SAFETY MANUAL Issued by the IARC Occupational Health and Safety

SAFETY MANUAL Issued by the IARC Occupational Health and Safety Committee (OHSC) Lyon, France, 2004 CONTENTS 1 Introduction Introduction from the Chairperson of the Occupational Health and Safety Committee (OHSC) Maintaining safety at IARC The OHS committee Committee members and their affiliations 2 Safety Procedures Evacuation Accidents First aid supplies and equipment Reporting an accident Fire General security procedures Theft Use of magnetic access cards EPIC and Latarjet Buildings Children Domestic animals Annex 3 Medical Surveillance IARC staff physician Periodic medical exams Individual medical surveillance 4 General Laboratory Safety General guidelines Presence in the laboratory outside normal working hours Unattended laboratory experiments Safety bulletin boards Instructions to cleaning or maintenance personnel Personal guidelines Drinking and eating in laboratories Storage of food and drink Cleanliness and laboratory tidiness Laboratory coats Gloves Safety glasses Protection against UV radiation Noise Safety procedures Safe use of lifts Safe use of fume cupboards Safe use of cold rooms Defrosting and decontamination of freezers or refrigerators Safe use of liquid nitrogen Safe use of gas cylinders Safe use of microwave ovens Laboratory services Glassware washing service (8th floor) Back-up freezers and refrigerators Storage of laboratory consumables Disposal of non-contaminated solid waste Disposal of contaminated solid waste Disposal of non-contaminated liquid waste Disposal of contaminated liquid waste Disposal of dangerous, out-of date or unidentified chemicals Handling carcinogens in the carcinogen room In case of an accident in the carcinogen room Transporting carcinogens within the building Handling carcinogens in the laboratory - dichloromethane, chloroform, benzene, formaldehyde, ethidium bromide, acrylamide Disposal of carcinogen-contaminated waste Solid carcinogenic waste Liquid carcinogenic waste Radioactive carcinogenic waste Destruction of waste carcinogens Decontamination of equipment and work area Annexes 8 Safe Use of Radioactive Substances Authorization to use radioisotopes and medical surveillance Registration Systematic surveillance by IRSN IARC laboratories authorized for radioactive materials Handling of sealed sources Ordering radioactive compounds Storing radioactive compounds Working safely with radioactive substances Personal protection Low-energy ß radiation High-energy ß radiation X and Υ radiation Safe handling of radioactive substances Radioactive waste Storage and disposal of radioactive waste Liquid radioactive waste Mixed radioactive waste Solid radioactive waste Very low radioactive waste Procedure in case of contamination Decontamination Decontamination of equipment Decontamination of work areas Personal decontamination References Annexes 9 Safe Use of Biological Substances Medical surveillance Confinement level for manipulating genetically modified organisms (GMO) and biological samples Handling of biological samples Blood, plasma and serum Tissue sections Cell lines Sources of contamination Aerosols Scalpels, needles and syringes Evaporation/lyophilization Centrifugation Pipetting Disposal of contaminated waste Decontamination 5 Laboratory Equipment Hazardous equipment Fragile and continuously operational equipment Unattended equipment Role of staff responsible for equipment 6 Storage and Use of Inflammable Substances Ordering of inflammable substances Storage of inflammable substances Authorised quantities of inflammable solvents Storage of waste solvents Authorised quantities and storage of inflammable radioactive substances Distribution of solvents Distribution of alcohols Use of inflammable gases 7 Safe Use of Carcinogens Ordering carcinogens Storage of carcinogens Working with carcinogens Personal protection Cell culture media Glassware Disposable "Nalgene" filters Scissors, tweezers and other instruments Inactivation of HIV Other precautions Accidents involving potentially contaminated biological fluids Biological samples archives Sasakawa basement Packaging samples for shipment Annexes 10 Working With Animals Access to the Animal Facility Animal Care and Use Committee 11 IARC Collections of Biological Specimens Access to the collections Biological Archives Committee 12 Working in the L3 laboratory Access to the L3 laboratory Biological Archives Committee L3 laboratory safety committee Chapter 1 Introduction 9 INTRODUCTION At the International Agency for Research on Cancer, INTRODUCTION considerable resources have been and will continue to be given to maintaining safety in our place of work; however, this can be jeopardized by a lack of knowledge of the general safety rules and unsafe work practices. The purpose of this Safety Manual, compiled by the Occupational Health and Safety Committee (OHSC), is to detail the safety procedures that are specific to IARC and to provide everyone working on the premises with sufficient information on good laboratory practice to avoid dangerous situations. Before starting work at the Agency, individuals must familiarize themselves with the relevant sections of the Manual and establish with their supervisors how experimental procedures will be undertaken safely. Visitors working in the laboratories should be assisted in this respect by IARC staff. Unit Chiefs and Group Leaders have a particular responsibility to ensure that all new staff and visitors are properly briefed and that all safety procedures are respected. Everyone, including those working in the Epidemiology Units and for the Administration, is required to complete a registration form, acknowledging that they have been informed of the safety procedures in place at IARC, and to attend a safety briefing given by the Laboratory Safety Officer. More detailed information on the procedures for working with animals and about the IARC Archives of Biological Materials can be found in the Standard Operating Procedures Manual and the Guidelines for the IARC Archives of Biological Material respectively, both of which are available from Unit secretaries. As circumstances change, this Safety Manual (web site http://ariane/labcom) will be updated accordingly. Staff members with questions or suggestions can contact any member of the Committee or the Laboratory Safety Officer. From the Chairperson of the Occupational Health and Safety Committee 10 Chapter 1 Introduction MAINTAINING SAFETY AT IARC General safety matters at IARC are under the supervision of the Administrative Services Officer (ASO), Mr G. Guillerminet (telephone number 8444) and his assistant the ASO Safety Officer, Mr M. Bazin (telephone number 8523). Mr Guillerminet is responsible on a day-today basis for ensuring that general safety rules are in place and respected. He coordinates the IARC security team and a team of staff with first-aid qualifications who can take immediate action in the event of an emergency anywhere on the IARC premises. The Laboratory Safety Officer, Miss B. Chapot (telephone number 8513), coordinates matters of laboratory safety and in particular those concerning the manipulation of biological samples, carcinogens and other toxic compounds. She is also the IARC’s Radiation Safety Officer and is thus responsible for all aspects of safety concerning the manipulation of radioisotopes in the IARC laboratories. She oversees the storage of dangerous compounds, the elimination of laboratory wastes, with particular responsibility for those that are carcinogenic or radioactive, and the monitoring of specialised laboratories – the carcinogen handling laboratory and the various laboratories authorised for the handling of radioisotopes. Mr Bazin aids her in many of these tasks and either may be contacted in case of an emergency. Miss Chapot works in close contact with the Chairman of the Occupational Health and Safety Committee (OHSC), Dr J. Hall, the IARC Staff Physician, Dr A. Robert, and ASO to ensure that all staff, and in particular new arrivals, are aware of and comply with the IARC Health and Safety rules. The IARC Staff Physician, Dr A. Robert, (telephone number 8426) is responsible for the medical surveillance of all staff, monitoring working conditions and accidents, and evaluating potential risks, if necessary in conjunction with the Occupational Health and Safety Committee. She is not present at IARC on a daily basis but can be contacted through the Personnel office (telephone number 8027). Chapter 1 Introduction 11 The interaction of these staff provides the basis of the health and safety programme at IARC but for it to be effective the input of all staff is necessary. The Director, Unit Chiefs and Group Leaders play an essential role in ensuring that the Administrative Services and the Laboratory Safety Officer are aware of changes in laboratory practices and the arrival of any new persons, visitors as well as staff members, and that all are properly briefed on safety issues. Finally, it is the duty of each person working at IARC, and in particular those working in a laboratory or those involved in any aspect of maintaining laboratory safety, to ensure that safe work practices are respected, and to avoid generating potentially hazardous situations. THE OHS COMMITTEE he OHS Committee is composed of a Chairman, nominated by the Director, and a variable number of members, chosen to represent each of the IARC laboratory floors, the epidemiology units, the EPIC and Latarjet buildings, the IARC Administrative Services (Administration, Technical Services, Safety), Common Laboratory Services (animal house, glass-washing room), the Chairman of the IARC Archives of Biological Material and the Staff Association. The IARC Staff Physician and the Radiation Safety Officer also serve on the Committee, as required by French law. The Committee considers all aspects of health and safety and in particular: • Emergency procedures • General laboratory and building safety • Medical surveillance • Safe use of laboratory equipment • Safe handling of inflammable substances • Safe handling of carcinogens • Safe use of radioactive substances • Safe use of biological materials and genetically modified organisms (GMO) • Working with animals T 12 Chapter 1 Introduction The Committee meets about once every 6 weeks but can be convened in case of an emergency by contacting the Chairman or the IARC Safety Officer. All practical aspects of laboratory safety and working conditions at IARC are discussed and anyone working at IARC may contact a member of the Committee if there is a health or safety issue they would like to be raised either at the Committee level or with the IARC administration. Practical safety recommendations are made to staff directly by the Committee, via the safety bulletin board on each floor and on the Intranet. The minutes of each meeting are accessible on the Committee's Web site: http://ariane/labcom/. When policy decisions are required, the Committee makes recommendations to the Director or the Unit Chiefs. Current committee members and their affiliations P. Barbieux M. Bazin A.M. Camus-Randon B. Chapot I. Deltour D. Galendo I. Gilibert G. Guillerminet Dr P. Hainaut Dr J. Hall Dr R. Kaaks M. F. Odefrey S.Pauly C. Piccoli Dr A. Robert Dr B. Sylla B. Vozar Technical Services Administrative Services Safety Officer Molecular Pathology Laboratory Safety Officer Radiation Safety Officer Radiation & Cancer Animal House Supervisor Endogenous Cancer Risk Factors Administrative Services Officer Chairman of the IARC Archives of Biological Materials Chairperson OHSC Hormones and Cancer Representative of Staff Association Genetic Cancer Susceptibility Gene Environment Interactions IARC Staff Physician Biological Substances Common Laboratory Services Nutrition & Cancer basement basement 11th floor 7th floor Latarjet 13th floor 8th floor 2nd floor 7th floor 7th floor EPIC 9th floor 10th floor 6th floor ground floor 6th floor EPIC Chapter 1 Introduction 13 Dr J. Hall, Chairperson P. Barbieux M. Bazin A.M. CamusRandon B. Chapot I. Deltour D. Galendo I. Gilibert G. Guillerminet Dr P. Hainaut Dr. R. Kaaks F. Odefrey S. Pauly C. Piccoli Dr A. Robert Dr. B. Sylla B. Vozar Chapter 2 Safety procedures 15 SAFETY PROCEDURES In an emergency in any of the IARC buildings, such as a fire, flood or earthquake, a siren, with a varying pitch, will be sounded. On hearing this warning, all staff in that building should immediately leave their office or laboratory and go as quickly as possible, at a walking pace, to the nearest exit. Only the stairs may be used. All cumbersome articles should be left behind. Emergency wardens have been appointed for each floor of all the IARC buildings. Their names are listed on notices posted on the door of the "Pater Noster" (the internal mail delivery system) in the Tower and in the corridors of the EPIC and Latarjet buildings. They will supervise the evacuation and ensure that everyone on their floor has heard and obeyed the warning. If you are on the telephone when the alarm is sounded, you should explain rapidly to your caller and hang up immediately. Each emergency warden should confirm the evacuation of his/her floor at the guard station in the main building and then proceed to the assembly point. Persons who panic, are disabled or collapse should be taken or carried to the safety of the stairwells, normally by not fewer than two and not more than four other staff members, and helped to descend to ground level. If they are incapacitated, their presence should be immediately signalled to the guard station so that the emergency services can take the necessary action. On reaching the ground floor, staff members should leave the building, go to the assembly point and check in. In case of an evacuation of staff working in the Tower, the assembly point is located next to the bicycle shed outside the Latarjet building. In the case of evacuation of staff working in the EPIC or Latarjet building, the assembly point is in the car park immediately behind the Tower (see Annex I). Instructions from the officials responsible for security must be obeyed. Staff EVACUATION Only the stairs may be used! 16 Chapter 2 Safety procedures members' cars must be left in the car park until the emergency is over. Cars should be parked correctly in the car park, only in the marked parking spaces, to ensure easy access for the Fire Brigade and other emergency vehicles. Staff are requested to familiarize themselves with the location of the fire extinguishers and the emergency exits, the security instructions and the names of the emergency wardens for each floor. In order to accustom staff members to the siren, it is sounded at noon for 15 seconds on the first working Monday of each month. ACCIDENTS Any person witnessing one of the following : • • • • explosion fire bodily injury, fainting, etc electrocution (in this case, first shut off the mains power) • leakage of suffocating, explosive or inflammable fluid • splashing by corrosive or biologically active or radioactive substances must break the cover of the red button to alert IARC security staff who have had first aid training. They will take immediate action to protect the individual(s) at risk and will, if necessary, call for help from the fire department or the S.A.M.U. (French medical emergency services). Before 8.30 a.m. and after 5 p.m., the alarm alerts guards who have been instructed how to proceed in case of an accident. The red button is situated opposite the Pater Noster (the internal mail delivery system) in the main building, opposite the stairwell in the EPIC building and next to the emergency exits in the Latarjet building (2 such exits per floor). Chapter 2 Safety procedures 17 If the use of the red button does not result in the arrival of IARC security staff, you should contact the switchboard or the security guards directly by picking up the red telephone located on each floor, near the red button. The switchboard has a direct line to the Lyon fire brigade. If an injured person requires medical attention, he/she will be accompanied by a qualified person to the hospital using appropriate transport. In exceptional circumstances and only if the first two procedures have not worked, you may call for help directly using one of the following numbers: • Lyon Fire Department • Emergency Medical Aid "S.A.M.U. " • Edouard Herriot Hospital - Emergencies • Poison Control Centre A list of all qualified first-aiders is displayed on the Safety Bulletin Board (located opposite the red lift in the Tower, on the ground floor in the EPIC and Latarjet buildings). 