1 0 0
room
SS11 UC/A/B 2 3 1 0 0 1 0 0
Change room-1 SS12 D 7 10 1 0 0 1 0 2
Change room -
SS13 C 4 5 1 0 0 1 0 1
2
SS14 B/A/B 2 3 1 0 0 1 0 0
Change room-3
SS15 B 2 3 1 0 0 1 0 0
Entry buffer SS16 B 2 3 1 0 0 1 0 0
Sterile passage SS17 B 2 3 1 0 0 1 0 0
Exit buffer SS18 C 4 5 1 0 0 1 0 0
Return Airlock SS19 D 7 10 1 0 2 1 0 2

Review of the environmental monitoring results indicate no excursion from the alert/ action limit in the core
aseptic processing area (Grade A and Grade B areas). However, 4 incidences of environmental monitoring

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results (by settle plate method as well as active air sampling method) equal to or greater that the alert
and action limited have been observed. Excursion from the action limit is observed in the change room –
1, which is not unusual due to the heavy personnel movement in this area. However, investigation for
these EM excursions have been performed according to the SOP #, “QA/EM/059”. Please refer
investigation report(s) IIR/17/079
9.12 Personnel Monitoring
Personnel monitoring of the personnel involved in the aseptic manufacturing process is performed during
the media fill batches and the reports are available with the corresponding batch records. A summary of
personnel monitoring results for all personnel that participated in the media fill batch manufacturing is
presented in the following Table 17.
Table 17: Personnel participates in the media fill
Department MF/01 Batch No. MF/02
Sr. No. Name Employee Code

1 Micro – 1 7404 Microbiology  

2 Prod – 1 7059 Production  
3 Prod – 2 3804 Production  
4 Prod – 3 4809 Production  
5 Prod – 4 9345 Production  
6 Micro – 2 8666 Microbiology  
7 IPQA – 1 4156 IPQA  
8 Micro – 3 4237 Microbiology  
9 Prod – 5 6880 Production  
10 Prod – 6 8674 Production  
11 Prod – 7 5909 Production  
12 QA – 1 8843 QA  
13 Prod – 8 8943 Production  
14 IPQA – 2 7593 IPQA  
15 QA – 2 6456 QA  
16 Eng. – 1 9676 Engineering  
17 Micro - 4 3948 Microbiology  
18 QA – 3 7975 QA  
19 Eng. – 2 1219 Engineering  
20 Prod – 9 1679 Production  
21 Micro – 5 1168 Microbiology  
22 Micro - 6 1835 Microbiology  
23 Prod – 10 4512 Production  
24 Prod – 11 2297 Production  
25 Prod – 12 1846 Production  
26 Prod – 13 3260 Production  
() participated
() Not Participated

Acceptance criteria: As per current version of SOP as “SOP on personnel qualification for aseptic
processing and sterility testing area SOP No. QA/QC/506” and “SOP on environmental monitoring of
sterile product manufacturing facility QA/QC/403” respectively.
Review of the personnel monitoring results in the Table 17 indicates all personnel who are
authorized to enter the aseptic processing room during manufacturing, including technicians and
maintenance personnel, microbiologist, production, and QA participated in at least one media fill
batch manufacturing. Their participation was consistent with the nature of each operator’s duties
during routine product batch manufacturing.

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9.13 Post Fill Visual Inspection

The media filled vials after filling and sealing operation were inspected manually by trained visual
inspectors according to the SOP # MF/PR/076-R10. Same procedure is followed for the inspection of the
routine product batch(s). The standard of inspection and criteria used for rejection applied to the media
filled vials is same as that being used in the routine product batch(s) visual inspection.
The vials observed with container/ closure integrity related defects, such as cracked vials, missing
stopper/ missing seal etc. were rejected and not considered for incubation. Such vials are internally
known as “non- integral vials”, which are identified as “Reject-1” in the Table 13. These vials are not
representative of the microbial quality of the product and hence all such units are discarded and
recorded in the corresponding media fill batch records. Summary of all such rejects is presented in Table
13.
All filled units identified and rejected as “cosmetic defects” in visual inspection step, such as deformed
containers, particulate matter, scratch, molding defects etc. are collected separately, incubated along
with the good vials and assessed as part of the process simulation test. Such units are internally known
as “cosmetic defects”, which are identified as “Reject-2” in the Table 13.
The standard and the process of visual inspection and criteria used for rejection applied to the media
filled vials is observed same as that being used for the routine product batch(s) visual inspection. This is
in accordance with: (a) USFDA guidance document, “Guidance for Industry Sterile Drug Products
Produced by Aseptic Processing — Current Good Manufacturing Practice”, Sep. 2004, and (b) PDA
Technical Report No. 22, “Process Simulation for Aseptically filled Products, 2011.
9.14 Vial Labeling
The media filled vials were passed through the labeling machine/ line to simulate the level and type of
handling that a routine product batch(s) vials would experience. However, labels were not applied to the
media filled vials. This step was not followed in any of the previously manufactured media fills batch(s),
which was not an exact simulation of the complete manufacturing process. However, considering the
possibility of inspected vials experiencing stress during handling and processing of the vials through the
rigor of the labeling and packaging step, SOP # QA/MF/010-R09, “Process Simulation by Media Fill
Study” has been revised to include instructions for accurately simulating the labeling and packaging
step. It has been implemented since these media fill batches and will be followed in the future as well.
9.15 Incubation
Each vial accepted for incubation were collected in a standard HDPE trays, each tray was labelled with a
printed label containing following details.
 Batch number
 Tray number
 Quantity
Each tray containing inspected vials is then transferred into qualified and calibrated temperature-controlled
rooms.
9.15.1 Orientation of media filled units during the incubation
All media filled units are stored/ incubated at 20 0C – 250C temperature range, in an inverted position for
the first 7 days of incubation, followed by storing/ incubating at 300C – 350C for the next 7 days, in an
upright position. Vial’s orientation, and loading details are recorded in the corresponding media fill batch
manufacturing record.
9.15.2 Incubation Conditions
All media filled units were incubated in qualified and validated incubation rooms for a minimum of 14
days. The units were incubated for 7 days at 20°C - 250C followed by further 7 days at 30°C - 350C.

