Download Hazardous activity identification process

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Date
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HSE section
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Replaces
15.12.2003
Hazardous activity identification process
Unit: IBT
Molgen
Date:
27.08.2013
Updated 02.02.2016 SHA
Participants in the identification process (including their function): You (M.student/PhD/PostDoc/researcher), Supervisor (Professor)
Student/employee/guest
Name (Capital letters)
Supervisor/leader
Signature
Name (Capital letters)
Signature
Short description of the main activity/main process: Molecular genetics and microbiology
ID
nr
Activity/process
Responsible
person
Laws,
regulations
etc.
Existing documentation
All lab work
1
Acrylamide gel, making of
Procedure must be written before acrylamide is used
2
Agarose gel, making,
running, Geldoc
Manual
3
Alginate isolation,
purification and nonradioactive alginate assay
Protocols
4
Antibiotic-stock solution,
make and use of
Lab-rules, relevant MSDSs
5
Work with liquid N2
Regulation for filling and transport. NTNU HSE
guidelines for work with chemicals and gasses
Existing safety measures
Comment
Nitrile gloves are to be used for all labwork, vinyl gloves can be used in order
to protect the sample (but does not offer
the same protection to the user).
Nitrile gloves, protective eyewear,
ventilation hood
Nitrile gloves, protective eyewear (with
UV filter), face shield when needed.
Gelred used for staining or alternatively
Gelgreen. Blue and white screens.
Nitrile gloves, protective eyewear when
using NaOH, Fume hood when using
ether
Procedure not
currently performed
Blue- and white
screens stops UVirradiation when
mounted properly
Risk evaluation
according to which
chemicals are used
Nitrile gloves, handle powder only
inside fume hood
Glasses, thick glowes. Easily removed
footwear. Avoid shoes with wide
opening to avoid spilling
N2 into the shoes. Suitable containers for
transport.
Training necessary.
See following point
regarding transport
of N2
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Hazardous activity identification process
6
Filling and transport of
liquid nitrogen
7
Autoclave
8
Bacteria class 1 and
recombinant bacteria
Manual
Operating instruction
Approval: (GMM regulation)(”Godkjenning av
laboratorier og anlegg for innesluttet bruk av
genmodifiserte mikroorganismer” (08.12.10
Helsedirektoratet)
9
Bacteria class 2
Approval letters from arbeidstilsynet (11.06.10 and
27.04.07).
10
Recombinant bacteria class
2
Approval ”Godkjenning av laboratorier og anlegg for
innesluttet bruk av genmodifiserte mikroorganismer”
(08.12.10 Helsedirektoratet)
Glowes, face protection, tank and carrier
for transport, easily removed suitable
shoes (not rubber boots). O2-meter by
the filler. Do not take the elevator with
the N2 (l)-tank.
Thermo-resistant gloves
Eye protection lab,
Instructions posted regarding not
opening autoclave before temperature
<80 °C, not overfilling bottles or closing
bottle lids completely (to avoid broken
bottles).
Autoclave accessible. Inactivation of
gene manipulated bacteria in
contaminated material and waste. Lab
coat mandatory. Lab bench surfaces
resistant to water, acid, alkali, solvents,
disinfective agents, decontaminating
agents and easy to clean. Transport
between labs only in closed containers.
Good microbiological practice as
defined in
http://www.lovdata.no/for/sf/ho/to20011221-1600-018.html
According to risk assessment: Risk
assessments: “Novel anti-infective
agents from marine bacteria (19.04.07)”
and “Sintef project no: 80117000”
(14.011.08) and “Risikovurderinger og
rutiner, arbeid med mikroorganismer i
smitterisikogruppe 2” (28.05.09).
