M AFETY ANUAL - IARC Governancegovernance.iarc.fr/ENG/Docs/safety_manual2009.pdf · The Laboratory...

2009

SAFETYMANUAL

SAFETY MANUAL

Issued by the IARC Occupational Health and Safety Committee (OHSC)

Lyon, France, 2009

SAFETY MANUAL

Foreword ...............................................3

1 Introduction.....................................5Introduction from the Chairpersonof the Occupational Health and Safety Committee (OHSC).......................5Maintaining safety at IARC........................6The OHS committee..................................7Committee members ................................8

2 Safety Procedures ...........................9Evacuation.....................................................9Accidents ....................................................10First aid supplies and equipment ...........11Reporting an accident ..............................11Fire ...............................................................12General security procedures..................12

Theft .......................................................12Use of magnetic access cards to control access to the agency ...........13Latarjet and BRC Buildings ................14Children .................................................14Domestic animals.................................14

Annex ..........................................................15

3 Medical Surveillance .....................17IARC Staff Physician .................................17Periodic medical examinations...............17Individual medical surveillance ...............18

4 General Laboratory Safety ..........19General guidelines ....................................19

Presence in the laboratories ............19Unattended laboratory experiments.19Safety bulletin boards..........................20Instructions to cleaning or maintenance personnel.........................20

Personal safety guidelines........................21Drinking and eating in laboratories .21Storage of food and drink ..................21Cleanliness and laboratory tidiness .21Laboratory coats..................................21Gloves.....................................................22Masks and safety glasses .....................22Protection against UV radiation .......22Noise ......................................................23

Safety procedures .....................................24Safe use of lifts ......................................24Safe use of fume cupboards...............24Safe use of cold rooms .......................24Defrosting and decontamination offreezers or refrigerators ....................25Safe use of liquid nitrogen..................25Safe use of gas cylinders .....................25Safe use of microwave ovens.............26

Laboratory services..................................26Glassware washing service (8th floor) ..............................................26Refrigerators and back-up freezers 26Storage of laboratory consumables.27Disposal of non-contaminated solid waste.............................................27Disposal of contaminated solid waste.............................................28Disposal of non-contaminated liquid waste ...........................................28Disposal of contaminated liquid waste ...........................................28Disposal of dangerous, out-of-date or unidentified chemicals ...................28

5 Laboratory Equipment.................29Hazardous equipment..............................29Fragile and continuously operational equipment...................................................30Unattended equipment............................31Role of staff responsible for equipment...................................................32

6 Storage and Use of Inflammable Substances...........................................33Ordering of inflammable substances ....33Storage of inflammable substances .......34

Authorised quantitites of inflammable solvents ...........................34Storage of waste solvents ..................34Authorised quantities and storage of inflammable radioactive substances..............................................34

Distribution of solvents...........................35Distribution of alcohols ..........................35Use of inflammable gases ........................35

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CONTENTS

2

CONTENTS

7 Safe Use of Carcinogens...............37Ordering carcinogens ..............................37Storage of carcinogens.............................38Working with carcinogens ......................39

Personal protection............................ 39Handling carcinogens in the carcinogen room................................. 39In case of an accident in the carcinogen room................................. 40Transporting carcinogens within the building........................................... 40Handling carcinogens in the laboratory................................................. 40Dichloromethane and chloroform.. 41Benzene ................................................ 41Formaldehyde ...................................... 41Ethidium bromide ............................... 41Acrylamide ........................................... 42

Disposal of carcinogen-contaminated waste .......................................................... 42

Solid carcinogen waste ...................... 32Liquid carcinogen waste.................... 43Radioactive carcinogenic waste....... 43

Destruction of waste carcinogens ....... 43Decontamination of equipment and work area .................................................. 44Annex ......................................................... 45

8 Safe Use of Radioactive Substances 47Authorization to use radioisotopes and medical surveillance ................................ 47

Registration.......................................... 48Systematic surveillance by IRSN ..... 48

IARC laboratories authorized for radioacative materials ............................. 49

Handling of sealed sources.............. 50Ordering radioactive compounds........ 50Storing radioactive compounds............ 50Working safely with radioactive substances ................................................. 51

Personal protection............................ 51Low-energy ß radiation..................... 51High-energy ß radiation .................... 52X and γ radiation ................................ 52Safe handling of radioactive substances ............................................ 52Radioactive waste ............................... 54Storage and disposal of radioactive waste ..................................................... 54Liquid radioactive waste.................... 55Mixed radioactive waste ................... 56Solid radioactive waste...................... 57

Procedure in case of contamination.... 58Decontamination .................................... 58

Decontamination of equipment ...... 58Decontamination of work areas ..... 58Personal decontamination ................ 59

References................................................. 60 Annexes I-X .............................................. 61

9 Safe Use of Biological Substances 71Medical surveillance ................................ 71Confinement level for manipulatinggenetically modified organisms (GMO)and biological samples ............................ 72Handling of biological samples.............. 73

Blood, plasma and serum .................. 73Tissue sections .................................... 74Cell lines ............................................... 74

Sources of contamination ...................... 75Aerosols................................................ 75Scalpels, needles and syringes .......... 75Evaporation/lyophilisation................. 76Centrifugation ..................................... 76Pipetting ................................................ 76

Disposal of contaminated waste .......... 77Decontamination ..................................... 77

Glassware ............................................. 78Disposal of ‘Nalgene’ filters.............. 78Scissors, tweezers and other instruments .......................................... 78Inactivation of HIV.............................. 78Other precautions.............................. 79

Accidents involving potentiallycontaminated biological fluids............... 80Annex ......................................................... 81

10 Working with Animals.................. 83

11 Collections of Biological Specimens ..................................... 85Description of the collections andinfrastructure ............................................ 85Handling biological specimens .............. 85Database .....................................................85

Basement laboratory.......................... 86Procedure for the transport of biological samples .....................................86

12 Working in the L3 Laboratory.... 83Access to the L3 laboratory ................. 83

Index ....................................................88

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Health and safety in the workplace are of fundamental importance to IARC. A commitmentin this area is not only a statutory requirement but presents an opportunity to demonstratethe priority that the Agency places on the well-being of its staff. This new edition of the Manualis therefore an important component supporting the IARC in its mission of cancer prevention.

Activities in laboratory sciences change rapidly, presenting new challenges to safety. The Agencytherefore needs to update and revise its Safety Manual on a regular basis. At the same time,health and safety in the workplace requires a partnership, with all staff and visitors requiredto play their part by reading this Manual carefully and following its recommendations. Finally Iwould like to acknowledge the specific contribution of the Occupational Health and SafetyCommittee in its vital role in this partnership.

Christopher P. Wild, PhDDirector

FOREWORD

INTRODUCTION

From the Chairperson of the Occupational Health and Safety Committee

At the International Agency for Research on Cancer (IARC), considerable resources have beenand will continue to be allocated to maintaining safety in our place of work; however, this canbe 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 everyoneworking on the premises with sufficient information on good laboratory practice in order toavoid dangerous situations.

Before starting work at the Agency, all individuals must follow a training course, which isavailable in English and French on the Intranet, concerning general safety procedures at IARC.They must then complete a questionnaire about this training course and have it validated byBrigitte Chapot, the Laboratory Safety Officer, who will then deliver a safety certificate whichenables new staff to obtain a personal badge giving access to the Agency. This procedure allowsthe Committee to ensure that all newcomers have familiarized themselves with safetymeasures upon arrival.

All newcomers working in the laboratories and/or handling biological samples must follow atraining course on safety and good working practices in the laboratory. They must also fill ina questionnaire which is subsequently checked by B. Chapot, who ensures that everything hasbeen fully understood.

Group Heads are responsible for making sure that all new staff and visitors areproperly briefed and that all safety procedures are respected.

