Step-by-step guide to designing standard a microbiology laboratory in pharmaceutical companies.pptx

1. Step-by-step guide to
designing standard a
microbiology laboratory in
pharmaceutical companies
Dr shawky

2. Dr/shawky
https://www.linkedin.com/in/shawky-abdel-fattah-
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3. The laboratory in the image is designed according
to pharmaceutical quality standards (GMP, ISO
14644, WHO, FDA) to prevent contamination
and ensure the accuracy of tests. I will explain
the layout in detail, from start to finish, and why
the rooms are arranged in this way.

4. designing
standard
a
microbiology
laboratory

6. Entry and Changing
01

7. 📍 Why start here?
This is where employees enter the lab and change clothes to
ensure they do not bring contaminants from the outside.
1️Primary Changing Room
Employees enter this room to change from regular clothes into
sterile garments (gowning area).
No one is allowed inside without proper attire.
2️
Airlocks (A/L-1, A/L-2, A/L-3)
After changing clothes, employees pass through these rooms,
which are controlled transition zones that prevent contaminants
from entering.
Each room regulates airflow to prevent particles from moving from
less sterile to more critical areas.

8. 3️ Buffer Zone (1M, 1F)
Employees pass through these areas to ensure no contamination
before entering critical zones.
These zones help protect the rest of the laboratory from external
contaminants.

9. Testing & Analysis
Laboratories
02

10. 📍
Why do these come after the changing area
?
Now that employees are sterile, they can enter the lab and
conduct tests. These rooms are used to test products and
ensure they are free from microbes and toxins
.
4️
LPC & BET Lab (1A) - Particle & Bacterial Endotoxin Testing
Tests particle counts in pharmaceutical liquids to ensure sterility
.
Detects bacterial endotoxins (LPS) that could harm patients
.
Applies global standards USP <85> for product testing
.
5️
MLT Lab (1N) - Microbial Limit Testing
Determines the acceptable microbial count in pharmaceutical
products to ensure their safety and quality
.
Applies global standards USP <61> & <62> for product testing
.

11. 6️
Culture Bioassay & Microbial Identification Lab (2C)
Microbial cultures are grown on media to identify bacteria
or fungi present in samples
.
Techniques such as PCR and MALDI-TOF are used for microbial
identification
.
7️
Sterility Lab (1H)
The most critical lab, testing whether products are 100% sterile
.
Operates under ISO Class 5 (the highest level of sterility)
.
Air pressure is controlled to ensure no contamination enters the
room
.
Applies global standards USP <71> for product testing
.

12. Storage &
Preparation Areas
03

13. 📍
Why do these come after the labs
?
These rooms support the laboratories by providing the
necessary tools and materials
.
8️
Media & Glassware Store (3B)
Stores culture media (e.g., Agar, Broth) used for microbial growth
.
Maintains sterile glassware for lab tests
.
9️
Media Preparation Room (3C)
Prepares nutrient solutions used for microbial cultivation
.
Equipped with an autoclave to sterilize these solutions before use
.
10️
Cold Room (1B)
Preserves microbial samples and chemical reagents at low
temperatures to maintain their quality
.

14. Equipment Support
& Cleaning Areas
04

15. 📍
Why do these come after storage
?
These rooms are responsible for cleaning and sterilizing the lab
equipment, preventing contamination
.
1️
Incubation Room (3F)
Incubates microbial plates at different temperatures
(
25°C, 30°C, 37°C
)
to monitor microbial growth
.
Contains incubators with controlled temperature and humidity
.
2️
Observation Room (3A)
Examines microbial cultures using a microscope and microbial
growth analysis devices
.

16. 3️
Drying Room (3E)
Dries sterilized glassware to ensure no moisture remains
that could cause contamination
.
4️
Washing Area (3G)
Equipped with sinks and ultrasonic cleaning devices to wash
lab tools before sterilization
.
5️
Waste Disposal for ETP Plant
Manages the safe disposal of hazardous biological waste
according to environmental safety standards
.

17. Safety &
Contamination
Control
05

18. 📍
Why is this the final step
?
These rooms support all laboratory operations and help prevent
contamination from spreading between different areas
.
6️
DPB - Dynamic Pass Box
Used to transfer materials between rooms without opening doors,
preventing contamination
.
7️
TPB - Triple Pass Box
Transfers materials from a lower sterility area to a highly sterile
area, minimizing contamination
.

19. 8️
HVAC & Clean Corridors (1, 3A)
The laboratory has an advanced ventilation system with
HEPA filters to maintain a sterile environment
.
Air pressure is carefully controlled to prevent contaminants from
spreading
.