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During all 14 days of incubation the incubator chamber temperature was monitored using dataloggers.
The incubation reports, incubator temperature monitoring records are available with the corresponding
media fill batch manufacturing record. Summary of media fill vials incubation is presented in the following
Table 18.
Table 18: Summary of Media Fill Vials Incubation
Incubation @ 200C - 250C Incubation @ 300C - 350C
Batch No. Date Temp. (0C) Date Temp. (0C)
Start End Min Max Start End Min Max
MF/01 01-04-2017 08-04-2017 08-04-2017 16-04-2017
21.9 24.9 32.0 34.7
MF/02 02-04-2017 10-07-2017 10-07-2017 17-04-2017

Review of the results in Table 18 indicates, incubation of media filled vials under conditions adequate to
detect microorganisms that might otherwise be difficult to culture. Incubation conditions are established
and followed in accordance with: (a) USFDA guidance document, “Guidance for Industry Sterile Drug
Products Produced by Aseptic Processing — Current Good Manufacturing Practice”, Sep. 2004, and (b)
PDA Technical Report No. 22, “Process Simulation for Aseptically filled Products, 2011.
9.16 Post Incubation
9.16.1 Inspection
Inspection of the incubated vials is performed twice during the incubation, after 7 days – at the end of the
incubation period at 200C - 250C and after 14 days – at the end of the incubation period at 30 0C - 350C.
All these inspections were done by the qualified microbiologists according to the SOP # QA/MF/010-
R09, “Process Simulation by Media Fill Study”.
The final inspection report documents accurate count of the number of units inspected and is part of the
corresponding media fill batch manufacturing record. Table 13 presents the post incubation visual
inspection summary including the number of contaminated unit(s), if observed.
One cracked vial, from the tray no. 68 was found during the inspection of the media filled vials on 7th
day of incubation. Picture of the subject, cracked vial is presented in the following Figure 2.
Figure 2: Picture of a cracked vial observed during the inspection of vials (MF/01) after 7 day’s
incubation

As this vial, can be representative of drug product released to the market and that, no growth/ turbidity/
contamination was noted in this vial, it was not excluded and allowed to remain for the rest of the
incubation period. This is in-line with the SOP # QA/MF/010-R09, “Process Simulation by Media Fill
Study” and USFDA guidance document, “Guidance for Industry Sterile Drug Products Produced by
Aseptic Processing — Current Good Manufacturing Practice”, Sep. 2004.
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An investigation was completed to find out the root cause and to identify appropriate CAPA for the
cracked vial – refer incident/ investigation report no. IIR/2017/021 Summary of the root cause
investigation and preventive actions is explained below:
Most Probable Root Cause – 1:
Hypothesis: Extensive handling and/ or transportation of the media filled vial between the labeling
packaging – incubation @ 200C to 250C – visual inspection (after 7th day of incubation) – incubation @
300C to 350C – visual inspection (after 14th day of incubation).
Media filled vial collected in a HDPE trays, as explained in section 9.15 experience more mechanical
shock and stresses as compared to a routine product batch(s), due to the following reason(s):
 Media fill vials remain unlabeled and un-packaged subsequent to the visual inspection stage, thus
vulnerable to mechanical shocks and stresses.
 They undergo comparatively more handling and mechanical stresses in unlabeled and un-
packaged condition than a routine product batch vial. The routine product batch(s) after visual
inspection are labelled and packaged in their final secondary and tertiary packs. The packaged
finished product is then neatly stacked on a pallet before it is moved to the finished product
quarantine/ store. However, the media filled vials are extensively handled and/ or transported in
unlabeled and unpackaged condition between following steps: labeling packaging – incubation @
200C to 250C (in inverted condition) – visual inspection (after 7th day of incubation) – incubation @
300C to 350C (in upright condition) – visual inspection (after 14th day of incubation).
 Most of the trays containing media filled vials are usually partially packed, due to identifying group
of media filled vials to the corresponding intervention. Vials in partially packed trays are unstable
and tend to fall and/ or collide with each other when moved and handled, thus experiencing more
mechanical shock and stresses as compared to tightly packed vials in same trays for a routine
product batch(s).
Most Probable Root Cause – 2:
Hypothesis: Torsional stresses usually do not induce a special failure mode in glasses. Fracture starts at
a flaw and the crack propagates normal to the plane of maximum tensile stress, which is aligned at 450
to the shear stress direction. In other words, the shear stress (τ) state is equivalent to one in which a
material element is pulled in tension (σ1) in one direction and compressed laterally (σ2) as shown in
Figure 4. Fracture primarily occurs due to σ1 tensile stress and fracture propagates perpendicular to σ1.
Reference: Footnote 15
Figure 318: Torsional loading causes fractures at an angle to the longitudinal axis of a part. The shear
stresses are τ. These generate equivalent principal stresses σ1 in tension and σ2 in compression as
shown in (a). Fracture occurs perpendicular to σ1. (b) shows a schematic of a glass rod fractured in
torsion. The origin is at the arrow. Note how branching occurs to either side of the origin, but that only
one branch in each case continues, giving rise to the curved fracture surface.

18 NIST practice guide, “Fractography of Ceramics and Glasses” – Sections 4.14,

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Cracks may grow stably when loaded at stresses and stress intensities less than that necessary for the
flaw to become unstable, glasses, oxide ceramics, and ceramics with glassy boundary phase are
susceptible. The mechanism of slow crack growth at ambient temperature is a stress corrosion
phenomenon whereby stressed cracks that are open to the environment are attacked by water (or
another polar molecule.) The water may be in the form of liquid or gaseous molecules. The water
molecules attack the strained silicate or oxide bonds at the crack tip causing them to rupture which in
turn leads to stable crack extension.
Facts and Discussion:
Each glass vial undergoes an over-sealing process in which, the vial is compressed from the top and is
rotated clock-wise to crimp the aluminum seal around the vial rim collar, thus experiencing the torsional
at an angle to the longitudinal axis of a vial. Fractographical analysis of the crack as shown in the
following Figure 4 shows torsional loadings that produced twisted fracture surfaces at 450 angle to the
part length.
Figure 4: Fractographic analysis of the crack observed in the media fill vial from batch # MF/01