Before starting work at the Agency, everyone should familiarize themselves withthe relevant chapters of the Manual which concern them in particular, andestablish with their supervisors how experimental procedures will be undertakensafely.

As circumstances change, this Safety Manual will be updated accordingly. Staff members withquestions or suggestions can contact any member of the Committee or the Laboratory SafetyOfficer.

The Committee has a website on the Intranet (http://intranet/labcom/) which contains a wealthof information about working in a laboratory. We encourage you to consult it regularly.

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6

1 INTRODUCTION

MAINTAINING SAFETY AT IARC

General safety matters at IARC are under the supervision of the Administrative ServicesOfficer (ASO), Mr G. Guillerminet (extension 8444) and the ASO Safety Officer (extension8523). Mr Guillerminet is responsible on a day-to-day basis for ensuring that general safetyregulations are in place and respected. He coordinates the IARC security team and a team ofstaff with first-aid qualifications who can take immediate action in the event of an emergencyanywhere on the IARC premises.

The Laboratory Safety Officer, Brigitte Chapot (extension 8513), is responsible for matters oflaboratory 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 andis thus responsible for all aspects of safety concerning the manipulation of radioisotopes in theIARC laboratories. She oversees the storage of dangerous compounds, the elimination oflaboratory wastes, with particular responsibility for those that are carcinogenic or radioactive,and the monitoring of specialised laboratories—the carcinogen handling laboratory and thevarious laboratories authorised for the handling of radioisotopes. The Safety Officer assistsher in many of these tasks, and either may be contacted in case of an emergency.

B. Chapot works closely with the Chairman of the Occupational Health and Safety Committee(OHSC), the IARC Staff Physician, Dr D. Cuche, and ASO to ensure that all staff, and inparticular new arrivals, are aware of and comply with the IARC Health and Safety rules.

The IARC Staff Physician, Dr Dorothée Cuche, (extension 8426) is responsible for the medicalsurveillance of all staff, monitoring working conditions and accidents, and evaluating potentialrisks. She is a member of the Occupational Health and Safety Committee and, although notpresent at IARC on a daily basis, she can be contacted through the Human Resources Office(extension 8027).

The interaction of these staff members provides the basis of the health and safety programmeat IARC, but for it to be effective the input of all staff is necessary. The Director and GroupHeads play an essential role in ensuring that the Administrative Services and the LaboratorySafety 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 isthe duty of each person working at IARC, and in particular those working in a laboratory orthose involved in any aspect of maintaining laboratory safety, to ensure that safe work practicesare respected, and to avoid generating potentially hazardous situations.

It is also very important that any incident or accident be made known immediately (StaffPhysician, Laboratory Safety Officer, IARC security team), so that the help of competentpersonnel can be brought in if necessary. Every incident, regardless of its seriousness, must bereported. This obligation concerns not only the staff involved but also their supervisors. Everyincident should be the object of an enquiry with the person involved. This could lead to amodification in procedure in order to prevent the incident happening again.

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INTRODUCTION 1

THE OHS COMMITTEE

The OHS Committee is composed of a Chairman, nominated by the Director, and variousmembers, chosen to represent each of the IARC laboratory floors, the epidemiology groups,the Biological Resource Centre (BRC) and Latarjet building, the IARC Administrative Services(Administration, Technical Services, Safety) and a representative of IARC Staff. The StaffPhysician and the Radiation Safety Officer also serve on the Committee, as required by Frenchlaw.

The Committee considers all aspects of health and safety and in particular:• Emergency procedures• General security measures in the laboratories and Agency buildings• Medical surveillance• Safe use of laboratory equipment• Safe handling of flammable substances• Safe handling of carcinogens• Safe use of radioactive substances• Safe use of biological materials, genetically modified organisms (GMO) and the L3

laboratory• Biological resources

The Committee meets every three months but can be convened in an emergency bycontacting the Chairman or the Laboratory Safety Officer. All practical aspects of laboratorysafety and working conditions at IARC are discussed, and anyone working at IARC may contacta member of the Committee if there is a health or safety issue they would like to have raisedeither at the Committee level or with the IARC Administration. Practical safetyrecommendations are made to staff directly by the Committee, via the safety bulletin boardon each floor and on the Intranet. The minutes of each meeting are accessible on theCommittee's Web site. When policy decisions are required, the Committee can makerecommendations to the Group Heads and to the Director.

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1 INTRODUCTION

COMMITTEE MEMBERS

P. Hainaut Chairperson OHSC

7th floor

D. BouchardSecretary7th floor

C. Cuenin8th floor

B. SyllaBiological

Substances9th floor

E. CabouxBiologicalResource

Manager, BRC

A.-M. Camus-Randon6th floor

P. BarbieuxTechnicalServices

Basement

A.-M. Aguelon 9th floor

B. ChapotLaboratory

Safety OfficerBRC

Dr D. CucheIARC StaffPhysician

Ground floor

J. FerlayLatarjet

N. Forey10th floor

G. GuillerminetAdministrativeService Officer

2nd floor

J.A. PedilASO Safety

OfficerBasement

J. McKayStaff

Representative4th floor

B. VozarBRC

SAFETY PROCEDURES

EVACUATION

Only the stairs may be used!

In an emergency in any of the IARC buildings (fire, flood,earthquake, etc.) a siren with a varying pitch will be sounded.On hearing this warning, all staff in that building shouldimmediately leave their office or laboratory and make theirway as quickly as possible, without running, to the nearestexit. Only the stairs may be used. All cumbersome articlesshould be left behind.

Emergency wardens have been appointed for each floor of all the IARC buildings. Their namesare listed on notices posted on the door of the “Pater Noster” (the internal mail deliverysystem) in the Tower and in the corridors of the BRC and Latarjet buildings. They will supervisethe evacuation and ensure that everyone on their floor has heard and obeyed the warning. Ifyou are on the telephone when the alarm is sounded, you should explain rapidly to your callerand hang up immediately. Each emergency warden should confirm that his/her floor has beenevacuated at the reception on the ground floor in the main building and then proceed to theassembly point.

Persons who panic, are disabled or collapse should be taken or carried to the safety of thestairwells, normally by not fewer than two and not more than four other staff members, andhelped to descend to ground level. If they are incapacitated, their presence should beimmediately signalled to the reception on the ground floor of the Tower so that the emergencyservices can take the necessary action.

On reaching the ground floor, staff members should leave the building and go to the assemblypoint. In the case of an evacuation of staff working in the Tower, the assembly point is locatednext to the bicycle shed outside the Latarjet building. In the case of evacuation of staff workingin the BRC or Latarjet buildings, the assembly point is in the car park immediately behind theTower (see Annex I, page 15). Instructions from those responsible for security must be obeyed.Staff 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 theemergency exits, the security instructions and the names of the emergency wardens on theirfloor.

In order to accustom staff members to the siren, it is sounded at noon for 15 seconds on thefirst working Monday of each month in the Tower building.

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2 GENERAL SECURITY PROCEDURES

ACCIDENTS

Any person witnessing one of the following accidents:• explosion• fire• bodily injury, fainting, etc.• electrocution (in this case, first shut off the mains power if possible)• splashing by corrosive or biologically active or radioactive substances

The red button is situated opposite the Pater Noster (the internal mail delivery system) in themain building, opposite the stairwell in the BRC building and near the emergency exits in theLatarjet building (2 exits per floor).

The security staff will take immediate action to protect the individual(s) at risk and will, ifnecessary, call for help from the fire brigade or the S.A.M.U. (French medical emergencyservices).

Before 8.30 a.m. and after 5.00 p.m., the alarm alerts the security guards at reception who havebeen instructed on how to proceed in case of an accident.

If the red button does not work, you should contact the switchboard or the security guardsdirectly 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.

It is forbidden to call outside emergency services directly as these services need to work withthe security team and those at reception (opening of barriers, shown place of accident, etc)If an injured person requires medical attention, he/she will be accompanied by a qualifiedperson to the hospital using appropriate transport.

