Clean Air Overview

What is clean air?

A space where particle concentration in the air
is controlled to the levels appropriate for working
on contamination-sensitive activities

Where is clean air products used?

Micro-electronics / Semiconductors
Optical manufacturing
Nuclear & Aerospace
Pharmaceuticals
Life sciences & laboratory
Food processing
Security (Anthrax mail)
Clean Air Devices

• Cleanrooms - Fan Filter Units

• Laminar Flow cabinets

• Class I, Class II and Class III Biohazard

Safety cabinets
Clean Air Standards

ISO 14644.1
Federal Standard 209E (obsolete)
British Standard BS 5295
German Standard VDI 2083
French Standard AFNOR X44101
Australian Standard AS 1386
Japanese Standard JIS B9920
Korean Standard KS27030.1
Particle Definitions

Definition of particle by ISO 14644:1999

solid or liquid object between 0.1 to 5 micron

Particle size measurement:

Discrete Particle Counter

Particle concentration:
number of individual particles per unit volume of air

Particle size distribution:

cumulative distribution of particle concentration with
respect to particle size
Particle Size Comparison
HEPA & ULPA Filter
HEPA: High Efficiency Particulate Air
ULPA: Ultra Low Penetration Air

Important definitions:
- Modern “American-convention” HEPA: 99.99%
at 0.3 microns (at Most Penetrating Particle Size)
- Modern “American-convention” ULPA: 99.999%
at 0.12 microns (at Most Penetrating Particle Size)

Note: The “classical” definition of HEPA filter is

99.97% at 0.3 microns, but nowadays all
BSC and LF in US use 99.99% at 0.3 µm
HEPA & ULPA Filter at Most
Penetrating Particle Size
HEPA/ULPA Capability
Removes a broad range of airborne
contaminants:
• Fine dust
• Smoke
• Bacteria (typical size: 500 to 0.3 micron)
• Soot
• Pollen
• Radioactive particles
• Impurity ion -> can affect Integrated Circuit
speed
Filter Construction
HEPA / ULPA filter
construction:
- Media pack of pleated
borosillicate glass fibers
- Pack is glued into a frame
- Frame is gasketed to form
final assembly
Separator VS Separatorless
Filters
Separator VS Separatorless
Filters
Modern minipleat separatorless:
• Compact size -> Contains more pleat per cm
• Increase usable area -> more dust holding -> more life
• Lower airflow resistance -> lower energy consumption
• Galvanized Steel frame->lighter,no swelling in moist
conditions
• Eliminates media damage by alum. Separator
• Handle some harsh environment that may attack alum.
separator

Conventional aluminium separator construction:

• Larger size -> Less pleat per cm -> shorter life
• Higher airflow resistance -> higher blower power
• Wood cardboard frame -> heavier, moist problem
• Possible media damage -> dangerous !
Filtration Principle

Larger particles
have too much
inertia and
hence
deviate from the
air stream
(which
normally curves
around the filter
fiber) thus
impacts on the
filter fiber
Filtration Principle

Medium sized
particles follow
the air stream
however have a
diameter larger
than the distance
between the air
stream and filter
fiber and are
hence intercepted
Filtration Principle

Sub-micron sized
particles vibrate
with Brownian
motion due to
bombardment by
air molecules.
When a particle
contacts the
fiber,
it is retained.
More significant effect for smaller particles
Laminar Flow
Cabinets
Definition of laminar / unidirectional flow:
Air flow straight, parallel pass in enclosed space

Principle of laminar flow cabinet:

Sterilize air though filter and blow it across work
surface as a particle-free laminar air stream

Typical laminar air flow velocity:

0.3 - 0.5m/s

Purpose of a laminar flow cabinet:

Product protection only (does not protect
operator)
Purpose of LF Cabinets
Sample applications of LF Cabinets:
- Pharmaceutical: sterile production of drugs
- Semiconductor: protect sensitive chip from
dust

Creating economical super-clean mini

environment:
Place a Class 10 or Class 100 LF cabinet in a
Class 1000 clean room to obtain a Class 1
mini-environment

Effectiveness of LF cabinet:
Room: 35 million/ft3
Inside LF cabinet: 35/ft3
Horizontal vs Vertical LF

• Risk of spillage on filter • No risk of spillage

• Large object -> blocking • Safer from blocking
• Blow air on face • No blow air on face
• Easier to put sensitive • Difficult to protect
Large Object:
Blocking

Major problem with Horizontal Laminar Flow:

Blocking: room air being induced due to
large object in airstream
The Need for
Biosafety
The need for biosafety
- High risk of laboratory-acquired infections
- First documented in the US in the 1940s

Purpose for BSC

- Operator protection by inflow (not in LF)
- Product protection (like LF ; only
BSC Class 2 and 3, but not Class 1)
Containment & Safety
Containment principles
- Primary containment (safety equipment)
- Secondary containment (facility design)

Standard microbiological practices

Principle of selecting which class of BSC

- Only operator or operator & product
protection
- Degree of operator protection (Biosafety
level)
Primary Containment
• All manipulations inside Biohazard Safety
Cabinets

• Clothing:wrap-around gowns, scrub suit, or

coverall

• Gloves: infectious materials / contaminated

equipment

• Frequent changing of gloves and hand wash

• If manipulations can not be done inside BSC:

- Personal protective equipment (respirator, face,
shield)
- Physical containment devices (centrifuge cup)
Secondary Containment (1)
Example: Biosafety Level 3 Lab:

• Access to lab is separated from general traffic flow

• Access is restricted - two self closing lockable door

• All windows are closed and sealed

• Interior surfaces (walls, floor, ceiling):
- Easy for cleaning and decontamination
- Smooth, impermeable to liquids & chemicals in lab
- Seams and openings must be sealed
- Floor: monolithic and slip-resistant
Secondary Containment (2)

• Bench top: resist moderate heat & chemicals

• Chairs: non-fabric material-> easy decontamination

• Waste decon: autoclave, chemical treat, incineration

• Ducted exhaust air ventilation - HEPA filtered
• Exhaust: away from occupied area and air intakes
• Directional room airflow:clean to contaminated area
• Audible alarm for HVAC failure

• Hands-free sink for hand washing - near exit door

• Eyewash station
• Personnel shower
Standard Microbiological Practice

• Access to lab limited by lab director during experimt

• Wash hand:
- Handling hazardous materials
- Change glove, exit from lab
• No eating, drinking, smoking, handling contact lens
• Contact lens --> wear goggles / face shield
• No mouth pipetting --> use mechanical pipet
• Safe handling of sharp objects
• Minimize aerosol generation
• Work surfaces decon per day or after spill
• Waste decon inside lab | outside with leakproof cont
• Insect & rodent control program
Biosafety Level 3 Germs (Air-borne)
Virus Cases (lab death)
Venezuelan equine encephalomyelitis 150 (1 death)
Rift Valley fever 47 (1 death)
Chikungunya 39
Yellow fever 38 (8 deaths)
Japanese encephalitis 22
Louping ill 22
West Nile 18
Lymphocytic choriomeningitis 15
Orungo 13
Piry 13
Wesselsbron 13
Mucambo 10
Oropouche 7
Germiston 6
Bhanja 6
Hantaan 6
Biosafety Level 4 Germs (air-borne)

According to US CDC in Atlanta:

Virus Cases (lab death)
Junin 21 (1 death)
Marburg 25 (5 deaths)
Russian Spring-Summer 8
Congo-Crimean hemorrhagic 8 (1 death)
Omsk hemorrhagic fever 5
Lassa 2 (1 death)
Machupo 1 (1 death)
Ebola 1
Sabia 3 (1 death)
Summary of Biosafety Levels

Lethality Medium Cure Ex

1 Safe Liquid Yes B.Subtilis
2 Some Liquid Some HIV
3 Serious Airborne Some TBC
4 Extreme Airborne None Ebola
Class I Biological Safety Cabinet
 Only operator protection
(no product protection).

 Used for Biosafety level I,

II, III Germs

 Inflow away from

operator.

 HEPA filtered exhaust

to environment.

 Current trend: Switch to

Class II
Class 2 Biological Safety Cabinet

• Both operator and product protection

• Biosafety level 1, 2, 3
• Inflow away from operator
• HEPA filtered exhaust to environment
• HEPA filtered laminar downflow
• Volatile Toxic Chemical in cell culture:
biological & chemical -> need ducting
Airstream® Class II Airflow
Class 3 Biological Safety
Cabinet

• Used for Biosafety level 1, 2, 3, 4 germs

• Offers both product and operator protection
• Gas leak tight 1x10-5 cc/sec leak rate
• Internal operations -> attached glove
• Material transfer -> 2 doors pass box
• Negative air pressure > 0.5 “WC (120 Pa)
• Supply is HEPA filtered
• Double exhaust HEPA filter in series or:
• Single exhaust HEPA and an incinerator
Class III: Double Exhaust
HEPA
Using the Biological Safety Cabinet
(1)
- Do not obstruct the front or back air grilles
- Work as far in to the cabinet as possible

- Minimize arm movement; make slow

movements to avoid disrupting cabinet
airflow

- When removing arms from cabinet be sure

to surface decontaminate first, and move
arms out slowly of the cabinet (in direction
perpendicular to plane of work zone
opening)
Using the Biological Safety Cabinet
(2)
- Work from “clean to dirty”
- Biohazard collection bags should be placed
inside the cabinet instead of outside
Using the Biological Safety Cabinet
(3)

- Use absorbent pads on the work surface

where appropriate to minimize splatter
and
aerosol generation in case of a spillage

- Surface decontaminate before removing

potentially contaminated items from the
interior

- Put air turbulence generating equipment such

as centrifuge, blender, or sonicator in the
back 1/3 of the cabinet
Using the Biological Safety Cabinet
(4)

- Clean materials should be at least 150mm

away from aerosol generating objects to
minimize the chance for cross contamination

- Hold lids / covers above dishes / sample

plates in order to prevent impingement of
downward air

- Do not use a gas flame whenever possible as

it interferes with airflow
Shutting down the Biological Safety
Cabinet
Note: operate the cabinet continuously
whenever possible to maximize protection

- Surface decontaminate all items

- Seal biohazard bags if used
- Surface decontaminate the cabinet inner
side walls, back wall, work surface, drain
pan and the inner side of the sliding sash /
hinged window with 70% IPA or other
agents
- Allow work zone air to purge
- Close sash and when available activate UV
lamp for 1 hour (using timer)