Compression Pressure

450
Torsional Stress

It appears, the subject vial had a flaw/ weakness as indicated by a “bright red dot” in the above Figure 4.
The vial when undergone the over-sealing process, the torsional stresses induced a microfracture at a
flaw that is difficult to identify visually in a vial visual inspection process. The subject vial when inverted
for incubation @ 200C to 250C the microfracture grown stably at ambient temperature. This is called as a
stress corrosion phenomenon whereby stressed cracks that are open to the environment are attacked by
water (or another polar molecule.) The water may be in the form of liquid or gaseous molecules. In this
case the water was available in the form of a media solution, water molecules attack the strained silicate
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or oxide bonds at the crack tip causing them to rupture which in turn led to propagation of a stable crack
normal to the plane of maximum tensile stress, which is aligned at 450 to the shear stress direction.
Perhaps, the mechanical stresses and shocks experienced by the subject vial during its extensive
handling and/ or transportation of the media filled vial between the labeling packaging – incubation @
200C to 250C
– visual inspection (after 7th day of incubation) – incubation @ 300C to 350C – visual inspection (after 14th
day of incubation) could have further augmented/ accelerated the process of crack formation as
explained in the “Most Probable Root Cause – 1” above.
The above discussion and assessment are further reinforced by the fact that, the shape, size and
location of the crack (running almost half length on the body of the vial) was highly visible and could
have been easily identified by the trained and qualified visual inspectors. These inspectors are capable
of identifying comparatively much smaller and finer defects.
Conclusion:
This crack appears to be an isolated incidence and resulted due to the combination of following factors:
(a) invisible/ dormant flaw in the subject glass vial, (b) torsional stresses it had experienced during the
vial over-sealing process, (c) subsequent mechanical shocks/ stresses it had experienced during the
post inspection handling and movement, and (d) stress corrosion phenomenon which came into effect
and further augmented the stressed cracks that are open to the environment are attacked by water
molecule (that was present in the form of media solution) when the subject vial was inverted for
incubation @ 200C to 250C for the first 7 days. Perhaps, the observed fracture did not result into a crack/
gap wide enough to breach the container integrity, thus protecting the contents inside the vial from
microbial contamination.
Preventive actions:
In order to avoid similar incidence in future batch(s) (routine product and/ or media fill batch) following
preventive actions have been recommended.
 Avoid partial packaging of HDPE trays. If unavoidable, then fill the empty space tightly with a
cardboard/ plastic bubble wrap packing.
 Do not incubate/ store the filled vials in an inverted position, as the vials are highly unstable in this
position and can fall and collide with each other resulting in a crack and/ or onset of a crack.
However, to ensure media solution contacts complete internal vial as well as rubber stopper
surface, each media filled vials would be inverted once in a controlled condition before its
incubation.
9.16.2 Stasis Test (Post Incubation Media Growth Promotion Test)
In order to demonstrate that, the media fill vials after subjecting to various processing and incubation
conditions still retains its capability to promote recovery of wide spectrum of microorganisms.
On completion of 14-days incubation, media growth promotion test (media fertility test) was performed
on actual media filled representative sample vials from each media fill batch, as per the Pharmacopeial
method. Results of the growth promotion test on media filled vials post incubation indicate that, each
media fill batch at the end of incubation period exhibited its growth promotion properties. The stasis test
reports are available with the corresponding batch records. Following table 18 presents the summary of
post incubation growth promotion test results.
Table 19: Summary of post incubation GPT (stasis test)
Total Media Fill batches Batches subject for post Batches passed in post
manufactured incubation GPT incubation GPT
76 76 76

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9.17 In-coming material and In-process Sampling:

Sampling of in-coming raw materials (sterile media and placebo powder – Mannitol) as well as the in-process materials was performed exactly in a manner it is
performed in a routine product batch manufacturing. However, sample of residual bulk media and sample vials after 14 days of incubation collected for GPT was
specific to the media fill batch manufacturing process. Additionally, the sample vials have been also tested for the “container-closure system integrity test” by microbial
suspension immersion test method to demonstrate suitability and performance of the container-closure system and vial over-sealing process respectively.
Table 20: Summary of incoming Media and Sterile Mannitol Sterility & Growth Promotion Test Results
Process Test to be performed Sample Acceptance
Stage Container No. Batch No. MF/01 A.R. No. Batch No. MF/02 A.R. No.
Variables / Reference quantity Criteria
1st Soluble. MB/17/0160 Soluble. MB/17/0197
Solubility in Medium 2 gm Shall be soluble. 2nd Soluble. MB/17/0168 Soluble. MB/17/0198
3rd Soluble. MB/17/0173 NA NA
1st Complies MB/17/0165 Complies MB/17/0193
Mannitol Shall be sterile 2nd Complies MB/17/0169 Complies MB/17/0194
Sterility from intact Complies MB/17/0173 NA. NA
Sampling Aseptic 3rd
bag before filling/ 6 gm
(Sample sampling 1st 1st RM/17/0069 1st RM/17/0111
GTP-205 each pouch
Pouch) Container’s taken RM/17/0069 RM/17/0111
2nd 2nd 2nd
3rd 3rd RM/17/0069 NA. NA
1st <0.0025 EU/mg MB/17/0164 <0.0025 EU/mg MB/17/0195
Bacterial Endotoxin / NMT 0.0025
2 gm 2nd <0.0025 EU/mg MB/17/0170 <0.0025 EU/mg MB/17/0196
GTP-206 each pouch EU/mg
3rd <0.0025 EU/mg MB/17/0174 NA. NA
1st Complies MB/17/0162 Complies MB/17/0184
2nd Complies MB/17/0162 Complies MB/17/0184
Should be sterile
Sterility and GPT as 3rd Complies MB/17/0175 Complies MB/17/0185
300+ Complies MB/17/0175 Complies MB/17/0185
Before Filling per 4th
Sterile Media 100 ml/
GTP-205 and 1st Growth observed GPT/BK/17/0163 Complies GPT/BK/17/0186
Containers
QA/QC/124-R07 Shall promote Growth observed GPT/BK/17/0163 Complies GPT/BK/17/0186
2nd
growth 3rd Growth observed GPT/BK/17/0176 Complies GPT/BK/17/0187
4th Growth observed GPT/BK/17/0176 Complies GPT/BK/17/0187

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 Table 21: Summary of liquid borne particulate count results for the sterile filtered media and machine parts rinse samples
A.R. No./ A.R. No./
Process Test to be performed / Sample
Stage Acceptance Criteria Batch No. MF/01 Ref. BMR Page Batch No. MF/02 Ref. BMR Page
Variables Reference quantity
No. No.

