1. Environmental Control & Support Processes

2. Environment & Control
Environment - the circumstances and conditions that surround
an organism or a group of organisms
Where product is being made
Control - To maintain desired conditions in a area /process/system by adjusting selected variables.

3. Environmental Control cont.
Biology, Chemistry, Engineering, Law, Sociology, Political Science, History, Literature, Art, Religion, Math, Physics
Nature, Culture and Society
Human Environment
Environmental Control – Control of environment or surroundings (which tend to influence the work place and products that are made there.

4. Environmental Control cont.
It involves the control of the levels of
Toxins
Chemical pollutants
Microbial contaminants
or other harmful substances
at a workplace

5. Get Familiar with ACRONYMS
CFR: Code of Federal Regulations
CFU: Colony Forming Unit
cGMP: Current Good Manufacturing Practice
EM: Environmental Monitoring
FDA: (US) Food and Drug Administration
ISO: International Organization for Standardization
cGDP: current Good Documentation Practices
RODAC: Replicate Organism Detection And Counting
SWOT: Strength, Weakness, Opportunities, Threats
USP: United States Pharmacopeia

6. AREAS RELATED TO ENVIRONMENT CONTROL
Air filters
Air-pressure monitors and controls
Airborne particle counters
Ceiling filter modules
Cleaning and disinfection equipment
Cleanrooms
Design and construction materials
Design and construction services
Doors, automatic and manual
Ducts
Environmental monitors
Environmental sensors
Exhaust systems and fans
Facility monitoring systems
Flooring/mats
Fume hoods
Furniture
Gloves
Humidity-control systems
HVAC systems
Isolators
Keyboards and keypads
Laminar-flow benches
Laminar-flow filters and test equipment
Laminar-flow units
Lighting
Maintenance services
Meters, airflow, humidity, and temperature
Microbial air samplers
Particle-control equipment
Swabs and wipers
Vacuum systems
Wall panels

7. Get familiar to ……………., What is a clean room?
A room or environment in which you move the air by way of supply and return locations to control the airborne particle levels and temperature and humidity.
What is Federal Standard 209E?
The Federal government's basic design and performance requirements for clean rooms.
Some of the information sets minimum and maximum levels, but in general it gives recommended guidelines.
What is a Class?
Defines the limit or measurement that a room will perform to particles per cubic foot at 0.5 micron or larger.

8. HEPA Filter High Efficiency Particulate Air
Class of air filters which meet a minimum performance level of 99.97% on 0.3 microns efficiency.
In the cleanroom market HEPA is normally rated at 99.99%
An additional face scan test is performed
to assure no pin holes or leaks are found.

9. Particulate Matter What is a particulate matter? What is Recovery?
Contamination found in the air or which is generated within a room from a process.
What is Recovery?
How quickly the room or area will clean up and return to normal?

10. World Health Organization
7 February, 2018
Specimens
Air samples
For Both viable & Non-viable & compressed air
Surface swabs
Floors, Walls, Equipment, etc.

11. For a known Quantity (1000L) of Air/cubic meter
Air Quality
Air Sampling
Passive Air Sampling
(Plate Exposure)
Open pre-incubated media plates (90mm) keep exposed for 4 hrs.
Active Air Sampling (Volumetric)
For a known Quantity (1000L) of Air/cubic meter

12. Clean Rooms Surface By contact plates By swab sampling (RODAC)
55 mm Plates to be fixed on a flat surface
By swab sampling
Swabs are rubbed over the test surface & tested for microbial contamination
Swabs are used for the surfaces that are not flat

13. 21 CFR 211.46 Ventilation, Air filtration, Air heating and Cooling:
Adequate control over microorganisms, dust, humidity and temperature
Air filtration systems including pre-filters and particulate matter air filters
for air supplies to production areas

14. Sample Sites Locations posing the most microbiological risk
Monitor critical area
Based on contamination risk
Easiness/difficulty in set up
Length of processing time
Impact of any interventions
Monitor filling/closing area
Air and surface samples near significant activity

15. Elements of SOP on Sampling
Frequency of sampling
Time of sampling
Duration of sampling
Sample size
- How much surface area?
- How much air volume?
- How many plates?
Specific equipment & technique
Alert & Action levels
- Appropriate response to deviations

