Standard Operating Procedure

Title: Aseptic Media Filling and Micro . Integrity Leak (Soup) Testing Procedure
______________________________________________________________________________________

Department Micro Laboratory Document no MICLAB 035

Prepared by: Date: Supersedes:

Checked by: Date: Date Issued:

Approved by: Date: Review Date:

Document Owner
Microbiology Laboratory Manager

Affected Parties
All Microbiology Laboratory and production colleagues

Purpose
One of the requirements of cGMP is a periodic evaluation of all aseptic processes by filling media into the
appropriate containers under normal production conditions . The media fill should reflect the sterility of the
entire process from the Sterilising filter to the filled primary container and should include all subsequent
manufacturing steps. This SOP outlines the procedures for both Media Fills and Microbiological Leak Tests .
For Validation purposes , a Microbiological Leak Test (Soup test) or a separate Protocol to verify the entire
process from the Bioburden Reduction Filter to the primary container may be required .

Scope
It is the responsibility of personnel as outlined in the Table below :
Media Fill Schedule Microbiology Manager
i Media runs are to be performed on all manufacturing shifts
for each aseptically filling machine process at least twice
per year (every 6 months). This is to be conducted after
significant modifications or maintenance . For terminally
sterilised lines and non-sterile process once per year/shift.
Preparation of Media Micro. Lab. Staff
Filling of Media Production Staff
Delivery to the 30pC (o 1.5pC) Incubation Room Micro. Lab Staff
Inspection of the containers at 7 & 15 days (Media Fill) Micro. Lab. Staff
Reporting - Media Fill Micro. Lab. Staff
Microbiological Leak (Soup) Test Micro. Lab. Staff
Inspection of the containers at 7 & 15 days (Soup) test Micro. Lab. Staff
Reporting - completed Protocol (Soup) test Micro. Lab. Staff

Definition
TSB Trypticase Soy Broth = Soybean Casein Digest Medium
BPN Batch Production Number
DR Deviation Report

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Standard Operating Procedure
Title: Aseptic Media Filling and Micro . Integrity Leak (Soup) Testing Procedure
______________________________________________________________________________________
The Process teams should their checklist for documentation control and all documentation is
to follow standard procedures and is to be sent to the QA Department . Form 670 is sent to
the Microbiology Laboratory with the media run , along with any media waste and rejected
media filled containers .
2.4. Level of fill for media filled containers and Soup tests, should be not more than 80% of the
volume of the container where possible , however some processes need correct volumes fill
for the inspection process.
2.4.1. Aseptic filling - containers to be at least 2/3 filled, to correct fill volume .
2.4.2. Terminally sterilised - containers to be at least 2/3 filled, to correct fill volume .

Note Sterile medium will be transferred to the filling machine as follo ws:

Method (a) Filter sterilisation of the medium from a Dispensing Pressure vessel directly
to the filling machine .
Method (b) Filter sterilisation of the medium from a Dispensing Pressure vessel into a
sterile 50L or 100L glass storage vessel and then transfer this vessel to the
sterile filling area for filling into containers .

When sterile filtering for any of the methods , install a pre-filter before the primary filter
to prevent it from clogging . If the pre-filter clogs during the run it can be chang ed to a new
filter, as long as the primary filter stays in place and sterility is not broken . This pre-filter
cannot withstand steaming in -place, so it is to be installed just before filling is to start .

3. Media Preparation
Prepare the required volume of Tryptone Soy Broth by dissolving the correct amount of dehydrated
powder in hot (80pC) Distilled Water. Simmer the medium on the hot plate and reconstitute if
necessary while transferring to either a 20 or 40 litre Dispensing Pressure Vessel .

Notes 1. medium may be heated in a double or triple strength form and then diluted with the
additional water required when being transferred to either a 20 or 40 litre Dispensing Pressure
vessel.
2. Filter sterilisation of the medium must be carried out within 4 hours of preparation of the
medium . The 4 hours may be extended if the media is sterile -filtered into a pre -
sterilised vessel.
3. The start time of the media run can be taken from the point when solution is sent down
the line to start the wetting of the Sterilisation filter to conduct the Pre -Integrity test.

4. The Aseptic Media Fill

The Aseptic Media Fill must simulate as closely as possible a normal production run . It should cover
worst case conditions or activities (however not to the point of failure ), e.g. filling rates, number and
activity level of Operators , temperature and should be run on different shifts where possible to cover
all operators, etc. The duration of the media run should be at least 4 hours or half a production shift
to allow for all routine interventions (see Appendix 1).
4.1. Environmental monitoring
During the media run environmental monitoring is to be conducted including the following :
i Settle plates (the number of settle plates will depend on the machine )
i Air sampling 1000L under the machine laminar flow and 200L in the filling room .
i Surface plates of the floor and machine .
i Personnel Monitoring for Sterile operators .
Note: After monitoring has taken place , all plates are to be sealed using tape and
delivered to the Microlab Sampling ca binet and logged into the appropriate logbook .