LBPC
Rinse Water Particulate 10 µ - NMT 60/ml
4.0 MB/17/0159 2.1 MB/17/0172
Before Matter and Endotoxin 25 µ - NMT 06/ ml
Machine parts 100 ml. 0.0 / Page No. 0.1 / Page No.
Sterilization Test as per QA/QC/080
and GTP-602 17 of 76 17 of 76
Endotoxin
< 0.125 EU/ml < 0.125 EU/ml
< 0.125 EU/ml

Page No. Page No.

0.2 µ Filter Pre 0.18 ml/min 0.31 ml/min
Filter Integrity Max. Flow Rate NMT 16 of 76 16 of 76
Integrity Pre & As per SOP
as per PI-SP-022 075ml/Min Page No. Page No.
Post Post 0.31 ml/min 0.29 ml/min
Before Media 17 of 76 17 of 76
Filling
Environment and
Environment and
Personnel Qualification Shall meet SOP acceptance
Personnel As per SOP Complies NA Complies NA
as per SOP No. criteria
Monitoring
QA/QC/403

Table 22: Summary of liquid borne particulate count results for the sterilized and dehydrogenated vials
Sampling details : Sterilized Vials on turn table received from tunnel after sterilization
Sample quantity : For Initial, Middle and End 36 numbers of vials each time

Batch No. MF/01 MF/02

LBPC Test Initial Middle End Initial Middle End
10 µ ( Limit NMT 60/ml ) 05 05 04 02 03 02
25 µ ( Limit NMT 06/ml ) 00 00 00 00 00 00
Visible Particles Initial Middle End Initial Middle End
Black Particles Nil Nil Nil Nil Nil Nil
White Particles Nil Nil Nil Nil Nil Nil
Fibers Nil Nil Nil Nil Nil Nil
Glass Pieces Nil Nil Nil Nil Nil Nil

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 Batch No. MF/01 MF/02
LBPC Test Initial Middle End Initial Middle End
Any other Particles Nil Nil Nil Nil Nil Nil
Endotoxin Test Initial Middle End Initial Middle End
Limit NMT 0.125Eu/ml Complies Complies Complies Complies Complies Complies
Sterility Test Initial Middle End Initial Middle End
Should be sterile Complies Complies Complies Complies Complies Complies
A.R. No. MB/17/0166 MB/17/0180 MB/17/0182 MB/17/0190 MB/17/0181 MB/17/0199

Table 23: Summary of liquid borne particulate count results for the sterilized rubber stopper samples
Sampling details : Rubber stopper after sterilization.
Sample quantity : For Initial, Middle and End (36 numbers of Bungs at each stage)

Batch No. MF/01 MF/02

LBPC Test Initial Middle End Initial Middle End
10 µ ( Limit NMT 60/ml ) 01 05 05 02 03 05
25 µ ( Limit NMT 06/ml ) 00 01 00 00 00 00
Visible Particles Initial Middle End Initial Middle End
Black Particles Nil Nil Nil Nil Nil Nil
White Particles Nil Nil Nil Nil Nil Nil
Fibers Nil Nil Nil Nil Nil Nil
Glass Pieces Nil Nil Nil Nil Nil Nil
Any other Particles Nil Nil Nil Nil Nil Nil
Endotoxin Test Initial Middle End Initial Middle End
Limit NMT 0.125Eu/ml Complies Complies Complies Complies Complies Complies
Sterility Test Initial Middle End Initial Middle End
Should be sterile Complies Complies Complies Complies Complies Complies
A.R. No. MB/17/0167 MB/17/0179 MB/17/0181 MB/17/0189 MB/17/0192 MB/17/0200

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 Table 24: Summary of GPT on the left-over bulk media solution
Sampling details Media after filling from each container (left over) 100ml.
Batch No
Details of
Test Acceptance Criteria Container No. MF/01 MF/02
Organism
Results A. R. No. Results A. R. No.
1st Complies MB/17/0177 Complies MB/17/0188
Growth Promotion 2nd Complies MB/17/0177 Complies MB/17/0188
Growth Should be observe
Test Complies MB/17/0178 Complies MB/17/0201
3rd
4th Complies MB/17/0178 Complies MB/17/0201

Table 25: Summary of GPT on the media filled vials after 14-days of incubation

MF/01 9 vials after 14 days of Incubation MF/02

Sampling details

Sample Qty 9 vials 9 vials

Growth should be observe Complies Complies
A. R. No. MB//17/0223 MB//17/0228

Table 26: Summary of container-closure integrity study by microbial ingress method performed on media filled vial samples collected from the media fill batches after 14-days
incubation
Sampling details : Vials after 14 days of Incubation
Sample quantity : 30 number of vials from each Initial, Middle and End of Batch Process.