20. HVAC SYSTEM
Provide a specific set of
environment condition required for
the manufacturing process.
Heating and cooling
Humidifying and dehumidifying
Cleaning the air
Regulate air flow
Pressurization
PURPOSE
FUNCTION
S
1) To prevent contamination
2) To provide comfortable working
conditions
USES

21. ● H.V.A.C–
Heating, Ventilation and Air Conditioning system
● The HVAC regulates
⮚ Room Temperature
⮚ Humidity
⮚ Air Quality
⮚ Air Flow

22. COMPONENTS:-
● Air conditioners
● AHUs air handling units
● Dehumidifiers/ Heater
● Filters (Pre & HEPA)
● Dust extractor
● Ducting (for delivery of controlled air)
● Supply fans
● Smoke detectors
● Damper
● Humidity/Temp./ Pressure sensor
● Heating & Cooling coils

23. Air handling
unit
Blower/Fan
Heating and cooling
coils
Humidifiers
Dehumidifiers
Air distribution
Network
Duct network
Insulators
Dampers or valves
Air filters
Pre-filter
Intermediates or low
efficiency filter
Terminal filter
(HEPA filter)
COMPONENTS OF HVAC SYSTEM

24. Schematic Representation of HVAC system

25. HVAC SPECIFICATIONS
Temperature 18-23 o
C
Relative humidity 45% ± 5%
Dry powder = 30% ± 5%
Moisture sensitive drug = 5% ±
5%
Air velocity 80- 120 ft/min
Air flow Laminar airflow
Pressure gradient 15 Pascal
Particulate count (Critical area) NMT 100 particles of 0.5 µm/ft3
Air system failure alarm (ASFA)

27. ENVIRONMENTAL MONITORING
(Schedule-m)
A Particulate monitoring in air Monthly
Daily
B HEPA filter integrity testing Yearly
C Air change rate Monthly
D Temperature and Humidity Daily
E Air pressure differentials Daily
F Microbiological monitoring Daily ( In aseptic areas)
Decreased frequency in
other areas

28. Validation of AHU/HVAC System
● Temperature control test
● Humidity control test
●
● Filter integrity test
● Air velocity test
● Air flow pattern(Smoke test)
● Microbial test

29. 1.Temperature control test:-
Equipment- Thermometer
Test procedure:-
•Environment is divided by a grid
•Size of square- 60cm×60cm or more
•Sampling location- work height
•Result compare with specification
60×60cm
Objective:- To demonstrate the ability of
the HVAC system to control temperature.
Acceptance criteria:- Temp. 20±2ºC

30. 2.Humidity control test:-
Equipment- Automatic humidity recorder
Test procedure:-
•Environment is divided by a grid
•Size of square- 60cm×60cm or more
•Sampling location- work height
•Result compare with specification
60×60cm
Objective:- To demonstrate the ability of
the HVAC system to control humidity.
Acceptance criteria:- Humidity: 45±5% Fig..
Digital
moisture
meter

31. Filter Integrity Test:-
Objective:- To provide evidence of the integrity of the HEPA
filter
Acceptance criteria:-
99.97 efficiency- 0.03% particles of 0.3 µm
99.99 efficiency- 0.01% particles of 0.3 µm
100%
0.03%
HEPA filter

32. DOP (Dioctyl phthalate) Test
HOT DOP
• Efficiency test
• Vaporization
• Mono-disperse aerosol
• 0.3 µm
COLD DOP
• Integrity test
• Pressurization
• Poly-disperse aerosol
• > 0.3 µm
• < 0.3 µm
• 0.3 µm (20-30 %)

33. Air flow velocity and uniformity test:-
Test procedure:-
• Environment is divided by grid (equal)
• Measure by Airflow meter
Fig.-
laminar
air flow
unit
Acceptance criteria:-
Vertical flows 0.30 m/sec ± 20%
Horizontal flows 0.45 m/sec ± 20%
Action: Deviation indicates blockage of filter
Solution : Alteration of fan speed HEPA filter replacement

34. AIR FLOW SYSTEM (SMOKE TEST ) :-
Generate visible smoke
upstream from the work zone
Establish the reference point
Videotape the direction of the
flow in both case
Determine the direction
Test procedure:- (smoke test)

35. MICROBIOLO
GICAL TESTS
later

36. classification of
clean areas for flow
rate of
airborne
particulate.