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Standard Operating Procedure
Title: Aseptic Media Filling and Micro . Integrity Leak (Soup) Testing Procedure
______________________________________________________________________________________
Reoccurring incidents of contaminated units in aseptic media fills for an individual line ,
regardless of the acceptance criteria , would be a signal of an adverse trend on the aseptic
processing line. This would lead to a DR being raised and an inve stigation started to look for
root cause, corrective action and possible revalidation .

Reject levels should be consistent with normal production reject levels .

Note: There may be a requirement as part of registered details for media runs to be
conducted on both terminally sterilised process or non -sterile manufacturing process .
Acceptance criteria will differ from aseptic process . If there is one positive unit found in the
process media fill, a DR will be raised and we will undertake an investigation an d determine
appropriate corrective actions .
4.6.1. The medium must be shown to be able to promote the growth of low levels of
microorganisms, etc.

If the medium fails to promote the growth of low levels of microorganisms and /or the
limits are exceeded , inform the Microbiology Manager, Quality Assurance Manager
and the Area Manager immediately , a DR is to be raised and an investigation
conducted. The Aseptic Media fill is invalid and must be repeated as soon as
practical.
4.6.2. Micro. Lab. personnel are to transfer the remaining media-filled containers to the
Security Rejects Bin in accordance with MICLAB 020.
4.6.3. A Validation Report is to be written by a Microbiology staff and approved by the
Microbiology Manager. The report is to detail the procedure , interventions conducted
both, routine and non-routine, results of the Media Fill run and is to be kept in the
Aseptic Media Fill Validations file in the Micro . Lab.
As part of the report , Micro. Lab. will receive back all documentation sent to the QA
Department concerning the media run, along with media run checklist .
When writing the report micro lab staff should review the for DRs related to the batch
for the media run .
4.6.4. The report is to be issued within 1 month of the completion of the test .

5. Microbiological Integrity Leak Test ( Soup Test)

5.1. The test is to be conducted as per the Protocol - (use Form 665).
The Protocol will change according to the particular circumstance .

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 Form 665
Issue date :
Microbiological Integrity (Soup) Test
(Ref. MICLAB 035)

Filling Machine and Number

Container Type and Size

Reason for Soup Test

Number of Units to be Tested

Initial Date of Test

Acceptance Criteria :

Not more than 1 in 1000 units to show evidence of microbial contamination .

Equipment Required:

In Soup Room :

44 Gallon drums on wheel trolleys

Circular mesh lids
Lead divers weights
Nylon netting bags
18mm hose with tap fittings
Drum Outlet valve hoses
Orange Crates

From Microbiology Laboratory :

3kg TSB powder per test drum (5 x 600gm bags / per drum)
1 bottle per test drum of 18hr E.coli culture in 250ml TSB
Rubber gloves
Rubber boots
Hibitane
Face-masks
Hair-nets
Thermometer

Soup Test Method: Day 1 Date Commenced : ____________

i Turn on hot water system, fan and heater.

Check that the Soup Room is at approximately 32 C.

i Prepare 1 x 250ml TSB bottle inoculated with an E.coli culture, per test drum.

Soup Test Method: Day 2 Date Commenced : ____________

(for each test drum )

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 Form 670
Issue date:
Aseptic Media Fill Information Sheet
(Ref. MICLAB 035)

Start Date _______________

Reason for Media run :

Micro to Complete

Machine Number
Batch number
Code
Media type
& Lot No. and Expiry Date
Volume of media prepared &
Storage Vessel size
Time media prepared
Media Prepared by
Manufacturing instruction sheet
filled up?
Container & Size
Volume fill size
Soup Test required YES / NO

Production to complete

Machine/Line No.
Line steamed and date
Machine Steaming and date
Holding Tank # & Steaming

File Location: Date Printed: Page 9 of 15

 Form 670
Issue date:
Aseptic Media Fill Information Sheet
(Ref. MICLAB 035)

Production to Complete

Number filled per shift :

Afternoon Night

Counter reading:
Number of Rejects :
Number of Filled units :

Intervention List :
The following routine interventions are to be included into the running of a media fills .
The number and types of interventions will vary depending on the process line, however we need to
cover off normal daily interventions that take place during the manufacturing of our production batches .

Type of Intervention Shift Time Person responsible

Weight and fill volume
adjustments

Component
replenishment

Filter Change

Filling needle change

Operator breaks
Personnel leaving the
room.

Component change

Operator Changes (Need

to note maximum number
of persons in room)

File Location: Date Printed: Page 11 of 15

 Form 670
Issue date:
Aseptic Media Fill Information Sheet
(Ref. MICLAB 035)

Incubation Details: Micro To Complete

Date Incubated at 30pC in

Microlab

Initials of person incubating

Media run

Media Check (as per MICLAB 035) Inspection Results :

Inspected of
Media fill Date Name of Inspectors Result
containers
7 days

On the 7th day inspection ensure that the containers are rotated and re -incubated in a new orientation , from
the first incubation period . Yes / No

15 days

Environmental Monitoring Results :

Fallout Plates : Include Gram-Stain and
times and shift Identification

Air sample: Include times

and shift

Contact Plates: Include

locations.

File Location: Date Printed: Page 13 of 15