Autoclave in building. Inactivation of
gene manipulated bacteria in
contaminated material and waste. Lab
coat and gloves mandatory. Lab bench
surfaces resistant to water, acid, alkali,
solvents, disinfective agents,
decontaminating agents and easy to
clean. Transport between labs only in
closed containers. Restricted access to
lab. Good microbiological practice
Room D4-106
(SINTEF). Separate
rules. Contact
SINTEF for further
arrangement
Room D4-106
(SINTEF). Separate
rules. Contact
SINTEF for further
arrangement
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Hazardous activity identification process
(http://www.lovdata.no/for/sf/ho/to20011221-1600-018.html). Biological
hazard sign on door. Measures to
minimize aerosols. Control of vectors
(closed windows, no food I lab, general
awareness of possible vectors),
11
12
Bioanalyzer
Use of open flames – (e.g.
sterilization with Bunsen
burners)
13
Cellcracker (Mini bead
beater)
General lab work
14
Refer to the appropriate section in lab-rules
Bunsen burner must not be left burning
Manual
Laboratory handbook NTNU, MSDSs, EcoOnline
system, signature for training/introduction to lab rules
Refer to respective kit protocols
No risk
Disposable
inoculation loops
will be tested as an
alternative to glass.
No risk
Safety rules according to risk
assessment
Use gloves and eye protection during
steps including NaOH. Use fume hoods
for procedures containing chloroform,
phenol or if it is indicated in the kit
manual
Use nitrile gloves or eye protection if
the kit contains irritating fluids or
proteases
15
DNA/RNA isolation and
purification
16
DNA/RNA kit for
enzymatic modifications or
clean up from reactions
Manuals
17
Electroporator, use of
Manual
18
Environmental samples for
cultivation and DNA
isolation, preparation of
Risk assessment (projects)
19
Enzyme-assays (i.e. betalactamase-assay,
phosphoglucomutase-assay,
epimerase assay...)
Relevant protocols
According to MSDSs of relevant
chemicals
20
Ethanol/dry ice bath
Lab-rules
21
FPLC, running of
Refer to instrument instructions. Precaution at
instrument.
Use thermo-resistant gloves and eye
protection, loose-fitting (e.g. rubber)
boots should not be used.
According to MSDSs of relevant
chemicals
Avoiding contact with high voltage by
following user instructions from manual.
According to risk assessment
No risk when used as
stated in manual.
Risk evaluation by
specific project
Potential damage to
eyes and frostbite
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Hazardous activity identification process
22
Luciferase measurements,
Promega Luciferase Assay
kit
Manuals
According to MSDSs of relevant
chemicals
No risk
23
Luminometer and
fluorometer, use of
Manual
According to MSDSs of relevant
chemicals
No risk
24
Mammalian cells, class 2
Risk evaluation and procedures must be written for
each experiment/new organism.
25
Preparation of medias for
growing bacteria
Recipe
According to MSDSs of relevant
chemicals
26
Mutagenesis using a
mutagenic agent (e.g.
Etylmethanesulfonate,
methylethanesulfonate,
nitrosoguanidine, UV)
Experimental procedure
Risk assessment and approval from project leader
Nitrile gloves, fume hoods, requires
written procedure and risk assessment
for the experimental procedure (typespecific) including detoxifying waste
and how to warn and protect others
working in the laboratory. Description
and risk assessments must be written
and approved by the responsible project
leader (signature). Approval is
mandatory not only for each
experimental procedure, but also for
each new person to perform that
procedure
27
Nano-drop measurements of
DNA/RNA
Manual
28
Northern and Southern
blotting
Manual
29
PCR
PCR-manual
Room D4-106
(SINTEF). Separate
rules for HTS/cell
lab. Contact SINTEF
for further
arrangement.
Hepatitis B vaccine
offered.
No risk
No risk
According to MSDSs of relevant
chemicals. Gloves and lab coat, work in
a fume hood if MOPS and/or
formaldehyde is used. Usual precautions
when working with UV light and DNA
staining.
Fume hood when DMSO is added
No risk
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Hazardous activity identification process
30
pH-adjustments
Manual
31
Protein isolation and
purification
32
Proteingel-gels. Running of
CBS – manual
http://cbsscientific.com/pdf/DCXInstructionsNov201
0.pdf
Sambrook and Russell, Molecular Cloning: A
laboratory manual. section A.8.42
33
Pulse field electrophoresis
Manual
Operating instruction
34
Radioactivity work
including use of scintillation
counter
http://www.nt.ntnu.no/ibt/innsidadokumentlager
/HMS/Veileder%20for%20arbeid%20med
%20radioaktivt%20materiale%20IBT.pdf
Risk evaluation and procedures must be written for
each experimental procedure
Operating instruction scintillation counter.