18 15 04 72 11 60 80 04 72 11 69 11 Having adequate first aid and safety equipment in a laboratory is the best way of ensuring quick help in the case of an accident. On each laboratory floor, safety showers are available for use by laboratory workers who have been splashed with toxic or corrosive chemicals. Showers are also available in the Tower basement. Eyewash stations are installed near the safety showers as well as in each laboratory. Emergency fire blankets are located in the corridors near laboratory entrances, on the 6th, 7th, 8th, 9th, 10th, 11th and 13th floors, as well as on the ground floor of the EPIC building. They should be used to extinguish clothing on fire. The first aid cabinets located on each floor are equipped only with basic supplies and can be replenished from the stock kept by ASO. More extensive first aid supplies can be obtained from the receptionist or security guards on the ground floor of the main building. Emergency first aid equipment can be found in First aid supplies and equipment 18 Chapter 2 Safety procedures the Staff Physician’s office (R13) on the ground floor, the key for which can be obtained from the switchboard. This equipment is for the use of trained personnel only. Reporting an accident Any person involved in an accident including visitors should report it to the Staff Physician and to the Laboratory Safety Officer. Any member of personnel who is a victim of an accident must complete a copy of form WHO 417 and the attending physician should complete a copy of form WHO 418 to report an accident in which bodily injury has resulted. These forms are available from the Personnel office on the 2nd floor (216). The staff member must forward form WHO 417 to his supervisor if the accident has occurred during working hours to report on the circumstances under which the accident took place. The supervisor must particularly specify: 1. whether the staff member was on duty 2. what were his/her particular duties at the time of the accident. The supervisor may write his report on a separate sheet of paper, to be attached to form WHO 417. The supervisor or the staff member must then send this completed form to Personnel. The form WHO 418 must be completed by the attending physician and then sent to Personnel who will take appropriate action. FIRE or obvious reasons of safety and health, smoking is forbidden everywhere in all IARC buildings. In laboratories, the greatest risk to personnel from fire comes from the use of inflammable solvents. Inflammable solvents must be stored in the laboratories, in limited amounts, in explosion-proof refrigerators. They must not be used near a hot electric element, an electric motor or a naked flame. The use of naked flames is prohibited in all the IARC build- F Chapter 2 Safety procedures 19 ings, except in certain exceptional circumstances (see chapter 6). A special room for the storage of inflammable solvents is located near the unloading area in the basement of the Tower and is open daily from 10.00 to 10.30 a.m. and from 2.00 to 2.30 p.m. (see chapter 6). There is an annual fire practice to ensure that all staff members know how to use correctly the fire extinguishers located on each floor. As safety is everyone's responsibility, it is important that staff members become familiar with the location, and the correct use, of these red fire extinguishers. (Green extinguishers are also located on each floor, they contain water and they should be used as safety showers). All newcomers at IARC are informed of the available safety equipment and procedures through a briefing by their Unit Chief or his representative and by a tour of their working place. They must also attend a general safety meeting within the first 3 months of their arrival at IARC, organised by the Laboratory Safety Officer. GENERAL SECURITY PROCEDURES IARC is not insured against, nor is it liable for, claims resulting from the loss or theft of personal property on its premises. Staff members should, therefore, not leave personal property of value, in particular mobile phones, handbags or wallets containing money or cheque books, in unattended offices and/or laboratories, or in unlocked desks or cupboards at any time. Similar precautions should be taken for any attractive items of office equipment such as calculators and portable computers, etc. Nothing of value should be left visible in cars, which should be locked when parked. If a valuable article disappears, the loss should immediately be reported to ASO. Theft 20 Chapter 2 Safety procedures Use of magnetic access cards ARC has a system of controlled access to ensure security. The controlled entrances are as follows: • entrance & exit to the car park • entrances & exits to Tower building • entrance to Latarjet building • entrance to EPIC building • entrance & exit gate cours Albert Thomas • loading area gate • bicycle shed • animal house • computer room 403 • radioactivity and carcinogen room 805 • L3 cell culture room 901 • Bacteriology room 919 • L2 cell culture room 920 • radioactivity storage room SS/SMH • radioactivity storage room S05 • storeroom S19 • room E01 EPIC • room E02 EPIC • room E03 EPIC A credit card sized badge, containing a miniature low frequency emitter/receiver programmed for each individual staff member, permits access to all or certain of the above-mentioned areas. The badge is issued by ASO to each person working at the Agency for periods of longer than a week. It can also be used as a "payment card" in the Cafeteria. At the controlled access point, the badge should be held in front of the card reader at a distance of about 20 to 30 cm, without excess movement. If the card’s code is accepted, the card reader’s signal light will change from red to green and access will be allowed. A central computer continuously records information on access points that have been activated. It is thus important that every time staff pass the main entry/exit points of the different IARC buildings, each person should use their badge so that in the event of an incident the number of staff in any building can be monitored. Care should be taken not to leave this badge in your car or office and any loss should be immediately reported to ASO’s office. I Chapter 2 Safety procedures 21 The Latarjet and EPIC buildings are protected by an alarm system which is activated: • at night (8.00 p.m. to 7.00 am), • during weekends, • on IARC public holidays. During these periods, access to the EPIC and Latarjet buildings is possible by going to see the guard in the entrance hall of the Tower. You must inform her/him when you leave the EPIC or Latarjet buildings so that the alarm system can be reactivated. Staff working in the EPIC or Latarjet buildings should also remember to close all windows when leaving the building each evening. Latarjet and EPIC buildings Children under 16 are not allowed access to any of the IARC buildings with the exception of the reception hall and the cafeteria. In this case, they should be taken directly to and from the cafeteria by their parents without going to any other location. o domestic animals are allowed in the Agency, to avoid contamination of laboratory animals by pathogenic agents. Children N Domestic animals 22 Chapter 2 Safety Procedures Annex I - Location of assembly points Chapter 3 Medical surveillance 23 MEDICAL SURVEILLANCE IARC STAFF PHYSICIAN IARC has a physician who is responsible for supervising the medical surveillance of all staff, who monitors working conditions and accidents, and who evaluates potential risks, if necessary in conjunction with the Occupational Health and Safety Committee. The physician advises the Director of the Agency on working conditions, in special cases of sickness or accident, on the need for a change of job or for an improvement in a work environment, with the understanding that all medical information is strictly confidential. The Agency Staff Physician examines staff members when they return to work after sick leave of more than 21 days and anyone who has had a work-related accident. The Staff Physician may also be asked to give an opinion to the Joint Medical Service in Geneva: - on special prescriptions (not routinely covered by the WHO Health Insurance) - on the follow-up of accidents incurred at work or during journeys to and from work. PERIODIC MEDICAL EXAMS For staff members, the Staff Physician carries out medical examinations prior to recruitment, at the end of service and periodically throughout the employee’s term of service. These examinations include a general physical examination, blood and urine tests, a lung X–ray and can include other tests as requested by the Staff Physician. 24 Chapter 3 Medical surveillance The frequency of these examinations is: • Staff members under the age of 40: A full examination every 5 years, with one intermediate general physical examination without biological tests or X-rays • Staff members aged 40 to 55: A full examination every two years • Staff members aged 55 and over: A full examination every year For new special training awardees with contracts for more than three months, a medical examination is required before recruitment. For other non staff members, there is no compulsory medical exam. All new laboratory personnel are required to present their vaccination certificates, showing immunological evidence of seroconversion, when starting work at IARC. The Staff Physician may be consulted on any issue and for an update of vaccinations, and he/she can ask for medical tests to be carried out if necessary. INDIVIDUAL MEDICAL SURVEILLANCE n addition to these routine medical exams, the Staff Physician is also responsible for supervising any additional medical surveillance that is required for specific categories of IARC personnel including: • The surveillance of all staff who work in laboratories: – Annual blood and urine tests – 6 monthly reporting of chemicals and biological agents handled • The surveillance of all staff who work with radioisotopes: - twice yearly radiotoxicological urine analysis - 3 monthly check of dosimeter results, - monthly reporting, by users, of the quantity of any radioisotopes handled - any other test required by the Staff Physician. • Ophthalmologic tests, carried out annually on staff members who work with visual display screens, I Chapter 3 Medical surveillance 25 • Medical examination of any person involved in an accident at work, • An examination of any person requesting one, either for themselves or a member of their family. • Vaccination programme for the prevention of infections such as hepatitis B, tetanus, polio, etc. Medical aid or prescriptions can be given in emergencies to all staff. N.B. All women working at IARC are required to inform the IARC Staff Physician as soon as they are aware that they are pregnant. Chapter 4 General laboratory safety 27 GENERAL LABORATORY SAFETY GENERAL GUIDELINES It is dangerous to work alone in a laboratory Only authorized persons are allowed access to the IARC buildings. To ensure better safety conditions for persons working in IARC laboratories, a Laboratory Presence Record is kept by the security guards on the ground floor (R01). This notebook enables the guards to identify any person working in a laboratory outside normal working hours: • On normal working days (before 8.00 am and after 5.00pm) • On weekends and IARC holidays During these periods, any person entering the building to work in a laboratory MUST fill in this notebook, in capital letters, with: 1. their first and last names 2. floor and laboratory number where they are going to work 3. time of arrival On normal working days, any person who enters the building between 8.00 a.m. and 5.00 p.m. to work in a laboratory and who decides to work late must also, from 7.00 p.m., inform the security guard on the ground floor (tel. 8800), of their presence and give him their name, floor and laboratory number. Upon leaving the building, he/she should not forget to record the hour of his/her departure in the record book, to ensure that the security guards know that he/she has left the building. For personnel who are classified as "non-staff members”, a form must be completed and signed by the Unit Chief or Group Leader which authorizes the person to work in Presence in the laboratory outside normal working hours 28 Chapter 4 General laboratory safety a laboratory outside normal working hours. This authorization needs to be renewed every 6 months. A copy of this authorization must be sent to ASO. Unattended laboratory experiments t can be tempting to leave unattended laboratory experiments that make use of specific pieces of equipment which if they malfunction could present a danger either to the operator or to others in the laboratory or which could result in damage to the instrument itself (see chapter 5 for full listings). This situation should be avoided if at all possible. However when it is necessary to leave such equipment running, the following procedure should be strictly adhered to: 1.During normal working hours, either the log-books of such equipment (e.g. ultracentrifuges, rotary evaporators, speed vacs, etc.) should be filled in noting the name of the operator or a clear label should be attached to the equipment itself (e.g. gel electrophoresis equipment, hybridisation ovens, etc.) giving this information. 2. Outside normal working hours, in addition to this requirement, the notice (see below) fixed on the door of the laboratory in which the equipment is located should be filled in giving the date, the name of the equipment functioning, the name of the operator and a telephone number at which this person can be contacted. This information will allow the night security personnel to contact the staff member who is using any piece of equipment if it develops a problem. It is important that when the equipment is subsequently switched off the notice is duly modified. I Laboratory Date Equipment Equipment running overnight Operator Contact telephone number Chapter 4 General laboratory safety 29 All equipment which is left on continuously (fridges, freezers, tissue culture incubators etc.) should carry a green label indicating the name and telephone number of at least 2 staff members who can be contacted in case of emergencies. It is the responsibility of the Unit Chief or Group Leader to ensure that these names are updated as and when necessary. afety bulletin boards are located on each floor of the Tower on the wall opposite the red lift, and on the ground floors of the EPIC and Latarjet buildings. Information is posted there concerning changes in safety regulations, dates of medical examinations and other information related to safety, including a list of qualified first-aid staff at IARC. S Safety bulletin boards The cleaning personnel who work in the evening are responsible for cleaning the floors, emptying the waste paper baskets and removing the cardboard bins used for the disposal of broken glass, which have been placed in front of the service lift by laboratory staff. Cleaning personnel present in the laboratories during the normal working hours are under the responsibility of laboratory staff who should ensure that they: • have taken any necessary health and safety precautions (wearing gloves, laboratory coats, choice of cleaning materials etc.) • know how and what in the laboratories should be cleaned • do not clean any piece of laboratory apparatus without getting prior authorization of laboratory staff Radioactive and carcinogenic waste containers must be clearly marked and, under no circumstance, are to be picked up or thrown away by the cleaning or maintenance personnel. To better ensure their safety, cleaning personnel should not work in laboratories where handling radioactive substances is authorized, tissue culture is carried out or in the carcinogen room (805). Instructions to cleaning or maintenance personnel 30 Chapter 4 General laboratory safety The cleaning of these laboratories is carried ot separately under the supervision of the responsible staff member who must ensure that there are no hazards present. PERSONAL SAFETY GUIDELINES Outlined below are some good laboratory practices that should be followed to reduce risks of exposure to potentially dangerous chemicals, equipment or situations. Drinking and eating in laboratories his rule also covers eating at desks in laboratories. The use of kettles and coffee machines is also forbidden outside of the kitchen on the ground floor of the Tower. Drink and snack distributors are located on the ground floor of the Tower. T It is strictly forbidden to drink or eat in the laboratories. Storage of food and drink It is strictly forbidden to store food and/or drinks in laboratory cold rooms, fridges or freezers. A fridge is available for this purpose in the kitchen on the ground floor of the Tower and may be used by all staff. Cleanliness and laboratory tidiness Elementary rules of cleanliness should be respected, i.e. wash your hands often, especially after handling biological substances, after wearing gloves and before leaving the laboratory. Laboratory coats and other pro- Chapter 4 General laboratory safety 31 tective clothing should not be worn in the cafeteria, the kitchen or other meeting rooms. The laboratory should be kept tidy and free of items that have no relation to laboratory work. For the safety of all staff members, fastened laboratory coats must always be worn during laboratory work, regardless of the type of work being carried out. Laboratory coats must not be worn in the cafeteria, in the kitchen and other meeting rooms. Protective laboratory clothing, marked with your own name, can be obtained from Supplies (SUP: ext. 8464). Dirty laboratory coats should be taken to the basement for washing and placed in the laundry bins. Once laundered they will be returned to your office or laboratory. Laboratory coats Latex, vinyl or rubber gloves must be worn when working with dangerous substances. To safely weigh carcinogens that are electrostatic, cotton gloves, available in room 805, should be worn. After use, contaminated gloves must be disposed of in special waste containers or "burn bins”. These bins should be clearly marked with the appropriate label: carcinogens, radioactive materials or biohazards. Gloves must not be worn in any of the 4 principal lifts, in the larger service lift or in the stairways. The wearing of gloves is tolerated in the red service lift, where food is forbidden. Always remember that after use, gloves may be contaminated. Special precautions should be taken to remove them before touching any objects, to avoid their contamination (laboratory instruments, pens, telephones, door knobs, electric light switches, laboratory notebooks or radiation dosimeters, etc.). Always remember to wash your hands after removing gloves. Gloves The necessity to wear safety glasses depends on the type of experiment and the work being carried out. Safety glasses 32 Chapter 4 General laboratory safety Safety glasses must be worn: All staff are provided with a pair of safety glasses which must be worn in all laboratory areas when there is manipulation at risk and near equipment which is judged to be dangerous as indicated by this blue adhesive sign. • hen handling any of the following substances: - Liquid nitrogen, - Strong acids such as hydrochloric acid, sulphuric acid, nitric acid, trichloroacetic acid, glacial acetic acid, etc. - Solutions of strong bases such as sodium hydroxide, potassium hydroxide, etc., - Corrosive agents such as phenol, - Halogenated solvents such as dichloromethane, chloroform, etc., - Organic solvents such as acetone, isoamyl alcohol, etc., - Radioactive isotopes, - Carcinogenic compounds, - Chlorine bleach. W • When working with any of the following equipment: - high performance liquid chromatography (HPLC), when injecting and when carrying out any operation involving disconnection of a solvent system under pressure, - rotary evaporators, - speed-vac evaporators, - ultracentrifuges, - lyophilizers, - refluxing or distilling apparatus, even when the operation is carried out in a fume cupboard, - autoclaves. • During any other hazardous operation (risk of projections, explosions or implosions) Contact lenses may not be worn in the situations cited above. Exceptions (those who find it impossible to wear glasses with prescription lenses) will be made by the IARC Staff Physician after examining the report of an ophthalmologist. Chapter 4 General laboratory safety 33 For situations in which the eyes are exposed to ultraviolet (UV) radiation, the wearing of special UV safety glasses or a UV visor is obligatory. This safety equipment is available near UV-emitting equipment. Protection against UV radiation Normal eyeglasses or safety glasses do not provide adequate protection to sources of UV radiation. In general, noise that exceeds the commonly accepted noise limit of 85 decibels at frequency on the "A" scale (85 db A) for 8 hrs exposure, will not occur in the laboratory. Ear protection can be obtained from ASO for working near noise sources such as ultrasound cleaners or sonicators. Noise SAFETY PROCEDURES he red service lift is the only one available for use by laboratory personnel for transporting