Batch No. MF/01 MF/02

Stage Initial Middle Initial Initial Initial End
No Growth No Growth No Growth No Growth No Growth No Growth
Test Vials
Observed Observed Observed Observed Observed Observed
No Growth No Growth No Growth No Growth No Growth No Growth
Negative Control (-Ve)
Observed Observed Observed Observed Observed Observed
Positive Control (+Ve) Growth Observed Growth Observed Growth Observed Growth Observed Growth Observed Growth Observed
A. R. No. MB/17/0220 MB/17/0221 MB/17/0222 MB/17/0225 MB/17/0226 MB/17/0227

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Summary of the results in:

 Table 20 indicates suitability of the sterile, dehydrated media and sterile placebo powder (Mannitol)
for their use in the media fill batch manufacturing.
 Table 21, Table 22, and Table 23 indicates accurate simulation of the in-process sampling which is
normally followed in a routine product batch manufacturing.
 Table 24 indicates, bulk media solution subjecting to the sterilizing conditions retained its growth
promotion capability as well as its sterility after experiencing various interventions and
manipulations necessary for connecting with the vial filling machine for the process simulation
study.
 Table 25 indicates, each lot of the media solution used in the media fill batch manufacturing did not
lose its growth promotion property at the end of 14-days incubation period.
 Table 26 indicates, suitability and satisfactory performance of the container-closure system and vial
over-sealing process respectively, when the representative sample vials were tested for the
“container-closure system integrity test” by microbial suspension immersion test method.
9.18 Aseptic Practices:
The media filling activities of batch No’s. MF/01 & MF/02 was carried out in two shifts (A+B). During each
media fill batch manufacturing, aseptic practices of the personnel were monitored by trained and
qualified personnel using a CCTV live feed on camera no’s 8, 9 & 10 according to the SOP #
MG/MF/050-R05- R06. Summary of objectionable aseptic practices observed during the manufacturing
of media fill batch numbers MF/01 and MF/02 is presented in the following Table 27 and Table 28
respectively. Details are available with the corresponding batch manufacturing record.
Table 27: Summary of objectionable aseptic practices during media fill batch # MF/01
Camera No. 1 Observation Camera No. 2 Observation Camera No. 3 Observation
Inadequate Aseptic Date 31.03.2017 Date 31.03.2017 Date 31.03.2017
Practice
Frequency/ Shift Frequency/ Shift Frequency/ Shift
(With respect to)
A B A B A B
Gowning Nil Nil Nil Nil NIL Nil
Posture 1 2 2 2 NIL Nil
Hand sanitization 7 3 10 1 1 3
Product handling Nil 1 1 Nil 1* Nil
Movement 1 4 Nil Nil NIL 3
Talking Nil Nil 1 Nil NIL Nil
Tool’s handling 1 1 2 4 NIL Nil
Machine Assembling Nil Nil Nil Nil NIL Nil
Powder charging Nil 1 Nil 2 NIL 3
Empty vial handling 1 Nil 1** + 2 3
Others (Details in the
9 2
following table)

Continued from above table, “Others” 01* Stopper

bowl not
Body touches machine door -- Swing conveyor not sanitize from B sanitized; rubber
02 times to A stopper bag not
sanitized during
Others Others Others
assembly
(A shift) (B shift) (A shift) 01** Hands not
sanitize before
Infeed TT door not sanitized --
Person enters completely in grade A plate exposure
02 times
at stopper bowl
location (Others)

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compressed air
Sampling bottle sanitization
sampler transfer
not done during transfer from
without
B to A grade
sanitization
Door not
In feed TT door opens for
sanitized after
longer duration
stopper transfer
Aluminum foil is
under the
Tray taken out from A to B remains under
Others LAF during bung
(B shift) sampling
Googles
Leaning over swing conveyor adjustment done
under LAF
Machine cleaning done with
mop exposed in grade B
Operator continue hold forceps
Others (B shift) after completion of activity--02
time

Table 28: Summary of objectionable aseptic practices during media fill batch # MF/02

Camera No. 1 Observation Camera No. 2 Observation Camera No. 3 Observation

Date 01.04.2017 Date 01.04.2017 Date 01.04.2017

Observation A shift B shift Observation A shift B shift Observation A shift B shift
Proper Proper
Proper gowning Nil Nil 01 time Nil Nil Nil
gowning gowning
0bservation
Posture 01 time 02 times Posture 01 time 03 times Posture Nil Nil
Hand Hand
Hand sanitization 02 times 03 times 02 times Nil Nil 03 times
sanitization sanitization
Product Product
Product handling 01 time Nil 01 time Nil Nil Nil
handling handling
Movement 01 time 02 times Movement Nil Nil Movement Nil 03 times
Talking Nil Nil Talking Nil Nil Talking Nil Nil
Tool’s Tool’s
Tool’s handling 01 time Nil handling 03 times 01 time handling Nil Nil
Filled vials Sanitization of
Assembling Nil Nil Nil Nil Nil Nil
handling m/c
Stopper
Powder charging Nil 01 time Others 02 times 03 times 1 03 times
loading
Empty vial
Nil Nil Others 5 01 times
handling
Others 2 02 times

New SS trays used for

stopper placement but
Infeed TT door not Sterile dress touches
these trays are not
sanitized properly inner side of curtain
validated in autoclave
Others (A load pattern
Others (A shift):
shift)
Others (A
shift)
Media plates exposed
Continue touches Curtain not sanitize
without hand
outfeed conveyor properly
sanitization

Person enters Door not sanitized

Persons enters class Others (B
Others (B shift) completely in grade A--2 after completion of
A shift) times stopper loading

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Camera No. 1 Observation Camera No. 2 Observation Camera No. 3 Observation

Date 01.04.2017 Date 01.04.2017 Date 01.04.2017

During compressed air

Not follow aseptic Air curtain disturb sampling, proper
practices completely aseptic practices not
followed

