QAS-032 - Quality Operations Procedures for Pharmaceutical, API, and Biotechnology - page 521

SOP No. QAS-032.00 Effective date: mm/dd/yyyy
Approved by:
Containers are filled with media, and stoppers are partially inserted. The containers are loaded
into the lyophilizer. A partial vacuum is drawn on the chamber and this level is held for a predeter-
mined time. The vacuum must not be so low as to permit the medium in the containers to boil over.
The chamber is then vented and the stoppers are seated within the chamber. The stopper units are
removed from the aseptic processing area and sealed.
Anaerobic condition : If there is a need for sterile inert gas to break the vacuum on the chamber
and remain in the container after sealing. The use of anaerobic media (e.g., alternative fluid thiogly-
colate medium) would be appropriated where the presence of anaerobic organisms has been con-
firmed in either environmental monitoring or, more likely, during end product sterility testing.
When anaerobic have not been detected in the environmental monitoring or sterility testing,
lyophilizer process simulation tests should utilize TSB and air.
Lyophilizer media fills will consist of a simulation of all filling and handling procedures that
would be required when processing a lyophilized product, such as, steam sterilization of lyophilizer
12 h (minimum) before loading the first tray of media, introduction of the vials into the lyophilizer,
holding vacuum for a minimum of 24 h and subsequent releasing of vacuum and activation of the
stoppering mechanism.
51.7.2.3 media filled vials/ampoule analysis
Media filled units will be incubated in an upright position for 21 days at two temperature ranges.
The units will be inspected for growth at two separate times, once at a minimum of 7 days of incu-
bation at 22.5 ± 2.5°C and 14 days of incubation at 32.5 ± 2.5°C. Record of these inspections will be
documented in QC registers. Any unit found which are positive for growth will be investigated as
detailed in the Acceptance Criteria section of the SOP.
51.7.2.4 growth Promotion of media-filled units
Growth promotion of the media filled units will be performed on representative units of each media
fill run after the completion of 21 days incubation period per instruction given in the Standard Test
Method MC-000. The media should be challenged after subjecting to the same condition of the
other media fill units.
Growth promotion ability of the medium in final filled containers must be demonstrated using 10
randomly selected containers for each challenge organism (more than 100 microorganisms per test
container).
The units used for growth testing must be subjected to the same processing steps (e.g., cleaning,
depyrogenation, sterilization, filtration, filling, lyophilization, and reconstitution) up to the point at
which they are placed in an incubator.
In case of growth promotion controls, growth must be observed in at least 95% of the test con-
tainers for all the challenge microorganisms.
If no growth is observed in all of the 10 challenged containers, a second stage test must be con-
ducted to rule out laboratory. In the second stage test both containers must support growth.
51.7.2.5 negative control
• Media from the bulk media container is incubated at a minimum of 7 days of incubation at
22.5 ± 2.5°C and at 32.5 ± 2.5°C for 14 days as negative control for liquid media fill to
check its sterility.
• No growth should be demonstrated in negative controls to indicate the acceptability
of media fill. If growth is observed the discrepancy should be investigated and
documented.