materials, chemicals, solvents, etc. Its use should be limited to these tasks. T Safe use of lifts Any operation which is likely to generate toxic fumes or even an unpleasant odour should be carried out in a fume cupboard. For maximum protection, close the sash completely and work with hands extended through the two round openings. For less hazardous operations, work can be carried out with the sash front positioned such that the two red arrows on the left of the sash coincide (air speed = 0.5 m/sec). When a fume cupboard malfunctions, an alarm bell and light automatically inform technical staff of the problem. Fume cupboards should not be used for storage of dangerous items. The efficiency of each fume cupboard is Safe use of fume cupboards 34 Chapter 4 General laboratory safety checked periodically with an anemometer and by the burning of a smoke bomb; the dates of the last checks are noted on the sash front of the fume hood. Problems with the fume cupboards should be immediately reported to ASO. Safe use of cold rooms Materials stored in cold rooms should be clearly marked with both the date of storage and the name of the person responsible for the material. To avoid the growth of undesirable microorganisms, paper cartons should not be stored in cold rooms and all bottled liquids must be placed in plastic or metal trays in case of breakage. Nothing should be stored on the floor. Two persons are responsible for the correct use of each of these cold rooms: • 6th floor: J. Michelon / C. Piccoli • 7th floor: B. Chapot / G. Martel-Planche • 8th floor: I. Gilibert / Dr H. Ohshima • 9th floor: Dr. M. Tommasino • 10th floor: Dr F. Canzian / Dr L. Yin • 11th floor: A.M. Camus-Randon / Dr H. Huang The cold rooms in the Agency are cleaned twice a year by an external company. Defrosting and decontamination of freezers or refrigerators Most freezers and some refrigerators are connected to a central alarm system that alerts security personnel if their temperature control fails. Thus, before undertaking the defrosting of a freezer or refrigerator, please inform P. Barbieux (ext. 8571). Short-term replacement freezers and refrigerators are available for storing material during such procedures; contact P. Barbieux. Freezers and refrigerators should be regularly emptied and defrosted to avoid accumulation of ice. They should be periodically cleaned using diluted detergent (Bactinyl) available from M. Bazin. In the event that a freezer or refrigerator is found to be contaminated with a biological sample or a Chapter 4 General laboratory safety 35 chemical agent, contact either B. Chapot or M. Bazin, if the freezer or refrigerator is located in the carcinogen handling room (805) or an IARC laboratory, or the IARC archive manager, if it is part of the archive collection, to inform them of the problem and to ensure that the correct decontamination protocol is followed (see Guidelines for the IARC Archives of biological materials for detailed protocols). IARC has a liquid nitrogen reservoir located in front of the EPIC building. Liquid nitrogen containers are located in certain laboratories and in the basement of the Sasakawa hall, as well as in the EPIC building. These containers are regularly filled by the staff members responsible for the distribution of liquid nitrogen. As liquid nitrogen can quickly freeze skin, it is necessary to wear safety glasses or a mask and suitable gloves when handling this substance. (Such equipment is available in the laboratory in the Sasakawa basement and can be used when removing samples from the liquid nitrogen tanks stored there; similar equipment is stored in the EPIC building). Liquid nitrogen should be transported only in suitable sealed containers that are not hermetically sealed allowing evaporation of any gases formed. (More details on handling liquid nitrogen and the safety procedures applicable can be obtained on the following website: www3.airproducts.com/productstewardship/ product-safety/safetygrams). Safe use of liquid nitrogen itrogen, carbon dioxide, oxygen, helium and argon cylinders are used in the IARC laboratories. Such cylinders must be securely attached to the wall with a safety chain. The users of such equipment should periodically verify the tubing to ensure the absence of leaks. The use of inflammable or toxic gases (H2, CH4, CO, etc.) is possible in the main building after prior agreement from ASO. N Safe use of gas cylinders 36 Chapter 4 General laboratory safety Safe use of microwave ovens icrowave ovens, often models supplied for kitchen and not specifically laboratory use, are being used increasingly to heat aqueous solutions in the laboratory. In addition to the widely known rule that metal objects should not be placed in a microwave oven, a serious risk of explosion exists if aqueous solutions contained in tightly closed glass bottles are heated in a microwave oven. Particular care should be taken when melting agarose: the container should not be closed with a fitted lid but plugged loosely with a folded paper handkerchief to allow gas to escape. Microwave ovens should not be used for heating organic solvents. M LABORATORY SERVICES Glassware washing service (8th Floor) he glassware washing service is common to all IARC laboratories, with each of the washroom personnel being responsible for the handling of glassware from several research groups or units. Only non-contaminated laboratory glassware should be sent for washing. Dirty glassware is placed, by the user, in soaking baths that are collected each morning by the washroom staff. The glassware is then washed, dried and sterilized if necessary and put back in the respective laboratories. Some other minor cleaning jobs can be asked of the washroom staff, for example the cleaning of small laboratory apparatus, incubators, etc., but only after it has been disconnected from the electricity and decontaminated by the laboratory staff. T Back-up freezers and refrigerators Three –80ºC, two –40ºC, two –20ºC back-up freezers and one 4ºC back-up refrigerator are available in the Sasakawa basement for use in emergencies. This equipment is kept locked; the pass number for the padlocks can be obtained from the Archive manager, the Agency security staff or the night guardians. When an alarm is activated on any of the freezers or refrigerators, the Chapter 4 General laboratory safety 37 Agency security staff or the night guardian will contact the staff member who is responsible for that piece of equipment. They should rapidly transfer the contents of the freezer or refrigerator that is malfunctioning, taking care to ensure that all biological samples are placed in sealed containers to avoid the risk of contaminating the back-up equipment (see the Guidelines for the IARC Archives of Biological Material for further details). A record of all breakdowns is kept, for which an incident sheet is completed by either the Archive Manager or the night guardians that must be countersigned by the Unit Chief the next day and returned to the Archive Manager. In order for this service to function well, storage time for samples in back-up freezers is limited to a maximum of one month. The contents of the back-up freezers will be checked regularly and any sample found in a back-up freezer that has not been reported to the Archive manager will be subject to immediate disposal. The back-up equipment may also be used for temporary storage of samples during the decontamination of freezers and refrigerators contact the Archive Manager for further details. One room located on the 1st basement level is reserved for this purpose: Since this room is relatively small and to avoid a potential fire hazard, the amount of laboratory consumables stored here should be limited. Small quantities of acids (acetic acid, hydrochloric acid, etc.) can also be stored in this room. Storage of laboratory consumables Several types of waste containers for non-hazardous wastes can be found in the Agency laboratories: • Black or brown plastic waste baskets: these bins are meant for the kind of waste generated in an office (i.e. paper, no sharp or dangerous objects); they are Disposal of noncontaminated solid waste 38 Chapter 4 General laboratory safety emptied daily by the cleaning personnel and are marked with the label: WASTE PAPER OR PLASTIC ONLY NO CHEMICALS OR GLASS • Plastic-lined cardboard bins: these bins are intended for the disposal of non-contaminated broken glass generated in a laboratory. They are marked with the label: Non-contaminated laboratory waste only (Pipettes, broken glass, etc.) No needles ! Déchets de laboratoire non-contaminés seulement (Pipettes, verre cassé, etc.) Pas d’aiguilles ! Filled bins should be left in front of the large service lift (monte-charge) to be taken to the basement by the cleaning staff for collection by the COURLY (Communauté Urbaine de Lyon). • Used glass container: this bin, located in the basement, provided by the COURLY, is meant for the disposal of clean glass bottles which are subsequently recycled. To ensure that glassware is clean, empty solvent bottles should be left open in the hood for 24 h to permit evaporation of any residual solvent before disposal. Bottles having contained toxic substances should be well rinsed before disposal. Glassware which is known to have contained carcinogenic or radioactive substances should not be recycled under any circumstances. • Burn-bins should be used for the disposal of small sharp objects such as needles and glass Pasteur pipettes. Chapter 4 General laboratory safety 39 All hazardous solid wastes should be disposed of in yellow burn-bins or plastic-lined paper boxes (available from the staff responsible for the operation of the incinerator) which have been clearly labelled to indicate the kind of waste they contain. • Radioactive waste: collected regularly by the ASO Safety Officer, for disposal by an outside organization (see chapter 8 for detailed protocols) • Carcinogenic waste: collected regularly by the staff responsible for operation of the incinerator (see chapter 7 for detailed protocols) • Biohazardous waste: collected regularly by the staff responsible for operation of the incinerator (see chapter 9 for detailed protocols) Disposal of contaminated solid waste ee chapter 6 for information on storage and use of inflammable substances. S Disposal of noncontaminated solvents Disposal of contaminated liquid waste ee chapters 7, 8 and 9 for information on disposal of contaminated liquid waste. S Each year, the Laboratory Safety Officer organizes a collection of all dangerous, out-of-date or unidentifiable chemicals that are no longer required. The Unit Chiefs and Group Leaders are informed of when this collection will take place and are asked to inform all members of their groups. Disposal of dangerous, out-of-date or unidentified chemicals Chapter 8 Safe use of radioactive substances 67 SAFE USE OF RADIOACTIVE SUBSTANCES Certain unstable isotopes spontaneously emit particles or rays as they decompose to a more stable state in a process called radioactivity. Radioisotopes are classified into different groups depending on their radiotoxicity. The authorised radioisotopes which may be handled in IARC laboratories, as open sources, are: Radioisotope Tritium 3H Carbon 14C Radiotoxicity Group Radiation type slight moderate 4 3 3 3 4 2 β β β β β γ Technical sheet in Annex III IV V VI VII VIII Phosphorus 32P moderate Phosphorus 33P moderate Sulfur 35S Iodine 125I slight high Any person at IARC who works with such radioactive substances must follow the appropriate procedures and security rules outlined in this chapter. Additional information is provided in the annexes to this chapter and can also be obtained from the Radiation Safety Officer, B. Chapot. She should also be contacted if the need arises to handle other radioisotopes. 68 Chapter 8 Safe use of radioactive substances AUTHORIZATION TO USE RADIOISOTOPES AND MEDICAL SURVEILLANCE Registration egistration with the French Ionising Radiation Protection Service (Institut de Radioprotection et de Santé Nucléaire IRSN) and notification of the Staff Physician. Any person at IARC required to work with radioactive substances must first obtain an authorization to handle radioactive isotopes by contacting B. Chapot who will register them with the IRSN. The list of radioisotope users is updated every 6 months. The list of persons authorised to handle radioisotopes is given to the IARC Staff Physician and all new registrations are brought to her attention. All registered users have to fill out each month a form noting the quantities of the different radioisotopes they have manipulated. These forms are sent directly to each user and should be returned to B. Chapot, even if no isotopes have been handled in the period concerned. This information is transmitted to the Staff Physician. All women are required to inform the Staff Physician as soon as they become aware that they are pregnant and are not normally allowed to handle radioisotopes. After discussion with the Staff Physician, exceptional authorization may be given on an individual basis. R Systematic surveillance by IRSN rinary bioassays This surveillance is carried out twice a year and is compulsory for all those registered to handle radioisotopes. It requires the collection of a twenty-four hour urine sample. The identification tag, attached to the specimen bottle, should be completed by the user before leaving it at the collection point (Annex IX). Each radioisotope user is expected to comply with the designated urine collection dates, which are organised to allow all the samples to be dispatched together to IRSN. U Chapter 8 Safe use of radioactive substances 69 In the case of an accidental internal exposure, the user should immediately contact B. Chapot who will organise, in consultation with the Staff Physician, the appropriate medical surveillance. External dose measurements Personal dosimeters (chest and wrist film badges) are obligatory for all staff who work with the radioisotopes 32P and 125I. They are provided and checked threemonthly by IRSN. Sufficient advance warning should be given to B. Chapot for any new registration, so that the necessary dosimeters are available when needed. Plastic storage boxes have been installed on each IARC floor where radioisotopes are handled. Each user has a named compartment where their IRSN chest and wrist dosimeters should be stored when not in use. Storing the dosimeters in this way facilitates their replacement. When working with quantities of radioisotopes greater than 37 MBq (1 mCi), a thermal luminescent finger dosimeter must be worn. These dosimeters are provided and checked by the French Institute of Nuclear Physics. They are available from B. Chapot as needed. IARC LABORATORIES AUTHORIZED FOR RADIOACTIVE MATERIALS All areas within IARC where radioisotopes are handled must have prior authorization from the Interministerial Commission. The handling of radioisotopes is only allowed in authorized areas defined by French legislation as: 70 Chapter 8 Safe use of radioactive substances ZONES UNDER SURVEILLANCE (blue radioactive label) CONTROLLED ZONES (green radioactive label) radioactive waste storage rooms where large amounts of radioisotopes are stored. The total amount of radioactivity (MBq) and the radioisotopes authorised to be stored and handled in each IARC laboratory are strictly controlled. The present authorised limits are listed below. The exact nature of the radioisotopes that can be handled and stored in each laboratory is listed on the door and must be complied with. Limit 3.7 MBq* 18 MBq 37 MBq (100 µCi)* (500 µCi) (1 mCi) Laboratory 713, 810 E06 (EPIC) 602, 609, 618, 714, 718, 809B, 812, 904, 906A, 914, 1009, 1018, 1101/1102, 1110, box 9 (13th) 719 607, 706 703/704, 805, 915, 1022 S05, SMHS3 (storage rooms in basement) 74 MBq 185 MBq 370 MBq 3700 MBq (2 mCi) (5 mCi) (10 mCi) (100 mCi) *A table for converting Ci to SI units (MBq) is given in Annex I Chapter 8 Safe use of radioactive substances 71 In general radioisotopes and radioactive waste should only be stored in the laboratories in which the handling of 37 MBq or more of radioactivity has been authorized. Laboratories 703/704, 805, and 1022 have absolute filters in the extraction system and should be used when handling volatile radioactive products such as 35S-methionine. Exceptionally, work may be carried out in non-authorised areas, but the following quantities must not be exceeded: 3H, 35S 5000 kBq (140 µCi) 14C, 32P, 33P 500 kBq (14 µCi) 125I 50 kBq (1.4 µCi) In such cases, B. Chapot must be informed so that the room can be checked for any possible radioactive contamination. Certain instruments (such as scintillation counters, gas chromatographs with electron capture) are equipped with detectors containing a sealed radioactive source (63Ni, 133Ba, 137Cs, 60Co). The acquisition and the elimination of such radioactive sources (when the instruments are taken out of service) necessitates special procedures. In both cases contact B. Chapot for information about the appropriate procedures to adopt. Handling of sealed sources ORDERING RADIOACTIVE COMPOUNDS Orders for radioactive compounds should be made on standard order forms. A special IRSN form is completed by the supplies office (SUP) for each new order sent to the supplier. An additional form is required for the importation or exportation of radioactive compounds from or to other laboratories. This form must be validated by IRSN before any movement between laboratories of the radioisotopes is allowed. Contact SUP for more details. 