Air sampler not

sanitized while transfer
B to A grade

Stopper bag touch the

B shift
bowl once

Review of the results in Table 27and Table 28 indicates, reasonable control of the aseptic area
personnel in following the aseptic practices. However, incidences of failure to follow/ inadequate aseptic
have been observed. The maximum frequency of the failure to follow/ inadequate aseptic practices
were; “hands sanitization after an intervention” followed by “usage of tools”. These have been identified
as training needs and all personnel who are authorized to enter the aseptic processing room during
manufacturing, including technicians and maintenance personnel, microbiologist, production, and QA
have been (re)trained on the aseptic practices. Training records are available with the corresponding
batch manufacturing record.
10.1 DISCUSSION, ANALYSIS AND INTERPRETATION
All results of the process simulation studies performed by manufacturing media fill batch numbers MF/01
and MF/02 in March and May 2017, have been thoroughly reviewed and the findings are summarized in
the preceding sections of this review document/ report. Following sections capture detailed discussion
and interpretation of all results and observations to arrive at a meaningful conclusion.
The conduct of process simulations (media fills) for aseptically produced sterile products at Model Life
Sciences Limited, Nagpur, Unit VI entails simulation of the process from the point of sterilization of the
product contact surfaces, including the container and closures, through over-sealing of the filled vials.
Simulation of aseptic activities/ processes performed in aseptically receiving the sterile drug powder in
the aseptic processing area, its sampling, and handling until connecting the sterile drug powder
container to the vial filling machine are also simulated as part of the process simulation test.
The aseptic process has been studied/ evaluated by simulating the normal, routine production
operations. In this process simulation study, a sterile placebo powder (Mannitol) and a sterile liquid
microbiological growth medium (SCDM) are substituted for filling in a routine container-closure system in
succession. The simulation process exposed the sterile placebo powder (Mannitol) to the product
contact surfaces of equipment, container closure systems, critical environments, and process
manipulations to closely simulate the same exposure that the routine product itself would undergo. The
sterile placebo powder (Mannitol) filled vials are then filled in quick succession with sterile liquid
microbiological growth medium (SCDM), stoppered and sealed like a routine product batch. Containers
filled with the sterile placebo powder and a growth medium were then incubated to detect microbial
contamination.
However, the simulated procedure has been adopted at some places in a manner which does not reduce
the effective challenge of the simulation and, as a result even improves the results of the simulation
relative to routine manufacturing process. The areas where adaptations done are as follows:
 Liquid Media Fill: The sterile placebo powder (Mannitol) filled vials were filled in quick succession
with sterile liquid microbiological growth medium (SCDM), stoppered and sealed like a routine
product batch. Containers filled with the sterile placebo powder and a growth medium were then
incubated to detect microbial contamination. As the liquid fill unit is used in the media fill batches
only, the vial filling speed lowers significantly as compared to the machine speed in routine product
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batch manufacturing and also that the routine product batch sizes are significantly higher, a
concept of alternate filling between media filled and empty/ powder filled vials has been followed.
The empty vials were filled at maximum speed to experience interventions that are usually
experienced due to running the filling machine at its maximum speed, immediately on correcting the
intervention vials were filled with media with an intent to evaluate the impact due to the intervention
and its correction. Such vials were appropriately identified and included into the subsequent
inspection as well as incubation.
Review of the results indicate that, the vial filling process followed is comparable with the routine
drug product compounding process(s) and its accurate simulation.
 Powder Filling: Sterile filtered inert/ a specified gas like Nitrogen/ Carbon Di-oxide respectively is
used for the flushing of empty vials and purging the powder from the dosing wheel in the
manufacturing of few products. As, the use and presence of inert gas in the manufacturing process
and in the vial, headspace could adversely impact the microbiological growth promotion, the inert
gas is replaced with sterile filtered compressed air in the manufacturing of media fill batches for an
accurate process simulation.
Review of the results indicate that, the vial filling process followed is comparable with the routine
drug product compounding process(s) and its accurate simulation.
 Interventions: Review of interventions data summarized in Table 12 indicates, interventions type as
well as the frequency simulated are > the number of times prescribed in the media fill protocol/
batch manufacturing record, thus providing higher challenge to the aseptic manufacturing process
and can be considered as a worst-case condition.
Review of the sterile vial filling step results indicate that, the sterile vial filling process followed in
both media fill batch manufactured is comparable and an accurate simulation of the routine drug
product filling process(s). The comparison is based on the filling speed, filling duration, filling
volume, inert gassing and interventions.
Following Table 29 summarizes the study observations, findings and results.
Table 29: Summary – media fill batch # MF/01 AND MF/02 manufacturing process
Parameter Observation/ Finding/ Result
Comparison of the manufacturing process followed for a routine product and media fill batch manufacturing
General Process
indicates; that the media fill manufacturing process closely simulates, sterile drug product manufacturing
Considerations
process.
The container and closure system are selected for the process simulation test according to the criteria
Container Closure
defined in SOP # QA/MF/010-R09, “Process Simulation by Media Fill Study” and is same as being used in
System
the manufacturing of routine product batches.
Comparison of the hold times of sterilized components and equipment before their use in the manufacturing
of media fill batch No’s MF/01 & MF/02 with the established/ validated hold time indicates; that the hold
Worst Case Situation times of the sterilized components were observed within the established hold time limits as per the “Hold
Time Study” report no. REP/MIS/005/00 & REP/MIS/006/00 and are comparable with the routine product
manufacturing batches.
Comparison of the routine product and media fill batch sizes indicates; that the batch sizes of the media fill
Process Simulation batch No’s. MF/01 & MF/01 vials meet the batch size criteria of SOP # QA/MF/010-R09, which is in line
Batch Size with the USFDA guidance for industry, “sterile drug products produced by aseptic processing – cGMP”,
Sep. 2004 and PDA technical report no. 22, revised 2011.
Appropriate nutrient media was used in the media fill batch No’s. MF/01 & MF/01 and that, it retained its
Media Selection
growth promotion property throughout the stages of manufacturing as well as incubation.
Media solution (3%) is prepared by dissolving the dehydrated media in Water for Injection at room
temperature and mixed for a nominal 30 minutes according to the SOP # QA/MF/056-R02. Before
Compounding
sterilizing the media, in-process sample was withdrawn to check media solution pH, which were observed
to meet its expected specification limit.