37. Sample locations and numbers
● The minimum number of sampling
Locations (N) is calculated as
N = A
√
● Where A = cleanroom area (m²).
● ✅ Example Calculation:
● For a 4m × 5m cleanroom:
● N = (4 × 5) = 20 = 4.47 5 locations
√ √ ≈
● 👉 At least 5 measurement locations are required.

38. ISO 14644-1 Cleanroom Classification
This document provides the maximum allowable airborne
particle concentration for each ISO cleanroom classification
according to ISO 14644-1.To ensure accurate measurement,
the minimum air sampling volume per location is calculated as:
ISO Class
≥ 0.1 µm
(particles/
m³)
≥ 0.2 µm
(particles/
m³)
≥ 0.3 µm
(particles/
m³)
≥ 0.5 µm
(particles/
m³)
≥ 1.0 µm
(particles/
m³)
≥ 5.0 µm
(particles/
m³)
ISO 1 10 2 — — — —
ISO 2 100 24 10 — — —
ISO 3 1,000 237 102 35 — —
ISO 4 10,000 2,370 1,020 352 83 —
ISO 5 100,000 23,700 10,200 3,520 832 29
ISO 6 1,000,000 237,000 102,000 35,200 8,320 293
ISO 7 — — — 352,000 83,200 2,930
ISO 8 — — — 3,520,000 832,000 29,300
ISO 9 — — — 35,200,000 8,320,000 293,000

39. How Much Air Should Be Sampled
at Each Location?
● To ensure accurate measurement, the minimum
air sampling volume per location is calculated as:
V = (20/C) × 1000
● Where : C = Maximum allowable particle concentration
for ISO Class 3 (1000 particles/m³).
● ✅ Example Calculation:
● V = (20/1000) × 1000 = 20 liters = 0.02 m³
● 👉 At least 20 liters of air must be sampled at each location.

40. A cleanroom is ISO Class 3 compliant if:
1️The average particle concentration at each
measurement location is below the class limit.
2️If fewer than 10 locations are sampled, the 95%
Upper Confidence Limit (UCL) must also be
below the class limit.

41. What if We Have Less Than 10
Measurement Locations?
● If N < 10, we must verify compliance using the 95%
Upper Confidence Limit (UCL) formula:
UCL = M + (F × S)
● Where:
● M = Average particle concentration across all locations.
● S = Standard deviation of particle concentrations.
● F = UCL factor (based on the number of locations).

42. 95% UCL Factors
Table of UCL Factors:
N = 2 → F = 6.3
N = 3 → F = 2.9
N = 4 → F = 2.4
N = 5 → F = 2.1
N = 6 → F = 2.0
N = 7-9 → F = 1.9

43. Example UCL Calculation
Given:
N = 5 locations
M = 800 particles/m³
S = 50 particles/m³
F = 2.1 (from table)
✅ Calculate UCL:
UCL = 800 + (2.1 × 50)
UCL = 800 + 105 = 905 particles/m³
✅ Since UCL (905) < 1000, the cleanroom meets
the ISO Class 3 requirement.

44. Example UCL Calculation
Given:
N = 5 locations
M = 800 particles/m³
S = 50 particles/m³
F = 2.1 (from table)
✅ Calculate UCL:
UCL = 800 + (2.1 × 50)
UCL = 800 + 105 = 905 particles/m³
✅ Since UCL (905) < 1000, the cleanroom meets
the ISO Class 3 requirement.

46. AIRFLOW DISTRIBUTION AND CONTROL
● Unidirectional
● Non-unidirectional airflow
● Mixed patterns
Clean room
Class Airflow Type
Av.Airflow
Velocity,
fpm
Air changes/hr
1 Unidirectional 70-100 350-650
10 Unidirectional 60-110 300-600
100 Unidirectional 50-90 300-480
1000 Mixed 40-90 150-250
10000 Mixed 25-40 60-120
100000 Mixed 10-30 10-40

47. Type of clean room classes

48. CREDITS: This presentation template was created
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infographics & images by Freepik and
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THANKS

49. References
● www.cedengineering.com.
● www05.abb.com
● A. Bhatia, B.E. PDH Online | PDH Center,5272 Meadow
Estates Drive,Fairfax, VA 22030-6658.
● www.who.int
● www.pharmaguideline.com
● en.wikipedia.org
● www.pharmamanufacturing.com
● Albert, M.J., Singh, K.V., et al., 1990. Molecular
epidemiology of shigella infection in
Central Australia. Epidemiol. Infect. 105, 51–57.
● https://doi.org/10.1016/j.ssci.2024.106458