72 Chapter 8 Safe use of radioactive substances STORING RADIOACTIVE COMPOUNDS n order to be in compliance with the French law concerning the traceability of radioactive sources in the building, every new compound is delivered together with a Radioisotope Source Tracing Sheet. This sheet indicates the date, radioisotope, quantity and name of the person who has ordered the product. It has to be affixed to the refrigerator or freezer where the compound is stored. After the source is discarded the sheet should be returned to B. Chapot. Refrigerators and/or freezers containing radioactive compounds should be clearly identified with the appropriate label. If they are also used to store other products, radioactive sources should be placed in clearly identified, closed boxes dedicated to this purpose. Radioactive compounds found on the list of carcinogenic substances (Chapter 7, Table 1) or having a longhalf life (3H and 14C) must be stored in room 805 (see Chapter 7; rules for the use of this room). A computer listing of all the radioactive carcinogens at IARC is maintained and checked yearly. Each person is responsible for the products they order and must keep up to date the paper copy of the listing kept in the room 906B. I WORKING SAFELY WITH RADIOACTIVE SUBSTANCES adioisotope users must understand the nature of the hazard and be aware of the characteristics of the particular isotope they are working with, especially the potential risks of internal or external contamination. Additional technical information is available in Annexes III to VIII. R You must never work with unprotected cuts or breaks in the skin, especially on the hands or forearms. Chapter 8 Safe use of radioactive substances 73 As in all laboratories, you are not allowed to eat, drink or apply cosmetics in an area where radioactive material is handled. The use of paper handkerchiefs is recommended and of course mouth pipetting of any solution is strictly forbidden. Laboratory coats and gloves must be worn when working with radioactive compounds, and where appropriate safety glasses and disposable cuff protectors should be used too. Ordinary latex gloves do not provide full protection against contamination with radioisotopes, especially when they are diluted in solvents. Gloves should be changed frequently. You must always store isotopes, sources or probes in appropriate closed containers, with clear labels indicating the isotope, activity, date and name of user. The container should be stored in a refrigerator or freezer dedicated to this purpose. Personal protection For the radioisotopes 14C, 3H, 35S, 33P, the risk of external exposure is practically non-existent. Latex gloves, if changed often, and safety glasses give sufficient protection from contamination in the majority of situations. For tritium, however, there is a potential risk of internal exposure by inhalation or through the skin. Work with this radioisotope should preferably be carried out in a fume cupboard or in a well ventilated room. The concentration of tritium in the working atmosphere should be kept below a derived air concentration of 800 Bq/l (0.02 µCi/l). For 35S, and in particular during the use of 35S-methionine where volatile breakdown products are formed, there is an increased risk of internal exposure, mainly from inhalation or through the skin. Work with 35Smethionine must be carried out in a fume cupboard. The concentration of 35S in the working atmosphere must be less than the derived air concentration of 200 Bq/l (0.005 µCi/l). It is also important to store all the waste containing these volatile degradation products under the hood in a closed bin. Low energy β radiation 74 Chapter 8 Safe use of radioactive substances High-energy β radiation For 32P, there is a risk of external exposure to the skin. Contact with the skin should be avoided and adequate protection (gloves, safety glasses, tweezers, protective screens) should be used. Gloves are useful for avoiding chemical contamination but they do not prevent external exposure from 32P radiation. The wearing of two pairs of gloves, one over the other, is recommended. For experiments using 37 MBq (1 mCi) or more, it is also necessary to use thermoluminescent finger dosimeters for hand exposure surveillance. These are available from B. Chapot. The use of both chest and wrist dosimeters (film badges) is obligatory when working with any concentration of 32P. X and γ radiation These types of radiation are emitted by 125I. The use of both chest and wrist dosimeters is compulsory when handling this isotope. The main use of this isotope at IARC is as the labelling component in diagnostic kits, a technique which uses low amounts of radioactivity. In this particular case the isotope may be handled on the bench with a minimal risk of internal contamination. Care should be taken when stocking wastes containing 125I: a leaded glass screen should be used. Safe handling of radioactive substances As a function of the radiotoxicity, activity and physical chemical nature of the radioisotope being used, work should be carried out • on the bench top for non-volatile compounds with low activity, • under a ventilated fume cupboard, or • with protective screens for 32P Plan ahead your experiment to minimise the time you spend handling radioactivity. The shorter the time, the smaller the dose to which you are exposed. Distance yourself from the sources of radiation: doubling the distance from the source quarters the radiation dose (inverse square law). Chapter 8 Safe use of radioactive substances 75 Monitor the work area before starting your experiment for contamination control, monitor the area and your hands frequently during manipulation and check after completion of work. Monitor your hands and wash them. The outer limits of the work zone on the bench should be marked using special yellow adhesive tape marked with the black radioactivity symbol. The work surface must be either plastic, stainless steel or an enamelled steel tray. The surface of the work area should be protected by plasticcoated paper (such as Benchkote); to better absorb spills, always place the Benchkote so that the plastic surface of the paper is in contact with the bench top or tray. Plastic trays can provide a limited work zone and should be used with a protective plastic liner which is easy to clean or to dispose of in case of a heavy contamination. Areas of the laboratory should be set aside for particular tasks: • the area reserved for clean equipment and accessories • the area for reagents • the work area itself • the area for equipment which has been contaminated and for radioactive waste • the area for the storage of radioactive sources Before beginning an experiment, you should have available: • sufficient clean equipment : it is highly recommended that only disposable plugged pipette tips and plastic pipettes are used. • reagents • a decontamination bath for glassware • a container for organic and/or aqueous waste • suitable containers for solid wastes, depending on which radioisotope is used and the quantity (plasticlined bags or burn-bins, Plexiglas waste bins ), • vermiculite, if necessary • safety glasses • protective gloves • paper tissues • Benchkote 76 Chapter 8 Safe use of radioactive substances When working with a high-energy radioisotope such as 32P, you will need to use Plexiglas screens, tube storage boxes, syringe guards and tweezers. Appropriate radiation detectors are available in each laboratory in which the handling of radioisotopes is authorised. Should any fault in their function be noticed, B. Chapot, who can organise a temporary replacement and repair of any defective equipment, should be contacted. Several scintillation counters located in different research units are also available. Extreme care should be taken if it is necessary to transport radioactive samples between different floors, and in particular for 32Plabelled samples, which must be placed in an appropriate sealed Plexiglas box to avoid any risk of external contamination. RADIOACTIVE WASTE All waste generated by radioisotope users must be separated from other, non-radioactive waste and must be disposed of in accordance with strict regulations as detailed below in Annex II. No radioactive waste may be disposed of in sinks or ordinary bins by laboratory personnel. Both solid and liquid radioactive waste is removed once a week, or on special request, by the ASO Safety Officer. The method of waste elimination depends on the radioisotope’s physical and chemical properties, decay half-life and activity. Thus, it is very important that radioactive wastes are not mixed. short radioactive decay half-life (< 100 days) 32P : 14.3 days 33P : 25.4 days 125I : 59.6 days 35S : 87.4 days Chapter 8 Safe use of radioactive substances 77 These types of waste are stored in the building, awaiting sufficient radioactive decay, before elimination as non-radioactive waste. long radioactive decay half-life (> 100 days) 3H : 12.35 years 14C : 5730 years 3H and 14C radioactive waste is stored until removal by the National Agency for the Processing of Radioactive Waste (Agence Nationale pour la Gestion des Déchets Radioactifs - ANDRA). The storage and disposal of radioactive waste must be done strictly in accordance with French regulations and is under the supervision of the ASO Safety Officer, M. Bazin and the Radiation Safety Officer, B. Chapot. Each waste container must have a special label (or adhesive tape) identifying the radioisotope it contains. When a waste container is full, it should be closed and left in an authorised location for collection by M. Bazin. Additional waste containers can also be obtained from him as required. Under no circumstances may radioactive biological infectious waste be eliminated without prior decontamination of the infectious agent. Three categories of waste have been defined, depending on the physical and chemical properties (French two-letter designations are given in parentheses): Storage and disposal of radioactive waste Aqueous liquid waste (LA) • 3H or 14C aqueous liquid waste is stored in 2 litre plastic bottles or in air-tight 30 litre containers. Liquid radioactive waste 78 Chapter 8 Safe use of radioactive substances or 33P very radioactive aqueous liquid waste should be placed in appropriately labelled clear plastic-covered bottles (green cap) stored in cylindrical plastic containers. 32P • • or 33P aqueous wash solutions should be placed in appropriately labelled 30 litre plastic containers. When the container is full, a 1 ml aliquot should be taken and counted and the result attached to the container; it will then be removed and replaced with an empty one. aqueous liquid waste should be placed in appropriately labelled clear plastic-covered 1 litre glass bottles. 35S 32P • • aqueous solutions should be placed in clearly identified 20 litre plastic containers. 125I Organic liquid waste (LS) • 3H or 14C organic liquid waste or solvents are collected in 2 litre plastic bottles. The contents of these containers are subsequently combined for storage in the basement in 30 litre containers by M. Bazin. Radioisotope concentrations are checked by B. Chapot before final disposal by ANDRA. • 32P, 33P, 35S and 125I organic liquid wastes should be collected in separate appropriately labelled clear plastic-covered 1 litre glass bottles. These containers are subsequently stored until the radioactivity has decreased sufficiently to permit disposal as nonradioactive waste. Chapter 8 Safe use of radioactive substances 79 Carcinogenic/radioactive liquid waste • 3H and 14C liquid wastes which are carcinogenic are collected in appropriately labelled plastic-covered glass 4 litre bottles which cannot be chemically attacked by the solvent or the carcinogen. The waste solution bottles must be labelled as either organic or aqueous. The contents of these bottles are subsequently combined into larger containers in the basement and, after measurement of the radioactivity levels, they are discarded, depending on the amount, as either carcinogenic or radioactive waste. Anyone handling any other radiolabelled carcinogen should contact B. Chapot before starting work to organise appropriate waste disposal facilities. This usually consists of scintillation vials and their contents (radioisotopes and scintillation fluids). Plastic and glass vials should be discarded in separate containers. • 3H or 14C waste are disposed of in appropriately labelled blue 30 litre barrels. Separate containers should be used for low (below 1000 cpm/ml) or high activity (above 1000 cpm/ml) waste. Indicate the total volume and total cpm or dpm of the vials disposed of on the ticket attached to the lid of the barrel. • 32P, 33P Mixed radioactive waste mixed wastes should be placed in the appropriately labelled blue barrels. These blue barrels are located in rooms 602, 703, 706, 915 and 1001. 125I • mixed wastes are placed in yellow plastic containers reserved for this purpose 80 Chapter 8 Safe use of radioactive substances Solid radioactive waste Solid biological waste (SO) Solid radioactive biological waste, such as animal carcasses and bedding, should be placed in double-thickness clear plastic bags for disposal. The bags should be clearly marked with both the quantity in kBq (or µCi) of the radioisotope they contain and the weight of their contents, which must not exceed 3 kg. These bags are stored in room 805 until collection by M. Bazin. The details of the contents of each bag should be noted in the record book located near the freezer. Solid non-biological waste (SP) Solid material contaminated with 3H, 14C, 35S or 125I such as disposable syringes, plastic tubes, pipette tips, gloves, filter paper, etc., should be placed in cardboard burn-bins. Each container should be labelled clearly to indicate the nature of its radioactive waste. Plexiglas plastic boxes lined with a plastic bag must be used for 32P and 33P contaminated waste. When full, the plastic bag inside the Plexiglas bin must be closed with the red ties and the bin labelled with radioactive tape. Very low radioactive solid waste Large metallic green bins have been placed in several laboratories where radioactivity is handled. These are intended for items of laboratory waste (such as gloves, filter paper…) with no or very low levels of radioactive contamination, which must not be put in the ordinary rubbish bins. M. Bazin checks and collects full bags, which are incinerated immediately. Solid wastes are sorted either by B. Chapot or M. Bazin before being: • disposed of as non-radioactive laboratory waste Chapter 8 Safe use of radioactive substances 81 when the radioactivity level is very low and if the total activity discarded per day, for the entire building, does not exceed the authorised limits. • burned in the IARC incinerator within the authorised limits. • disposed of as ordinary waste after radioactive decay (in the case of radioisotopes with a short half-life) if the residual activity is less than the authorised limits • disposed of by the company ANDRA (in the case of long half-life radioisotopes). All containers (bottles, boxes, bags) which contain radioactive waste must, at the end of each day, be clearly labelled and stored in one of the laboratories authorised for the use of radioactive chemicals, to avoid their inadvertent removal by the cleaning staff. It is extremely important that all radioactivity warning labels are removed from any packaging, containers or other rubbish before they are discarded as normal non-radioactive waste. IMPORTANT PROCEDURE IN CASE OF CONTAMINATION In the event of a spill: - verbally warn all people around - isolate the contaminated zone and restrict unnecessary movement into and through the area - call B. Chapot or M. Bazin or break the cover of the red emergency button. - treat contaminated personnel first. In case of skin contamination, the contaminated part of the body should be rinsed abundantly. Use the eye wash or the shower located in the corridor (see Annex X) - In case of persistence and/or internal contamination, the person should be taken to the nearest hospital (Hôpital Edouard Herriot) Any incident, even minor, when working with radioactivity should be reported to B. Chapot. 82 Chapter 8 Safe use of radioactive substances DECONTAMINATION Decontamination of equipment At the end of each experiment, equipment and the workbench should be checked for possible contamination, using an appropriate detector for the radiation emitted (medium or high X, β or γ), or by carrying out a wipe test with a piece of filter paper (4-5 cm2), soaked in ethanol, which is then measured in a scintillation counter. To decontaminate equipment and glassware, use a solution of 10% RBS-35 in demineralized water. Contaminated glassware or equipment should be soaked in two successive decontamination baths for at least 12 hours and rinsed, before being allowed to dry, (decontamination bath and rinse liquids should be saved as LA wastes, see page 77). Decontamination of work areas Any radioactive contamination should be eliminated without delay. If solutions have been accidentally spilt on the floor or on the benchtop, use an absorbent powder such as vermiculite which should then be disposed of as radioactive waste. Surfaces should then be washed with a 10% RBS-35 solution. A check should be made to see if decontamination is complete using either the appropriate radiation detector or a swab soaked in ethanol, which is then measured in a scintillation counter. To prevent contamination by radioactive aerosols, which pose a particularly high risk, volatile radioactive substances should only be handled in the fume cupboards fitted with special absolute filters in the authorised laboratories. Personal decontamination he correct use of gloves, safety glasses, laboratory coats or special clothing should be sufficient to avoid bodily contamination. However, in case of accidental contamination, the contaminated part of the body should be washed abundantly (refer to Annex X) T Chapter 8 Safe use of radioactive substances 83 It is strongly advised to wear two pairs of gloves: the top pair can be disposed of immediately when the handling of the radioisotope source is finished or in case of an accidental contamination. Care should be taken to remove gloves before touching any non-radioactive object or material (pen, telephone, doorknob, note-book, detector…) to prevent spreading contamination. B. Chapot should be immediately contacted whenever heavy contamination occurs. In addition to the publications listed below, you can find on the web page of the OHSC (http://ariane/ labcom/) links to relevant sites on this subject. 1. Décret No. 86-1103 du 2 octobre 1986 relatif à la protection des travailleurs contre les dangers des rayonnements ionisants. JO, 12 oct. 1986 2. Décret No. 88-52 du 18 avril 1988 modifiant le décret 66-450 du 20 juin 1966 relatifs aux principes généraux de protection contre les rayonnements ionisants, JO, 6 mai 1988. 3. Avis aux utilisateurs de radioéléments soumis au régime d’autorisation prévu par le code de la santé publique, relatif à l’élimination des déchets radioactifs (sources non scellées exclusivement). JO du 6 juin 1970. 4. Protection contre les rayonnements ionisants. JO No. 1420 (ed 1992) 5. La sécurité dans l'emploi des radionucléides en sources non scellées. Institut National de Recherche et de Sécurité (1978) REFERENCES 6. Les mesures de radioactivité à l'aide de compteurs à scintillation. G. Simonnet, M. Oria (1986) 7. Radioprotection et laboratoires de Recherche. M. Dupeyroux (1990) Technique et Biologie 4 , 103-109 8. Radioprotection et personne compétente. D. Dugrillon (1990) R.G.S. 96 , 67-71 9. Radioprotection. Formation de la personne compétente Institut de Physique Nucleaire de Lyon/CNRS, Edition 1995. 10. Some safety procedures for handling 32P during postlabelling assays. M. Castegnaro, H. Brésil and J.P. Manin, in: Postlabelling Methods for Detection of DNA Adducts, Ed. D.H. Phillips, M. Castegnaro & H. Bartsch, Lyon, International Agency for Research on Cancer, 1993. 