In exceptional circumstances and only if the first two procedures have not worked, you maycall for help directly using one of the following numbers :

must break the glass of the red button to alert IARC securitystaff who have had first aid training.

Lyon Fire Department 18Emergency Medical Aid ‘S.A.M.U.’ 15Hôpital Edouard Herriot - Emergencies 04 72 11 60 80Poison Control Centre 04 72 11 69 11

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GENERAL SECURITY PROCEDURES 2

A list of all qualified first-aiders is displayed on the Safety Bulletin Board (located on the floorsopposite the red lift in the Tower, on the ground floor in the BRC and Latarjet buildings).

FIRST AID SUPPLIES AND EQUIPMENT

Having adequate first aid and safety equipment in a laboratory isthe best way of ensuring quick help in the case of an accident. Oneach laboratory floor, safety showers are available for use bylaboratory workers who have been splashed with toxic orcorrosive chemicals. Showers are also available in the Towerbasement. Eyewash stations have also been installed near the safetyshowers as well as in each laboratory. Emergency fire blankets,located in the corridors on the 6th, 7th, 8th, 9th and 10th floors, aswell as on the ground floor of the BRC building, should be used toextinguish clothing which has caught fire.

The first aid cabinets located on each floor are equipped only withbasic supplies and can be replenished from the stock kept by ASO.More extensive first aid supplies can be obtained from thereceptionist or security guards on the ground floor of the mainbuilding.

Emergency equipment (defibrillator and oxygen bottle) can be found on the ground floor ofthe Tower and BRC. A defibrillator only has also been installed on the ground floor of theLatarjet building. This equipment is available to those staff trained in its use.

REPORTING AN ACCIDENT

Any person involved in an accident, including visitors, should report it to the Staff Physician andto the Laboratory Safety Officer.

Any staff member who has an accident must complete a copy of form WHO 417 and have theattending doctor complete a copy of form WHO 418. These forms are available from theHuman Resources Office on the 2nd floor (216).

The staff member must forward form WHO 417 to his Group Head if the accident occurredduring working hours. The supervisor must report on the circumstances of the accident, andin particular:1. Whether the staff member was on duty2. What his/her particular duties were at the time of the accident, the circumstances, the date

and the location.

The Group Head may write his report on a separate sheet of paper, to be attached to formWHO 417. The Group Head, or the staff member as the case may be, must then send thiscompleted form to HRO.

Form WHO 418 must be completed by the attending doctor and then sent to HRO, who willtake appropriate action.

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FIRE

In laboratories, the greatest risk to personnel from fire comes from the use of flammablesolvents. Flammable solvents must be stored in the laboratories, in limited amounts, inexplosion-proof refrigerators. They must not be used near a hot electric element, an electricmotor or a naked flame. The use of naked flames is prohibited in all the IARC buildings, exceptin certain exceptional circumstances (see Chapter 6). A special room for the storage ofinflammable solvents is located near the unloading area in the basement of the Tower and isopen 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 drill to ensure that all staff members know how to use the fireextinguishers located on each floor correctly. As safety is everyone's responsibility, it isimportant that staff members become familiar with the location, and the correct use, of thesered fire extinguishers. It is important to remember that a red extinguisher should never beused on a person as the pressure is too high. However, green extinguishers contain water andcan be used as safety showers.

All newcomers at IARC should be informed of the available safety equipment and proceduresthrough a briefing by their Group Head or his representative and by a tour of their workplace.

GENERAL SECURITY PROCEDURES

Theft

IARC is not insured against, nor is it liable for, claims resulting from the loss or theft of personalproperty on its premises. Staff members should therefore not leave personal property of value,in particular mobile phones, handbags or wallets containing money, or chequebooks, inunattended offices and/or laboratories, or in unlocked desks or cupboards at any time. Similarprecautions should be taken for any office equipment such as calculators and laptops, etc.

Nothing of value should be left visible in cars, which should be locked when parked. If a valuablearticle disappears, the loss should immediately be reported to ASO.

Smoking is forbidden everywhere in all IARC buildings and in the IARC grounds

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Use of magnetic cards to control access to the Agency

IARC has a system of controlled access to ensure improved security.

The controlled entrances are as follows:

• entrance & exit of the car park• entrances & exits of the Tower building• entrance & exits of the Latarjet building• entrance & exists of the BRC building• loading area gate• bicycle shed• computer room (403)• radioactivity and carcinogen room (805)• L3 laboratory (901)• Bacteriology laboratory (919)• L2 laboratory (920)• radioactivity storage room (SS/SMH)• radioactivity storage room (S05)• storeroom (S19)• room E01 BRC building• room E02 BRC building• room E03 BRC building

A credit card-sized badge, containing a miniature low frequency emitter/receiver programmedfor each individual staff member, permits access to all or certain of the above-mentioned areas.

All newcomers at the Agency, whatever their level or status, must follow a training courseconcerning general safety procedures at IARC. They must then complete a questionnaire aboutthis training course and have it validated by the Laboratory Safety Officer in order to obtaina personal badge giving access to the Agency which is issued by ASO.

This badge gives access to certain parts of the Agency and can also be used as a means ofpayment in the Cafeteria. It can be clipped to a piece of clothing or worn around the neck. Itshould not be left in the car, the office or in a pocket but worn permanently.

At any controlled access point, each badge holder should present their badge in front of thecard reader at a distance of about 20 to 30 cm. If the card’s code is recognised, the cardreader’s signal light will change from red to green and access will be given. A central computercontinuously records information on access points that have been activated.

It is thus important that every time staff go through the main entry/exit points of the differentIARC buildings, each person should use their badge so that in the event of an incident thenumber of staff in any building can be monitored.

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Any visitors in the Agency, even for a short period should be registered at the Reception andwill receive a visitors badge which should be worn visibly. Every visitor should always beaccompanied by an IARC staff member.

If, for any reason, someone does not have their badge (left at home, etc.), they should sign inat the Reception as a visitor.

Any loss of a badge should be immediately reported to ASO’s office.

Latarjet and BRC buildings

The Latarjet and BRC buildings are protected by an alarm system which is activated:• at night (8.00 p.m. to 7.00 a.m.),• during weekends• on IARC public holidays

During these periods, access to the BRC and Latarjet buildings is possible by going to see theguard in the entrance hall of the Tower. You must inform her/him when you leave the BRC orLatarjet buildings so that the alarm system can be reactivated.

Staff working in the BRC or Latarjet buildings should also remember to close all windowswhen leaving the building each evening.

Children

Children under 16 are not allowed access to any of the IARC buildings with the exception ofthe reception hall and the cafeteria (mealtimes only). They can eat with their parents andshould then be accompanied back to Reception.

Domestic animals

No domestic animals are allowed in any Agency building,.

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Annex : Position of IARC Assembly Points

AssemblyPoint 1

For people working inthe tower building

Assembly Point 2

For people working inthe EPIC and Latarjetbuildings

MEDICAL SURVEILLANCE

IARC STAFF PHYSICIAN

The IARC Staff Physician is responsible for the medical surveillance of all staff, monitoringworking conditions and accidents, and evaluating potential risks, if necessary in conjunctionwith the Occupational Health and Safety Committee.

The Physician advises the Director of the Agency on working conditions, in special cases ofsickness or accident, on the need for a change of job or for an improvement in a workenvironment, with the understanding that all medical information is strictly confidential.

The Staff Physician examines staff members when they return to work after sick leave of morethan 21 days, anyone who has had a work-related accident and women staff members on theirreturn from maternity leave. The files are sent under confidential cover to the Joint MedicalService at WHO in Geneva.

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 EXAMINATIONS

These examinations are carried out as follows:

• Staff members under the age of 40: A full examination every 5 years, with one intermediate general physical examination withoutbiological 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

The frequency of these examinations can be decided by the Staff Physician depending on theperson’s health.

