HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE

1. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Welcome to an on-line health and safety awareness package intended for staff and students working within UofE buildings on the Little France campus. Information contained within these pages is intended for use by University of Edinburgh staff and students only.

2. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE This on-line awareness package is intended for laboratory workers using and disposing of ethidium bromide in Little France. Last updated: February, 2018

3. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE This presentation is supplementary to general guidance provided by The University of Edinburgh, and which is available at: http://www.docs.csg.ed.ac.uk/estatesbuildings/waste/Ethidium_Bromide_briefing.pdf

4. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Please take time to view the following material, and direct any urgent questions to your H&S Advisor, Laboratory Manager, or the Little France Buildings H&S Manager, the contact details for which are shown on the last pages of this presentation. Thank you.

5. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Ethidium bromide (EtBr) is a nucleic acid stain used in techniques such as agarose gel electrophoresis. When exposed to ultraviolet light, EtBr fluoresces with a red-orange colour, intensifying almost twenty-fold after binding to DNA.

6. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE EtBr is mutagenic, and possibly also carcinogenic and teratogenic. EtBr is highly toxic to the respiratory system, and irritating to the eyes, skin, mucous membranes and upper respiratory tract.

7. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Not to put too fine a point on it, if EtBr intercalates so readily with DNA in vitro, it’ll intercalate with your DNA too, in vivo, given half a chance! And you certainly don’t want that, do you?

8. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE The fact that EtBr is rapidly absorbed through even intact skin, and also that it is highly toxic by inhalation, strongly recommends control measures that include local exhaust ventilation (e.g. use of a fume hood) and suitable laboratory glove material.

9. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Before using EtBr, you must do a suitable and sufficient COSHH risk assessment, develop a Safe System of Work, communicate these effectively to all users, and ensure that work is carried out accordingly.

10. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE The principles of effective risk management, and the hierarchy of controls, dictate that, wherever possible, a less hazardous substance must be identified and used. Alternatives to the use of EtBr are listed at: https://www.ed.ac.uk/health-safety/guidance/hazardous-substances/ethidium-bromide

11. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE If no safer alternative to ethidium bromide can be identified for a specific experimental application, consideration must be given to reducing the risks of storage, handling and disposal of EtBr to levels that are as low as reasonably practicable.

12. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE So, until a safer alternative can be identified and adopted … • Minimise your stock holdings of EtBr. • Use EtBr tablets in preference to powders, and solutions in preference to any form of solid EtBr. • Always use spill containment when working with EtBr.

13. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE • If use of EtBr in solid form really is unavoidable, make up working solutions by adding a small amount of diluent to the solid form, and then adjust to the final desired concentration. • Always wear the personal protective equipment specified in the relevant Safe System of Work.

14. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Also, if weighing powder or crystals of ethidium bromide really is unavoidable, do so in a fume hood to prevent inhalation of EtBr dust or aerosols, and use disposable pads to protect surfaces and help contain spillages.

15. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE But it is critically important to realise that there is no acceptable level of contamination with ethidium bromide.

16. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Design experiments before commencement to minimise concentrations of EtBr and contamination of equipment (e.g. Try using EtBr in agarose gel, and not in a running buffer).

17. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE • If you must transport solutions of EtBr, use double- containment (i.e. with a secondary container around the primary vessel). • When finished working with EtBr, discard your gloves immediately and in the correct waste bin/bag.

18. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE • EtBr-stained gels are usually visualised under ultraviolet light, which is associated with its own safety-related risks. • Care should be taken to comply with all relevant Safe Systems of Work related to use of ultraviolet light sources such as transilluminators

19. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE If you are pregnant, or are planning a family, you should contact the site’s Heath & Safety Manager or the University’s Occupational Health Unit for more information regarding maternity risk assessment. But certain work with EtBr may not be appropriate during pregnancy.

20. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE UofE’s Occupational Heath Unit can be contacted on a confidential basis for further information and advice by telephoning 0131 650 8190 or by emailing: Occupational.Health@ed.ac.uk

21. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE • Nitrile gloves, or something similar that is non-allergenic and affords at least equal protection, should be worn when working with EtBr (including EtBr waste) • Double-gloving may be desirable in some circumstances to further reduce risks.

22. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE First Aid • Skin and Eyes – Rinse splashes with cool clean water for fifteen minutes, then seek medical attention. • Inhalation or Ingestion – Seek medical attention immediately. • In all cases, supply product safety information (MSDS and COSHH Risk Assessment etc) to attending physicians.

23. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Response to Accidents • Replenish spill management kits and correctly dispose of contained waste and contaminated PPE, etc. • EtBr, and materials contaminated with the substance, should be managed as cytotoxic waste and disposed of accordingly* * Further guidance is contained in Section 18 (Waste Disposal) of this site’s safety Manual.

24. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Report All Accidents • Complete and submit an accident report using the form at: https://www.ed.ac.uk/health-safety/accident-reporting • Investigate the occurrence, review all relevant factors and, if necessary, revise contingency plans.

25. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Waste Treatment & Management Since EtBr comprised mostly of carbon, it will be completely inactivated by incineration (the waste management treatment of choice for substantial concentrations of the chemical).

26. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE • Basically, if in doubt, on any aspect of laboratory waste disposal, ask your lab’s Waste Management Adviser. • Further advice may be sought from the University’s Waste & Environment Manager (contact details for whom are towards the end of this presentation). • The following, however, summarises current policy:

27. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE • The preferred method of decontaminating or deactivating solutions of EtBr is to use an appropriate type of activated carbon filter or “destaining bags”. • Spent filters and destaining bags are to be managed as Special Waste.

28. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE • Solutions of less than 0.1% Decontaminate and then dispose of to drain. • Gels of less than 0.1% Use the orange bag waste stream. Label bags “Ethidium bromide waste: Below hazardous threshold”.

29. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE • Powder and crystals of EtBr Manage as Special Waste. • Solutions and gels of greater than 0.1% Manage as Special Waste. • Activated carbon filters and “destaining bags” Manage as Special Waste.

30. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Ensure that all Special Waste is contained securely within the laboratory pending disposal to an authorised cytotoxic waste incinerator via the site’s appointed clinical waste removal contractor (arranged through the University’s Waste & Environment Manager).

31. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Waste Management: Further Information To arrange for an uplift of cytotoxic (special) waste, for label templates, and any further information that may be required regarding waste management, contact: Waste & Environment Manager Tel: 0131 651 4287 or Email: waste@ed.ac.uk

32. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Health & Safety-Related Information Relevant policy and guidance notes are available for reference at: https://www.ed.ac.uk/medicine-vet-medicine/staff-and-current-students/cmvm-health-and-safety/little-france These cover subject matter including: * Ethidium bromide * Laboratory coats and gloves

33. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE Health & Safety-Related Information For any further information that may be required regarding health and safety considerations related to the handling or disposal of EtBr, contact: H&S Manager Tel: 0131 242 6390 Little France internal extension: 26390 Email: lgm@staffmail.ed.ac.uk

34. HEALTH & SAFETY @ LITTLE FRANCE ETHIDIUM BROMIDE You have now completed this on-line awareness package summarising key aspects of safety in respect of using and disposing of ethidium bromide in Little France. Thank you