84 Chapter 8 Safe use of radioactive substances Annex I INTERNATIONAL SYSTEM OF UNITS n 1975, the International Commission of Radiation Units and Measurements (ICRU) recommended that the becquerel be adopted as the unit for radiation measurement and be included in the International System of Units (SI). Many countries have already adopted the becquerel (Bq) as the basic unit of measurement of radiation. Conversion Tables : Curies to Becquerels µCi to kBq mCi to MBq Ci to GBq atto femto pico nano micro milli kilo mega giga tera penta exa a f p n µ m k M G T P E 10-18 10-15 10-12 10-9 10-6 10-3 103 106 109 1012 1015 1018 1 2 3 4 5 6 7 8 9 10 15 20 25 30 37 74 111 148 185 222 259 296 333 370 555 740 925 1110 35 40 45 50 55 60 65 70 75 80 85 90 95 100 µCi to MBq mCi to GBq Ci to TBq 1.29 1.48 1.66 1.85 2.03 2.22 2.40 2.59 2.77 2.96 3.14 3.33 3.51 3.70 I Prefixes for SI Units Prefix Symbol Factor Converting SI Units / non-SI units To convert : From becquerel curie (Ci) gray (Gy) rad (rad) sievert (Sv) rem (rem) To curie becquerel rad gray rem sievert Multiply by 2.7 x 10-11 3.7 x 1010 100 0.01 100 0.01 Chapter 8 Safe use of radioactive substances 85 Annex II DISPOSAL OF CARCINOGENIC AND/OR RADIOACTIVE WASTE HALF-LIFE short ( < 100 days) long ( >100 days) Storage awaiting decay 2 Specific activity 1 < 74 kBq/kg Remaining activity < 74 kBq/kg ACTIVITY weak strong Specific activity 1 > 74 kBq/kg and if Class 1 2 3 4 Total activity < 3.7 kBq < 37 kBq < 370 kBq < 3700 kBq Storage in approved containers Treated as carcinogenic waste Treated as carcinogenic waste Pick-up by LABOSERVICES or INCINERATION Pick-up by ANDRA 1 2 per day for entire building Before deciding to store radiolabelled compounds, it is important to consider fully the merits of each case 86 Chapter 8 Safe use of radioactive substances Annex III Working Safely With Tritium Radioactive half-life (T1/2 ) Principal emission Monitoring for contamination Biological monitoring Annual limit on intake (ALI) by ingestion or inhalation Maximum range in air Maximum range in water Shielding required 3 H 12.4 years 8.6 keV beta (maximum) Swabs counted by liquid scintillation Urine samples 3x109 Bq (~80 mCi)** (tritiated water) 6 mm 6 x 10-3 mm None Special considerations ue to its low beta-energy, tritium cannot be monitored directly, and therefore regular swabbing and counting of the work area is advisable. • Tritium compounds can be absorbed through the skin therefore gloves must always be worn. • Although external contamination does not lead to significant radiation dose, it can lead to hazardous internal contamination. • ALIs can vary considerably eg DNA precursors such as tritiated thymidine are regarded as more toxic than tritiated water partly because the activity is concentrated in cell nuclei. • D ** Based on occupational effective dose equivalent limit of 50 mSv for stochastic risks. Chapter 8 Safe use of radioactive substances 87 Annex IV 14 C Working Safely With Carbon-14 5730 years 0.156 MeV beta (maximum) Thin end-window Geiger-Müller detector Urine samples 9 x 107 Bq (~2.4 mCi) ** (labelled organic compounds) 24 cm 0.28 mm 1 cm Perspex/Plexiglas. Thinner Perspex/Plexiglas down to 3 mm although adequate to reduce dose, does not have good mechanical properties. Radioactive half-life (T1/2) Principal emission Monitoring for contamination Biological monitoring Breath measurements (CO2) Annual limit on intake (ALI) by ingestion Maximum range in air Maximum range in water Shielding required organic compounds may be absorbed through surgical gloves. • Avoid the generation of CO2 which could be inhaled. ** Based on occupational effective dose equivalent limit of 50 mSv for stochastic risks. • Some Special considerations 88 Chapter 8 Safe use of radioactive substances Annex V Working Safely With Phosphorus-32 Radioactive half-life (T1/2 ) Principal emission Monitoring for contamination Biological monitoring Annual limit on intake (ALI) by inhalation Maximum range in air Maximum range in water Dose rate from 1 MBq in 1 ml Shielding required 14.3 days 1.709 MeV beta (maximum) Geiger-Müller detector Urine samples 1 x 107 Bq (~0.3 mCi) ** 790 cm 0.76 cm 210 mSv/h (21 rem/h) at surface 2.5 µSv/h (0.25 mrem/h) at 1 m 1 cm Perspex (Plexiglas) stops betas and minimizes production of Bremsstrahlung 32 P Special considerations • void unnecessary exposure to this high energy radionuclide, use tube racks or holders. • Finger dosimeters should also be used when handling quantities of 30 MBq (~1 mCi) and above. • Lead shielding is required when handling quantities above 300 MBq (~10 mCi) due to the production of high energy Bremsstrahlung when the beta particles are absorbed. A ** Based on occupational effective dose equivalent limit of 50 mSv for stochastic risks. Chapter 8 Safe use of radioactive substances 89 Proportion of a quantity of 32P remaining with time HOURS: DAYS: 0 4 8 12 16 20 24 28 32 36 40 44 48 52 1.000 0.824 0.679 0.559 0.460 0.379 0.312 0.257 0.212 0.175 0.144 0.119 0.098 0.080 0.976 0.804 0.662 0.546 0.449 0.370 0.305 0.251 0.207 0.170 0.140 0.116 0.095 0.078 0.953 0.785 0.646 0.533 0.439 0.361 0.298 0.245 0.202 0.166 0.137 0.113 0.093 0.077 0.930 0.766 0.631 0.520 0.428 0.353 0.291 0.239 0.197 0.162 0.134 0.110 0.091 0.075 0.908 0.748 0.616 0.507 0.418 0.344 0.284 0.234 0.192 0.159 0.131 0.108 0.089 0.073 0.886 0.730 0.601 0.495 0.408 0.336 0.277 0.228 0.188 0.155 0.127 0.105 0.086 0.071 0.865 0.712 0.587 0.483 0.398 0.328 0.270 0.223 0.183 0.151 0.124 0.102 0.084 0.070 0.844 0.695 0.573 0.472 0.389 0.320 0.264 0.217 0.179 0.147 0.121 0.100 0.082 0.068 0 12 24 36 48 60 72 84 90 Chapter 8 Safe use of radioactive substances Annex VI Working Safely With Phosphorus-33 Radioactive half-life (T1/2 ) Principal emission Monitoring for contamination Biological monitoring Annual limit on intake (ALI) by inhalation Maximum range in air Maximum range in water Shielding required 33 25.4 days 0.249 MeV beta (maximum) Geiger-Müller detector Urine samples 1 x 108 Bq (~3 mCi) ** (Class D) P 49 cm 0.6 mm 1 cm Perspex (Plexiglas). Thinner Perspex/Plexiglas down to 3 mm although adequate to reduce dose, does not have good mechanical properties. Special considerations • Phosphorus-33 does not require special precautions over and above those necessary for any beta-emitting radionuclide of this energy of emission. ** Based on occupational effective dose equivalent limit of 50 mSv for stochastic risks. Chapter 8 Safe use of radioactive substances 91 Proportion of a quantity of 33P remaining with time DAYS 0 10 20 30 40 50 60 70 80 90 100 0 1.000 0.761 0.579 0.441 0.336 0.256 0.195 0.148 0.113 0.086 0.065 1 0.973 0.741 0.564 0.429 0.327 0.249 0.189 0.144 0.110 0.084 0.064 2 0.947 0.721 0.549 0.418 0.318 0.242 0.184 0.140 0.107 0.081 0.062 3 0.921 0.701 0.534 0.406 0.309 0.236 0.179 0.136 0.104 0.079 0.060 4 0.897 0.683 0.520 0.395 0.301 0.229 0.174 0.133 0.101 0.077 0.059 5 0.872 0.664 0.506 0.385 0.293 0.223 0.170 0.129 0.098 0.075 0.057 6 0.849 0.646 0.492 0.374 0.285 0.217 0.165 0.126 0.096 0.073 0.055 7 0.826 0.629 0.479 0.364 0.277 0.211 0.161 0.122 0.093 0.071 0.054 8 0.804 0.612 0.466 0.355 0.270 0.205 0.156 0.119 0.091 0.069 0.053 9 0.782 0.595 0.453 0.345 0.263 0.200 0.152 0.116 0.088 0.067 0.051 92 Chapter 8 Safe use of radioactive substances Annex VII Working Safely With Sulfur-35 Radioactive half-life (T1/2 ) Principal emissions Monitoring for contamination Biological monitoring Annual limit on intake (ALI) by ingestion Maximum range in air Maximum range in water Shielding required 35 S 87.4 days 0.167 MeV beta (maximum) Thin end-window Geiger-Müller detector Urine samples 4x108 Bq (~10mCi)**(inorganic compounds) 2 x108 Bq (~5 mCi) * (elemental sulfur) 26 cm 0.32 mm 1 cm Perspex/Plexiglas. Thinner Perspex/Plexiglas down to 3 mm although adequate to reduce dose does not have good mechanical properties. Special considerations should be opened and used in ventilated enclosures. • Avoid generation of sulfur dioxide or hydrogen sulfide which could be inhaled. • Radiolysis of 35S-amino acids may lead to the production of volatile breakdown products resulting in an increased risk of internal exposure, mainly from inhalation or through the skin. • Vials ** Based on occupational dose equivalent limit of 50 mSv for stochastic risks. * Based on occupational dose equivalent limit of 500mSv for deterministic risks. Chapter 8 Safe use of radioactive substances 93 Proportion of a quantity of 35S remaining with time DAYS: WEEKS: 0 1 2 3 4 5 6 7 8 9 10 11 12 1.000 0.946 0.895 0.847 0.801 0.758 0.717 0.678 0.641 0.607 0.574 0.543 0.514 0.992 0.939 0.888 0.840 0.795 0.752 0.711 0.673 0.636 0.602 0.569 0.539 0.510 0.984 0.931 0.881 0.833 0.788 0.746 0.705 0.667 0.631 0.597 0.565 0.534 0.506 0.976 0.924 0.874 0.827 0.782 0.740 0.700 0.662 0.626 0.592 0.560 0.530 0.502 0.969 0.916 0.867 0.820 0.776 0.734 0.694 0.657 0.621 0.588 0.556 0.526 0.498 0961 0.909 0.860 0.814 0.770 0.728 0.689 0.652 0.616 0.583 0.552 0.522 0.494 0954 0.902 0.853 0.807 0.764 0.722 0.683 0.646 0.612 0.579 0.547 0.518 0.490 0 1 2 3 4 5 6 94 Chapter 8 Safe use of radioactive substances Annex VIII Working Safely With Iodine-125 Radioactive half-life (T1/2 ) Principal emissions 125 I Monitoring for contamination Biological monitoring Annual limit on intake (ALI) by inhalation Dose rate from 1 GBq point source at 1m First half value layer 59.6 days 35 keV gamma (7% emitted, 93% internally converted) 27-32 keV X-ray (140% Te K X-rays) Scintillation detector Thyroid scans (scintillation detector) 2 x 106 Bq (~55 µCi) * 41 µSv/h (4.1 mrem/h) 0.02 mm lead Special considerations • • • • • reezing or acidification of solutions containing iodide ions can lead to formation of volatile elemental iodine. Active aerosols can be produced by opening a vial of high radioactive concentration of 125I. Some iodo-compounds can penetrate surgical rubber gloves, two pairs are recommended. In the event of a suspected contamination of personnel, block the thyroid by the administration of potassium iodate (170 mg) or potassium iodide (130 mg). Spills of 125I should be stabilized with alkaline sodium thiosulfate solution before commencing decontamination. F * Based on occupational dose equivalent limit of 500 mSv for deterministic (non-stochastic) risks to the thyroid. Chapter 8 Safe use of radioactive substances 95 Proportion of a quantity of 125I remaining with time DAYS 0 20 40 60 80 100 120 140 160 180 200 220 240 0 1.000 0.793 0.628 0.498 0.394 0.313 0.248 0.196 0.156 0.123 0.098 0.077 0.061 2 0.977 0.774 0.614 0.486 0.385 0.305 0.242 0.192 0.152 0.120 0.095 0.076 0.060 4 0.955 0.756 0.600 0.475 0.377 0.298 0.236 0.187 0.149 0.118 0.093 0.074 0.059 6 0.933 0.739 0.586 0.464 0.368 0.292 0.231 0.183 0.145 0.115 0.091 0.072 0.057 8 0.911 0.722 0.572 0.454 0.359 0.285 0.226 0.179 0.142 0.112 0.089 0.071 0.056 10 0.890 0.706 0.559 0.443 0.351 0.278 0.221 0.175 0.139 0.110 0.087 0.069 0.055 12 0.870 0.689 0.546 0.433 0.343 0.272 0.215 0.171 0.135 0.107 0.085 0.067 0.053 14 0.850 0.673 0.534 0.423 0.335 0.266 0.211 0.167 0.132 0.105 0.083 0.066 0.052 16 0.830 0.658 0.521 0.413 0.328 0.260 0.206 0.163 0.129 0.102 0.081 0.064 0.051 18 0.811 0.643 0.509 0.404 0.320 0.254 0.201 0.159 0.126 0.100 0.079 0.063 0.050 96 Chapter 8 Safe use of radioactive substances Annex IX Instructions for twenty-four-hour urine specimens INSTITUT DE RADIOPROTECTION ET DE SURETE NUCLEAIRE Boîte postale no 35-78110 LE VESINET LABORATOIRE D’ANALYSES DE BIOLOGIE MEDICALE Numéro d’enregistrement : 78-69 • UPON RECEIPT store collection bottle away from any possible source of radioactive contamination. • TWENTY-FOUR-HOUR URINE SPECIMENS COLLECTION (see instructions below) • DISPATCH OF THE SPECIMEN - make sure the bottle is securely closed (check both cap and package). • FILL OUT IDENTIFICATION FORM 478 (English translation overleaf ) and include with the sample in the enclosed white bag. DO NOT WRITE ANYTHING ON THE BOTTLE COLLECTION OF TWENTY-FOUR-HOUR URINE SPECIMENS Day 1 DO NOT COLLECT ON WORKING PREMISES Day 2 Day 2 1. Empty your bladder at a chosen time on day 1, for example 8:00 AM. DO NOT COLLECT 2. Then collect ALL urine excreted over the following 24-h period: 8:00 AM to 8:00AM STORE IN THE COLD 3. Deliver filled sample bottle to the lab as soon as possible ATTENTION. • DO NOT DISCARD PRESERVATIVES THE BOTTLE MAY CONTAIN • DO NOT RINSE • DO NOT FREEZE Chapter 8 Safe use of radioactive substances 97 Annex X The following procedures can be carried out on the Decontamination premises in case of external contamination with a radioisotope. There are emergency showers located on each laboratory floor and in the basement of the tower building. techniques for external contamination Take a warm shower and after extensive soaping, with Shower using light rubbing if necessary, the contaminated area should be abundantly rinsed. Care should be taken not to spread the contamination beyond its existing limits, in particular avoid the eyes and nose. Each shower should last a maximum of 5 minutes and if necessary, repeat this procedure up to three times before drying the washed skin, without rubbing, using a towel. soap and water for extensive contamination inse and soap the contaminated area. The skin must never be rubbed so that it is broken and care should be taken that all folds of skin and the areas around and under fingernails are thoroughly washed and rinsed, for at least one minute, if necessary using a soft nail brush. Each rinse cycle should last a maximum of three minutes and may be repeated up to three times: in the event of contamination persisting, medical help must be sought immediately. R Washing with warm water and soap or detergent for a localised contamination This technique should be reserved for situations where Washing with the above procedures can not be used, for instance where there is a cut in the skin. In this case, the skin should be abundantly rinsed for a maximum of 3 minutes with fast-flowing water or physiological saline, such that any dirt or other particles are rinsed away. In a situation where mucous membranes have been contaminated, the following procedures should be followed: water or physiological saline 98 Chapter 8 Safe use of radioactive substances The mouth Rinse the mouth repeatedly with water, taking extreme care not to swallow the washing solution. The nose Blow the nose gently using paper handkerchiefs, which should be kept to check the level of contamination by the medical services. The eyes Rinse eyes abundantly using an eye wash containing water or physiological saline. Physiological saline: 9 g of sodium chloride (NaCl) per 1 litre water: the solution must not be kept for longer than one month. Chapter 9 Safe use of biological substances 99 SAFE USE OF BIOLOGICAL SUBSTANCES In a research institute such as IARC, where biological agents or compounds are stored and used, the risk of infection or contamination exists not only for laboratory personnel but also for the other people working around them. It is therefore essential that a certain number of health and security regulations be strictly followed when working with biological agents (bacteria, fungi, viruses) or biological samples (cells, blood or blood-derived components, body secretions or organs of human or animal origin) to minimise these risks. All biological agents and samples must be handled in laboratories providing an appropriate level of confinement and wherever possible special precautions must be taken to inactivate potential disease-causing agents (viruses, bacteria, fungi) which may be present in biological samples. It is strictly forbidden for any person to work with viruses, bacteria, cell cultures or other biological samples without having first been informed of the special procedures applicable for decontamination or disposal of waste generated in the laboratory. The purpose of this chapter is to draw your attention to the circumstances in which an accidental infection may occur and to outline the measures to be taken to minimise risks. In addition to adopting good working habits, which are essential for accident prevention and safety, vaccination against infectious agents can provide an extra measure of protection for laboratory staff. Vaccination against hepatitis B virus is compulsory for all labora- MEDICAL SURVEILLANCE 100 Chapter 9 Safe use of biological substances tory personnel (except in cases where there are medical contra-indications). A vaccination certificate (showing immunological evidence of seroconversion) must be provided by all new personnel when starting work in the IARC laboratories. If the vaccination is no longer valid, a repeat vaccination can be given by the Staff Physician. All staff are strongly advised to inform the Staff Physician of any potentially dangerous laboratory activities that are carried out on a regular basis. It is compulsory to inform the Staff Physician of any work accident, however minor. CONFINEMENT LEVEL FOR MANIPULATING GENETICALLY MODIFIED ORGANISMS (GMO) AND BIOLOGICAL SAMPLES nfortunately, it is impossible to evaluate rigorously the risks linked to the genetic manipulation of DNA, often called genetic engineering or biotechnology. Measures and recommendations have been formulated in almost all countries to try to limit potential dangers linked to this technology. Four different levels of laboratory confinement (L1-L4) for the handling of biological agents have been established. Any project involving the use of genetically modified organisms must be authorised and registered with the French Ministry of ‘Jeunesse, Education nationale et Recherche. The authorization is given for 5 years. The projects, depending on the agents manipulated, are classified into different groups. This grouping will determine the necessary level of confinement and thus the laboratory in which the experiments can be performed. In addition, other biological samples, not necessarily genetically modified but for instance contaminated with viruses, other infectious agents or carcinogens, must also be handled in laboratories with an appropriate level of confinement. For instance any biological sample contaminated with hepatitis B or C viruses, (HBV or HCV) U Chapter 9 Safe use of biological substances 101 or human immunodeficiency virus (HIV) must be handled in an L2 laboratory. Full details about the level of confinement required for the handling of samples contaminated with different viruses and other infectious agents are available from B. Chapot. • Laboratory Level L1: used for handling of biological agents in group I: "not susceptible to causing a disease in humans". Such a laboratory has no specific restrictions and experiments are carried out on the open bench. Most laboratories at the IARC are of this type. • Laboratory Level L2: used for handling of biological agents in group II: "could cause a disease in humans and might represent a risk for working personnel". Such a laboratory has restricted access, as well as adequate equipment for decontamination and disposal of biohazard waste. Laboratories 603, 604, 708, 717, 809A, 914, 920, 