This document summarizes a presentation given by an FDA inspector on regulatory perspectives of aseptic processing. It provides an overview of the FDA's guidance and applicable regulations on aseptic processing. It discusses the objectives of aseptic processing validation and common observations cited in FDA inspections, such as deficiencies in facility design, equipment qualification, environmental monitoring, and personnel training. It concludes by emphasizing the importance of understanding limitations of the aseptic process and applying a science-based approach to validation to address risks in a manner that can be scientifically defended.

1. Regulatory & Compliance
Perspectives on
Aseptic ProcessingIIR Aseptic Filling Conference, Philadelphia, PA November 4, 2004
FOI Releasable 11/2004
Robert Darius
Inspector & Review Microbiologist
FDA/CBER
Office of Compliance & Biologics Quality
Division of Manufacturing & Product Quality
Darius@cber.fda.gov
301-827-4471

2. The views expressed during this
presentation are my own and may
not be those of the FDA.

3. Overview
• Regulatory Tool Box
– Revised Guidance & Link
– Applicable Regulations
• Objectives of Aseptic Processing &
Validation
• FDA 483 Inspection Observations
• Conclusions

4. Revised Guidance for Industry
Sterile Drug Products Produced by
Aseptic Processing
• Released September 30, 2004
•http://www.fda.gov/cder/gui

5. 21 CFR 10.115 (d)(1)
• Are you or FDA required to follow a
guidance document?
– No. Guidance documents do not establish
legally enforceable rights or responsibilities.
They do not legally bind the public or FDA.
– Alternate approaches are acceptable if they
apply to the relevant statutes and regulations.
– Guidance documents represent current thinking.

6. 21 CFR 211.113 (b)
Control of microbial
contamination
• Appropriate written procedures, designed to
prevent objectionable microorganisms in
drug products purporting to be sterile, shall
be established and followed.
• Such procedures shall include validation of
any sterilization process.

7. Barr Decision
• USA v. Barr Laboratories, Inc.
February 4, 1993, Decided
Civil Action No. 92-1744
812 F. Supp. 458; 1993 U.S. Dist. LEXIS 1932
www.gmp1st.com/barr1.htm
Includes discussion on:
-Failures & Failure Investigations;
-Sampling Sites & Sizes;
-Cleaning & Methods Validation; and
-Mixing Times.

8. Objectives of Aseptic Processing
(or, What’s Really Important)
• What is the fundamental objective of
aseptic processing?
– To keep sterile “things” sterile.
• What are the fundamental objectives of
validating an aseptic process?
– To demonstrate that the process, environment,
and personnel are capable of maintaining
sterility of the final “assembled” product.

9. Selected FDA 483 Observations
on
Aseptic Processing

10. Design
• The filling machine hopper is located
outside the Class 100 curtained area in the
filling room. Sterilized stoppers are
charged into an unsterilized stopper hopper
located in a Class 10,000 environment.

11. Design
• Design & construction of the facility did not
prevent ingress of rainwater into the aseptic
processing areas.
• Steam condensate back-up from the double
door autoclave supporting the aseptic filling
suite repeatedly flooded the Class 100
aseptic filling area.

12. Design & Personnel
• Personnel interrupt first flush laminar flow HEPA
filtered air directly over unprotected vials on
unscrambler table.
a. Gowned personnel were viewed brushing against vial
unscrambler table on numerous occasions while
moving to the other side of the filling equipment to
perform fill line maintenance, view non-viable
monitoring devices, and exchange microbiological
sampling plates.
b. Vial unscrambler is not protected by barrier and area
around unscrambler is too narrow to allow for
minimization of personnel contact with table edges.

13. Design & Personnel
• Fill line operator entered the stopper bowl cabinet
several times using tools that were non-sanitized,
nor sterilized, prior to use. No requirements
specifying sterilization/sanitization of tools prior
to use.
• Autoclave wrap covering the sterilized stopper
bowl was dislodged during set-up, exposing the
stopper bowl to gowned operator’s arm and
unsanitized tools.

14. Qualification
• Velocity testing and smoke studies have not
been performed to demonstrate laminarity
of the air supplying the curtained Class 100
filling areas.