For non-staff members working at the Agency in an epidemiology group and holding a contractof more than three months, a medical examination is required before recruitment.

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3 MEDICAL SURVEILLANCE

For non-staff members working in a laboratory group, a medical examination is required beforerecruitment whatever the length of the contract. Moreover, when starting work at IARC, theperson should present their vaccination certificates, showing that they are immunized againsthepatitis B.

The Staff Physician should be consulted on any medical issue or update of vaccinations. He/shecan ask for further medical tests to be carried out if necessary.

INDIVIDUAL MEDICAL SURVEILLANCE

In addition to these routine medical examinationss, the Staff Physician is also responsible forsupervising any additional medical surveillance that is required, including:

• Analysis of annual biological examinations of laboratory personnel including blood tests and urinary cytopathological examinations

• Analysis of questionnaires concerning the use of dangerous chemical products and biologicalagents handled in the laboratories

• The specific monitoring of all staff who work with radioisotopes (6-monthly radiotoxicological urine analysis, 3-monthly check of dosimeter results, etc) and any othertest required by the Staff Physician

• Medical examination of any person involved in an accident at work

• Medical examination of any person requesting one, either for themselves or a member oftheir family

• Vaccination programme for the prevention of infectious risks such as hepatitis B, tetanus, polio, influenza, etc

• Medical aid or prescriptions can be given in emergencies to all staff

All women working at IARC are required toinform the IARC Staff Physician as soon as they

are aware that they are pregnant.

GENERAL LABORATORY SAFETY

GENERAL GUIDELINES

Presence in the laboratories

Only authorized persons who possess a magnetic badge are allowed access to the IARCbuildings.

It is very important that each person use their badge to enter and leave the Agency. This meansthat it is possible to know who is on site if necessary.

If, for any reason, staff do not have a badge (left at home, etc.), they should sign in as a visitorat the Reception.

It is especially important to use one’s badge when present in any of the Agency buildingsoutside normal working hours:

• On normal working days: BEFORE 8.00 a.m. and AFTER 5.00 p.m.

• Outside working days: at weekends and during IARC holidays

Unattended laboratory experiments

It is sometimes necessary to leave unattended certain laboratory experiments that make useof specific pieces of equipment which, if they malfunction, could present a danger either to theoperator or to others in the laboratory or which could result in damage to the equipmentitself (see chapter 5). This situation should be avoided if at all possible. However, when it isnecessary to leave such equipment running, the following procedure should be strictly adheredto:

1. During normal working hours, either the log-books of such equipment (e.g. ultracentrifuges, speed-vacs, etc.) should be filled in noting the name of the operator, or a clear label should beattached 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 examplebelow) fixed on the door of the laboratory in which the equipment is located should becompleted in giving the date, the name of the equipment functioning, the name of the operatorand a telephone number at which this person can be contacted. This information will allowthe night security personnel to contact the staff member who is using any piece of equipmentif it develops a problem. It is important that when the equipment is subsequently switched offthe notice is duly modified.

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IT IS DANGEROUS TO WORK ALONE IN A LABORATORY

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4 GENERAL LABORATORY SAFETY

All equipment that 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 staffmembers who can be contacted in case of emergencies. It is the responsibility of the GroupHead to ensure that these names are updated as and when necessary.

Safety bulletin boards

Safety bulletin boards are located on each floor of the Tower on the wall opposite the red lift,and on the ground floors of the BRC and Latarjet buildings. Information concerning changesin safety regulations, dates of medical examinations and other information related to safety,including a list of qualified first-aid staff at IARC is posted on these boards .

Instructions to maintenance or cleaning personnel

The cleaning personnel who work in the evening are responsible for cleaning the floors,emptying the waste paper baskets, removing the cardboard bins used for the disposal of brokenglass, which have been placed in front of the service lift by laboratory staff, and taking thecardboard bins containing recycled paper down to the basement.

Cleaning personnel present in the laboratories during normal working hours are under theresponsibility of laboratory staff who should ensure that:

• Cleaners have taken any necessary health and safety precautions (wearing gloves, laboratory coats, choice of cleaning materials, etc.); and

• Cleaning procedures in these laboratories are the right ones.

No cleaning of equipment should be carried out without firstobtaining permission from laboratory staff.

Radioactive and carcinogenic waste containers must be clearly marked and under nocircumstance are to be picked up or thrown away by the cleaning personnel.

To better ensure their safety, cleaning personnel should not work in laboratories where thehandling of radioactive substances is authorized or tissue culture is carried out, or in thecarcinogen room (805).

Equipment running overnight

Date Equipment Operator Contact telephone number

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GENERAL LABORATORY SAFETY 4

The cleaning of these laboratories is carried out separately under the supervision of the staffmember responsible who must ensure that there are no hazards present.

PERSONAL SAFETY GUIDELINES

Outlined below are some good laboratory practices that should be observed to reduce therisk of exposure to potentially dangerous chemicals, equipment or situations.

Drinking and eating in laboratories

This rule also covers eating at desks in laboratories!

The use of kettles and coffee machines is also forbidden outside of the kitchen on the groundfloor of the Tower. Drink and snack distributors are located on the ground floor of the Tower.

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 inroom 1201 on the 12th floor. They may be used by all staff.

Cleanliness and laboratory tidiness

Elementary rules of cleanliness should be respected, i.e. wash your hands often, especially afterhandling biological substances, after wearing gloves and before leaving the laboratory. Thelaboratory should be kept tidy and free of items that have no relation to laboratory work.

Laboratory coats

For the safety of all staff members, fastened laboratory coats must always be wornin the laboratories, regardless of the type of work being carried out.

It is strictly forbidden to eat or drink in thelaboratories

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However, it is strictly forbidden to wear laboratory coats in the cafeteria, the kitchen or othermeeting rooms.

A system of renting and washing protective laboratory clothing is in place at IARC. Laboratorycoats marked with your own name, can be obtained from Supplies (ext. 8464). Dirtylaboratory 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.

IARC short-term personnel should obtain a laboratory coat from the supplies store in thebasement. The person responsible for the supplies store will when necessary replace soiledcoats in exchange for clean ones.

Gloves

Latex, vinyl or rubber gloves must be worn when working with dangerous substances. Tosafely weigh carcinogens that are electrostatic, cotton gloves, available in room 805, should beworn.

After use, contaminated gloves must be disposed of in special waste containers. These binsshould be clearly marked with the appropriate label: carcinogens, radioactive materials orbiohazards.

Gloves must not be worn in any of the four principal lifts, in the larger service lift or in thestairways. The wearing of gloves is tolerated in the red service lift. However, food is strictlyforbidden in this lift.

Always remember that after use, gloves may be contaminated. Special precautions shouldtherefore be taken to remove them before touching any objects, to avoid their contamination(laboratory instruments, pens, telephones, door knobs, electric light switches, laboratorynotebooks or radiation detectors, etc.).

Masks and safety glasses

Certain laboratory experiments require the wearing of safety glasses.Standard safety glasses are made available wherever experiments thatare judged to be dangerous are carried out. These places are indicatedby the blue adhesive sign (see right) indicating that the wearing ofsafety glasses is compulsory. Safety glasses are also provided to allnew staff in laboratory groups.

Always remember to wash your hands afterremoving gloves and before leaving the laboratory

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Safety glasses must be worn:

When handling any of the following substances:• Liquid nitrogen (mask recommended)• Strong acids such as hydrochloric acid, sulphuric acid, nitric acid, trichloroacetic acid, glacial

acetic acid, sulfochromic acid, etc.• Solutions of strong bases such as sodium hydroxide, potassium hydroxide, etc.• Corrosive agents such as phenol, etc.• Halogenated solvents such as dichloromethane, chloroform, etc• Organic solvents such as acetone, isoamyl alcohol, etc.• Radioactive isotopes• Carcinogenic compounds• Chlorine bleach

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 • Ultra-centrifuges• Lyophilizers• Autoclaves

During any other operation which involves an increased risk (projections,explosions, implosions)

Contact lenses may not be worn in the situations cited above. Staff members with a particularproblem (those who find it impossible to wear glasses with prescription lenses) should consultan ophthalmologist and send the results to the IARC Staff Physician.