1006, 1007 and 1303 at IARC are L2 laboratories, most of them being cell culture laboratories. Access to laboratory 920 is controlled by magnetic card and, as there is an autoclave on the same floor, this room has been designated for the manipulation of viruses. • Laboratory Level L3 and Level L4: used for handling of biological agents in groups III and IV: "could cause a serious disease in humans and represents a major risk for working personnel". In addition to the characteristics necessary for an L2 laboratory, more rigorous rules in term of access and equipment apply in order to better protect the laboratory worker and to avoid the dispersal of agents in the environment. Laboratory 901 is a L3 laboratory (see Chapter 12 for details about its use). Each L2 laboratory has a person responsible for its safe running. This person also supervises its cleaning, which is not carried out by the general cleaning staff. The IARC Laboratory Safety Officer, B. Chapot, must always be informed: - before starting any new project involving GMO (e.g., plasmid preparation). Forms have to be completed by the scientist in charge of the project 102 Chapter 9 Safe use of biological substances describing the nature of the samples which will be handled. - at the end of the project. Laboratory workers should follow the same rules and take the same precautions when working with DNA as when working with infectious agents or materials for which the danger to humans is better understood, such as HIV or human leukaemia virus. HANDLING OF BIOLOGICAL SAMPLES Blood, plasma and serum Handling blood products must always be considered as a hazardous manipulation. Samples, even control samples, might be contaminated with hepatitis viruses or HIV and often the status of the samples is unknown. It is therefore obligatory to wear a lab coat and gloves to handle all blood samples. Working in a laminar flow hood, class II is highly recommended. When this equipment is not available, a fume cupboard should be used instead, which allows the work area to be well defined. All work surfaces must be thoroughly cleaned with a diluted bleach solution at the end of every experiment. Any biological samples that might be contaminated with an infectious agent must be handled in a laboratory which provides the appropriate level of confinement (contact B. Chapot for details). If tubes appear to be contaminated on the outside, they must be wiped with a tissue impregnated with diluted bleach before opening them. The dirty tissue must be thrown away in a burn-bin for incineration. It is recommended, whenever experimental procedure allows, that tubes containing blood products be heated at 56°C for 45 min before any manipulation to inactivate HIV. Chapter 9 Safe use of biological substances 103 One major risk of contamination is due to aerosol formation when opening tubes. One should be careful not to open tubes full-face, even under a hood. Contamination can also occur by projection of biological liquids onto mucous membranes (eyes, nose, and mouth) or onto broken skin. It is highly recommended that disposable plastic items are used for all such experimental procedures and these be disposed of in either burn-bins or boxes for incineration. If the use of glassware cannot be avoided, it must be decontaminated with a diluted bleach solution before being given to the glassware cleaning service. Always wash your hands after finishing your work, even if you have been wearing gloves. Various laboratory techniques are carried out using tissue sections where the tissues have usually been fixed with formalin. Although the potential risks of contamination are quite low, wearing a laboratory coat and gloves is compulsory and working under a hood is highly recommended. Handling frozen tissue has a higher risk of contamination, as viruses and other infectious agents are not inactivated. A protocol specific for the experiment and the samples being handled should be prepared before starting any manipulation, taking into account any potential contamination of the samples and thus the level of confinement necessary. Tissue sections ell lines from human and animal samples are routinely established and cultured in many IARC laboratories. They must be handled in laminar flow hoods that meet the European guidelines for safety and which are located in L2 laboratories. Disposable gloves and laboratory coats should be worn at all times. Particular care should be taken when establishing lymphoblastoid cell lines by infecting lymphocytes with Epstein-Barr virus (EBV). It is highly recommended that anyone routinely establishing or C Cell lines 104 Chapter 9 Safe use of biological substances working with such cell lines should be aware of their own EBV immunological status so that all necessary precautions are taken when manipulating such cultures. SOURCES OF CONTAMINATION he principal causes of individual contamination are negligence and overconfidence. Remember that, whatever experience you have, accidents can happen anytime. The main sources of contamination are: T Aerosols nhalation of aerosols constitutes one of the principal causes of laboratory infection. As mentioned previously, the handling of blood samples can generate very contaminating aerosols. Urine and faeces of laboratory animals inoculated with infectious agents are also an important source of aerosols. • Aerosols can be formed during laboratory experiments, for example, by trying to save the last drop at the end of a pipette or by opening a bottle which has been shaken. • Filtration of an infectious liquid without taking adequate precautions can lead to the contamination of vacuum pump tubing. To minimise this, a disinfectant-containing trap should be placed between the filtration system and the vacuum pump, and should be regularly emptied and cleaned. • The sonication and maceration of infected tissue can also produce an aerosol. Only blenders or mixers which have been designed to minimise aerosol formation should be used. These risks can be further reduced by working in a tissue culture hood. • The inoculation of egg embryos with infectious agents or the collection of infectious material can also lead to the formation of an aerosol and should also be carried out in a fume cupboard. I Chapter 9 Safe use of biological substances 105 It should be noted that any sharp or cutting item can be a source of contamination. Scalpel blades should be used only where absolutely necessary and must be used attached to the appropriate handle. The use of disposable scalpels is very highly recommended: changing a scalpel blade can be dangerous. Accidents frequently involve the use of syringes and needles. Therefore, work very carefully when handling them and use them only when necessary. Try to avoid drops of liquid at the end of a syringe needle just before injecting: many accidents are due to accidental injection and a deep injection could result in tissue contamination. Never try to put needles back in their initial protective packaging but dispose of them of in appropriate containers: burn-bins or boxes labelled 'Jet Aiguilles'. Scalpels, needles and syringes Needles must never be disposed of in an ordinary waste-paper basket efore handling a lyophiliser, you must know how to operate the machine correctly (if in doubt ask the staff member responsible for the instrument before starting). Contamination can occur both during the operation of the instrument and when the machine is being stopped. When carrying out a lyophilisation, always verify that the vacuum tubes are correctly connected and always work carefully with infectious, lyophilised material. Containers of lyophilised, infectious material which are broken upon reception are often sources of laboratory contamination. Reception and unpacking of tubes containing infectious material should be carried out in a laminar flow hood. B Evaporation/ lyophilisation Make sure you know how to operate the centrifuge before beginning to use it. Keep centrifuges and buckets clean and decontaminate them regularly. Contamination often occurs when buckets are filled, Centrifugation 106 Chapter 9 Safe use of biological substances when bottle caps or liquid layers are removed or when tubes break during centrifugation. The use of caps fitted to the buckets is recommended and obligatory when centrifuging blood samples; this will ensure that the sample remains confined in case of a problem and minimises the risk of contamination. Transport of samples between two different laboratory floors should be avoided whenever possible. All L2 cell culture rooms should be equipped with a centrifuge. Pipetting isposable plastic pipettes should be used whenever possible. Mouth pipetting is absolutely forbidden. Use pipette bulbs or automatic aspirators. Never leave a dirty pipette lying on the benchtop. Decontaminate pipettes by immersing them in a disinfectant bath (see below). Limit the use of glass Pasteur pipettes and handle them carefully; accidents often happen when putting on or taking off the pipette bulb. D DISPOSAL OF CONTAMINATED WASTE All biological waste should be incinerated. Solid waste must be disposed of in the yellow cardboard burn-bins or the plastic-lined paper bags (provided by the staff responsible for operation of the incinerator). These bins should be clearly labelled: The waste bins are collected twice a week and incinerated. Chapter 9 Safe use of biological substances 107 DECONTAMINATION Decontamination of biologically-contaminated cell cultures and other material is the best means of protection against infection for laboratory personnel. Decontamination is also necessary to protect the environment, i.e. to avoid spreading environmentally dangerous substances. In general: - Work benches should be disinfected regularly; do not ask cleaning staff to carry out this task. - Glassware, plastic containers, solutions and other materials which have been directly in contact with biological agents or substances should be systematically decontaminated. Never leave any glassware or contaminated material on the bench tops. Medium which has served for cell culture, whether contaminated or not by viruses, should be decontaminated with a bleach solution before being discarded down the sink. Sufficient diluted bleach solution should be added such that the colour of the medium becomes yellow. Blood samples should be decontaminated by adding diluted bleach. NEVER add a concentrated solution of bleach to blood samples; it produces a vigorous chemical reaction which generates chlorine vapours. Contaminated Pasteur pipettes, disposable tips and other glass or plastic-ware should be disposed of in burn-bins for incineration. Never autoclave glassware covered in chlorine bleach, as there is risk of toxic chlorine vapours being formed: thoroughly rinse all bleach-decontaminated material before autoclaving. Whenever possible all culturing of bacteria should be carried out in laboratory 919 on the 9th floor, which has been set up for this purpose. Media and flasks which have been used for bacterial cultures must be autoclaved before being discarded. Contaminated petri dishes should be put into plastic-lined paper bags or boxes (supplied by the staff responsible for operation of the incinerator) before autoclaving or incineration. Cell culture media 108 Chapter 9 Safe use of biological substances Glassware Erlenmeyer flasks, pipettes, tubes, test tubes and glass bottles are soaked after use, completely immersed, in a detergent bath. Glassware which has been used for cell cultures should be decontaminated by total immersion in detergent solution for a minimum of 3 hours. Disposable "Nalgene" filters isposable filters which have been used for filtering non-contaminated liquids can be discarded into the normal waste. Those which have been used for filtering viral solutions or contaminated liquids must be thrown away in boxes destined for incineration. D Scissors, tweezers and other instruments issection instruments must be soaked in a concentrated detergent solution immediately after being used. They should then be washed and rinsed in distilled water, dried and autoclaved. Syringes, needles, razor blades, Pasteur pipettes and pieces of glass must never be thrown in ordinary waste baskets meant for waste paper or plastic, but disposed of in cardboard burn-bins. D Inactivation of HIV (human immunodeficiency virus) HIV, the etiological agent of acquired immune deficiency syndrome (AIDS) belongs to a group of RNAcontaining viruses, called retroviruses. Transmission of HIV occurs either by sexual intercourse (semen, vaginal secretion) or by blood or blood products. However, the virus has also been isolated from saliva, tears, breast milk, cerebrospinal fluid, amniotic fluid, urine and other body fluids and secretions. These biological components should be considered as potential sources of contamination, even though epidemiological evidence indicates that blood, semen, vaginal secretion, and possibly breast milk are the principal routes of transmission. The prevalence of HIV infection is increasing, so the risk that health care, medical and clinical laboratory workers will be exposed to blood and other body fluids and secretions from persons infected with HIV is also increasing. It is important for these workers to Chapter 9 Safe use of biological substances 109 be aware of and to take all the necessary precautions to avoid contamination with HIV or with other human retroviruses. The recommendations described in this chapter, if followed, are largely sufficient to reduce the risk of contamination by many infectious agents, including HIV. In addition, HIV is an enveloped virus and therefore very sensitive to most of the usual detergents used in the laboratory. HIV is inactivated rapidly if exposed to commonly used chemical disinfectants at concentrations much lower than those normally used. A 1 to 10% solution of sodium hypochlorite (chlorine bleach), depending on the amount of material to be treated, is effective. Detergents like SDS (lauryl sulfate), Triton and Nonidet are also effective. The following disinfectants have also been proven effective in inactivating HIV: • • • • • • • 10% RBS HBS 10 at 10% at 37ºC 2% Chloramine 7% Isopropyl alcohol 4% Formaldehyde 2% Glutaraldehyde 6% Hydrogen peroxide Heating at 56°C for 45 min is effective to inactivate HIV in biological fluids, although some investigators suggest longer incubation. • 25 to 27% ethanol or 1% glutaraldehyde should be sufficient to disinfect medical and laboratory instruments. ench surface contamination can be avoided by working on sheets of absorbent plastic ("Benchkote"). After use, these sheets should be thrown away in boxes destined for incineration. The use of trays inside the hood or on the bench to contain the products or the samples reduces the spreading of contamination in case of an incident. B Other precautions 110 Chapter 9 Safe use of biological substances Reminder Never bring, store or eat any food or drink in a laboratory. A buttoned laboratory coat and gloves should be worn at all times when working in situations where there is a potential risk. Wash your hands frequently even if you have been wearing gloves. ACCIDENTS INVOLVING POTENTIALLY CONTAMINATED BIOLOGICAL FLUIDS (See Annex I) n case of an injury while working with potentially contaminated biological fluids, immediately clean the cut. Do not make it bleed. Wash with soap and water and rinse. Disinfect by immersing the cut for 5 minutes at least in a solution of DAKIN, available in the medicine cabinet on each floor (or a 1/10 diluted bleach solution or 70% ethanol). In case of splashes onto mucous membranes (eyes, nose) rinse thoroughly with water. Evaluate rapidly the risk of potential infection: if a biological sample was involved in the accident consult immediately Dr Robert, the IARC Staff Physician (ext. 8426) or B Chapot, (ext. 8513). In their absence the IARC security team, who can be summoned by pressing the red alarm button, will ensure that the necessary action is taken. The HIV and HCV status of blood samples can be assessed rapidly by the "Laboratoire d’analyses medicales” located opposite IARC, at 184, av. des Frères Lumière (tel. 04 78 74 03 58). 