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Parameter Observation/ Finding/ Result

Component, Machine
Review of the sterilization records indicate using same sterilization cycle(s) that are being used for
Parts & Accessories
sterilization of machine parts and components in the routine product batch manufacturing.
Sterilization
Filling
Review of the vial filling machine speeds indicate close simulation of the normal operating machine
speeds that is used for the filling of routine product batch(s). The vial filling machine speed range
simulated were adequate to mimic commercial production conditions, accurately assess the potential for
Filling Speed commercial batch contamination and simulate activities that are representative of the manufacturing
process. Simulation of;
(a) high line speed helped in evaluating a significant degree of manual manipulation due to the frequent
interventions, and (b) slow line speed helped in evaluating prolonged exposure of the sterile drug product
and containers/closures in the aseptic processing area.
Review of the vial filling duration indicate appropriate simulation of the vial filling duration and that the
routine product batch filling duration and the media fill batch duration are comparable. The duration of the
media fill runs was adequate to incorporate manipulations and interventions, as well as appropriate
Filling Duration
consideration of the duration of the actual aseptic processing operation. All interventions that commonly
occur in a routine product batch manufacturing were simulated in the entire duration of filling each media fill
batch.
The sterile placebo powder (Mannitol) and sterile, liquid media fill weight and volume respectively indicates,
Filling weight
the fill weight and volume are comparable with the values suggested in the SOP # QA/MF/010-R09 and
and volume
closely simulates the sterile powder filling step/ process.
The use of inert gas in routine product manufacturing process was substituted with the sterile filtered
Inert Gassing compressed air without changing/ modifying the normal control parameters, thus closely simulating
a routine product manufacturing process.
Interventions type as well as the frequency simulated are > the number of times prescribed in the media fill
protocol/ batch manufacturing record, thus providing higher challenge to the aseptic manufacturing process
Interventions and can be considered as a worst-case condition. Performing each intervention, that normally and routinely
occur in a routine product batch manufacturing process, is a close and accurate simulation of a routine
product manufacturing process.
Review of the vial reconciliation indicates, the vials reconciliation for the media fill vial batch nos. MF/01 &
MF/02 is done in a manner it is done in a routine product batch. The only exception is, the integral rejects
of vial visual inspection step, such as visible particulate matter and other cosmetic defects are also
accounted and incubated along with the good vials. This is in accordance with: (a) USFDA guidance
document, “Guidance for Industry Sterile Drug Products Produced by Aseptic Processing — Current Good
Vials Reconciliation Manufacturing Practice”, Sep. 2004, and (b) PDA Technical Report No. 22, “Process Simulation for
Aseptically filled Products, 2011.
Vial quantities for the individual media fill batches are accurately reconciled and no discrepancies
observed. This fact is confirmed by reconciliation of the overall sum total numbers, as well and that no
contaminated unit was found in the batch nos. MF/01 and MF/02 after a 14-day incubation period.
No excursion observed from the alert/ action limit in the core aseptic processing area (Grade A and Grade
B areas). However, 4 incidences of environmental monitoring results (by settle plate method as well as
Environmental active air sampling method) equal to or greater than the alert and action limited have been observed.
Monitoring Excursion from the action limit is observed in the change room – 1, which is not unusual due to the heavy
personnel movement in this area. EM excursions have been performed according to the SOP #,
“QA/EM/059”. Please refer investigation report(s) IIR/17/079
All personnel who are authorized to enter the aseptic processing room during manufacturing, including
technicians and maintenance personnel, microbiologist, production, and QA participated in at least one
media fill batch manufacturing. Their participation was consistent with the nature of each operator’s duties
Personnel
during routine product batch manufacturing. The manner in which personnel participated in the media fill
batch manufacturing process, is a close and accurate simulation of a routine product manufacturing
process.
The media filled vials after filling and sealing operation were inspected manually by trained visual
inspectors according to the SOP # MF/PR/076-R10. Same procedure is followed for the inspection of the
Post Fill Visual routine product batch(s). The standard of inspection and criteria used for rejection applied to the media
Inspection filled vials is same as that being used in the routine product batch(s) visual inspection. The only exception
is, the integral rejects of vial visual inspection step, such as visible particulate matter and other cosmetic
defects were documented, but not rejected like in a routine product batch manufacturing, but were actually

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Parameter Observation/ Finding/ Result

accounted and allowed for incubation along with the good vials. This is in accordance with: (a) USFDA
guidance document, “Guidance for Industry Sterile Drug Products Produced by Aseptic Processing —
Current Good Manufacturing Practice”, Sep. 2004, and (b) PDA Technical Report No. 22, “Process
Simulation for Aseptically filled Products, 2011.
The media filled vials were passed through the labeling machine/ line to simulate the level and type of
handling that a routine product batch(s) vials would experience. However, labels were not applied to the
media filled vials. This step was not followed in any of the previously manufactured media fills batch(s),
which was not an exact simulation of the complete manufacturing process. However, considering the
Vial Labeling
possibility of inspected vials experiencing stress during handling and processing of the vials through the
rigor of the labeling and packaging step, SOP # QA/MF/010-R09, “Process Simulation by Media Fill Study”
has been revised to include instructions for accurately simulating the labeling and packaging step. It has
been implemented since these media fill batches and will be followed in the future as well.
Incubation
Orientation of media All media filled units are stored/ incubated at 200C – 250C temperature range, in an inverted position for the
filled units during first 7 days of incubation, followed by storing/ incubating at 300C – 350C for the next 7 days, in an upright
the incubation position.
Incubation of media filled vials was done under conditions adequate to detect microorganisms that might
otherwise be difficult to culture. Incubation conditions are established and followed in accordance with: (a)
Incubation Conditions USFDA guidance document, “Guidance for Industry Sterile Drug Products Produced by Aseptic Processing
— Current Good Manufacturing Practice”, Sep. 2004, and (b) PDA Technical Report No. 22, “Process
Simulation for Aseptically filled Products, 2011.
Post Incubation
Inspection of the incubated vials is performed twice during the incubation, after 7 days – at the end of the
incubation period at 200C - 250C and after 14 days – at the end of the incubation period at 300C - 350C. All
these inspections were done by the qualified microbiologists according to the SOP # QA/MF/010-R09,
“Process Simulation by Media Fill Study”.
Inspection The final inspection report documents accurate count of the number of units inspected and is part of the
corresponding media fill batch manufacturing record. Table 13 presents the post incubation visual
inspection summary including the number of contaminated unit(s), if observed.
One cracked vial, from the tray no. 68 was found during the inspection of the media filled vials on 7th day of
incubation. However, investigation for this cracked vial has been performed according to the SOP #, “QA”.
On completion of 14-days incubation, media growth promotion test (media fertility test) was performed on
Stasis Test (Post
actual media filled representative sample vials from each media fill batch, as per the Pharmacopeial
Incubation Media
method. Results of the growth promotion test on media filled vials post incubation indicate that, each media
Growth Promotion
fill batch at the end of incubation period exhibited its growth promotion properties.
Test)
Sterile, dehydrated media and sterile placebo powder (Mannitol) used in the media fill batch manufacturing
were observed suitable for their intended use.
In-process sampling performed in the media fill batch manufacturing was observed comparable and
accurate simulation of in-process sampling performed in a routine product batch manufacturing.
Bulk media solution subjecting to the sterilizing conditions retained its growth promotion capability as well
In-coming material as its sterility after experiencing various interventions and manipulations necessary for connecting with the
and In-process vial filling machine for the process simulation study.
Sampling: Each lot of the sterile media solution used in the media fill batch manufacturing did not lose its growth
promotion property at the end of 14-days incubation period.
The container-closure system as well as the vial over-sealing process suitability and performance
respectively were observed satisfactory based on a successful the “container-closure system integrity test”
performed on a representative media filled vials after 14-days incubation using a microbial suspension
immersion test method.
A reasonable control of the aseptic area personnel in following the aseptic practices was observed.
However, incidences of failure to follow/ inadequate aseptic have been observed. The maximum frequency
of the failure to follow/ inadequate aseptic practices were; “hands sanitization after an intervention” followed
Aseptic Practices: by “usage of tools”. These have been identified as training needs and all personnel who are authorized to
enter the aseptic processing room during manufacturing, including technicians and maintenance personnel,
microbiologist, production, and QA have been (re)trained on the aseptic practices. Training records are
available with the corresponding batch manufacturing record.

