Faculty of Biological Sciences Assessment Title SDS-PAGE Gel Electrophoresis preparation of gels Area/Service Protein Production Facility

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1 Faculty of Biological Sciences Assessment Title SDS-PAGE Gel Electrophoresis preparation of gels Area/Service Protein Production Facility COSHH Assessment Brief Description of Activity, Experiment or Process PI(s) or Manager(s) in charge Dr Brian Jackson Assessment Code PPF03 Date: July 2016 Preparation of SDS PAGE gel. Acrylamide gel mixtures prepared and then cast between glass plates. Gel running buffer prepared and apparatus assembled. Safety Overview In addition to Good Lab Practice the following controls are required within this experiment (refer to details below for further information) Control Fume Hood Other Ventilation Spill Tray Control of aerosols Gloves* Glasses/visor Protective Clothing* Other (give brief details) Tick *additional to measures taken or PPE worn when working within Good Lab Practice e.g. thick chemical resistant gloves, apron etc. Emergency Information detail how foreseeable emergencies based on risk from chemical would be dealt with (fill this section in once assessment is complete) Examples: Flammable chemicals extinguishing if the quantities involved in the work caught fire Corrosive chemicals first aid measures/actions if there is contact with eyes or skin Chemicals dangerous by inhalation fire aid/actions if worker accidentally inhaled Chemicals dangerous if swallowed first aid/actions if worker ingests accidently Chemical Action in foreseeable emergency In case of skin contact: Take off contaminated clothing and shoes immediately. Wash off with soap and plenty of water. Take casualty to a shower if severity warrants it and contact H&S for advice and potential treatment with Diphoterine. In case of eye contact: Rinse thoroughly with plenty of water for at least 20 minutes If swallowed: Do NOT induce vomiting. Rinse mouth with water if conscious. If inhaled: Move person into fresh air. If not breathing, give artificial respiration. Summon ambulance or take casualty to A&E if effects of any contact are not relieved by simple first aid measures. Small Lab Fires: Contain any small lab fire with a fire blanket or a CO2 fire extinguisher. If fire is not under control within 1 minute initiate building evacuation. SDS powder Acrylamide/bisacrylamide solution Hydrochloric acid 37% Ammonium persulphate powder TEMED stock solution Brief Overview No symbol assigned (+0) Fire; use water, foam, CO2 or powder extinguisher. First aid; standard first aid procedures. Spillage; collect with an electrically protected vacuum cleaner or by wet brushing, place in a suitable container and dispose of via Healthcare waste / orange bag route. First aid; standard first aid procedures + if skin contact wash off with soap & plenty of water, take victim immediately to hospital; take MSDS if possible. Fire; use appropriate extinguisher for surrounding environment. First aid; standard first aid procedures. Spillage; collect with an electrically protected vacuum cleaner or by wet brushing, place in a suitable container and dispose of via Healthcare waste / orange bag route. First aid; standard first aid procedures + if skin contact wash off with soap & plenty of water, take victim immediately to hospital; take MSDS if possible. Labelling/Pictograms of chemicals involved in Process (+1) (+3) (+3) (+3) Signal Words Danger Warning (+0) Refer to Guidance Tick all that Apply No Hazard Health Hazard Corrosive Flammable Oxidiser Explosive Serious Health Hazard Toxic Gas Cylinder Harmful in Environment

2 Chemical Name and CAS no. Acrylamide / bis-acrylamide % solution Sodium Dodecyl Sulphate (SDS) Sodium Dodecyl Sulphate (SDS) 10% solution Ammonium Persulphate (APS) Tetramethylethylenediamine (TEMED) Tris(hydroxymethyl)aminomethane (Tris base) Hazard Labels and Signal Words and specific health effect (if applicable) e.g. Damage to respiratory system if inhaled Toxic (if swallowed). Serious Health Hazard. Danger of serious damage to health by prolonged exposure through inhalation, in contact with skin and if swallowed. May cause cancer & heritable genetic damage. (Danger ) Highly Flammable. Toxic (in contact with skin); causes skin, eye & respiratory irritation. Harmful if swallowed. Corrosive. Causes skin irritation & serious eye damage. (Danger) Oxidiser. Serious Health Hazard. Harmful if swallowed. Causes serious eye irritation. May cause allergic skin reaction. Highly Flammable (liquid & vapour). Corrosive. Causes severe skin burns & eye damage. Health hazard. Harmful if ingested/inhaled. Health Hazard: Skin, eye & respiratory irritation. (Warning). Chemicals Hazards Classification (1-5) Scoring of hazards presented within experiment - Does the experiment pose a hazardous exposure? A B C D E F G H Hazardous vapour or fumes (+3) Dusty/light powders Aerosols generated Splashes or drips likely (+1) Skin or eye contact likely Ignition source may cause fire Carcinogen, mutagen, toxin or teratogen (+5) Other Hazard (assign score 1-3 based on severity) (e.g. working above WEL*) 5 Yes Yes Yes Yes Yes) 18 5 Yes Yes Yes 11 4 Yes Yes Yes 9 5 Yes Yes Yes Yes (Oxidiser) 4 Yes Yes Yes Yes Yes 14 1 Yes Yes 5 Total Hazard (1-25) 12

3 Hydrochloric acid 37% solution (HCl) Corrosive. May cause respiratory irritation. (Danger) Delete unused cells as needed. Add new cells by adding row. *WEL = Work place exposure limit. Detailed in HSE document EH Yes Yes Yes Yes 12 Hazardous vapour or fumes (+3) Dusty/light Aerosols Splashes or Skin or Ignition Carcinogen, powders generated drips likely eye source may mutagen, (+1) contact likely cause fire toxin or teratogen (+5) A B C D E F G H Other Hazard (assign score 1-3 based on severity) (e.g. working above WEL*) Control identification and Specification Give details of controls/procedures required in addition to Good Lab Practice to avoid or reduce exposure to hazards (e.g. identify stages where a control is needed, specific controls for one chemical etc). Be as specific as possible e.g. perform process A using chemical X in fume hood; weight out chemical Y powder in chemical cabinet; pipette chemical W very gently to reduce aerosol; pipette (do not pour) chemical Z from stock bottle. s are reduced by deducting hazard score allocated above where exposure is successfully controlled. is only partially reduced for control by PPE only (+1 to remain). Record final residual score in table below. Total Hazard (1-25) Chemical: Acrylamide/bis-acrylamide solution Controlled? Sc. Chg. Chemical: SDS powder Controlled? Sc. Chg. Hazards: A C D E G Hazards: B E F All work with Acrylamide stock solution to be carried out in fume cupboard. (A) Yes, (G) 6 Weigh out stock powder and make up solutions to desired volume in a fume cupboard. Yes 2 (A&G). Once diluted gel mix can be poured on the bench. Partially (B). SDS solutions can be used on the bench once powder diluted. Where possible do not pour acrylaminde soluctions, pipette gently to avoid (C&D) Yes, 4 Work in a fume cupboard; wear lab coat, gloves and safety glasses to avoid skin/eye Yes 2 generating aerosols, splashes or drips. (C,D&G) (G) Partially contact. (E) Wear lab coat, gloves and safety glasses to avoid skin/eye contact. (E&G) Partially 2 Weigh out in fume cupboard and isolate from sources of ignition. (F) Yes 2 Total Changes 12 Total Change 6 Chemical: SDS 10% solution Controlled? Sc. Chg. Chemical: Ammonium persulphate (APS) Controlled? Sc. Chg. Hazards: C D E Hazards: B D E Where possible do not pour, pipette gently to avoid generating aerosols, Yes 3 Weigh out stock powder and make up solutions to desired volume in a fume cupboard. Yes 2 splashes or drips. (C&D) (B). APS solutions can be used on the bench. Wear lab coat, gloves and safety glasses to avoid skin/eye contact. (E) Partially 1 Aliquot stock to 1 ml tubes to reduce amount that can be spilled. Where possible do Yes 1 not pour, pipette gently to avoid generating splashes or drips. (D) Work in a fume cupboard when using powder; wear lab coat, gloves and safety glasses Yes 2 to avoid skin/eye contact. (E) Measure out in fume cupboard & isolate from sources of ignition (F) Yes 2 Total Change 4 Total Change 7

4 Chemical: TEMED Controlled? Sc. Chg. Chemical: Tris base Controlled? Sc. Chg. Hazards: A C D E F Hazards: B E Aliquot from stock bottle in fume cupboard to provide small quantity for bench Yes 3 Spoon powder out gently to reduce dust. Tris solutions can be made and used on the Yes 2 use. Keep lid on stock tube until needed. Pipette the small amount needed using good lab practice. (A) bench. (B) Where possible do not pour, pipette gently to avoid generating aerosols, splashes or drips. (C&D) Yes 3 Wear lab coat, gloves and safety glasses to avoid skin/eye contact. (E) Partially 1 Wear lab coat and gloves to avoid skin contact. Very small quantity in use at any Partially 1 one time to reduce possibility of splashes and drips (E) Measure out in fume cupboard & isolate from heat/sparks/open flames. (F) Yes 2 Total Change 9 Total Change 3 Chemical: Hydrochloric acid 37% Controlled? Sc. Chg. Chemical: Controlled? Sc. Chg. Hazards: A C D E Hazards: A B C D E F G H (delete as required) All work with concentrated stock solution to be carried out in fume cupboard. Yes 3 (A). Diluted solutions for conditioning can be used on bench. Where possible do not pour, pipette gently using a dropping pipette to avoid Yes 3 generating aerosols, splashes or drips. (C&D) Wear lab coat, gloves and safety glasses to avoid skin/eye contact. (E) Partially 1 Total Change 7 Total Change Residual Risk from Chemicals in Experiment If a hazardous exposure is adequately controlled then Hazard from chemical hazard identification above can be reduced accordingly and recorded below. cannot be reduced below Classification Highlight scores greater than 12 in Red or Bold/Underline) Chemical Haz. Sc. Change Residual Risk Chemical Haz Sc. Change Residual Risk Acrylamide/bis-acrylamide solution TEMED SDS powder Tris base SDS 10% solution Hydrochloric acid 37% Ammonium persulphate If residual risk score is still greater than 12 then Appendix A must be completed to ensure specific controls for the chemical are specified and put in place Waste Disposal Detail how waste resulting from this work will be disposed of; Yes Solid Waste: Can all solid waste be disposed of into Healthcare waste/autoclave waste as appropriate or to biological agent contamination? If no, give details below Liquid Waste: Can all liquid waste be disposed of into clinical waste/autoclave waste as appropriate or to biological agent contamination? Yes If no, give details below Specific measures required for waste containing chemicals not suitable for disposal to healthcare waste or to the drain. Environmental Hazards: Are any chemicals used in this work labelled as environmental hazards? These must not be placed into the drain. No If YES, detail below Chemical (e.g. Chloroform) State of waste (e.g. liquid, contaminated solids etc) Disposal Route (chemical waste (solvent) route for liquid and cin bin for solids) Notes about disposal (e.g. remove solvent waste to Miall waste store) Polymerised gel (waste) Solid gel Orange bag, still within pot solution was made in. Do not try to get solid out of tube, place directly in orange bag.

5 Risk assessment Approval COSHH Assessment must be approved by the Academic(s)/Manager(s) in charge of the work before it becomes valid The risk assessment approver must ensure that staff and students carrying out the work assessed here are provided with adequate supervision to ensure they are carrying out the procedures stipulated. Name of Manager/Academic in charge Signature Date Dr Brian R Jackson 18/07/2016 Further Actions Table note action details and when completed Review Dates note assessment review dates below (date and sign) Action Detail Person responsible Date of Completion Date of Review Signature of reviewer