0.5ml of the blood sample involved in the accident is necessary and should be taken directly to the laboratory. It is important to note that if a treatment against HIV is to be considered, its efficiency will be much greater if started within 4 hours. But as it is a complicated I Chapter 9 Safe use of biological substances 111 therapy, with a lot of potential side-effects, it should be prescribed only by a qualified physician. Depending on the seriousness of the injury and the results of the blood analysis, further help can be obtained by contacting: - Hôpital E. Herriot : tel. 04 72 11 02 29 Pavillon P Service du Pr. Touraine - Hôpital de l'Hôtel Dieu : tel. 04 72 41 30 68 Service du Pr. Trepo - Hôpital de la Croix Rousse : tel. 04 72 07 17 54 Service maladies infectieuses All accidents should be reported to Dr Robert or B. Chapot, however minor. BIOLOGICAL SAMPLE ARCHIVES (See chapter 11 for additional details) A database of biological samples stored at IARC has been created, under the responsibility of the Archive Manager. This database lists all biological samples stored at the Agency either as part of collaborative studies with other institutes or those generated at IARC, such as cell lines. Any new samples to be stored as part of this archive must be clearly identified and their details submitted to the Archive Manager. small laboratory has been fitted out in the Sasakawa basement for the reception, sorting and aliquoting of biological samples. The laboratory is equipped with a laminar flow hood in which these operations should be carried out. This laboratory is kept locked. The key can be obtained at anytime from the IARC reception after completing the logbook. A Sasakawa basement 112 Chapter 9 Safe use of biological substances PACKAGING OF SAMPLES FOR SHIPMENT Samples must be placed in tightly closed tubes or bottles which are then placed in a second leak-proof plastic container, strong enough to withstand the conditions of transportation. (See United Nations recommendations on the transport of dangerous goods: "Packing instructions P650" in annex II) If, before packaging, the outside of a tube or flask has been contaminated by blood or biological compounds, such contamination must be removed with a dilute chlorine bleach solution. Samples must be clearly identified with an indelibly marked label, for example "attention blood". Chapter 9 Safe use of biological substances 113 Annex I 114 Chapter 9 Safe use of biological substances Annex II PACKING INSTRUCTIONS FOR THE TRANSPORT OF DANGEROUS GOODS General provisions iagnostic specimens shall be packed in good quality packagings, which shall be strong enough to withstand the shocks and loadings normally encountered during transport, including trans-shipment between transport units and between transport units and warehouses as well as any removal from a pallet or overpack for subsequent manual or mechanical handling. Packagings shall be constructed and closed so as to prevent any loss of contents when prepared for transport which might be caused under normal conditions of transport, by vibration, or by changes in temperature, humidity or pressure. Primary receptacles shall be packed in secondary packagings in such a way that, under normal conditions of transport, they cannot break, be punctured or leak their contents into the secondary packaging. Secondary packagings shall be secured in outer packagings with suitable cushioning material. Any leakage of the contents shall not substantially impair the protective properties of the cushioning material or of the outer packaging. For transport each package shall be clearly and durably marked with the words "DIAGNOSTIC SPECIMENS ". The completed package shall be capable of successfully passing the drop test as specified in 6.3.2.5, 6.3.2.3 and 6.3.2.4 except that the height of the drop shall not be less than 1.2 m. D For liquids he primary receptacle(s) shall be leakproof and shall not contain more than 500 ml. There shall be absorbent material placed between the primary receptacle and the secondary packaging; if several fragile primary receptacles are placed in a single T Chapter 9 Safe use of biological substances 115 secondary packaging, they shall be either individually wrapped or separated so as to prevent contact between them. The absorbent material, such as cotton wool, shall be in sufficient quantity to absorb the entire contents of the primary receptacles and there shall be a secondary packaging which shall be leakproof. The primary receptacle or the secondary packaging shall be capable of withstanding without leakage an internal pressure producing a pressure differential of not less than 95 kPa (0.95 bar). The outer packaging shall not contain more than 4 litres. The primary receptacle(s) shall be siftproof and shall not contain more than 500g. If several fragile primary receptacles are placed in a single secondary packaging, they shall be either individually wrapped or separated so as to prevent contact between them and there shall be a secondary packaging which shall be leakproof. The outer packaging shall not contain more than 4 kg. Provided that diagnostic specimens are packed in accordance with this packing instruction, no other requirements of these regulations shall apply. For solids Source: P650 Packing instruction. In:Recommedations on the Transport of Dangerous Goods. Model Regulations (twelfth revised edition). United Nations, ST/SG/AC.10/1/Rev.12, p.362. Chapter 5 Laboratory equipment 41 LABORATORY EQUIPMENT There are two main categories of laboratory equipment which require particular care because their use could either result in a health risk (burns, electrocution etc.) to the operator or because the instrument itself could be damaged by improper or inappropriate use. Some instruments may belong to both categories. It is absolutely essential that anyone using equipment in either category for the first time familiarises themselves with the operating procedures and potential dangers which could be encountered, by consulting the person responsible for that piece of equipment in the planning stages of any experimental procedure. Under certain circumstances a specific training must be completed under the supervision of either the manufacturer or a trained operator of the machine. HAZARDOUS EQUIPMENT This category includes laboratory equipment whose use could result in a health risk (burns, electrocution, etc.) to the operator. The following information should be clearly displayed for such equipment: • the name of the person responsible for the piece of equipment • a list of instructions for its use, in both English and French • a users' logbook in which each user should record their name, the date and the time of use. Other more specific information may also be requested for certain pieces of equipment (e.g. temperature of operation), • a maintenance logbook Certain items of equipment in this category are used on a routine basis by laboratory staff, e.g. fluorescent UV tables, power supplies and microwave ovens. It is recog- 42 Chapter 5 Laboratory equipment nised that for such items completing a users' logbook for very short and repeated periods of time can be a constraint; however, all users must be aware of the potential dangers associated with each item of equipment and they must use the appropriate safety equipment (e.g. eye shields for fluorescent UV tables etc.). Likewise, it is impractical to have a users' logbook for a liquid nitrogen container, but the correct procedures for handling must be respected and the dangers relating to its use must be fully understood by all users. For some items of equipment (e.g. ultracentrifuges), the logbook is absolutely essential and must be completed every time the equipment is used. Hazardous equipment at IARC Apparatus Animal cage washing machine Autoclaves Boiler Centrifuges Fluorescent table HPLC Liquid nitrogen containers Lyophiliser Microtomes Microwave oven Power supplies Rotary evaporator Speed vac concentrator Ultracentrifuge Ultrasonic processor UV/vis spectrophotometer Risks Burns Burns Burns Projections UV exposure Projections Burns Implosion Cuts Explosion Electrocution Implosion Implosion Projections Noise UV exposure FRAGILE AND CONTINUOUSLY OPERATIONAL EQUIPMENT Equipment in this category can be easily damaged if not used correctly or could harm or destroy their contents if malfunctioning (e.g. cold-rooms, refrigerators, freezers, Chapter 5 Laboratory equipment 43 CO2 incubators). The following information should be clearly displayed for such equipment: • the name of the person responsible for the piece of equipment • a list of instructions for its use, in both English and French • a users' logbook in which each user should record their name, the date and the time of use. Other more specific information may also be requested for certain pieces of equipment (e.g. temperature of operation), • a maintenance logbook In addition all equipment left on continuously, such as refrigerators, freezers and CO2 incubators, must have a green label attached that indicates the name of the person to be contacted if the equipment malfunctions. Unit Chiefs and Group Leaders should make sure that the names of the persons listed is kept up to date. Apparatus Autoclaves * Automatic injectors CO2 incubators* Cold room +4°C* DNA extractor DNA sequencer FACS machine Film developer Fluorimeter Freezers –20°C * Freezers –80°C * Gas chromatograph Gel dryer Hybridization ovens * HPLC/DHPLC Ice-making machines Irradiator * Luminometer Mass spectrometer Paraffin inclusion equipment PCR systems Permuted water station Phosphor-imager analyzer Refrigerators * Scintillation counter * Spectrophotometer TEA * Fragile and continuously operational equipment at IARC *Piece of equipment with a green label shown below: Personne responsable - Responsible person Nom / Name: Poste / Ext.: Unité / Unit: 44 Chapter 5 Laboratory equipment UNATTENDED EQUIPMENT (See chapter 4 for more details) hen it is necessary to leave equipment running unattended, the following procedure should be carefully followed: 1. During normal working hours, either the equipment logbooks (e.g. ultracentrifuges, rotary evaporators, speed vacs etc.) should be filled in noting the name of the operator or a clear label should be attached to the equipment itself (e.g. gel electrophoresis equipment, hybridisation ovens etc.) giving this information. 2. Outside normal working hours, in addition to this requirement, the notice (see below) fixed on the door of the laboratory in which the equipment is located should be filled in giving the date, the name of the equipment functioning, the name of the operator and a telephone number at which this person can be contacted. This information will allow the night security personnel to contact the staff member who is using any piece of equipment if it develops a problem. It is important that when the equipment is subsequently switched off the notice is duly modified. Laboratory Date Equipment Equipment running overnight Operator Contact telephone number W ROLE OF STAFF RESPONSIBLE FOR EQUIPMENT hose in charge of any piece of equipment must: 1.Ensure that the user and maintenance logbooks exist and are routinely filled in. 2.Ensure that a user’s guide, in English and French, is T Chapter 5 Laboratory equipment 45 clearly on display and that all new users are aware of how the instrument functions. 3.Keep informed of any problems encountered with the apparatus, so that the necessary action can be taken (i.e. repair, suspend the use of the apparatus when damaged, etc.). 4.Inform IARC technical services (and eventually the OHS Committee) if a potential hazard exists. Chapter 6 Storage and use of inflammable substances 47 STORAGE AND USE OF INFLAMMABLE SUBSTANCES ORDERING OF INFLAMMABLE SUBSTANCES A non-exhaustive list of inflammable substances for which special precautions must be taken and which must be labelled with a label similar to that shown here, is given below. Normally, containers of these substances, when purchased in France, carry a label with the word "inflammable" and an image of a flame. However, certain solvents are purchased in bulk and transferred to smaller containers. It is the responsibility of the person receiving a shipment of unlabelled containers or transferring inflammable liquid to smaller containers to mark each such container with a suitable label, available from the ASO Safety Officer, M. Bazin. Amines Ethylamine Triethylamine Ethers Dimethyl ether Diethyl ether Ethylene oxide Hydrocarbons Pentane Isopentane Hexane Cyclohexane Toluene Xylene Miscellaneous Acetonitrile Dichloroethane Carbon disulfide Pyridine Scintillation fluids Alcohols Butanol Ethanol Methanol Propanol Aldehydes & Ketones Formaldehyde (gas) Acetaldehyde Acetone Esters Ethyl acetate 48 Chapter 6 Storage and use of inflammable substances STORAGE OF INFLAMMABLE SUBSTANCES Authorised quantities of inflammable substances rench regulations require that a maximum of 150 litres of inflammable solvents may be in use at any one time in the laboratories of the main building (see table below for maximum quantities per floor). F Floor 13th 11th 10th 9th 8th 7th 6th total Maximum quantity 10 litres 40 litres 10 litres 10 litres 25 litres 30 litres 25 litres 150 litres Requests for changes in these limits should be directed to the IARC Safety Officer. During periodic checks to see if these limits have been respected, M. Bazin will transfer excess solvents to the underground storage room. When not in use, any inflammable solvents should either be transferred to the basement storage room or be kept in explosion-proof refrigerators. Storage of waste solvents Non-carcinogenic waste solvents (both inflammable and non-inflammable) should be stored prior to disposal in plastic containers (5 litres capacity) clearly labelled with the following label: Chapter 6 Storage and use of inflammable substances 49 Carcinogen-containing waste solvents (both inflammable and non-inflammable) should be stored prior to disposal in 4 litre, transparent, plastic-covered glass bottles (see Chapter 6) clearly labelled to indicate whether they contain either aqueous or organic solutions. Please note that where formaldehyde is used in large quantities such as in the animal house, it should be put in containers specific for this purpose carrying a label "used formaldehyde”. Waste solvents are collected each Wednesday morning by the staff responsible for operation of the incinerator. If your experimental protocol involves the use of inflammable radioactive substances, contact B. Chapot to organise appropriate waste containers which will depend on the amount of radioactivity and the isotope involved BEFORE starting any experiment. The waste containers must be marked with labels specific for this purpose which are available from M. Bazin and may only be stored in limited amounts in the authorised laboratories, from where they will be collected . Authorised quantities and storage of inflammable radioactive substances DISTRIBUTION OF SOLVENTS nflammable solvents which are in excess of the 150-litre limit authorised in the laboratories (up to the maximum of 600 litres authorised for the site) are stored under lock and key in a basement storage room. To respect this limit, all orders for inflammable solvents must be cleared by the ASO Safety Officer, M. Bazin before being sent to SUP. Inflammable solvents may be obtained daily from M. Bazin during the following time periods: I • 10.00 to 10.30 a.m. • 2.00 to 2.30 p.m. 50 Chapter 6 Storage and use of inflammable substances M. Bazin may be contacted exceptionally for an additional distribution, but only if his workload permits. Staff are reminded that solvents should not be transported by hand, but by means of a cart or bottle carrier. Only the red lift should be used for transport of such materials. DISTRIBUTION OF ALCOHOLS ax laws in France strictly regulate the use of certain alcohols. A special form, CIRC 10, available in the basement solvent storage room, must be completed and given to M. Bazin to obtain methanol, ethanol or propanol. T USE OF INFLAMMABLE GASES nder certain circumstances, inflammable gases such as hydrogen, oxygen, butane and propane may be used in the laboratory but only after clearance has been obtained from ASO. All gas bottles and lines should be clearly labelled so that there is no possible confusion over their contents. Hydrogen for gas chromatography-flame ionization detectors must be produced in the laboratory through the use of a hydrogen generator; hydrogen bottles are not permitted in IARC laboratories. The use of open flames (bunsen burners, etc.) is prohibited in the main building. The careful use of small portable burners such as camping gas for certain essential operations is, however, tolerated. U Chapter 7 Safe use of carcinogens 53 1.The workbench and any equipment used have been cleaned and, if necessary, decontaminated 2.The carcinogen container has been tightly closed, to give an airtight seal, and has been stored in its proper place 3.The users ‘logbook’ has been completed with their name, the product name, code number and amount used. When a carcinogen container is empty, the last user must cross out the line corresponding to it and indicate their name on the listing. Then they must decontaminate the container, following procedures described at the end of this chapter, before disposing of it in the container reserved for carcinogenic waste. In addition, if a compound is older than its maximum use date, the person responsible for the compound should inform B. Chapot, who will have the compound disposed of. After dilution of pure compounds, each solution container should be marked with the following information: • name of carcinogen • dilution solvent • concentration • user’s name • date CARCINOGEN................................................ CONC........................IN.................................... USER..........................DATE.............................. 54 Chapter 7 Safe use of carcinogens Diluted carcinogen solutions must always be placed in well identified sealed boxes carrying the following labels (which are available in the carcinogen room). All equipment where any solutions of carcinogens are stored (refrigerators, freezers, etc.) must be marked with the following label, available from M. Bazin. WORKING WITH CARCINOGENS Personal protection See also 'Handling Chemical Carcinogens in the Laboratory', IARC Scientific Publication No.33, chapter II, pages 7-8. It is extremely important when handling pure carcinogenic compounds that the user avoids exposing himself or those around him to these compounds. The use of protective clothing when working with carcinogens is absolutely essential. Disposable gloves, overshoes and facemasks are always available in room 805 and can also be obtained from M. Bazin. If carcinogens are routinely handled it is advisable to reserve a laboratory coat for this purpose. In certain circumstances trousers and caps should also be worn and are available in room 805 or from SUP. Protective clothing must be changed frequently, and immediately after Chapter 7 Safe use of carcinogens 55 accidental contamination. In the latter case, protective clothing must be decontaminated by appropriate methods or if necessary destroyed. If in doubt, contact the Laboratory Safety Officer. Remember that gloves do not provide complete protection and that certain carcinogenic substances such as nitrosamines, aflatoxins and aromatic amines can diffuse through gloves, especially when the compounds are in solution. It is preferable, in these cases, to use two pairs of gloves, one on top of the other. In the case of an accidental contamination, gloves should be removed immediately and thrown into the waste receptacles (burn-bins) provided for this purpose, labelled with a "carcinogenic waste" sticker: It is absolutely forbidden to eat or drink in any IARC laboratory Reminder ee also 'Handling Chemical Carcinogens in the Laboratory', IARC Scientific Publication No.33, chapter II, pages 8-10. Any synthesis or purification of chemical carcinogens must, without exception, be carried out in this room. This type of procedure is forbidden elsewhere in the building. Room 805 has 2 hoods: one fume cupboard exclusively reserved for handling radioisotopes and a laminar flow hood equipped with HEPA and charcoal filters for the handling of carcinogens. The 2 hoods must not be in operation at the same time, as they will not function properly. It is therefore necessary to reserve a time slot before working in this laboratory. Handling powders, whether electrostatic (aflatoxins, bromomethyl benzanthracene, etc.) or not, raises a S Handling carcinogens in the carcinogen room 56 Chapter 7 Safe use of carcinogens serious problem. Although no method is entirely satisfactory, one should avoid working in draughts or using latex or vinyl gloves. When handling such compounds, the use of cotton gloves and face masks is recommended. One method for handling such compounds is to dissolve all the preweighed carcinogen in an appropriate solvent. The stock solution can then be treated as if it were a non-volatile carcinogen. In case of an accident n case of an accident either in room 805 or anywhere else in the building, when working with carcinogens, do not spread the contamination, take any contaminated clothes off and break the cover of the red button opposite the “Pater Noster’. Isolate the contaminated zone and decontaminate it as quickly as possible. In case of a skin contamination, rinse abundantly with water (use the eye wash or the