Included Chemicals, Mixtures and Solutions

See Instructions in EMV´s HMS-page

Risk Assessment of

Coomassie Blue Staining

Department Group Date

EMV Muscle Biology 2013-05-13

Tasks included in the risk assessment  Rinse gel with fixing solution

 Prepare solutions  Cover gel with methanol/acetic acid destaining solution

 Handle polyacrylamide gels Sample preparation is not included in this risk assessment.

 Precipitate protein from gel with fixing solution For SDS-PAGE see separate risk assessment: SDS-PAGE according to the standard Laemmli gel method.

 Cover gel with Coomassie staining solution

Included chemicals, mixtures and solutions

Chemical, mixture or solution CAS Number Indication of danger (hazard statement and limit value)

Fixing solution for Coomassie blue and 500 ml silver staining, 50% methanol and 10% acetic acid Methanol 67-56-1 H225 Highly flammable liquid and vapour. H301 Toxic if swallowed. H311 Toxic in contact with skin. H331 Toxic if inhaled. H370 Causes damage to organs Respirable Level limit value Ceiling limit value Short-term value Total dust dust [ppm] [mg/m3] [ppm] [mg/m3] [ppm] [mg/m3] [mg/m3] [mg/m3] 200 250 250 350 Chemical, mixture or solution CAS Number Indication of danger (hazard statement and limit value)

Remark H: Skin penetrating substance according to The Swedish Work Environment Authority's provisions and on occupational exposure limit values, AFS 2011:18

Acetic acid, conc 64-19-7 H226 Flammable liquid and vapour. H314 Causes severe skin burns and eye damage Respirable Level limit value Ceiling limit value Short-term value Total dust dust [ppm] [mg/m3] [ppm] [mg/m3] [ppm] [mg/m3] [mg/m3] [mg/m3] 5 13 10 25

Coomassie blue staining solution, 50% 500 ml methanol, 0,05% Coomassie Brilliant Blue R-250 and 10% acetic acid

Coomassie Brilliant Blue R-250 6104-59-2 Not classified

Methanol 67-56-1 H225 Highly flammable liquid and vapour. H301 Toxic if swallowed. H311 Toxic in contact with skin. H331 Toxic if inhaled. H370 Causes damage to organs Respirable Level limit value Ceiling limit value Short-term value Total dust dust [ppm] [mg/m3] [ppm] [mg/m3] [ppm] [mg/m3] [mg/m3] [mg/m3] 200 250 250 350 Remark H: Skin penetrating substance according to The Swedish Work Environment Authority's provisions and on occupational exposure limit values, AFS 2011:18

5 13 10 25

Methanol/acetic acid destaining solution, 500 ml 5% methanol and 7% acetic acid Methanol 67-56-1 H225 Highly flammable liquid and vapour. H301 Toxic if swallowed. H311 Toxic in contact with skin. H331 Toxic if inhaled. H370 Causes damage to organs Respirable Level limit value Ceiling limit value Short-term value Total dust dust [ppm] [mg/m3] [ppm] [mg/m3] [ppm] [mg/m3] [mg/m3] [mg/m3] 200 250 250 350 Remark H: Skin penetrating substance according to The Swedish Work Environment Authority's provisions and on occupational exposure limit values, AFS 2011:18 Acetic acid, conc 64-19-7 H226 Flammable liquid and vapour. H314 Causes severe skin burns and eye damage Respirable Level limit value Ceiling limit value Short-term value Total dust dust [ppm] [mg/m3] [ppm] [mg/m3] [ppm] [mg/m3] [mg/m3] [mg/m3] 5 13 10 25 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 matrix Estimated risk after preventive measure

Consequence Probability Prepare solutions Potential for exposure via Serious Possible Use labcoat, safety goggles and disposable nitrile gloves. Low risk inhalation and direct contact Work in fume hood or on ventilated bench with Plexiglas with eyes and skin. hood. Write in handling and safety instruction. Potential for exposure to The limit values for methanol and acetic acid are not exceeded highly corrosive vapours, and at any occasion. for direct contact of corrosive All staff participate in mandatory fire training every 5 years.. solution with eyes and skin Labs are equipped with fire extinguishers. Potential fire hazard Handle acrylamide gels Small amount of Serious Possible Use labcoat, safety goggles and double, disposable nitrile Low risk unpolymerised acrylamide gloves. Work in fume hood or on ventilated bench with potential for exposure via Plexiglas hood. Write in handling and safety instruction. direct contact with skin.

Precipitate protein from gel Potential for exposure via Serious Possible Use labcoat, safety goggles and disposable nitrile gloves. Low risk by cover with fixing solution inhalation and direct contact Work in fume hood or on ventilated bench with Plexiglas with eyes and skin. hood. Write in handling and safety instruction. Potential fire hazard. The limit values for methanol and acetic acid are not exceeded at any occasion. All staff participate in mandatory fire training every 5 years.. Labs are equipped with fire extinguishers. Stain gel by cover with Potential for exposure via Serious Possible Use labcoat, safety goggles and disposable nitrile gloves. Low risk Coomassie staining solution inhalation and direct contact Change gloves frequently, especially if you suspect with eyes and skin. contamination. Remove gloves carefully without touching Potential fire hazard. skin. Work in fume hood. Write in handling and safety instruction. The limit values for acetic acid is not exceeded at any occasion. All staff participate in mandatory fire training every 5 years.. Labs are equipped with fire extinguishers. Task Identified risk/problem Assessment of risk, see risk matrix Estimated risk after preventive measure

Consequence Probability Rinse gel with fixing solution Potential for exposure via Serious Possible Use labcoat, safety goggles and disposable nitrile gloves. Low risk inhalation and direct contact Work in fume hood or on ventilated bench with Plexiglas with eyes and skin. hood. Write in handling and safety instruction. Potential fire hazard. The limit values for methanol and acetic acid are not exceeded at any occasion. All staff participate in mandatory fire training every 5 years.. Labs are equipped with fire extinguishers. Destain gel by cover with Potential for exposure via Serious Possible Use labcoat, safety goggles and disposable nitrile gloves. Low risk destaining solution inhalation and direct contact Work in fume hood or on ventilated bench with Plexiglas with eyes and skin. hood. Write in handling and safety instruction. Potential fire hazard. The limit values for methanol and acetic acid are not exceeded at any occasion. All staff participate in mandatory fire training every 5 years.. Labs are equipped with fire extinguishers. Clean and take care of waste Potential for exposure via Serious Possible Same personal protective equipment as during the task Low risk inhalation and direct contact Dispose of as chemical waste. with eyes and skin Potential fire hazard. Potential environmental risk. At power failure or Fumes evaporate to lab. Serious Possible Close containers and close fume hood. Write in handling and Low risk evacuation Potential for exposure via safety instruction. inhalation. Potential fire hazard.

Cleaners Potential for exposure via Serious Possible Before sending glass ware to be washed it must be properly Low risk direct contact with eyes and rinsed. Write in handling and safety instruction. skin. Cleaning must only be performed in free areas and cleaners must not wipe spill. Write in instruction for cleaners.

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

 Protocol for Coomassie Blue Staining

 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