Protection against UV radiation

For all work that exposes eyes to ultra-violet radiation, the wearing of special UV safety glassesor, even better, a UV visor is compulsory. This safety equipment is made available near UV-emitting equipment.

Noise

In general, noise that exceeds the commonly accepted noise limit of 85 decibels on the "A"scale (85 db A) for eight hours exposure, will not occur in the laboratory. However, ifnecessary, ear protection can be obtained from ASO when working near noise sources suchas ultrasound cleaners or sonicators.

Normal glasses or safety glasses do not provideadequate protection against sources of UV radiation

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SAFETY PROCEDURES

Safe use of lifts

The red service lift is the only one authorised for use by laboratory personnel totransport chemical materials, solvents and any other laboratory product. It istherefore restricted to laboratory staff for this use.

Safe use of fume cupboards

Any operation which is likely to generate toxic fumes or even an unpleasant odour should becarried out in a fume cupboard. For maximum protection, close the sash completely and workwith hands extended through the two round openings. For less hazardous operations, workcan be carried out with the sash front positioned in such a way that the two red arrows onthe left of the sash coincide (air speed = 0.5 m/sec).

When a fume cupboard malfunctions, a red light and an alarm bell automatically indicate aproblem.

Do not work in a fume cupboard that is not working properly; call the technical staff.

Fume cupboards should not be used to store dangerous items.

The efficiency of each fume cupboard is checked periodically with an anemometer and by theburning of a smoke bomb; the dates of the last checks are noted on the sash front of the fumehood.

Safe use of cold rooms

Materials stored in these rooms should be clearly marked with both the date of storage andthe name of the person responsible for the material. To avoid the growth of undesirable micro-organisms, cardboard boxes should not be stored in cold rooms, and all bottled liquids mustbe placed in plastic or metal trays in case of breakage. Nothing should be stored on the floor.One person per floor is responsible for the correct use of each of these cold rooms:

• 6th floor : A.M. Camus-Randon• 7th floor : G. Martel-Planche• 8th floor : C. Cuenin• 9th floor : A.M. Aguelon• 10th floor : F. Le Calvez

The cold rooms in the Agency are cleaned and decontaminated regularly.

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Defrosting and decontamination of freezers and refrigerators

Most refrigerators and freezers are connected to a central alarm system that alerts technicalstaff or security personnel if their temperature control fails. Thus, before undertaking thedefrosting of a freezer or refrigerator, please inform P. Barbieux (ext. 8571). Short-termreplacement freezers and refrigerators are available for storing material during suchprocedures.

Refrigerators and freezers should be regularly emptied and defrosted to avoid accumulationof ice. They should be periodically cleaned using diluted detergent (Bactinyl).

In the event that a refrigerator or freezer is found to be contaminated with a biological sampleor a chemical agent, contact either the Laboratory Safety Officer or the ASO Safety Officer, ifit is located in an IARC laboratory, or the Biological Resources Manager if it is located in thefreezer location in the basement to inform them of the problem and to ensure that the correctdecontamination protocol is followed. (See ‘Guidelines for IARC Archives of BiologicalMaterials’ for detailed protocols).

Safe use of liquid nitrogen

IARC has a liquid nitrogen reservoir located in front of the BRC building. Liquid nitrogencontainers are located in the basement of the Sasakawa Hall, as well as in the BRC building.These containers are regularly filled by the staff members responsible for the distribution ofliquid nitrogen. As liquid nitrogen is very cold (-196°C) and can quickly freeze skin, it isnecessary to wear safety glasses or a mask and suitable gloves when handling this substance.Such equipment is available in the Sasakawa basement and in the BRC building, and should beworn when removing samples from the liquid nitrogen tanks.

Liquid nitrogen should only be transported in suitable sealed containers that are nothermetically sealed allowing evaporation of any gases formed. It should not be transported incompletely open containers either, as liquid could be spilled onto someone in the event of afall or accidental contact with another person.

Safe use of gas cylinders

Nitrogen, carbon dioxide, oxygen, helium and argon cylinders are used in the IARClaboratories. Such cylinders must be securely attached to the wall with a safety chain. Theusers of such equipment should periodically verify the tubing to ensure the absence of leaks.The use of inflammable or toxic gases (hydrogen, methane, etc.) is only possible using smallcapacity cylinders after prior agreement from ASO.

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Safe use of microwave ovens

Microwave ovens, often models purchased for kitchen and not specifically for laboratory use,are increasingly being used to heat aqueous solutions in the laboratory. In addition to thewidely known rule that metal objects should not be placed in a microwave oven, a serious riskof explosion exists if aqueous solutions contained in tightly closed glass bottles are heated ina microwave oven. Particular care should be taken when melting agarose: the container shouldnot be closed with a fitted lid but plugged loosely with a folded paper handkerchief to allowgas to escape. Microwave ovens should not be used to heat organic solvents.

LABORATORY SERVICES

Glassware washing service (8th Floor)

The glassware washing service is common to all IARC laboratories, with each of the washroompersonnel being responsible for the handling of glassware from several research groups.

Only non-contaminated laboratory glassware should be sent for washing. Dirty glassware isplaced by the user into 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 respectivelaboratories.

The washroom staff can take care of other minor cleaning jobs, for example the cleaning ofsmall laboratory apparatus, boilers or incubators, but only after they have been unplugged anddecontaminated by the laboratory staff.

Refrigerators and back-up freezers

Should the temperature of a freezer which is protected by the alarm system go over a certainlimit, the security staff are alerted.

During normal working hours, the technical staff are informed and can intervene if necessary.In the case of a breakdown, the contents of the freezer are transferred to a back-up freezerby the staff of the group concerned.

Outside normal working hours, a rotating system has been put in place to monitor the freezers.Each group wishing to include its freezers in the communal system should provide the nameor names of those responsible. These people would be nominated for a week and would becontactable via a provided mobile phone which is provided. In the case of a breakdown and/oran alarm going off, the person responsible is contacted by the Security staff. This person shouldbe capable of coming to the Agency and, if necessary, transferring the contents of themalfunctioning freezer to a back-up freezer.

In all cases, samples that have been transferred must be placed in closed containers which areclearly identifiable to avoid the risk of contaminating the back-up equipment (see “Guidelinesfor IARC Archives of Biological Material” for further details). Back-up freezers are available(four -80ºC, one -40ºC and two –20ºC back-up freezers).

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All breakdowns must be reported on the Incident Report form, which must be counter-signedby the Group Head and returned to the Biological Resources Manager.

In order for this service to function well, storage time for samples in back-upfreezers is limited to a maximum of one month. The contents of the backupfreezers will be checked regularly, and any non-identified samples found in a back-up freezer will be subject to immediate disposal. The back-up equipment may alsobe used for temporary storage of samples during the decontamination of freezersand refrigerators. Contact the person responsible for the BRC for further details.

Storage of laboratory consumables

One room located on the 1st basement level is reserved for this purpose. Since this room isrelatively small, the amount of laboratory consumables stored here should be limited to avoida potential fire hazard.

Disposal of non-contaminated solid waste

Several types of waste containers for non-hazardous wastes can be found in the Agencylaboratories:

• Black or brown plastic wastepaper baskets: These bins are meant for the kind of waste generated in an office (i.e. paper, plastic, etc., but no sharp or dangerous objects); they are emptied daily by the cleaning personnel.

• Cardboard boxes: These boxes are intended to collect paper for recycling only. These boxes are emptied by the cleaning staff.