35
Centrifugation (Sorvall +
table)
Manual
Operating instruction
36
Sonication
Manual
Operating instruction
Nitrile gloves, eyewear, shoes/shoe bags
with protection against acid/base spill,
sufficient ventilation (hood/”cap”)
According to MSDSs of relevant
chemicals
Nitrile gloves (e.g. protection against
acrylamide), eyewear, staining and
de-staining in fume hoods
Protection against voltage (lid etc.) to
avoid electrical shock. Avoid contact
with de-staining reagents
(methanol/acidic acid).
According to relevant MSDSs.
Protection against voltage (lid, gloves
etc.) to avoid electrical shock.
Gloves, eye protection, lab coat, hood.
Testing of work area for contamination
after work.
Accurate balancing, accurate attachment
of rotor, not exceed maximal G-forces
for each type of tube
Ear protection. Pregnant women are not
allowed to stay in the room during
sonication. Sonication cabinet with
sound absorbing material. Shut doors of
the room where sonication is taking
place, and post warning sign on door.
No risk (confer
bacteria, sonication
and chemicals used
when applicable)
Department rules
state that the local
radiation- protection
coordinator must preapprove any users of
radioactive
compounds and
experimental
procedures must also
be approved. The
scintillation liquid
used may contain
dangerous solvents
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Hazardous activity identification process
37
Supercompetent or
electrocompetent cells,
making of
Eye protection, gloves and protective
shoes/shoe bags when handling liquid
nitrogen or ethanol/dry ice bath. Shoes
must be easy to take off in case of spill
into (never use rubber boots)
38
Transformation
39
Ultrasound waterbath, use of
40
UV/Vis Spectrophotometer
41
Ventilation hoods, use of
Operating instruction
42
Washing machine, use of
Manual
43
Waterbaths and shaking
waterbaths, (heat and water
quality)
Manual
44
Western blotting
No risk
Operating instruction
Ear protection. Pregnant women are not
allowed to stay in the room during
sonication, close the door and post
warning sign.
Avoid skin/eye exposure to UV light.
Never disassemble the instrument.
Follow instructions/manual when
changing light bulb.
Opening minimized when not in use,
keep the hood-opening small when in
use (for efficient protection).
No risk
No risk
Timer on water baths. Large water baths
(> 1 L) are not to be used to boil small
samples. Put up notices if heating the
water baths at temperatures > 70°C.
Chemicals. Avoid contact with buffers
(e.g. methanol).
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7 out of 6
Risk assessment
Unit:
IBT -molgen
Line manager: Kjetil Rasmussen
Date
Date: 27.08.2013 Updated 02.02.106 SHA
Participants in the identification process (including their function): You (M.student/PhD/PostDoc/researcher), Supervisor
(Professor)
ID
nr
Activity from the
identification process form
2
Agarose gel electrophoresis
Potential undesirable incident/strain Likelihood Consequence:
Risk
:
value
Likelihood Human Environment Economy/ Reputa
(1-5)
(A-E) (A-E)
material tion
(A-E)
(A-E)
Unknown A
Unknown
Long term health risk from GelRed
1
A
B
(unknown effect)
UV-light: skin or eye damage
2
2
B
A
A
B
2B
5
Work with liquid N2
Frostbite
2
C
A
A
B
C2
5
Work with liquid N2
Suffocation
1
E
A
A
E
E1
6
6
Filling/transport of liq N2
Filling/transport of liq N2
Frostbite due to spurt of liquid N2
Low oxygen atmosphere
2
1
B
B
A
A
A
A
B
D
2B
1D
Comments/
status
Suggested
measures
Gelred and
Gelgreen is said not
to penetrate cell
membranes, and
thus should not act
as mutagen even if
it is DNA-binding.
Gloves also
minimize the risk
for exposure.
UV damages on
unprotected
skins/eyes if
instructions not
followed
Follow regulations
for filling/transport.
Oxygen
measurement?
Follow regulations
for filling/transport.
Oxygen
measurement?
O2-meter by
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Risk assessment
tank/filler. Do not
ride the elevator
together with the
N2 (l)-tank!
Will not happen if
instructions are
followed
Rapid pressure fall due to opening the
2
autoclave to soon my cause hot liquid burns
on eye or skin
Bacteria class 1 and recombinant Release of GMO to environment
2
bacteria
Bacterial infections
C
A
A
B
2C
A
A
A
B
2A
9
Bacteria class 2
Bacterial infections. Release of infective,
pathogen bacteria to environment.
2
C
B
A
C
2C
10
Recombinant bacteria class 2
Bacterial infections. Release of infective,
pathogen gene manipulated bacteria to
environment.
2
C
B
A
C
2C
12
Bunsen burners
Skin burns
3
B
A
A
A
3B
12
Bunsen burners
Initiation of fire.
3
C
A
B
A
3C
14
General lab work
Deviation from established safety rules and -routines
--
--
--
--
--
15
DNA/RNA isolation and
purification
Exposure for dangerous for health
chemicals: Phenol-chloroform mix
C
A
A
B
7
8
Autoclave
Date
2
2C
Low risk because of
strict regulations
and good lab
practise.
Low risk because of
strict regulations
and good lab
practice.
New rules how to
handle burners+
ethanol baths
already installed.
New rules how to
handle burners +
ethanol baths
already installed.
No injuries
requiring more than
simple first aid in
these laboratories
for the past 5 years,
the present routines
seem sufficient
Phenol-chloroform
mix requires the
work in a
ventilation hood
only. All waste
should be placed in
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Risk assessment
unknown
18
Environmental samples for
cultivation and DNA isolation,
preparation of
Bacterial infections, production of
1
toxic/infectious compounds from cloned
DNA. Usage of hazardous compounds (e.g.
chloroform) for DNA isolation.
20
Dry ice/ethanol
Eye injury. Frostbite
1
B
22
FPLC, running of
Leakage of buffer
23
Luminometer and fluorometer,
use of
24
Mammalian cells, class 2
26
28
Date
a special box for
hazardous
materials.
In general mild
1unknown injuries, that can be
avoided by present
routines and
sensible carefulness
during lab-work.
1B Eye-protection and
Gloves for handling
the Dry ice
A
A
C
A
A
B
2
See
A
MSDS
A
Depends
on injury
1A
Possible exposure health injurious
chemicals.
2
A
A
A
A
2A
See MSDS for
relevant chemicals
Exposure to biological agents, infections.
2
D
A
A
D
2D
Mutagenesis using a mutagenic
Exposure to mutagens
agent (e.g. Etylmethanesulfonate,
methylethanesulfonate,
nitrosoguanidine, UV)
1
E
A
A
E
1E
Applying aseptic
procedures.
Good
microbiological
practices.
Probably too small
amounts to ever
damage the
environment.
Because of the
known risk of
cancer, these
experiments are
always conducted
with extreme
caution, and using
gloves and fume
hoods.
Northern/Southern
1
C
A
A
A
1C
Exposure to injurious chemicals. Northern
blot may involve formaldehyde and
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Risk assessment
29
PCR
30
pH-adjustments
32
Protein gel-eletrophoresis
33
Pulse field electrophoresis
acrylamide which are toxic, thus fume hood
and protective clothing must be used. Due
to the use of toxic chemicals special
disposal methods are required.
Exposure to DMSO when used
1
Date
A
A
A
A
1A
Exposure to corrosive chemicals: Skin and 2
eye damage
Electrical shock
1
B
A
A
B
2B
E
A
A
C
1E
Electrical shock and depending on method
used for DNA preparation :)
1
E
A
A
A
1E
33
Exposure to hazardous chemical:
phenylmethanesulphonyl fluoride (toxic,
corrosive)
1
C
A
A
B
1C
34
Radioactivity work including use Health injury as result of radiation
of scintillation counter
(external, internal as result of uptake).
Radioactive contamination of working
environment.
Centrifugation (Sorvall + table)
Damage caused by loose rotor
Sonication
Damage to hearing (including foetus)
Creation of injurious aerosols (infectious
microorganisms, toxic agents)
Supercompetent or
Exposure of skin and eyes to extremely
electrocompetent cells, making of cold liquid (nitrogen or ethanol)
1
E
A
A
E
1E
1
1
C
C
A
A
B
A
C
B
1C
1C
2
C
A
A
B
2C
35
36
36
37
PCR-machine
should be in
ventilation hood
when DMSO is
added, use lab coat
and gloves
Eye-protection and
lab coat
Equipment for
electrophoresis will
only work when lid
is closed, thus
minimizing the
possibility of
suffering an electric
shock.
