See Instructions in EMV´s HMS-page
Risk Assessment of
SDS-PAGE according to the standard Laemmli gel method
Department
Group
Date
EMV
Muscle Biology
2013-04-22
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Tasks included in the risk assessment
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Prepare buffers; separating gel buffer, stacking gel buffer, electrophoresis running
buffer and 5 x loading sample buffer
Prepare and cast separating gel (2 full size 10% gels)
Prepare and cast stacking gel
Prepare sample and load gel
Run gel (electrophoresis)
Sample preparation is not included in this risk assessment.
For adjustment of pH see separate risk assessment: pH-justering i vattenlösning.
If ready made gels are used, skip “Prepare and cast separating gel” and “Prepare and cast
stacking gel”.
Included chemicals, mixtures and solutions
Chemical, mixture or solution
CAS Number
Separating gel buffer: Tris-Cl/SDS pH 8.8
(1.5 M Tris-Cl containing 0.4% SDS)
Amount
Indication of danger (hazard statement and limit value)
500 ml
Tris(hydroxymetyl)aminometan (s)
77-86-1
90 g
H315 Causes skin irritation.
H319 Causes serious eye irritation.
H335 May cause respiratory irritation.
SDS (s)
151-21-3
2g
H228 Flammable solid.
H302 Harmful if swallowed.
H311 Toxic in contact with skin.
H315 Causes skin irritation.
H319 Causes serious eye irritation.
H335 May cause respiratory irritation.
Stacking gel buffer: Tris-Cl/SDS pH 6.8 (0.5 M Tris-Cl
containing 0.4% SDS)
500 ml
Chemical, mixture or solution
CAS Number
Amount
Indication of danger (hazard statement and limit value)
Tris(hydroxymetyl)aminometan (s)
77-86-1
30 g
H315 Causes skin irritation.
H319 Causes serious eye irritation.
H335 May cause respiratory irritation.
SDS (s)
151-21-3
2g
H228 Flammable solid.
H302 Harmful if swallowed.
H311 Toxic in contact with skin.
H315 Causes skin irritation.
H319 Causes serious eye irritation.
H335 May cause respiratory irritation.
Electrophoresis running buffer:
25 mM Tris, 192 mM glycine, 0.1% SDS
4 litre
Tris(hydroxymetyl)aminometan (s)
77-86-1
60.5 g
H315 Causes skin irritation.
H319 Causes serious eye irritation.
H335 May cause respiratory irritation.
SDS (s)
151-21-3
20 g
H228 Flammable solid.
H302 Harmful if swallowed.
H311 Toxic in contact with skin.
H315 Causes skin irritation.
H319 Causes serious eye irritation.
H335 May cause respiratory irritation.
Glycine
56-40-6
288 g
H319 Causes serious eye irritation.
Loading sample buffer: 2 M Tris-HCl,15% SDS,
β-Mercaptoethanol and Bromphenol blue
Tris(hydroxymetyl)aminometan (s)
or
2 M Tris-HCl pH 6.8
77-86-1
SDS (s)
or
151-21-3
10 ml
2.4 g
1.63 ml
3g
H315 Causes skin irritation.
H319 Causes serious eye irritation.
H335 May cause respiratory irritation.
H228 Flammable solid.
H302 Harmful if swallowed.
Chemical, mixture or solution
CAS Number
30%
Amount
Indication of danger (hazard statement and limit value)
5 ml
H311 Toxic in contact with skin.
H315 Causes skin irritation.
H319 Causes serious eye irritation.
H335 May cause respiratory irritation.
β-Mercaptoethanol
60-24-2
0.5 ml
H301 Toxic if swallowed.
H310 Fatal in contact with skin.
H315 Causes skin irritation.
H317 May cause an allergic skin reaction.
H318 Causes serious eye damage.
H330 Fatal if inhaled.
H400 Very toxic to aquatic life.
H410 Very toxic to aquatic life with long lasting effects.
Bromphenol blue
62625-28-9
0.5 mg
R36/37/38 Irritating to eyes, respiratory system and skin.
Sucrose, 2M
57-50-1
2.88 ml
Not classified
20 ml
H301 Toxic if swallowed.
H315 Causes skin irritation.
H317 May cause an allergic skin reaction.
H319 Causes serious eye irritation.
H330 Fatal if inhaled.
H334 May cause allergy or asthma symptoms or breathing difficulties if inhaled.
H340 May cause genetic defects .
H350 May cause cancer .
H361F Suspected of damaging fertility.
H372 Causes damage to organs through prolonged or repeated exposure
Acrylamide solution Ready made
30% acrylamide/ 0.8% bisacrylamide
Nivågränsvärde
Takvärde
Korttidsvärde
Totaldamm Resp.damm
[ppm] [mg/m3] [ppm] [mg/m3] [ppm] [mg/m3] [mg/m3]
0,03
[mg/m3]
0,1
Medical examination required, according to AFS 2011:18, read here
CMR-substances: Substances which, if possible, should be phased out, or otherwise require
special investigation (§ 38-44 AFS 2011:19). Those who are exposed to these substances (if it
may involve risk of illness) should be registered. (the list includes substances with any of the
phrases R45, R46, R49, R60, R61, H340, H350, H360)
Ammonium persulfate, 10 %
300 µl
Chemical, mixture or solution
CAS Number
Amount
Indication of danger (hazard statement and limit value)
Ammonium persulfate, (s)
7727-54-0
10 mg to 10 ml
H272 May intensify fire; oxidiser.
H302 Harmful if swallowed.
H315 Causes skin irritation.
H317 May cause an allergic skin reaction.
H319 Causes serious eye irritation.
H334 May cause allergy or asthma symptoms or breathing difficulties if inhaled.
H335 May cause respiratory irritation.
Miljöskyddsåtgärder: Förhindra utsläpp i avloppssystemet.
TEMED (N,N,N’,N’Tetrametyletylendiamin) (l)
110-18-9
20 µl
H225 Highly flammable liquid and vapour.
H302 Harmful if swallowed.
H314 Causes severe skin burns and eye damage.
H332 Harmful if inhaled..
Isobutyl alcohol, 100%
78-83-1
5 ml
H226 Flammable liquid and vapour.
H315 Causes skin irritation.
H318 Causes serious eye damage.
H335 May cause respiratory irritation.
H336 May cause drowsiness or dizziness.
NB! Skin penetrating substance according to The Swedish Work Environment Authority's
provisions and on occupational exposure limit values, AFS 2011:18
Level limit value
Ceiling limit
value
Short-term value
Total
dust
Respirable
dust
[ppm] [mg/m3] [ppm] [mg/m3] [ppm] [mg/m3] [mg/m3] [mg/m3]
50
150
75
250
Identification of risks of accident or ill-health, assessment of risks and suggestion of preventive measures
Task
Identified risk/problem
Assessment of risk, see risk rating
Preventive measures
Estimated
risk after
preventive
measure
Consequence
Probability
Potential for exposure via
inhalation and direct contact
with eyes and skin.
Potential fire hazard.
Serious
Possible
Very high
risk
Use lab coat, safety glasses and double, disposable nitrile
gloves. Change gloves frequently, especially if you suspect
contamination. Remove gloves carefully without touching
skin. Weigh and dissolve in fume hood.
Write in handling and safety instruction.
All staff participates in mandatory fire training every 5 years.
Labs are equipped with fire extinguishers.
Low risk
Prepare and cast separating
gel including preparing H2Osaturated isobutyl alcohol
Potential for exposure via
inhalation and direct contact
with eyes and skin.
Potential for contamination
of bench and other areas.
Potential fire hazard.
Serious
Possible
Very high
risk
Use lab coat, safety glasses and double, disposable nitrile
gloves. Change gloves frequently, especially if you suspect
contamination. Remove gloves carefully without touching
skin. Work in fume hood. Coat work bench in fume hood with
absorbent bench paper. Change this paper layer frequently.
Write in handling and safety instruction.
The limit values for acrylamide and isobutyl alcohol are not
exceeded at any occasion.
All staff participates in mandatory fire training every 5 years.
Labs are equipped with fire extinguishers.
Medium risk
Prepare and cast stacking gel
Potential for exposure via
inhalation and direct contact
with eyes and skin.
Potential for contamination
of bench and other areas
Potential fire hazard
Serious
Possible
Very high
risk
Use lab coat, safety glasses and double, disposable nitrile
gloves. Change gloves frequently, especially if you suspect
contamination. Remove gloves carefully without touching
skin. Work in fume hood. Coat work bench with absorbent
bench paper. Change this paper layer frequently.
Write in handling and safety instruction.
The limit values for acrylamide and isobutyl alcohol are not
exceeded at any occasion.
All staff participates in mandatory fire training every 5 years.
Labs are equipped with fire extinguishers.
Medium risk
Prepare buffers
For adjustment of pH see
separate risk assessment: pHjustering i vattenlösning.
Assessed
Risk
Task
Identified risk/problem
Assessment of risk, see risk rating
Consequence
Probability
Preventive measures
Estimated
risk after
preventive
measure
Assessed
Risk
Prepare the sample and load
the gel
Potential for exposure via
inhalation and direct contact
with eyes and skin
Potential for needle stick
injury.
Serious
Possible
Very high
risk
Use lab coat, safety glasses and double, disposable nitrile
gloves. Change gloves frequently, especially if you suspect
contamination. Remove gloves carefully without touching
skin. Work in fume hood.
Handle needles with care, do not re-sheathe.
Write in handling and safety instruction.
The limit values for acrylamide and isobutyl alcohol are not
exceeded at any occasion.
Low risk
Run the gel (electrophoresis)
Potential for serious electrical
shock or electrocution due to
leaking chamber, faulty or
corroded electrode cables, or
faulty power supply.
Very serious
Possible
Very high
risk
Ensure all equipment is in good working order, and that lid is
always on electrophoresis tank when in operation. Turn off
power before disassemble.
Write in handling and safety instruction.
Medium risk
Chipped glass plates.
Potential for causing
lacerations or deep cuts to
skin. May also result in
leaking of unpolymerised
acrylamide solution.
Serious
Possible
High risk
Ensure all equipment is in good working order and discard
any chipped or damaged plates.
Low risk
Clean and take care of waste
Potential for exposure via
direct contact with eyes and
skin
Environmental risk.
Serious
Possible
High risk
Same personal protective equipment as during the task
Dispose of as chemical waste.
Low risk
In case of power or
ventilation failure or if
evacuation is necessary
Fumes evaporate to lab
Serious
Possible
High risk
Close containers and close fume cupboard. Write in handling
and safety instruction.
Low risk
Cleaners
If glass ware not rinsed
before sending to be washed,
potential for exposure via
direct contact with eyes and
skin
Serious
Possible
High risk
Before sending glass ware to be washed it must be properly
rinsed. Write in handling and safety instruction.
Cleaning must only be performed in free areas and cleaners
must not wipe spill. Write in instruction for cleaners.
Low risk
Have hazardous tasks been defined and rules for working alone been considered?
Yes
Have risks during power/water failure etc or because of evacuation been considered?
Yes
Do you have preparedness in case of spillage, eg first aid, absorption material?
Yes, a spillage kit will be available in the chemical storage room
Do you have a written handling and safety instruction for this work?
Yes
Do you have a written instruction for cleaners and other maintenance personal?
Task in the Improvement plan for EMV 2013
References:
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Protocol for SDS-PAGE according to the standard Laemmli gel method
SDS for all chemicals
LU´s Handling Regulations for Management of hazardous waste
EMV:s HMS-webpage
Participants: Katarina Johansson and Madeleine Durbeej-Hjalt
Reviewed by: Åsa Gustavson, Hans Hovenberg and Kristin Holmberg
Approved by Research Group Leader
Name in printing
Madeleine Durbeej-Hjalt