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Model Report

A thorough review and critical analysis of all observations and study findings described in the above
sections confirms that, in the manufacturing of the media fill batch(s) MF/01 and & MF/02:
 The steps preceding filling and sealing have been designed and executed in manner to
incorporate all conditions, product manipulations, and interventions that could impact on the
sterility of the product.
 All preparation and sterilization procedure followed were same as those being followed in
manufacturing of a routine production batch
 The vials preparation, filling, stoppering and sealing machine and aseptic manufacturing area used
in the media fill batches were used in exactly the same manner as normally done in the routine
manufacturing batch
 The studies incorporated all product manipulations, additions, and procedures involving
exposure of product contact surfaces to the environment.
 The media fill manufacturing process was comparable and accurately simulated the routine product
manufacturing process at Model Life Sciences, Nagpur, Unit VI.
Based on a thorough review, analysis and above discussions it is confirmed that, the process simulation
study results derived from the media fill batch numbers MF/01 and MF/02 manufacturing are reliable and
demonstrate that process controls are adequate to protect the product during manufacturing. Thus,
can be utilized for drawing meaningful conclusion of the study as well as identifying further improvement
areas, as applicable.
It is understood that, a successful completion of a process simulation test(s) cannot be considered as a
sole-criteria for validation of aseptic manufacturing processing in same sense that a performance
qualification effort involving biological challenge and temperature measurement can support a steam
sterilization process. Aseptic processing relies heavily on personnel intervention practices, equipment
features, facility design/ control and procedures that in combination serve to exclude micro-organisms
from sterile components and products. These elements of aseptic processing cannot be as rigorously
controlled as a sterilization process, resulting in a lingering risk of contamination. Thus, the aseptic
process simulation is only a demonstration of the capability of the process to produce sterile products
aseptically at the time of its execution using the defined process, materials, facility, equipment and
personnel.
11.0 CONCLUSION
Based on the above discussions, it is concluded that: (a) the aseptic manufacturing process/ techniques/
technology, (b) manufacturing and containment controls, (c) environmental conditions and controls, (d)
personnel involved/ engaged, (e) applicable corresponding SOP’s and procedures available and/ or
being followed at Model Life Sciences Limited, Nagpur, Unit VI demonstrates the on-going capability of
producing sterile drug product by aseptic processing.
However, a thorough review and critical analysis has resulted in identification of few improvement areas
as described in the “Recommendations” section.
12.1 RECOMMENDATIONS
A. In order to avoid the incidence of a cracked vial as explained in the above section 9.13 in future
batch(s) (routine product and/ or media fill batch) following preventive actions are recommended.
 Avoid partial packaging of HDPE trays. If unavoidable, then fill the empty space tightly with a
cardboard/ plastic bubble wrap packing.
 Do not incubate/ store the filled vials in an inverted position, as the vials are highly unstable in
this position and can fall and collide with each other resulting in a crack and/ or onset of a
crack. However, to ensure media solution contacts complete internal vial as well as rubber
stopper
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 Process Simulation Test (Media Fill)
Model Report

surface, each media filled vials would be inverted once in a controlled condition before its
incubation.
In order to eliminate even a remote possibility of developing crack into the vial during post
inspection handling/ transport in a last loosely packed tray, it is recommended to extend the same
requirement for the routine drug product batches as well.
B. Although a reasonable control of the aseptic area personnel in following the aseptic practices was
observed. However, incidences of failure to follow/ inadequate aseptic have been observed. The
maximum frequency of the failure to follow/ inadequate aseptic practices were; “hands sanitization
after an intervention” followed by “usage of tools”. These have been identified as training needs and
Training records are available with the corresponding batch manufacturing record..
It is recommended to (re)train and qualify all personnel who are authorized to enter the aseptic
processing room during manufacturing, including technicians and maintenance personnel,
microbiologist, production, and QA have been (re)trained on the aseptic practices. Establish a
mechanism to verify effectiveness of this training and integrate review of aseptic practices as part
of the Quality Management Review process/ meetings.
C. Revise the following media fill documentation for simplification and/ or presentation improvement.
 Tags/ identifiers to be provided for all rejects for ease of cross-reference and accountability
 PFD of the manufacturing process, indicating the points where rejects are generated, their
tag/ identifier.
 Vis-à-vis comparison as of the rejection method (auto/ manual) and disposition of the rejects
in routine product and media fill batch manufacturing

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RedLotus Pharmtech Pvt. Ltd.