• Plastic-lined cardboard bins: These bins are intended for the disposal of non-contaminated broken glass only. Full boxes should be placed in front of the large service lift (monte-charge) to be taken by the cleaning staff down to the basement where they will be disposed of by an outside company.

• Used glass container: This bin, located in the basement, is provided by the COURLY and is intended for the disposal of clean glass bottles, which are subsequently recycled. Toensure that glassware is clean before being thrown away, empty solvent bottles should beleft open in the hood for 24 hours to permit evaporation of any residual solvent before disposal.

Office waste Paper to be recycled

Non-contaminatedbroken glass

Clean glass to be recycled

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Glassware that is known to have contained toxic, carcinogenic or radioactivesubstances should NOT be recycled under any circumstances!

‘Sharpsafe’ plastic boxes must be used for the disposal of small sharp objects suchas needles and glass Pasteur pipettes.

Disposal of contaminated solid waste

All hazardous solid waste should be placed in the appropriate waste bins, which have beenclearly labelled to indicate the kind of waste they contain:

Biohazardous waste: collected regularly by the staff responsiblefor waste collection (See chapter 9 formore details)

Carcinogenic waste: collected regularly by the staff responsiblefor waste collection (See chapter 7 formore details)

Radioactive waste: collected regularly by the ASO SafetyOfficer for disposal by an outsideorganization or stored until the radioactivehas decreased sufficiently to permit disposalas non-radioactive waste (See chapter 8 formore details)

Disposal of non-contaminated liquid waste

See chapter 6 for information on storage and use of inflammable substances.

Disposal of contaminated liquid waste

See chapters 7, 8 and 9 for information on disposal of contaminated liquid waste.

Disposal of dangerous, out-of-date or unidentified chemicals

Each year, the Laboratory Safety Officer organizes a collection of all dangerous, out-of-date orunidentifiable chemicals that are no longer used.

LABORATORY EQUIPMENT

At IARC, there are two main categories of laboratory equipment that require particular carebecause their use could either result in a health risk (burns, electrocution, radiation, etc.) tothe operator or because the instrument itself could be damaged by improper or inappropriateuse. Some instruments may belong to both categories.

It is absolutely essential that anyone using equipment in either category for the first timefamiliarises themselves with the operating procedures and potential dangers which could beencountered, by consulting the person responsible for that piece of equipment in the planningstages of any experimental procedure. Under certain circumstances specific training must becompleted under the supervision of either the manufacturer or a trained operator of themachine.

Some equipment can only be used by a restricted number of operators who have receivedspecific training from the manufacturer (autoclaves).

HAZARDOUS EQUIPMENT

This category includes laboratory equipment whose use could result in a health risk (burns,electrocution, radiation, etc.) to the operator. The following information should be clearlydisplayed 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, for example)

• 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 clear that for such itemscompleting 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 ofequipment and they must use the appropriate safety equipment (e.g. glasses or eye shields forfluorescent tables).

Similarly, it is impractical to have a users' logbook for a liquid nitrogen container, but the correctprocedures for handling must be respected and the dangers relating to its use must be fullyunderstood by all users as well as the wearing of the correct protective clothing (see chapter 4).

For some items of equipment (e.g. ultra-centrifuges), the logbook is absolutely essential andmust be completed every time the equipment is used.

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30

5 LABORATORY EQUIPMENT

Hazardous equipment at IARC

FRAGILE AND CONTINUOUSLY OPERATIONAL EQUIPMENT

Equipment in this category can be easily damaged if not used correctly (power supplies, HPLC,robots, pumps...). Their contents may be damaged or even destroyed if not functioning correctly(cold-rooms, fridges, freezers, CO2 incubators). The following information should be clearlydisplayed for this type of 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 be requested for certain pieces of equipment

(e.g. temperature of operation)• a maintenance logbook

Concerning freezers, please refer to Chapter 4.

Apparatus RisksAutoclaves BurnsCentrifuges ProjectionsFluorescent tables UV burnsHPLC ProjectionsLiquid nitrogen containers Burns, frostbiteLyophilisers Implosions, projectionsMicrotomes CutsMicrowave ovens Explosions, burnsPower supplies ElectrocutionSpeed vac concentrators Implosions, projectionsUltracentrifuges ProjectionsUltrasonic processors NoiseUV/vis spectrophotometers UV exposurWater-baths Burns

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LABORATORY EQUIPMENT 5

Fragile and continuously operational equipment at IARC

UNATTENDED EQUIPMENT

(See chapter 4 for more details)When it is necessary to leave equipment running unattended, the following procedure shouldbe carefully followed:

1. During normal working hours, either the equipment logbooks should be filled in (e.g.ultracentrifuges, rotary evaporators, speed vacs, etc.) noting the name of the operator, or aclear label should be attached to the equipment itself (e.g. gel electrophoresis equipment,hybridization ovens, etc.) giving this information.

2. Outside normal working hours, in addition to this requirement, the notice (see next page)fixed on the door of the laboratory in which the equipment is located should be completedgiving the date, the name of the equipment functioning, the name of the operator and atelephone number at which this person can be contacted. This information will allow the nightsecurity personnel to contact the staff member who is using any piece of equipment if itdevelops a problem. It is important that when the equipment is subsequently switched off thenotice is duly modified.

ApparatusAutoclaves* Hybridization ovensAutomatic injectors HPLC/DHPLCCO2 incubators* IrradiatorCold room +4°* Mass spectrometerDNA extractor Vacuum pumpDNA sequencer Water bathElectrophoresis apparatus Paraffin inclusion equipmentFACS machine PCR systemsFilm developer Permuted water stationFluorimeter Phosphor-imager analyzerFreezers -20°C* Refrigerators*Freezers -80°C* Scintillation counterGas chromatograph SpectrophotometerGel dryer TEA 502 (Thermo-electron)*Piece of equipment with a green label:

Personne responsable – Responsible personNom / Name :Poste / Ext. :Groupe / Group :

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5 LABORATORY EQUIPMENT

ROLE OF STAFF RESPONSIBLE FOR EQUIPMENT

Those in charge of any piece of equipment falling into either of these categories must:

1. Ensure that the user and maintenance logbooks are available and are routinely completed.2. Ensure that a user’s guide, in English and French, is 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 possibly the OHS Committee) if a potential hazard

exists.

Equipment running overnightDate Equipment Operator Contact telephone number

STORAGE AND USE OF INFLAMMABLESUBSTANCES

ORDERING OF INFLAMMABLE SUBSTANCES

Inflammable substances for which special precautions must be taken should carry a label similarto that shown opposite. Normally, containers of these substances, when purchased in France,carry a label containing the word "flammable" and showing the picture of a flame. However,certain solvents are purchased in bulk and transferred to smaller containers. It is theresponsibility of the person receiving a shipment of unlabelled containers or transferringinflammable liquid to smaller containers to mark each such container with a suitable label.These labels are available from the ASO Safety Officer.

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Non-exhaustive list of inflammable substances used at the Agency

Amines EthersEthylamine Dimethyl etherTriethylamine Diethyl ether

Ethylene oxideAlcoholsButanol HydrocarbonsEthanol PenteneMethanol IsopentanePropanol Hexane

CyclohexaneAldehydes & ketones TolueneFormaldehyde (gas) XyleneAcetonitrileAcetone Miscellaneous

AcetaldehydeEsters Dechloroethaneacetate Carbodisulfide

PyridineScintillation fluids

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6 STORAGE AND USE OF INFLAMMABLE SUBSTANCES

STORAGE OF INFLAMMABLE SUBSTANCES

Authorised quantities of inflammable substances

French regulations require that a maximum of 150 litres of inflammable solvents may be in useat any one time in the laboratories of the main building. The table below lists maximumauthorised quantities per floor.