Use of gloves/fume
hoods for handling
of hazardous
chemicals will
minimize risk.
Conducted with
extreme caution,
and using eyeprotection and
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Risk assessment
Date
gloves.
39
Ultrasound waterbath, use of
Damage to hearing (including foetus)
1
C
A
A
B
1C
41
Ventilation hoods, use of
1
C
A
A
B
1C
43
Waterbaths and shaking
waterbaths, (heat and water
quality)
Exposure to hazardous chemicals due to
insufficient airflow (effects on local hood
or other hoods)
Electrical hazards, Burns
1
B
B
A
A
1B
44
Western blotting
Exposure to hazardous chemicals
1
See
A
MSDS
A
Depends
on injury
1A
Likelihood, e.g.:
1. Minimal
2. Low
3. Medium
4. High
5. Very high
Consequence, e.g.:
A. Safe
B. Relatively safe
C. Dangerous
D. Critical
E. Very critical
Risk value (each one to be estimated separately):
Human = Likelihood x Human Consequence
Environmental = Likelihood x Environmental consequence
Financial/material = Likelihood x Consequence for Economy/materiel
Can be prevented
by maintaining
sufficient airflow
Regular service by
trained personnel.
Heat-proof gloves
must be used during
handling of hot
items.
Hot surfaces must
be labelled with
warning signs.
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Risk assessment
Date
Potential undesirable incident/strain
Identify possible incidents and conditions that may lead to situations that pose a hazard to people, the environment and any materiel/equipment involved.
Criteria for the assessment of likelihood and consequence in relation to fieldwork
Each activity is assessed according to a worst-case scenario. Likelihood and consequence are to be assessed separately for each potential undesirable
incident. Before starting on the quantification, the participants should agree what they understand by the assessment criteria:
Likelihood
Minimal
1
Once every 50 years or less
Low
2
Once every 10 years or less
Medium
3
Once a year or less
High
4
Once a month or less
Very high
5
Once a week
Consequence
Grading
Human
E
Very critical
D
Critical
May produce fatality/ies
C
Dangerous
B
Relatively safe
Serious personal injury
A
Safe
Permanent injury, may produce
serious serious health
damage/sickness
Injury that requires medical
treatment
Injury that requires first aid
Environment
Financial/material
Very prolonged, non-reversible
damage
Prolonged damage. Long
recovery time.
Shutdown of work >1 year.
Minor damage. Long recovery
time
Minor damage. Short recovery
time
Shutdown of work < 1 month
Insignificant damage. Short
recovery time
Shutdown of work < 1day
Shutdown of work 0.5-1 year.
Shutdown of work < 1week
The unit makes its own decision as to whether opting to fill in or not consequences for economy/materiel, for example if the unit is going to use particularly
valuable equipment. It is up to the individual unit to choose the assessment criteria for this column.
Risk = Likelihood x Consequence
Please calculate the risk value for “Human”, “Environment” and, if chosen, “Economy/materiel”, separately.
About the column ”Comments/status, suggested preventative and corrective measures”:
Measures can impact on both likelihood and consequences. Prioritise measures that can prevent the incident from occurring; in other words, likelihoodreducing measures are to be prioritised above greater emergency preparedness, i.e. consequence-reducing measures.
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side
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Erstatter
09.02.2010
Risk assessment matrix
CONSEQUENCE
Risk assessment matrix NTNU
Very
serious
E1
E2
E3
E4
E5
Serious
D1
D2
D3
D4
D5
Moderate
C1
C2
C3
C4
C5
Small
B1
B2
B3
B4
B5
Very
small
A1
A2
A3
A4
A5
Very small
Small
Moderate
High
Very high
PROBABILITY
Color-coding in the risk assessment matrix
Color
Description
Red
Yellow
Green
Unacceptable risk. Measures must be implemented to reduce risk.
Evaluation area. Evaluate whether measures need to be implemented.
Acceptable risk. Measures may be considered based on other factors.