1. USP <1116> and its impact on Microbiology
Dr. Tim Sandle
Pharmaceutical Microbiology:

2. Introduction EM guidance Background to USP <1116>
Main changes and debates
Method limitations
Incident rates
Frequencies of monitoring
Locations of monitoring
Other changes
Regulatory issues
Rapid methods
Pharmaceutical Microbiology:

3. Sources of EM Guidance
EU GMP - last EM revision 2009; new annex ?
FDA aseptic filling guide (2004)
PDA Technical Report (2014, 3rd revision)
ISO (1998). Future update?
USP <1116> (2011, published 2012)
Pharmig Current Review (2010)
PHSS Biocontamination control guide 2015
Pharmaceutical Microbiology:

4. Sources of EM Guidance ISO 14698
Under long-term review
Possible development to a viable cleanroom classification standard (like ISO 14644)
No other standard is likely to be reviewed in the short-term
Pharmaceutical Microbiology:

5. ISO 14644 ISO 14644 Parts 1 and 2 revised in December 2015
A standard for cleanroom classification.
Some detail about on-going monitoring.
Does not cover viable monitoring at all.
Changes:
New look-up tables
Increase in counter locations
Changes to sampling volumes
Each individual location must pass.
More risk based.
Pharmaceutical Microbiology:

6. USP <1116> (2012) USP <1116> Revision began in 2005
Objectives of USP committee:
Focus the chapter on environmental monitoring only, removing information relating to aseptic process validation.
Focus the document exclusively on the monitoring of aseptic environments.
Reconsider the alert and action level (limit) concept.
Effective 1st May 2012, 35th edition of the USP
Pharmaceutical Microbiology:

7. Main changes Former title:
“Microbial Control and Monitoring Environments
Used for the Manufacture of Healthcare Products”
Revised title:
“Microbiological control and monitoring of aseptic processing environments”
Pharmaceutical Microbiology:

8. Main changes Scope: Environments Pharmaceutical sterile products
Bulk sterile drug substances
Sterile intermediates
Excipients
Environments
Conventional clearoom with UDAF
Blow-fill-seal
RABS
Isolator
Pharmaceutical Microbiology:

9. Main changes
The emphasis on the word “aseptic” in the introduction implies that the chapter is not applicable to all “sterile” products.
This means that terminally sterilised products are outside the scope of the chapter.
Pharmaceutical Microbiology:

10. Main changes
By “aseptic” a low level of contamination is acknowledged:
“an expectation of zero contamination at all locations during every aseptic processing operation is technically not possible and thus is unrealistic”
Pharmaceutical Microbiology:

11. Main changes
USP notation system dropped: M3.5 etc. And old FDA 209E classes e.g. Class 100, Class 10,000)
Replaced by ISO classes in the operational state
Difference with EU GMP:
Class 5 = EU GMP Grade A
Class 7 = EU GMP Grade B
Class 8 = EU GMP Grade C
Pharmaceutical Microbiology:

12. Main Changes Relative risks
Picture showing on how process separation and product protection interact (adopted from Bioquell U.K Ltd).
Pharmaceutical Microbiology:

13. Main changes
The design and construction of clean rooms and controlled environments are covered in ISO
ISO stipulates the total particulate counts required for a clean environment to meet the defined air quality classifications. USP accepts this standard verbatim.
Pharmaceutical Microbiology:

14. Main Changes New guidance for cleanroom operations:
ISO class 8 = 20 air changes per hours
ISO class 7 = 50 air changes per hour
ISO class 5 = 100 air changes per hour
Isolators = can have a different justification for air changes and air velocity
An EM programme should only be constructed and executed once airflow mapping and HVAC dynamics have been optimised.
Pharmaceutical Microbiology:

15. Environmental monitoring
USP places emphasis upon particle and viable monitoring
Particle counting most important for isolators
Viable monitoring is regarded as semi-quantitative
Trending is the most important aspect of the monitoring programme
Isolated counts “a normal phenomenon in conventional cleanrooms” which do not require specific corrective action and there is the possibility of a false positive
Pharmaceutical Microbiology:

16. Environmental monitoring
USP requires monitoring of:
Surfaces
Air (room and enclosure)
Compressed gas
Changes in trend must be investigated, and include assessment of:
Maintenance
Disinfection
Unusual events and activities
Physical changes e.g. temperature and humidity
Staff training
Pharmaceutical Microbiology:

17. Environmental monitoring
SCDM (TSA) is a suitable medium, incubation at ‘low’ and ‘high’ temperatures (20-35oC for not less than 72 hours)
Consideration given to fungal medium e.g. SDA
Certain conditions may require micro-aerophilic monitoring e.g., certain gasses or sterility test failure using anaerobic media
Pharmaceutical Microbiology:

18. Methods Settle plates: EU GMP:
Semi-quantitative measurement (CFU / 4hours)
Require desiccation study.
USP 1116: “The exposure of open agar-filled Petri dishes, or settling plates, is not to be used for quantitative estimations of the microbial contamination levels of critical environments.”
Pharmaceutical Microbiology:

19. Monitoring methods No EM programme can prove sterility
Environmental control is the most important, supported by EM and media simulations
EM can demonstrate that a clean room is operating with in a consistent state of control
However, “EM” requirements have evolved in a manner that did not fully consider analytical capability and metrology
Pharmaceutical Microbiology:

20. Monitoring methods All monitoring methods are flawed:
Require people to take them
Variability in technique
Risk of false positives
No one method can detect all types of contamination
All methods have relatively poor recoveries (“insensitive”)
Air-samples are particularly weak
Surface methods have poor recoveries
Settle plates are not considered quantitative
All methods are poor at recovering damaged or stressed microorganisms
Pharmaceutical Microbiology:

21. Monitoring methods Therefore, numerical targets (as CFU) should:
Not be used as limits (they are “levels”)
Not be considered specifications
Be seen as informational only
Low or zero counts are not, by themselves, guarantees for microbial control;
Equally, excursions beyond numerical limits are not necessarily and indication of loss of control.
Instead:
Count non-zero events.
Use a contamination recovery rate metric based on historical findings
Inference that these should be stable over time with little variation.
Pharmaceutical Microbiology:

22. Plate counting debate DEBATE
Should not simply count CFU due to the inaccuracies of current environmental monitoring methods
It is more important to count the incidence rates and investigate out of trend.
The limit of quantitation (the number of cfu that can be reported accurately) is 15.
If CFU below 15 cfu , do not worry if trend is OK.
If above 15 cfu, investigate.
Pharmaceutical Microbiology:

23. What does this mean?
At very low recovery levels there is no way to establish Alert or Action Levels statistically-the counts are simply too low to make statistical analysis useful.
Instead, emphasis should be on incidents.
“Hits” in ISO 5 aseptic environments should be infrequent.
Pharmaceutical Microbiology:

24. What was the pre-USP 2012 situation?
Take active air-samples:
Microbiological cleanliness levels ‘In Operation’ cfu/m3
Area
EU GMP
FDA Guide
USP
Aseptic core
<1
<3
Support for aseptic filling
<10
<20
Controlled process area
<100
Controlled support area
<200
Not specified
Pharmaceutical Microbiology:

25. USP contamination rates
Class
Active air sample
Settle plate
Contact plate
Swab
Isolator or
Closed RABS (ISO 5 or better)
<0.1%
ISO 5
<1%
ISO 6
<3%
ISO 7
<5%
ISO 8
<10%
Pharmaceutical Microbiology:

26. USP contamination rates
NOTE: Contamination recovery rates should be based upon actual monitoring data and should be re-tabulated monthly.
When contamination recovery rates are observed that exceed the recommendations in the table or are greater than established process capability corrective actions should be taken.
Pharmaceutical Microbiology:

27. USP contamination rates
For example
Grade B cleanroom
Active air-sampling
One year of data reviewed
Pharmaceutical Microbiology:

28. USP contamination rates
Count (cfu)
Frequency
Cumulative percentage
1240
80.78% 1
191
93.22% 2 52
96.61% 3 20
97.92% 4 8
98.44% 5 7
98.89% 6
99.02%
99.22%
99.35% 9
99.41% 10
Pharmaceutical Microbiology:

29. USP contamination rates
Pharmaceutical Microbiology:

30. USP contamination rates
Corrective actions may include but are not limited to:
Sanitization program including types of disinfectants, application methods, and frequencies.
Personnel practices by supervisory staff.
Microbiological sampling methods and techniques.
Training on gowning practices.
Pharmaceutical Microbiology:

31. Frequencies of sampling
Isolators
Active Air Sampling-once/day
Surface sampling-at end of each campaign
Glove sampling-left to the users discretion
Note differences to EU GMP for continuous monitoring.
Pharmaceutical Microbiology:

32. Frequencies of sampling
RABS
Open RABS and closed RABS are different regarding contamination risk.
Open RABS is more similar to a conventional clean room.
Closed RABs are advanced aseptic processing systems.
Pharmaceutical Microbiology:

33. Frequencies of sampling
Cleanrooms
Unchanged from the previous version of <1116> appearing in PF31(2).
ISO class 5 = Each operating shift
ISO class 7 = Each operating shift
ISO class 8 = Twice per week
Other areas = Once per week
Pharmaceutical Microbiology:

34. Sampling locations
ISO grid approach for particles discussed, but dismissed
“Microbiological sampling sites are best selected with consideration of human activity during manufacturing operations.”
From careful observation and mapping of the clean room
The most likely route of contamination is airborne
Pharmaceutical Microbiology:

35. Other changes / main features
Strong emphasis upon staff training, including those who take microbiological samples
Need for a qualified site microbiologist
Staff health checks and control of entry to critical areas
Importance of correct gowning
Importance of risk assessment and risk mitigation
Pharmaceutical Microbiology:

36. Other changes / main features
USP is strongly supportive of the use of “RMMs” in Environmental Monitoring and all forms of microbiological analysis.
All references to validation have been removed from the chapter.
Pharmaceutical Microbiology:

37. Regulatory differences
Results:
EU GMP Annex 1 discusses averaging of micro data.
FDA mandates response to individual excursions.
USP focuses on incident rates.
Disinfection residues
FDA and EU GMP suggest microbial resistance to sanitizers is possible.
USP does not cover this.
Media fills.
All guidances use the same media fill criteria.
USP provides no details for large fills.
Pharmaceutical Microbiology:

38. Future developments
Development of a chapter on Microbiological Control & Monitoring of Non-Aseptic Processing Environments <1111> has been discussed by USP MSA
Problems:
Operations vary much more widely than in aseptic processing.
No widely accepted standards for the various facility designs.
Significant differences in approach for the same product types.
A clear path?
Pharmaceutical Microbiology:

39. Summary
The challenge in aseptic processing is always personnel: as a source of microbial and particle contamination.
Environmental monitoring continues to be especially contentious.
The contamination rate concept is a significant departure from the alert and action level system that has been used in industry since the early 1980’s.
Pharmaceutical Microbiology:

40. Pharmaceutical Microbiology: http://www.pharmamicroresources.com
Thank you
Pharmaceutical Microbiology:
Pharmaceutical Microbiology: