HEALTH SAFETY Eb Q ETHIDIUM BROMIDE Welcome to

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Welcome to an on-line health and safety awareness package intended for staff and students working within Uof. E buildings on the Edinburgh bio. Quarter campus. Information contained within these pages is intended for use by University of Edinburgh staff and students only.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE This on-line awareness package is intended for laboratory workers using and disposing of ethidium bromide within Uof. E buildings on the Eb. Q campus. CM&VM @ Eb. Q Last updated: May, 2021

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE This presentation is supplementary to general guidance provided by The University of Edinburgh, and which is available at: https: //www. ed. ac. uk/healthsafety/guidance/hazardoussubstances/ethidium-bromide

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Please take time to view the following material, and direct any urgent questions to your H&S Advisor, Laboratory Manager, or the H&S Manager for Uof. E buildings on the Eb. Q campus, the contact details for which are shown on the last pages of this presentation. CM&VM @ Eb. Q Thank you.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Ethidium Bromide (Et. Br) is Mutagenic, and possibly also Carcinogenic and Teratogenic. Et. Br is Highly Toxic to the respiratory system, and Irritating to the eyes, skin, mucous membranes and upper respiratory tract.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Et. Br is a nucleic acid stain used in techniques such as agarose gel electrophoresis. When exposed to ultraviolet light, Et. Br fluoresces with a redorange colour, intensifying almost twenty-fold after binding to DNA.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Not to put too fine a point on it, if Et. Br 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?

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE The fact that Et. Br 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.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Safety and Risk Phrases • • R 22 – Harmful if swallowed R 26 – Very toxic by inhalation R 36 – Irritating to eyes R 37 – Irritating to respiratory system • R 38 – Irritating to skin • R 68 – Possible risk of irreversible effects

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Hazard and Precautionary Statements Hazard Statements: • H 302 Harmful if swallowed • H 331 Toxic if inhaled • H 341 Suspected of causing genetic defects Precautionary Statements: • P 261 Avoid breathing dust/ fume/ gas/ mist/ vapours/ spray • P 281 Use personal protective equipment as required • P 311 Call a Poison Centre CENTER or Doctor/ Physician

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Before using Et. Br, 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.

HEALTH & SAFETY @ Eb. Q 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 Et. Br are listed at: https: //www. ed. ac. uk/healthsafety/guidance/hazardoussubstances/ethidium-bromide

HEALTH & SAFETY @ Eb. Q 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 Et. Br to levels that are As Low As Reasonably Practicable.

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

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE • If use of Et. Br 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.

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

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE But it is critically important to realise that there is No Acceptable Level of Contamination with ethidium bromide.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Where Et. Br has escaped containment, and perhaps contaminated surfaces and equipment, inform colleagues and evacuate the area, take steps to ensure that others do not enter the area, inform laboratory managers, and only then take time to plan a carefully considered approach to safely decontaminating the affected area.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Where items, such as weighing equipment, is judged to be irredeemably contaminated, label the item(s) accordingly to prevent further use, and obtain special advice regarding the most appropriate route of disposal.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Design experiments before commencement to minimise concentrations of Et. Br and the potential to contaminate equipment (e. g. Try using Et. Br in agarose gel, and not in a running buffer).

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE • If you must transport solutions of Et. Br, use doublecontainment (i. e. with a secondary container around the primary vessel). • When finished working with Et. Br, discard your gloves immediately and in the correct waste bin/bag.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE • Et. Br-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

HEALTH & SAFETY @ Eb. Q 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 Et. Br may not be appropriate during pregnancy.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Uof. E’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

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

HEALTH & SAFETY @ Eb. Q 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.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Response to Accidents • Replenish spill management kits and correctly dispose of contained waste and contaminated PPE, etc. • Et. Br, 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.

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

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Waste Treatment & Management Since Et. Br is comprised mostly of carbon, it will be completely inactivated by incineration (the waste management treatment of choice for substantial concentrations of the chemical).

HEALTH & SAFETY @ Eb. Q 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 Manager (contact details for whom are towards the end of this presentation). • The following, however, summarises current policy:

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE • The preferred method of decontaminating or deactivating solutions of Et. Br 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.

HEALTH & SAFETY @ Eb. Q 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”.

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE • Powder and crystals of Et. Br 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.

HEALTH & SAFETY @ Eb. Q 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 Manager).

HEALTH & SAFETY @ Eb. Q 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 Manager Tel: 0131 651 4287 or Email: waste@ed. ac. uk

HEALTH & SAFETY @ Eb. Q ETHIDIUM BROMIDE Health & Safety-Related Information Relevant policy and guidance notes are available for reference at: https: //www. ed. ac. uk/medicine-vetmedicine/staff-and-current-students/cmvmhealth-and-safety/edinburgh-bioquarter These cover subject matter including: * Ethidium bromide * Laboratory coats and gloves CM&VM @ Eb. Q

HEALTH & SAFETY @ Eb. Q 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 Et. Br, contact: H&S Manager (Uof. E CM&VM @ Eb. Q) Tel: 0131 242 6390 Internal extension: 26390 Email: lgm@staffmail. ed. ac. uk CM&VM @ Eb. Q

HEALTH & SAFETY @ Eb. Q 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 within Uof. E buildings on the Edinburgh bio. Quarter campus. CM&VM @ Eb. Q Thank you