16. 483 – Sample, Warning Letter
Regarding the increased non-routine surveillance monitoring performed to further evaluate the Bldg. 37 Flu manufacturing
There was no plan in place
specifying the locations to be tested
Method of sampling
Actions to be taken in case of microbial contamination
Samples on CFU were evaluated for morphological characteristics
Only Gram negatives were stained and identified

17. 483 – Sample, Warning Letter cont.
The method as written and used for increased surveillance monitoring of the environment has not been QUALIFIED.
A qualification study with more data
On location
Frequency
# of sample sites was needed

18. Number of Sites
International Organization for Standardization (ISO)
Describes a method to determine the number of sampling sites for site qualification
NL = √A
Where NL is the minimum number of sampling locations (rounded up to a whole number)
A is the area of the clean room or the zone in meters2
This works well for non-viable particulate measures

19. Classification of Clean Rooms - Federal Standard 209
Quality Control in Biomanufacturing
Monday July 23rd, 2007
Classification of Clean Rooms - Federal Standard 209
≥ 0.1µm
Particles/ft3
≥ 0.2µm Particles/ft3
≥ 0.3µm Particles/ft3
≥ 0.5µm Particles/ft3
≥ 5.0µm
Class 1 35
7.5 3 10
350 75 30
100
750
300
1000
1,000 7
10,000 70
100,000
700
BIOMAN Portsmouth NH 19

20. Selected Equivalent Classes
FS Classes
Class 1 10
100
1,000
10,000
100,000
ISO Classes 3 4 5 6 7 8

21. Particle Detection The validation of a clean room is ongoing
The air quality of a clean room must be monitored
An optical particle counter is used to monitor air quality
“Real-time” test results 21

22. Types of Particle Counters
Facilities Maintenance System
Portable Particle Counter

23. Active Air sampling
Active sampling involves the collection of air to be evaluated through a sampling tube or probe using a pump.
The compounds of interest accumulate on the sorbent material in the tube or the nutrient plate and are returned to the laboratory for analysis, giving a result as a “mass per tube”. (i.e., micrograms per tube)
The volume of air passed through the sorbent or collection device is also accurately determined. (i.e. cubic meters)
The “mass per tube” is then divided by the volume of air to give the airborne concentration. (i.e. micrograms per cubic meter).

24. Passive Air Sampling
It is a frequently used measure of cleanroom (controlled zone) monitoring.
Settle plates are exposed for specified time period.
Advantages:
Ability to remain in continuous exposure for at least 4 hours
(Extended hours need to be validated)
Not disruptive to the immediate environment
Air sample are collected very near to product exposure
Not prone to variation among different vendors
Disadvantages:
Handling, transport and lab contamination

25. Plates Before and After exposure
Settling Plate- Before Exposure
Settling Plate; After Exposure & After Incubation

26. Inspection of Settle Plate and Count
Total microbial count
Bacteria
Mold
The colonies are counted and reported as colony forming units (CFU) per cubic meter of air

27. Microbial Evaluation and Classification of Clean Rooms and Clean Zones
Establishment of Alert and Action limits
Suggests limits for airborne, surface and personnel contaminant levels.
Methods and equipment for sampling
Identification of isolates
Aseptic media fills

28. Action/Alert/Acceptable Levels
Action Level at 118
Alert Level at 88
Passing level at 4

29. Setting Alert and Action Limits
Action limits can be established in a variety of guidance documents
Alert limits
Lower than action limits
Reflect actual historical results under normal processing conditions

30. Exceeding Limits 1/3
Alert limits are designed to provide some warning that environmental quality is approaching action limit and allow you time to correct.
Exceeding alert limit triggers a warning response
Exceeding multiple alerts - triggers action level response

31. Exceeding Limits 2/3 Action limit excursions require investigations
Speciation of organism(s)
Review batch records from date of excursion
Review other recent EM data (trends)
Review cleaning records
Interview personnel
Product impact - must quarantine until determined

32. Exceeding Limits 3/3
Excursions from action limits require corrective actions that may include:
More rigorous or additional monitoring
More rigorous cleaning
Retraining of personnel
Procedural changes - change to or addition of disinfection procedures, for example
HVAC maintenance

33. Water for Pharmaceutical Use
Water is the most widely used substance / raw material
Used in production, processing, formulation, cleaning, quality control
Different grades of water quality available

