Svp Media Fill Process Validation Protocol 89-2-16 653862

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 Confidential Document

MEDIA FILL PROTOCOL

Equipment/system name:

6000/hr SVP Ampoule Filling Machine

Protocol number :

Val 01-4

Asset number:

As per finance

Model:

F97/22-K-TDB Liquid filling machine

Serial number:

Filler: RA3051

Department:

Multi product filling suite

Addendum:

N/A

Effective Date:

Page 1 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 Contents

Page

1.

OBJECTIVE

3

2.

SCOPE

3

3.

INTRODUCTION

3

4.

PROTOCOL CHANGES

4

5.

REFERENCES

4

6.

DEFINITIONS

4

7.

SPECIAL PRECAUTIONS

5

8.

9.

RESPONSIBILITIES  Validation Department  QA Manager / QA Release Pharmacists  Microbiological Laboratory  S.V.P Department WORST CASE RATIONALE  Environmental Exposure & Filling Room Complement  Container Size and Line Speed  Filling Medium & Interventions

5

6

10.

FILLING METHODOLOGY

8

11.

MEDIA FILL TEMPERATURE AND MEDIA SELECTION RATIONALE

12

12.

ACCEPTANCE CRITERIA

14

13.

NUMBER OF RUNS REQUIRED

15

14.

REPORT

16

15.

INVESTIGATION OF FAILURES

16

Page 2 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 Completion of the above signature block signifies that the appropriate parties have reviewed this protocol and agree with the methodology and acceptance criteria. A complete report will be issued for separate approval. 1. OBJECTIVE The objective of this SVP Aseptic Media Fill Process Simulation Testing Protocol is:     

To demonstrate the capability of the aseptic process to produce sterile drug products; To contribute to confidence in the sterile status of product manufactured by the specific operator since last media fill the operator was involved with; To promote sterility assurance of future products manufactured employing aseptic technique; To comply with current Good Manufacturing Practice requirements; To provide a measure of aseptic technique of each operator involved in the media fill

Note: This study is supplemented by Val 133 - Aseptic Fill Challenge for Vial Filling Operators, which is intended as a certification tool for operators assigned to filling aseptic product, on either an Ampoule or a Vial filling line. In cases where an operator is not available to take part in a media fill, such exposure may be used to qualify the operator in absence of Media Fill. However, each operator shall be exposed to at least one run of a conventional media fill per year.

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Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 2. SCOPE This study addresses the validation of the aseptic processing during:  compounding and  filling activities. It describes methods and procedures for the conduct of the process simulation tests, as described below. The scope of this protocol relates to the filling equipment currently used for filling of all Vials as well as all Aseptically filled Ampoules within the current SVP Filling Suite. A special section deals with Aseptic Compounding. 3. INTRODUCTION The aseptic fill validation is based on the fill of bacterial culture medium as a substitute for product and in this respect it is essential that validation be conducted as a normal production run. All equipment should be cleaned, sanitized, sterilized, handled and assembled as per standard operating procedure with all conditions prevailing as normal. Similarly all compounding and filling operations must also be carried out as per standard operating procedure, while introducing interventions as experienced during day-to-day filling operations. 4. PROTOCOL CHANGES  Inclusion of Incubation Temperature selection rationale  Inclusion of Media Selection rationale  Inclusion of the Aseptic Compounding process (XXXXXXX® manufacture) 5. REFERENCES  PDA Journal of Pharmaceutical Science and Technology Technical Report 22: Process Simulation Testing for aseptically Filled Products (1997)  Journal of Parenteral Science and Technology 1987 Current Practices in the Use of media Fills for the Validation of Aseptic Processing  Journal of Parenteral Science and Technology 1990 Validation and Environmental Monitoring of Aseptic Processing  Practical Aspects of Pharmaceutical Validation – David Begg Lecture 12: Validation of Aseptic Processing using Sterile medium Simulations Page 4 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5    

Validation of Aseptic Pharmaceutical Processes (Carelton & Agallaco) Chapter 24: Validation of Aseptic Filling Operations The Validation Dictionary – Institute of Validation Technology SA GMP Guide EU GMP Guide

6. DEFINITIONS Growth Sustaining Media:A substance, which contains low levels of micronutrients, and satisfy the minimum requirements for sustaining microbial life. Growth Promoting Media: A substance which contains optimal levels of the correct nutrients, maintained at ideal pH and osmolality in order to actively promote proliferation of the specific microorganisms it was designed to . Aseptic Filling:

Part of aseptic processing where a pre-sterilized product is filled and/or packaged into sterile containers and closed.