safety shower in the corridor), wash with a mild soap and rinse again. In case of a persistence and/or internal contamination, the person should be taken to the nearest hospital (Hôpital Edouard Herriot). I Chapter 7 Safe use of carcinogens 57 ure chemical carcinogens must never be removed from room 805, which is reserved for their storage, weighing and dilution. Only diluted solutions can be taken into the other laboratories. Each user must bring all equipment and solvents necessary to perform such dilutions to room 805. Before transporting diluted solutions of carcinogens within the building, flasks and bottles should be tightly closed and placed inside the metal transport containers. These containers, which contain an absorbent, are designed to withstand accidental dropping or chemical attack by the solvents used for dilution. All solutions must be labelled with appropriate sticker (see p.54). The cylindrical metal containers are available in the room (805) and must be returned to this room after use. When transporting diluted chemical carcinogens from one floor to another, use only the red lift. In case of an accident during transportation, push the red button across from the "Pater Noster”, isolate the contaminated zone and proceed as quickly as possible to decontaminate the zone. If the accident occurs while you are in the red lift, call for the emergency team directly from the lift and redirect the lift to the ground floor where you will be met by the emergency team. P Transporting carcinogens within the building should be taken when working with diluted solutions of carcinogens. A general rule to be followed is that solutions of volatile carcinogens should only be handled in a fume cupboard, equipped with an activated carbon absolute filter and kept at a pressure lower than that in the laboratory. Solutions of non-volatile carcinogens may be handled in trays on uncluttered workbenches. Your attention is also drawn to the proper use of certain halogenated solvents such as dichloromethane and chloroform for which sufficient proof is available Precautions Handling carcinogens in the laboratory Dichloromethane and chloroform 58 Chapter 7 Safe use of carcinogens of their carcinogenicity in animals. The stock of these solvents must be kept in rooms S16 and S12 in the basement and only a limited quantity for everyday use may be stored elsewhere on the IARC premises. These solvents must always be handled in a well-ventilated fume cupboard. Empty containers should be left under the fume cupboard to evaporate residual solvent before the containers can be safely discarded. Benzene Formaldehyde The use of benzene, a human carcinogen, should be avoided. Toluene or xylene are suitable replacement solvents. Solutions of formaldehyde, used to fix tissues, pose a special problem as formaldehyde vapours have been shown to be carcinogenic to animals. It is strongly recommended that persons who handle these solutions wear gloves as well as safety glasses and work under a fume cupboard especially adapted for use with formaldehyde, such as in laboratory 1303. Formaldehyde is also used in the histology laboratory and in certain molecular biology laboratories: it is often used as a component in gels for Northern blotting, which should be poured and run in a fume hood to reduce the risk of personnel exposure. Formaldehyde liquid waste is put in plastic containers for "solvants usés" which are collected once a week. thidium bromide is widely used in molecular biology laboratories as a fluorescent stain. It is a DNA intercalating agent which inhibits or suppresses RNA and protein synthesis in cells. As it is a genotoxic agent, it should be handled with extreme care to avoid any risk of exposure, and stored in closed and clearly identified containers. Storage at 4°C limits bacterial contamination. Agarose gels containing ethidium bromide should be discarded in waste boxes for incineration. Concentrated ethidium bromide solutions used to stain gels should be disposed of only in the aqueous carcinogen bottles (see below). It is also important to note that all plastic items (gel holders, combs, elec- Ethidium bromide E Chapter 7 Safe use of carcinogens 59 trophoresis tanks) become, with time, impregnated with ethidium bromide. They should therefore be handled only when wearing gloves. Acrylamide solutions are widely used in the labora- Acrylamide tory for the analysis of nucleic acids and proteins (separation of DNA or RNA fragments, sequencing, Western blot, polypeptide separation, etc.). Acrylamide is a carcinogen and potential neurotoxin which is absorbed through the skin. Its effects are cumulative. Thus, certain precautions are called for when working with this compound: - Always wear gloves when working with acrylamide powders or solutions of acrylamide or bis acrylamide. - Wear a mask when weighing these compounds and work in a fume cupboard, if possible. Acrylamide polymerizes to polyacrylamide which is considered to be non-toxic, but polyacrylamides should also be handled with care as traces of non-polymerized acrylamide may be present. It is highly recommended that preweighed acrylamide or acrylamide solutions be purchased to avoid having to weigh the powder. DISPOSAL OF CARCINOGEN-CONTAMINATED WASTE Waste which is or may have been contaminated by carcinogens must be separated from other wastes and placed in appropriately labelled containers: 60 Chapter 7 Safe use of carcinogens Solid carcinogenic waste Solid carinogenic waste such as disposable objects, paper, filters and gloves should be placed in "burnbins” or for larger items, in plastic-lined paper boxes marked with the label CARCINOGENIC WASTE. Liquid carcinogenic waste iquid carcinogenic waste is stored in clear plasticcovered, brown glass bottles which are resistant to both solvents and carcinogens. These containers should be marked with the appropriate label depending on whether aqueous or organic solvents have been used: L Both solid and liquid carcinogenic wastes are collected twice a week by the staff responsible for operating the incinerator, or under exceptional circumstances upon request. Radioactive carcinogenic waste iquid or solid carcinogenic wastes which have been contaminated by radioactive carcinogens must be collected in special containers (either glass bottles or burn-bins) marked with both labels, "Carcinogenic waste” and "Radioactive waste”. L Chapter 7 Safe use of carcinogens 61 Such radioactive waste is kept either until there has been sufficient radioactive decay for it to be disposed of as ordinary waste or it is removed and disposed of by the Agence Nationale pour la Gestion des Dechets Radioactifs (ANDRA). These radioactive/carcinogenic wastes are collected once a week by the ASO Safety Officer, or under exceptional circumstances upon request. DESTRUCTION OF WASTE CARCINOGENS When a carcinogenic compound (pure, in concentrated solution or in dilute solution) is of no further use, it should be destroyed by the principal user or the person responsible for the compound (see the computerized user register in the carcinogen room). Such destruction should be carried out chemically whenever a suitable technique is available. Decontamination techniques for many classes of carcinogens are described in the IARC scientific publication series covering: 'Laboratory Decontamination and Destruction of...', which can be consulted in the IARC library, or obtained from IARC Press (ground floor). The following documents have been published: • No. 37 : Aflatoxins • No. 43 : N-Nitrosamines • No. 49 : Polycyclic aromatic hydrocarbons • No. 54 : Hydrazines • No. 55 : N-nitrosamides • No. 61 : Haloethers 62 Chapter 7 Safe use of carcinogens • • • • No. 64 : No. 73 : No. 113 : No. 114 : Aromatic amines and 4-nitrobiphenyl Antineoplastic agents Mycotoxins Heterocyclic hydrocarbons Chemical destruction methods can be dangerous and instructions should be followed carefully and precisely to avoid accidents. Pure carcinogens or carcinogen solutions for which no adequate chemical destruction method is described in the scientific publications listed above should be disposed of as carcinogenic waste either by incineration or by removal by the French company responsible for the treatment of carcinogenic wastes. An inventory of carcinogenic substances is made every year by B. Chapot. On this occasion, unused or outof-date products can be discarded. Any elimination of a carcinogenic product should be done in collaboration with the safety officer. DECONTAMINATION OF EQUIPMENT AND WORK AREA Carcinogen-contaminated glassware must be kept separately from ordinary glassware and be decontaminated as soon as possible by the user before being sent to the central glassware washroom. The decontamination methods mentioned above can be used for this purpose. Glassware contaminated with radioactive and carcinogenic substances is first washed with a suitable solvent. Rinse solvent is collected in clear plastic-covered glass bottles marked "carcinogenic waste” and "radioactive waste” and one of the following two labels: for an organic solution or an aqueous solution. The glassware is then placed in trays provided for the decontamination of radioactive glassware (chapter 8). Chapter 7 Safe use of carcinogens 63 Glassware which has contained a pure carcinogen should be decontaminated as indicated above, if possible, or placed in a burn-bin for incineration. Cleaning of benchtops and fume cupboards is carried out by an outside company under the supervision of laboratory staff, according to a schedule organised by ASO in collaboration with laboratory technicians. If necessary, laboratory bench tops and fume cupboards should be decontaminated by laboratory personnel before such cleaning and no containers in which carcinogenic or toxic compounds have been stored are to be handled by the cleaning personnel. 64 Chapter 7 Safe use of carcinogens Annex I Proven human carcinogensa Chemicals 4-Aminobiphenyl Arsenic and arsenic compounds Benzene Benzidine Beryllium and beryllium compounds Bis(chloromethyl)ether and chloromethyl methyl ether Cadmium and cadmium compounds Chromium[VI] compounds Coal-tar pitches Coal-tars Ethylene oxide Mineral oils (unrefined) Mustard gas 2-Naphthylamine Nickel compounds Shale oils 2,3,7,8-Tetrachlorodibenzo-para-dioxin (TCDD) Vinyl chloride Chronic infections Epstein-Barr virus Helicobacter pylori Hepatitis B and C viruses Human immunodeficiency virus 1 Human T-cell lymphotropic virus I Human papillomavirus types 16, 18 Liver flukes (Opisthorchis viverrini) Schistosoma haematobium Industrial processes Aluminium production Auramine, manufacture of Boot and shoe manufacture and repair Coal gasification Coke production Furniture and cabinet making Haematite mining with radon exposure Iron and steel founding Isopropanol manufacture (strong-acid process) Magenta, manufacture of Painter (occupational exposure) Rubber industry Strong-inorganic-acid mists containing sulfuric acid Substance abuse Alcoholic beverages Betel quid with tobacco Smokeless tobacco products Tobacco smoke Dust and mineral fibres Asbestos Erionite Silica, crystalline Soots Talc containing asbestiform fibres Wood dust Foods and food contaminants Aflatoxins Salted fish (Chinese-style) Radiation Radon and its decay products Solar radiation Chapter 7 Safe use of carcinogens 65 X-radiation and γ-radiation Neutrons Radium-226 and -228 Plutonium-239 and its decay products Radioisotopes of iodine including iodine-131 Internally deposited α-particle emitting radionuclides Internally deposited β-particle emitting radionuclides Medical drugs and treatments Analgesic mixtures containing phenacetin Azathioprine N,N-Bis(2-chloroethyl)-2-naphthylamine (chlornaphazine) 1,4-Butanediol dimethanesulfonate (busulphan) Chlorambucil 1-(2-Chloroethyl)-3-(4-methylcyclohexyl)-1-nitrosourea (methyl- CCNU) Ciclosporin Cyclophosphamide Diethylstilboestrol Etoposide in combination with cisplatin and bleomycin Melphalan 8-Methoxypsoralen plus ultraviolet A radiation MOPP and other combined chemotherapy Oestrogen replacement therapy Oestrogens, non-steroidal, steroidal Oral contraceptives, combined Oral contraceptives, sequential Phosphorus-32b Radium-224b Tamoxifen Thiotepa Thorium-232 and its decay products (Thorotrast)b Treosulfan aGroup 1 carcinogens, as classified in IARC Monographs Vols 1–82. bRadionuclides with specific medical uses. Chapter 10 Working with animals 117 WORKING WITH ANIMALS ACCESS TO THE ANIMAL FACILITY The animal facility is located on the 13th floor of the IARC tower and is currently supervised by Dr ZhaoQi Wang, Chief of the Unit of Gene-Environment Interactions. The health of the animals housed here is ensured by following the European (FELASA) guidelines. All the rules and regulations concerning access to the animal facility and working with animals are described in the Standard Operating Procedures (SOP) Manual, a copy of which is available from each Unit secretary. This manual must be read before authorization can be given by the Animal Facility Supervisor to enter the animal facility, which is strictly limited to people actively involved in animal research projects. It should be noted that in order to maintain the specific-pathogen-free (SPF) status of the animals and to respect ethical rules, the standard operating procedures must be scrupulously followed and that working in the animal facility necessitates the wearing of certain items of protective clothing (specific laboratory coats, masks, hairnets, shoe covers, etc.). Staff working in the animal boxes after 7p.m. should leave the outer door of the box open so that the night security staff know that someone is present . Daily work in the animal facility is carried out by the animal facility personnel: • Mrs Dominique Galendo, Animal Facility Manager • Mrs Marie-Pierre Cros, Laboratory Technician • Mr José Cardia Lima, Animal Technician • Mr Roland Dray, Animal Technician • Mr José Garcia, Animal Technician • Dr Lionel Zenner, Veterinary Surgeon, Consultant to the IARC animal facility (extension 8013) (extension 8550) (extension 8550) 118 Chapter 10 Working with animals ANIMAL CARE AND USE COMMITTEE he Animal Care and Use Committee (ACUC) was created in 1992 and is currently chaired by Dr Robert Baan. This Committee is responsible for examining all study proposals involving the use of animals at IARC. The Committee is composed of three people nominated by the Director, in addition to the Chairman, the Supervisor of the animal facility and the IARC Veterinary Surgeon. The serving members of the Committee are listed on the Intranet. A detailed experimental protocol must be submitted to this Committee for approval before any animal experiment is started. The forms are available on the Intranet. Requests for importation of animals must be submitted to the Committee, on the appropriate forms available on the Intranet. A copy of the completed form must also be sent to Mrs D. Galendo. T Chapter 11 IARC collection of biological specimens 119 IARC COLLECTION OF BIOLOGICAL SPECIMENS ACCESS TO THE COLLECTIONS All biospecimens (including human tissues, animal tissues, cell lines) kept at IARC are indexed in a central database. Each specimen is identified by a unique IARC identification number. The specimens (frozen or paraffin-embedded) are kept either on Unit’s floors or in common storage spaces. The main storage location is in the basement of the Sasakawa building (room S1). This location contains small liquid nitrogen tanks, –80°C and –40°C deep freezers, and –20°C freezers. This room also houses a small laboratory dedicated to the manipulation of incoming or outgoing specimens. Larger liquid nitrogen tanks are kept in the EPIC building (E03). The rules for access to the common storage space and to the laboratory are described in the Guidelines for Archives of Biological Materials, a copy of which is available from each Unit secretary. These guidelines also provide instructions on how to use the central database. The common storage space and the database are currently supervised by Dr P. Hainaut, Head of the Unit of Molecular Carcinogenesis. Daily maintenance of the infrastructure is performed by an Archive Manager. It should be noted that biospecimens stored at IARC may represent an important source of biological contamination. The guidelines must be scrupulously followed and all manipulations of biospecimens necessitate the wearing of personal protective clothing (laboratory coats and gloves). 120 Chapter 11 IARC collection of biological specimens BIOLOGICAL ARCHIVES COMMITTEE he Biological Archives Committee was created in 2000 and is currently chaired by Dr Pierre Hainaut. This Committee is responsible for overseeing all practical, technical, logistic and computer aspects of the organization of storage facilities. The Committee is composed of three staff members nominated by the Director, in addition to the Chairman, the Archive Manager, the Chairman of the Occupational Health and Safety Committee, the Administrative Services Officer and the Technical Service Officer. The serving members of the Committee are listed on the Intranet. Requests for acquisition of new storage equipment and for the creation of new tissue collections must be submitted to the Committee Chairman. T Chapter 12 Working in the L3 laboratory 121 WORKING IN THE L3 LABORATORY ACCESS TO THE L3 LABORATORY The L3 laboratory has a controlled access and is located in room 901 on the 9th floor. Its use is currently supervised by Dr Massimo Tommasino, Chief of the Unit of Infection and Cancer. All rules and regulations concerning access to the L3 laboratory are described in the Instructions for handling infectious material and genetically modified organisms (GMO) in the L3 laboratory. Anyone planning to perform experiments requiring L3 confinement level must with the agreement of his Unit Chief, submit an application form requesting authorisation to work in the L3 laboratory to the L3 Laboratory Safety Committee. It should be noted that there are medical restrictions to working in such a laboratory; the application will be proved only with the agreement of the Staff Physician. After approval and before starting work, the candidate must be trained in safety precautions and experimental procedures. L3 LABORATORY SAFETY COMMITTEE The L3 Laboratory Committee is currently chaired by Dr Massimo Tommasino, Chief of Unit of Infection and Cancer. The Committee has an advisory, regulatory and teaching role. Its mandate is to ensure that the persons working in the L3 laboratory at the Agency do so under conditions of maximum security and safety. The Committee is composed of the Unit Chief of INC, one representative of a research unit at IARC, one representative from ASO’s office, both of whom are nominated by the Director, in addition to the Staff Physician and the OHSC chairman. Serving members of the Committee are listed on the Intranet.