34. Water Quality Different Water System Water by Reverse Osmosis
Water for Injections (WFI)
Potable water

35. Micro Contamination of water
Microorganisms
Protozoa
Cryptosporidium
Giardia
Bacteria
Pseudomonas
Gram negative, non-fermenting bacteria
Escherichia coli and coliforms

36. Environmental Trending
Should trend monitoring results: (environmental and water)
Periodic (quarterly or monthly) review by QA and others
Re-evaluation of action and alert limits on an annual basis
This trending information is generally included in the Annual Product Review

37. Insect and Rodent Control
Great number of illnesses are associated with animal and insect contact.
Encephalitis
Lyme Disease
Rocky Mountain spotted fever
Bubonic plague
West Nile virus

38. Microbial Monitoring Devices
Slit-to-Agar (STA):
Air taken in through a slit
Below which is a slowly revolving plate
Sieve impactor:
Vacuum draws in air through perforated cover
That is impacted onto petri dish
That contains nutrient agar

39. Microbial Monitoring Devices cont.
Surface contaminant monitoring devices:
Contact Plates –
plates filled with nutrient agar; for regular surfaces
Swabs –
useful for hard to reach or irregular surfaces;
swab placed in suitable diluent and inoculated onto microbiological plate

40. What is Bioburden Testing?
It is microbial testing performed on medical equipment and products.
It measures the number of living organisms on a surface prior to final sterilization and use.
Certain microbial levels are allowed during handling of a product, and Bioburden testing makes sure those limits are in force.

41. How is Bioburden Testing Regulated?
All Bioburden tests must conform to the procedure outlines in ISO (International Organization for Standardization)
Tests must fit the basic FDA criteria which include testing for the following:
Yeast Candida albicans
Bacteria Escherichia coli
Bacteria Pseudomonas aeruginosa
Bacteria Staphylococcus aureus
Fungus Aspergillus niger

42. How is Bioburden Testing Performed?
Swabs of the outside of a product are collected and grown in petri dishes.
Once the samples have grown, they are tested for various bacteria, fungi, and other contaminants.
Bioburden is measured in colony forming units (CFU) per gram of product.
A sterilization procedure is determined.

43. Endotoxin Testing Which products are tested?
Injectable drugs and medical devices
which will contact blood or spinal fluid
Includes
Raw materials
Water
In process materials 43

44. The Pyrogen Test conducted with rabbits
Two tests – USP
The Pyrogen Test conducted with rabbits
Bacterial Endotoxins Test
Also called
Limulus Amebocyte Lysate (LAL) Test.

45. Pyrogen Assay USP considers a solution to be pyrogenic when:
10 ml/kg is injected into a rabbit and there is a rise of temperature of 0.6 °C or more for any rabbit or
a total rise of more than 1.4 °C for three rabbits in a three rabbit test group.

46. LAL Test Limulus Amebocyte Lysate test: Developed in 1960’s Faster
Based on clotting reaction of horseshoe crab (Limulus polyphemus) blood cell (Amebocyte) lysate to endotoxin
Developed in 1960’s
Drs. Bang and Levin
Faster
more economical
more sensitive than rabbit Pyrogen test All methods accepted by the FDA, USP, EP,& JP 46

47. Types of LAL Tests Gel Clot Turbidimetric Colorimetric
All accepted by the FDA, USP, EP,& JP

48. LAL Method 1. Gel Clot: Original method The official “reference test”
The specimen is incubated with LAL of a known sensitivity.
Formation of a gel clot is positive for endotoxin.
2. Turbid metric: In the presence of endotoxin, LAL becomes turbid

49. 3. Chromogenic Method
Endotoxin concentration is measured as a function of color intensity
LAL contains enzymes that are activated in a series of reactions in the presence of endotoxin.
The last enzyme activated in the cascade:
Splits the chromophore, Para-nitro aniline (pNA), from the chromogenic substrate
Producing a yellow color.

50. Documents vs. Records
Documents
Records
Offer guidance or explain actions to be carried out
Can be changed
Provide verification of things that have happened
Cannot be changed

51. Documents vs. Records cont.
Documents are materials that provide management directions:
environmental policy
internal standards and operating procedures
process information
organization charts
emergency plans
Records include
training records
incident reports
product information
complaints and responses
audit results
management review meeting minutes