Aseptic Processing:

Handling sterile materials in a controlled environment, in which the air supply, materials, equipment and personnel are regulated to control microbial and particulate contamination to acceptable levels. This term encomes aseptic compounding and filling activities.

Aseptic Compounding:

A process whereby two or more materials are brought together and mixed in an aseptic environment. The finished product is not subject to sterilization by any means, either prior or post filling.

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Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 7. SPECIAL PRECAUTIONS As indicated above the validation activity must mimic production as closely as possible. The only extraordinary attention that must be paid, is in containing and attending to spillages or breakages, since media so introduced into the clean room is by design intended to promote proliferative microbial growth, which is totally contradictory to clean room principles. 8. RESPONSIBILITIES Undertaking of a Protocol of this nature requires the participation and assistance of: 

Validation Department 8..1. To guide and co-ordinate the execution of this protocol. 8..2. Acquisition of raw material and componentry. 8..3. To confirm that the HVAC System is functioning within specifications. 8..4. Perform monitoring of non-viable particulates using either remote scanning or a handheld scanner. 8..5. Control and report on incubation conditions of the Media Fill. 8..6. Issuing of a report at completion of the Media Fill.



QA Manager / QA Release Pharmacists 8..1. To ensure that conditions simulated had been a true reflection of production conditions 8..2. Official Approval of the Media Fill Challenge Results



Microbiological Laboratory 8..1. To assist in planning of Media Fill, and acquisition of materials required. 8..2. To demonstrate that the media and the incubation period and temperatures employed are effective for the promotion of growth of low levels of normal microbial contaminants as encountered in microbiological monitoring as well as standard challenge microorganisms. In addition, special attention should be given to growth promotion of media post homogenization and filling as performed in the Aseptic Compounding Challenge. 8..3. To monitor and control the fill process in conjunction with Production. 8..4. To evaluate the filled containers. 8..5. To conduct routine microbial monitoring of the filling and media introduction areas during the validation run. 8..6. To assist in compilation of the final report, inclusive of environmental results.

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Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 

S.V.P Department 8..1. To plan for provision of equipment and personnel at times convenient to Production, microbiology and Validation departments. 8..2.

To carry out filling under control and supervision of a technically competent person who must be responsible for ensuring that all components and equipment are correctly treated and handled and that all assembly and process operations are correctly carried out.

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Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 9. WORST CASE RATIONALE As per standard Validation approach, “worst case” conditions should be used when performing a Media Fill. However, care must be taken that this is not overdone; i.e. inadvertently moving from “Worst Case” to “Ridiculous Case”. Standard operations should be performed (e.g. standard machine setup, volume adjustments, etc.) The following criteria “Worst Case” criteria should be considered: 

Environmental Exposure In the course of normal operation, equipment is routinely sterilized prior to use. In other words, equipment may be exposed to the Clean Room environment for some time prior to usage. This exposure is normally restricted to 72 hours; however standing time normally does not exceed 24 to 48hours. In addition, although a truly sterile Clean Room Environment is pursued, it must be demonstrated (by validation) that the environment does not contribute to loss of sterility of any stored product equipment. In order to justify the maximum allowable window though, a standing time in the vicinity of 24 to 48hours should be pursued for a media fill, in order to demonstrate the maintenance of equipment sterility. If a standing time of longer than the norm can be demonstrated, it may be accepted that the product equipment is not at risk during normal staging activities. Standing time should therefore be made as long as is practically possible. However, as it is neither practical nor possible to stage equipment for each run three days prior to filling, this requirement is only enforced for the first run of the Media Fill, on each machine. The completion time for the initial Staging Time Validation study will therefore be three Media Fills (1.5 years).



Filling Room Complement Standard staff complement in the SVP Filling Room is as follows:  Four Filling Machine Operators (1 per machine)  One additional operator ()  One Technician (enters when required)  One Cleaner Page 8 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5  One representative from the Microbiological Laboratory (sampling; enters when required)  XXXXXXX Compounder (enters only when aforementioned product is being compounded) Once again, worst case conditions must apply, therefore, the following is recommended:  Filling Machine Operators as per norm (four)  One additional Operator  Technician / representative to be present in the area during the filling process (exits thereafter, but re-enters upon starting the next run)  One Cleaner (permanently present)  One representative from the Microbiology Laboratory (to be permanently present)  XXXXXXX Compounder (to be present when the aforementioned is being filled)  One representative from the Validation Department (to be present for entire duration of filling) 

Container Size and Line Speed Container selection has a two-fold impact: Exposed Aperture and Filling Line Speed. Therefore two different size containers are to be filled:  Smallest Size filled on each specific machine This increases the filling rate (which should be set to maximum used for the specific size container), while causing handling difficulty (risk).  Largest Size filled on each specific machine This represents the slowest operational (filling) speed (and should be set to the minimum speed normally used for the specific size container). This also represents the worst-case contamination risk, as these containers tend to have the largest filling aperture, which increase the risk of aerial contamination entering.



Filling Medium Normal product (which may be growth inhibiting, sustaining or promoting) is substituted with a medium, which is ideal for maximal proliferation of microorganisms.



Interventions As per normal filling operation, interventions must take place. This is to include (but may not be limited to):  At least one needle change per run  At least one simulated (or actual) machine breakdown per run Page 9 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5

10. FILLING METHODOLOGY Below describes the methods for both products sterilized by filtration and aseptically compounded product. 

General 10..1. For process as well as filling machine details, refer to the attached Batch Documentation (Appendices 1 – 5). 10..2. The entire process (from compounding through filling) is to be videotaped for review and problem resolution. Ensure moving of the Video Camera as to capture operations of all compounding and filling lines, as far as practical. 10..3. All containers must comply to the following: 10..3.1. Must be of clear glass, of the quality as per in-house specification. 10..3.2. As a cost-saving measure, non-approved containers may be used in a media fill, provided that such use does not compromise the integrity of the Media Fill (i.e. units rejected on the basis of dimensional nonconformity) may be used. 10..3.3. Bungs must comply to all quality requirements and must be approved by Quality Assurance 10..3.4. All vessel types used during standard production must be used in the Media Fill, e.g. direct filling will not be allowed if transfer and collecting vessels are used as norm, and visa versa. 10..3.5. No deviation from standard operations will be allowed during the Media Fill operation. 10..3.6. Target is 3000units per run. At least 3200 units must be filled and incubated in order to allow for any breakages that may occur. 10..3.7. Media Fill Volume and Media Batch Sizes have been specified in the attached Batch Books (Appendix 1 –5). This must be strictly adhered to. 10..3.8. Environmental Monitoring shall be performed as per standard operation, although additional monitoring may be indicated. This shall include Settle Plates, Plates, Air counts (both viable and nonviable), Finger Dabs, etc.



Aseptically Filled Product, Sterilized by Filtration 10..1. Identify the largest, as well as the smallest, routinely filled container for each filling machine/process. 10..2. Prepare components and equipment as per control document and procedural instruction (refer attached Media Fill Batch Books). Page 10 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 10..3. Prepare sufficient media (refer to point 11.2 for selection criteria) as per manufacturer’s instructions (refer attached Media Fill Batch Books as well as point 10.2 – Aseptic Compounding). 10..4. Carry out standard machine set up, observing all necessary precautions and aseptic technique. 10..5. All compounding and filling operations must be carried out as per standard operating procedure (Refer SVP9209157 – SVP Mixing Room Operations; SVP9209159#9809405 – Filter Integrity Testing; SVP9609336 – Bulk Product Transfer of SVP Product from Mixing to Filling; SVP9209151 – General Rules and Precautions for working in the SVP Filling Room). 10..6. Each Batch of Media shall be continuously sparged with Nitrogen throughout the filling process. 10..7. Included in the Media Fill Challenge, for each machine, must be at least one needle change intervention (Refer procedure SVP9208126 – Filling Needle Change) as well as one stoppage per run to simulate production breaks. All media filled containers specific to and around these interventions must be clearly identified. 10..8. Sufficient volume of media is to be filled into each container as to be able to coat the entire internal surface upon being swirled. A minimum volume of half the volume of the container is recommended. 10..9. Fill a minimum of 3000 (target of 3200 to 3500) ampoule/vials of the required size to standard fill volume, and submit the containers (in suitable packaging) to the Validation Department for incubation. 10..10. An additional two runs will be required, however refer to point 13 for rules guiding the reduction of number of runs required. 10..11. Incubate all containers, as specified under point 11.3 & 11.4. At the end of the incubation period, all containers are inspected for growth by the Microbiology Laboratory. Staff from other areas may be used to assist in the inspection process, however this will only be allowed once the Microbiology Laboratory Manager is satisfied with such staff’s ability of distinguish sterile and compromised containers. 10..12. Any ampoules showing growth shall be crack detected to rule out process damage. Any vials showing growth shall be visually checked for any signs of damage. 

Aseptically Compounded Product 10..1. XXXXXXX is only filled into one vial size, which is 2ml. As the purpose of this challenge is only to demonstrate method suitability, the run is only to be executed once.

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Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 10..2. Prepare components and equipment as per control document and procedural instruction (refer attached Media Fill Batch Book – Appendix 5). 10..3. All compounding and filling operations must be carried out as per standard operating procedure (Refer SVP9209157 – SVP Mixing Room Operations; SVP9209159#9809405 – Filter Integrity Testing; SVP9609336 – Bulk Product Transfer of SVP Product from Mixing to Filling; SVP9209151 – General Rules and Precautions for working in the SVP Filling Room; SVP9305193 – XXXXXXX Aseptic Compounding). 10..4. Media (refer to point 11.2 for selection criteria) is prepared using Water-forInjection, as per standard media fill practice, and is transferred to the XXXXXXX Compounding Area. Sterilization is by filtration. 10..5. In the XXXXXXX Compounding Area, pre-sterilized (by Gamma-irradiation) Cake Flour is used to mimic the normal compounding procedure. In other words, the XXXXXXX compounding method is used, substituting Medroxyprogesterone for Cake Flour (in a ratio of 1:4.5; Flour : Medroxyprogesterone Acetate). For compounding instructions, refer to procedure SVP9305193 – XXXXXXX Aseptic Compounding). 10..6. Carry out standard machine set up, observing all necessary precautions and aseptic technique. 10..7. As the challenge is only for actual Compounding step, needle changes and breakdowns need not be simulated. 10..8. As the filling operation is independently challenged, actual fill into final dosage form is not required as part of this study. All aseptic compounding and sampling procedures are however carried out as per normal. 10..9. Fill a minimum of 3 x 5L, pre-sterilized containers (Schott Bottles are suitable) aseptically, and submit the containers (in suitable packaging) to the Validation Department for incubation. 10..10. An additional two runs will be required, however refer to point 13 for rules guiding the reduction of number of runs required. 10..11. Incubate all containers, as specified under point 11.3 & 11.4. At the end of the incubation period, all containers are inspected for growth by the Microbiology Laboratory. Staff from other areas may be used to assist in the inspection process, however this will only be allowed once the Microbiology Laboratory Manager is satisfied with such staff’s ability of distinguish sterile and compromised containers. NOTE :

a A Control Report document must be drawn up for each validation . and appropriately completed with all necessary details. a. b. Each validation run requires the protocol (Media Fill Batch Book) to be completed in its entirety. All equipment must be rePage 12 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 prepared, sterilized and the process started afresh for next validation run. A minimum of three runs of both smallest and largest unit routinely filled on each specific machine must be carried out on that machine. c. c. Details of any problems experienced during runs must be recorded in the Control Report document. d. d It is critical that careful attention be given to post validation . cleaning and sanitization of equipment and area. It is recommended that a Fumigation of the filling area be carried out at completion of the Media Fill (Refer procedure VAL 0107538). 11. MEDIA FILL TEMPERATURE AND MEDIA SELECTION RATIONALE  GENERAL In the case of Aseptic Challenges / Media Fills, intent is to establish the presence of viable organisms in the finished product. As the organisms may be damaged or weakened by the preparatory steps, conditions optimum for bacterial proliferations should be pursued within the container to be challenged. Not only will this speed up bacterial multiplication, making it easier to detect the presence of the organisms with the naked eye, but also serves to speed up organism recovery and regeneration post preparation and filling activities. Although this would generally appear to be over compensating or excessive/unrealistic “worst-case”, it must be considered the organism may have two years (or even longer) to recover from the trauma should it be present in finished product destined for the market place. These optimum conditions are supplied by: 1. Providing a suitable, nutritious medium at the optimum pH, etc. for the organisms to proliferate 2. Ensuring that the containers are maintained at the optimum temperature required to ensure ideal growth rates 