15. Sterilizing Grade Filters
• Concerning Sterilizing Grade Filters:
a. Post-use integrity testing of 0.2 µm vent filters on
autoclaves, WFI holding tanks, purified water systems,
and product flowpath assemblies have never been
assessed, nor performed.
b. There are no data available to support the 12-month
replacement frequency for the sterilizing grade vent
filters on the WFI holding tanks maintained at 80 °C.
The same filters installed on purified water systems
have a six-month replacement frequency.

16. Equipment Suitability
• Malfunction of the stopper feed system
required numerous entries into the stopper
bowl cabinet by fill line operators and
engineers. Stopper bowl was alternately
either partially covered with sterile paper
sheets or left entirely uncovered during
repairs performed prior to aseptic filling
operation.

17. Training
(Believe it or Not!)
• House fly (Musca
domestica) in Class
100 filling room was
not gowned per
procedure.

18. Filling Room Tools
• Non-sterilized tools and lyophilizer loading
trays stored in Class 100 areas used fill line
set-up,repairs and vial transport have not
been microbially assessed.

19. Equipment Transport
• Climet data loggers are transported between
Class 100 and unclassified areas to the QC
Microbiology department for data
downloading. These Climets are wiped
down with alcohol prior to their return to
the Class 100 areas but the potential impact
of this practice has not been assessed
microbiologically.

20. Fill Line Set-Up Operations
• Concerning Fill Line Set-Up Activities:
a. Procedure does not specify how to perform set-up of
the filling line. Variability in aseptic technique was
noted between different personnel performing fill line
set-up.
b. Sterilized filling equipment (stoppering tubes and pins)
are completely unwrapped in Class B areas during fill
line set-up operations and transferred to the Class A
areas for line set-up.
c. Non-sterilized tools are used during fill line set-up and
have never been microbiologically evaluated, or
monitored.
d. Viable microbial monitoring has not been evaluated,
nor performed, during fill line set-up.

21. Aseptic Technique
• Personnel were observed having lengthy
discussions while holding the door to the
cleanroom fully open.
• Personnel were viewed speaking to each
other across the room in close proximity to
the aseptic operations near open vials.

22. Aseptic Technique
• Personnel were observed righting unfilled
sterilized vials that were inverted and had
contacted the sanitized surface of the unscrambler
table prior to filling. These vials were filled, not
discarded.
• Operators loading depyrogenated vials onto the
unscrambler table were observed sanitizing hands
with 70% alcohol in spray bottle directly over the
the open, sterilized vials.

23. Aseptic Technique
• Personnel filling the stopper hopper allowed
the exterior of the stopper bag to contact the
sterilized surfaces of the stopper hopper.
Equipment described has not been
evaluated during the environmental
monitoring qualifications performed.

24. Personnel Monitoring
• Microbial evaluation of thumbs has not
been considered, nor monitored, as part of
the personnel monitoring program.

25. Personnel Monitoring
• Personnel monitoring procedures direct
filling personnel to perform microbial
monitoring on themselves during aseptic
filling operations. Additionally, QA
oversight of personnel monitoring is not
specified.

26. Air Flow Dynamics
• Smoke studies performed did not clearly
demonstrate laminarity of air flow over the
exposed product pathway in that the
direction of air flow swept over the
operators and downwards toward the filling
equipment when the Lexan doors at the
filling needles were opened.

27. Air Flow Dynamics
• Smoke studies were not conducted under
dynamic conditions following the addition
of a Lexan barrier to improve airflow
laminarity.

28. Air Flow Dynamics
• Dynamic smoke studies demonstrated that
swirling and turbulence were observed in
front of the dry heat sterilizer supplying the
Class 100 aseptic filling area and vial
accumulator table.

29. Time Limits
• Maximum allowable time limits for aseptic
filling operations have not been established.
Media fills do not duplicate the longest
time used in production for aseptic filling.

30. Sterility Testing
• Sterility samples were stored at – 40 °C
prior to testing. In addition, suitability of
the containers & closures to maintain
integrity at – 40 °C has not been assessed.

31. Procedures
• The batch production record and associated
procedures are not available to operators in
the aseptic filling areas where processing
occurs and manufacturing data are recorded.
Operators must leave the room to record
data in the batch record.

32. Change Control
• Incorrectly sized parts were installed on the
filling line pump which resulted in rubbing
of metal filling pistons against the metal
support blocks during filling operations.
Metallic particles were noted in final filled
product. QC/QA departments did not
assess potential impact of the changes
made.