Requests for changes in these limits should be forwarded to the Laboratory Safety Officer.During periodic checks to see if these limits have been respected, the ASO Safety Officer willtransfer excess solvents to the basement storage room. When not in use, any inflammablesolvents 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 plasticcontainers (5 litres capacity) clearly labelled.

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) clearlylabelled to indicate whether they contain either aqueous ororganic solutions (see label opposite).

Waste solvents are collected on each floor at the same time as contaminated waste. They aresubsequently collected by an outside company.

Authorised quantities and storage of inflammable radioactive substances

If your experimental protocol involves the use of inflammable radioactive substances, contactB. Chapot before starting any experiment to organise appropriate waste containers. These willdepend on the amount of radioactivity and the isotope involved. The waste containers mustbe marked with labels specific for this purpose which are available from the ASO Safety Officerand may only be stored in limited amounts in the authorised laboratories, from where they willbe collected.

Floor Maximum quantity10th 10 litres9th 10 litres8th 25 litres7th 30 litres6th 40 litresTotal 115 litres

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STORAGE AND USE OF INFLAMMABLE SUBSTANCES 6

DISTRIBUTION OF SOLVENTS

Inflammable solvents which are not in the laboratories are stored under lock and key in thebasement storage room (up to the maximum of 600 litres authorised for the site).

To respect this limit, all orders for inflammable solvents must be cleared by the ASO SafetyOfficer before being sent to SUP.

Inflammable solvents may be obtained daily from the ASO Safety Officer at the following times:

10.00 to 10.30 a.m. 2.00 to 2.30 p.m.

The ASO Safety Officer may be contacted exceptionally for additional distribution only if hisworkload permits.

Staff are reminded that solvents should not be transported by hand, but by means of a cart orplastic bottle carrier. Only the red lift should be used for the transport of such materials.

DISTRIBUTION OF ALCOHOLS

Tax laws in France strictly regulate the use of certain alcohols. A special form, CIRC 10, availablein the basement storage room, must be completed to obtain methanol, ethanol or propanol.

USE OF INFLAMMABLE GASES

Under certain circumstances, inflammable gases such as hydrogen, oxygen, butane and propanemay be used in the laboratory, but only after clearance has been obtained from ASO. All gasbottles and lines should be clearly labelled so that there is no possible confusion over theircontents. Hydrogen for gas chromatography-flame ionization detectors must be produced inthe laboratory through the use of a hydrogen generator; hydrogen bottles are not permittedin IARC laboratories.The use of open flames (bunsen burners, etc.) is prohibited in the main building. The carefuluse of small portable burners such as camping gas for certain essential operations is, however,tolerated.

SAFE USE OF CARCINOGENS

This chapter does not pretend to contain all the information which is available on chemicalcarcinogen safety, but it attempts to provide the basic rules to which any person working withcarcinogenic compounds should adhere. For more detailed information, we recommend thefollowing references:

• Handling Chemical Carcinogens in the Laboratory: Problems of Safety, edited by R. Montesano, H. Bartsch, E. Boyland, G. Della Porta, L. Fishbein, R.A. Griesener, A.B. Swan and L. Tomatis. IARC Scientific Publications No. 33).

• NIH Guidelines for Laboratory Use of Chemical Carcinogens: US Department of Health and Human Sciences (this document can be consulted in the library).

• Méthodes d’élimination des déchets mutagènes et cancérogènes applicables aux laboratoires de recherche, M. Castegnaro and H. Brésil, 12èmes journées de l’ADHYS, 22 and 23 September 1994.

• Manipulation des substances génotoxiques utilisées au laboratoire, edited by X. Rousselin,J. Dayan-Kenigsberg, C. Pleven, M. Castegnaro, A. Picot and F. Zajdela, Institut National de la Recherche Scientifique, Paris, (1994).

These documents contain a wide list of references pertaining to this subject. You can also findrelevant information on the OHSC home page: http://intranet/labcom/index.htm.

The list of compounds evaluated by the IARC Monographs for their carcinogenic activity isavailable on the IARC Monographs website: http://monographs.iarc.fr. Those that have beenclassified as human carcinogens (Group I) are listed in Annex 1 at the end of this chapter.

ORDERING CARCINOGENS

The purchase or the synthesis of carcinogenic chemicals must always be limited to theminimum amount necessary to carry out the proposed experiment.All chemical carcinogens, whether obtained commercially or in any other way, must be includedin the IARC carcinogen inventory: upon reception, the compound must be added to the listof carcinogens, which can be found in room 805. Indicate the name of the person responsiblefor the compound, their group, the name of the compound, the source, the reference, thequantity, the location, the date of reception. A code number must be assigned to the newcarcinogen (Code No. column) and marked on the sample bottle(s). The inventory is regularlyupdated.

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7 SAFE USE OF CARCINOGENS

STORAGE OF CARCINOGENS

Upon delivery, carcinogenic compounds should be unpacked and must be stored only in room805, either in the freezer or the refrigerator, or in a plastic box for those compounds thatrequire storage at room temperature. It should be noted that this laboratory is also used forthe handling of radioactive isotopes.

For security reasons, access to room 805 is restricted, and only those staff who have anappropriately programmed magnetic card may enter this room. Each time the card is used, itis automatically recorded by computer. Any new staff member who needs to enter this roomregularly must ask the Laboratory Safety Officer, to activate their magnetic card.

When users have finished their work in this laboratory, they should ensure that:• The workbench and any equipment used have been cleaned and, if necessary,

decontaminated,• The carcinogen bottle has been tightly closed, and has been stored in its proper place,• The users logbook located in the “sas” 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 toit. Then they must, if possible, decontaminate the container, following procedures described atthe 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 thecompound should inform the Laboratory Safety Officer, who will have the compound disposed of.

After dilution of pure compounds, each solution container should be marked with the followinginformation:

• name of carcinogen • dilution of solvent• concentration• user’s name• date

Diluted carcinogen solutions must always be stored in well identifiedsealed boxes in refrigerators or freezers in the laboratories carrying thefollowing labels which are available in the carcinogen room:

All areas where any solutions of carcinogens are stored (refrigerators,freezers, etc.) must be marked with the following label, available fromthe ASO Safety Officer.

CARCINOGEN....................................................CONC .................................... IN .......................................USER ....................................... DATE ..................................

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SAFE USE OF CARCINOGENS 7

WORKING WITH CARCINOGENS

Personal protection

It is extremely important when handling pure carcinogenic compounds that the user avoidsexposing himself or those around him to these compounds. The use of protective clothingwhen working with carcinogens is absolutely essential. Disposable gloves, shoe covers andfacemasks are always available in room 805 and can also be obtained from the ASO SafetyOfficer. If carcinogens are routinely handled, it is advisable to reserve a laboratory coat for thispurpose. In certain circumstances trousers and caps should also be worn for greaterprotection; these are available in the room itself or from SUP.

Protective clothing must be changed frequently, and immediately after accidental contamination.In the latter case, protective clothing must be decontaminated by appropriate methods or ifnecessary destroyed. If in doubt, contact the Laboratory Safety Officer. Remember that glovesdo not provide complete protection and that certain carcinogenic substances such asnitrosamines, aflatoxins and aromatic amines can diffuse through gloves, especially when thecompounds are in solution. It is preferable, in these cases, to use two pairs of gloves, one ontop of the other. In the case of an accidental contamination, gloves should be removedimmediately and deposited in the plastic waste receptacles provided for this purpose, labelledwith a "carcinogenic waste" sticker.

Handling carcinogens in room 805

Any synthesis or purification of chemical carcinogens must, without exception, be carried outin this room. This type of procedure is forbidden elsewhere in the building.

Room 805 has 2 hoods: one fume cupboard exclusively reserved for handling radioisotopesand a laminar flow hood equipped with HEPA and charcoal filters for the handling ofcarcinogens. This last hood ensures both the protection of the handler and that of theenvironment. The 2 hoods must not be in operation at the same time, as they will not functionproperly. It is therefore necessary to reserve a time slot before working in this laboratory.