NUTRIENT PROVISION Tryptone Soy Broth (TSB) is used as medium for various reasons: 1. It is versatile, all-purpose medium, that s growth of many fastidious organisms 2. s growth of aerobes, facultative anaerobes, certain fungi 3. No serum addition required 4. Has a high nutrient value 5. Maintained with a pH of 7.2 by inclusion of a phosphate buffer (which mimics human physiological pH) Page 13 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 Main shortcoming is lack of for strict anaerobes NOTE :

a. e. b. f. c.



Growth promotion studies are to be performed on each lot of media used in the media fill. In the case of Aseptic Compounding, growth study should be performed after the media has been sterilized. In addition, it must be demonstrated that the sterilized Cake Flour does not negatively influence bacterial proliferation.

TEMPERATURE SELECTION Temperature is one of the most important environmental factors influencing the growth and survival of microorganisms. It can affect living organisms in either of two opposing ways. As temperature rises, chemical and enzymatic reactions in the cell proceed at more rapid rates and growth becomes faster. However, above a certain temperature, proteins, nucleic acids, and other cellular components are sensitive to high temperatures and may be irreversibly denatured. Usually, therefore, as the temperature is increased within a given range, growth and metabolic function is increased to a certain point where inactivation reactions set in. Above this point cell functions fall sharply to zero. Microorganisms are usually divided into five groups according to their optimum growth temperatures: Organism Type Psychrophiles Psychrotroph Mesophiles Thermophiles Extreme Thermophiles

Growth Range <0C - <20C 0C - >25C 15C - 45C 50C - 70C 70C - >100C

Historically, the bacteria encountered in the clean room have been identified as mesophiles (15C - 45C) and facultative psychrophiles (0 - 20C) and are usually grown at temperatures from 20C - 37C. Various recommendations and requirements are available as to incubation temperature selection. According to USP24 (<71> - Sterility Testing) incubation time Page 14 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 for microbial recovery are given as 32.5C±2.5C for bacteria (with an incubation time of 7 days) and at 22.5C±2.5C for Yeasts and Molds (with an incubation time of 7 days). Therefore, assessment is to be performed after a minimum of 14 days. The SA GMP Guide s this requirement. As it is impractical to duplicate aseptic challenge trials (in addition to the introduction of an additional variable) the same containers shall be incubated at both temperature ranges. Note :

a b



Incubation temperature is given as a target. Although it is recommended that above temperatures be followed, some degree of variation will be acceptable. Because of the risk of bacterial protein denaturation, temperatures above 37C should be avoided, even for brief periods of time.

INCUBATION METHOD Incubation is to take place in the “Media Fill Incubation Chamber”, located in the Warehouse. As discussed under point 11.3, a two-tier incubation method is used. Total incubation period not less than 14 days (as per standard practice for all aseptically filled products). Week 1: 25C (22.5C±2.5C; i.e. 20-25 C) (“Room Temperature”) Incubation at 25C for 7days will allow for optimal cultivation of molds and yeasts, while ing growth of bacterial contaminants Week 2: 35C (32.5C±2.5C; i.e. 30-35C) Incubation at 35C for 7days will allow for optimal cultivation of bacterial contaminants, while ing growth of mold or yeast contaminating organisms. Upon loading of the filled containers into the Incubation Chamber, each package is to be gently inverted, as to assure coating of all internal surfaces of the container with media.