33. Environmental Monitoring
• Viable air monitoring is performed by
sampling one-half the volume of air and
multiplying by two in the Class 10,000 and
100,000 areas.

34. Environmental Monitoring
• Microbial monitoring of the stopper hopper
and stopper feed tracks are not performed.
This equipment is sanitized, not sterilized
prior to use.
• Non-viable particulate monitoring is not
performed in the area near the stopper bowl,
where operators manually charge the bowl
with sterile stoppers from breather bags.

35. HEPA Filters
• Integrity testing of the HEPA filters in the
Class 100 aseptic filling area have never
been performed.

36. HEPA Filters
• 85% of HEPA filters in the aseptic filling
suite were determined to be non-integral
during the scheduled six month HEPA filter
integrity testing performed by an outside
contractor.

37. Culling of Media Fill Units
• Visual inspection personnel remove media
filled bottles which are damaged or
defective after the incubation period is
completed. The turbidity status of these
bottles is not always recorded in the batch
record and the damaged or defective bottles
are not sent to QC Microbiology for
evaluation prior to destruction.

38. Media Fills
• Not all filled units are incubated in that
eight units per batch are removed and
inoculated as positive controls.

39. Media Fills
• Media fill operations have not used
container/closure systems representative of
those used routinely during aseptic filling
operations.

40. Media Fills
• Media fills do not include simulations of all
aseptic operations such as aseptic additions
of buffers and formulation components
transferred into the mixing vessels
supplying the filling line.

41. Media Fills
• There is no assurance that adequate
volumes of media have been filled into each
bottle to assure that it contacts all interior
surfaces of the filled containers.

42. Media Fills
• Growth promotion testing of media used for
aseptic media fill simulations has not been
evaluated or performed.

43. Particulate Monitoring
• Non-viable particulate monitoring
performed is not representative of actual
conditions in that the monitoring probes are
located in the return air ducts of the Class
100 aseptic filling area.

44. Particulate Monitoring
• Non-viable particulate monitoring is not
performed during performance of aseptic
connections made for the sterile filtration
step of buffer solutions.

45. QA/QC Oversight
• Non-sterile filtered nitrogen is used to
pressurize the sterile formulated bulk tank
supplying the aseptic filling line.
– Both the QA and QC Microbiology units
approved the process and deemed the practice
acceptable.
– Anaerobic monitoring of the process has not
been evaluated or performed.

46. Isolators
• Operator standing on stool in order to reach half-
suite fell during performance of a sterility test and
damaged isolator.
• No method available to remove accumulated
empty vials from inside isolator after a five-day
filling operation. Vials blocked air return grills.
• Paper edged HEPA filters installed in isolator
deteriorated after several VHP sterilization cycles.

47. Lyophilization
• Product pathways for partially stoppered,
and unprotected, vials conveyed from the
filling area to the lyophilizers are monitored
according to Class 10,000 rather than Class
100 environmental standards.

48. Lyophilization
• Lyophilizer chamber is vented directly to a
Class 100,000 environment. A sterilizing
grade vent filter was not installed to protect
the chamber. In addition, there are no data
available to support the maximum
allowable seven day hold on the sterilized
lyophilizer prior to use.

49. Lyophilization
• Non-viable particulate monitoring is
performed at a location eight inches below
the ceiling HEPA filter face (or four feet
above the vial traying operations) for
laminar flow air supplying the accumulator
table where partially stoppered vials are
loaded onto trays prior to lyophilization.

50. Novel Approach for Determination
of Media Fill & Sterility Failures

51. Parting Thought
• The most frequently issued observations
begin with:
There are no data available to support…

52. Conclusions
• Understand your process and its limitations.
– Be alert to the practical realities of people and the
process. Strive to minimize identified or perceived
risks.
• Maintain a science-based evaluation of the
processes to develop validation approaches that
can be scientifically defended.
– Question “why?” and “how?” to stimulate internal
dialogue on the process, risks, and validation
approaches.
• There are solutions to all problems.
– The solutions are waiting to be discovered.
• Apply good common sense.

53. “The obscure we see eventually.
The completely apparent takes
longer.”
E.R. Murrow
Robert Darius, FDA/CBER
Darius@cber.fda.gov
301-827-4471