11.5 USE OF CAKE FLOUR IN ASEPTIC COMPOUNDING CHALLENGE Cake Flour is used in this instance due to its characteristics of: 1. Similar consistency and particle size to Medroxyprogesterone

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Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 2. Difficult wetability 3. Clumping 4. Insolubility 12. ACCEPTANCE CRITERIA  Less than 0.1% contamination rate.  A minimum of three runs for each Line / process in both largest and smallest container size normally process on each specific machine. For details on reduction of the number of runs, refer to point 13.  All environmental results must comply with standard specification. For non-viable particulate limits, refer to the Bodene Airclass Monitoring Policy and Federal Standard 209E / ISO14644. NOTE:

a . b .

NOTE :

c. d .

In the event of any unsatisfactory results a full investigation must be carried out, assignable causes determined and necessary corrective action taken before attempting validation repeat. In addition, all operators and equipment involved in the affected run shall be restricted to the filling of Terminally Sterilized product until such investigation has been completed and ratified by the QA Manager. Any containers showing growth must be further cultured to identify the contaminant microorganism. The growth promoting ability of the medium must be demonstrated by means of controlled challenge with low levels of suitable microorganisms.

13. NUMBER OF RUNS REQUIRED  Although practice is to perform trials in triplicate, under certain conditions, the number of runs for a Media Fill may be reduced.  In order for such reduction, the following criteria must be satisfied: 13..1. No process failures for the previous three consecutive media fills. 13..2. Failures may only by classed as assignable and non-process related when:

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Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 13..2.1. They have been demonstrated to be personnel related (operator to be removed from Filling Room immediately and only reitted upon successful qualification – Refer Val 133); 13..2.2. Process deviation from normal practice (to be corrected immediately and reported on. Refer point 14.2); 13..2.3. Filter Failures, total equipment failure or other type on compromise of the area; 13..2.4. Sterilization failure of componentry (full autoclave investigation and requalification required).  

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Where so indicated, number of runs required may be reduced to two. Upon completion of two consecutive Media Fills, without any process failures (barring assignable failures as discussed under point 13.2.2), the Media Fill requirement may be reduced to one run per line, for both largest and smallest containers filled on that line. Should a process failure be recorded at any time, revert back to three runs per line, for both smallest and largest containers filled.

14. REPORT  Ensure timeous recovery of data, record sheets and signatures for report compilation and presentation.  The report is to include Investigations into run failures (if any), which are to be signed off by the Production Manager, QA Manager and Managing Director. Such investigation is to include assignable cause, as corrective action taken, as well as the date of the repeat Media Fill. In addition, the endorsement is to include the timeframe of the moratorium on Aseptic Processing on the affected lines / processes as well as status of product already filled.  Incubation Temperature Report is to be issued and attached to the final document.  Results for each Filling Line to be kept on file by the Validation Department and records are to be readily available to the Department, Production and the Inspectorate.  Report all results to the Validation and Production Departments. 15. INVESTIGATION OF FAILURES  In the event of any unsatisfactory results a full investigation must be carried out, assignable causes determined and necessary corrective action taken before attempting validation repeat.  Such investigation shall include: 15..1. Container integrity verification Page 17 of 17

Title: SVP Aseptic Media Fill Simulation Testing Protocol (Aseptically Filled Products) Number: Val 01-4 Copies to: Distributed upon request, as per procedure Val 01-4 Supercedes: Val 01-3 Date: June 1999 Compiled by: Checked by: Approved by: Original: Kept as a working copy in the validation file Issue Date: 4 February 2002 Brief details amendment/update: Inclusion of Media Fill Rationale and Processing Changes Validation Interval: Bi-annual Appendices: 5 15..2. Contaminant identification (to at least genus level) 15..3. Identification of possible source of the organism (wherever possible) 15..4. Batch Documentation review 15..5. Review of environmental results 15..6. Review of the Media Fill Video record 15..7. Review of any breakdowns, interventions or deviations  Issue a Failure Investigation Report, containing at least the following: 15..1. A summary of the occurrence 15..2. A listing of all systems investigated, including outcome, regardless whether positive or negative 15..3. A conclusion as to cause of the failure 15..4. Any corrective action taken 15..5. List of recommendations 15..6. Outcome of the repeat study 15..7. This document is to be signed by the Validation committee The Investigation report is to be kept on file with the Media Fill results.

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