Handling powders, whether electrostatic (aflatoxins, bromomethyl benzanthracene, etc.) ornot, raises a serious problem. Although no method is entirely satisfactory, one should avoidworking in draughts or using latex or vinyl gloves. When handling such compounds, the useof cotton gloves and face masks is recommended.

One method for handling such compounds is to buy preweighed quantities and to dissolve allthe carcinogen in an appropriate solvent. The problem is then limited to the handling of liquidstock solutions.

CARCINOGENIC WASTES

DECHETS CANCEROGENES

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In the case of an accident

In the case of an accident either in room 805 or anywhere else in the building, when workingwith carcinogens, do not spread the contamination. Take any contaminated clothes off, isolatethe contaminated zone and sound the alarm using the red button opposite the ‘Pater Noster’in order to receive help from security staff and begin decontamination as quickly as possible.

In the case of skin contamination, rinse abundantly with water (use the eye wash or the safetyshower in the corridor), wash with a mild soap and rinse again.

Should this not be sufficient and/or in the case of internal contamination, the person shouldbe taken to the nearest hospital (Hôpital Edouard Herriot).

Transporting carcinogens within the building

Pure chemical carcinogens must never be removed from room 805, which is reserved for theirstorage, weighing and dilution. Only diluted solution can be taken into the other laboratories.Each user must bring all equipment and solvents necessary to perform such dilutions to room805.

Before transporting diluted solutions of carcinogens within thebuilding, flasks and bottles should be tightly closed and placedinside the metal transport containers. These containers, whichcontain an absorbent, are designed to withstand accidentaldropping or chemical attack by the solvents used for dilution.These metal containers are available in room 805 and must bereturned to this room after use. All solutions must be labelledwith appropriate stickers.

When transporting diluted chemical carcinogens from one floor to another, use only the red lift.

In the case of an accident during transportation, isolate the contaminated zone, push the redbutton opposite the “Pater Noster” to obtain help from security staff, and proceed as quicklyas possible to decontaminate the zone. If the accident occurs while you are in the red lift, callfor the emergency team directly from the lift and redirect the lift to the ground floor wherethe emergency team will be waiting.

Handling carcinogens in the laboratory

The precautions which should be taken when working with diluted solutions of carcinogensdepend on the type of experiment being carried out. A general rule to be followed is thatsolutions of volatile carcinogens should only be handled in a fume cupboard, equipped withan 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.

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Dichloromethane and chloroform

It is also important to ensure the proper use of certain halogenated solvents, such asdichloromethane and chloroform for which sufficient proof is available of their carcinogenicityin animals. These solvents must always be handled in a well-ventilated fume cupboard. Emptycontainers should be left under the fume cupboard to evaporate residual solvent before thecontainers can be safely discarded.

Benzene

The use of benzene, a human carcinogen, should be avoided. Toluene or xylene are suitablereplacement solvents.

Formaldehyde

Solutions of formaldehyde, used to fix tissues, pose a special problem, as formaldehyde hasbeen shown to be carcinogenic; it is classified in Group I of the IARC Monographs. It istherefore strongly recommended that persons who handle these solutions wear gloves as wellas safety glasses, and work under a well ventilated fume cupboard especially adapted for thisuse. Formaldehyde is used in the histology laboratory and in the molecular biology laboratories;it is often a component in gels for Northern blotting, which should be poured and run in a fumehood to reduce the risk of exposure.

Formaldehyde liquid waste is put in plastic containers for "solvants usés" which are collectedonce a week.

Ethidium bromide

Ethidium bromide is widely used in molecular biology laboratories as a fluorescent stain. It isa 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 bacterialcontamination.

The preparation of agarose gels containing ethidium bromide should respect the followingprecautions:

• reduce to a minimum the transport of melted agarose that is very hot• leave to cool for a few seconds (not necessarily under running water)• add the ethidium bromide under a fume cupboard in order not to breathe in the vapours• avoid transporting gels after electrophoresis for reading on a UV table. If necessary, put

them in a container in order to avoid spilling in the corridors! Agarose gels containing ethidium bromide should be disposed of in waste boxes for incineration.

42


Concentrated ethidium bromide solutions used to stain gels must be disposed of only in theaqueous carcinogen bottles (see next section).

It is also important to note that all plastic items (gel holders, combs, electrophoresis tanks)become, with time, impregnated with ethidium bromide. They should therefore be handledonly when wearing gloves.

Ethidium bromide, even if used in very small quantities, is a toxic product whichis also genotoxic, mutagenic and teratogenic. Its regular and repetitive use canlead to us forgetting how dangerous it can be.

Acrylamide

Acrylamide solutions are widely used in the laboratory for the analysis of nucleic acids andproteins (separation of DNA or RNA fragments, sequencing, Western blot, polypeptideseparation, etc.). Contarary to agarose, acrylamide is a carcinogen and potential neurotoxinwhich is absorbed through the skin. Its effects are cumulative. Thus, certain precautions arecalled 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 (which is toxic) may be present.

DISPOSAL OF CARCINOGEN-CONTAMINATED WASTE

Solid carcinogenic waste

Waste that is or may have been contaminated by carcinogens must be separated from otherwastes and placed in appropriately labelled containers:

It is highly recommended that preweighedacrylamide or acrylamide solutions be purchased

to avoid having to weigh the powder.

43


Solid carcinogenic waste such as paper, filters, gloves, etc.,should be placed in “sharpsafe” bins or in plastic-linedcardboard boxes marked with the label CARCINOGENICWASTE.

Liquid carcinogenic waste

Liquid carcinogenic waste is stored in clear plastic-covered glass bottlesthat are resistant to both solvents and carcinogens. These containersshould be marked with the appropriate label depending on whetheraqueous or organic solvents have been used.Both solid and liquid carcinogenic wastes are collected by those personsresponsible for managing waste products.

Radioactive carcinogenic waste

Liquid or solid carcinogenic wastes that have beencontaminated by radioactive carcinogens must bestocked in special containers marked with labels,“CARCINOGENIC WASTE” and “RADIOACTIVE WASTE”.

Such radioactive waste is kept either until there has been sufficient radioactive decay for it tobe disposed of as carcinogenic waste or it is removed and disposed of by the Agence Nationalepour la Gestion des Déchets Radioactifs (ANDRA) under the supervision of the Institut deRadioprotection et de Sûreté Nucléaire (IRSN) (see Table I, Chapter 8).These radioactive/carcinogenic wastes are collected once a week by the ASO Safety officer, orunder exceptional circumstances upon request.

Before beginning any experiment involving carcinogenic or radioactive products, contact theLaboratory Safety Officer to ensure that the procedures are adapted to the products handled.

DESTRUCTION OF WASTE CARCINOGENS

All disposal of carcinogenic products (pure, in concentrated solution or in diluted solutionsmust be carried out in collaboration with the Laboratory Safety Officer.

Pure carcinogens or carcinogen solutions can be conditioned to be disposed of in specific binswhich will be taken away to be destroyed by the company responsible for collecting waste.

Decontamination methods exist but they can be dangerous and must be followed scrupulouslyin order to avoid any incidents. Decontamination techniques are described in the IARCScientific Publications series. 'Laboratory Decontamination and Destruction of...'which can be consulted in the IARC library.

44


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• No. 64: Aromatic amines and 4-nitrobiphenyl• No. 73: Antineoplastic agents• No. 113: Mycotoxins• No. 114: Heterocyclic hydrocarbons

DECONTAMINATION OF EQUIPMENT AND THE WORK AREA

Carcinogen-contaminated glassware must be kept separately from ordinary glasswareand be decontaminated as soon as possible by the user before being sent to the centralglassware washroom. The decontamination methods mentioned above can be used for thispurpose.

Glassware contaminated with radioactive and carcinogenic substances is firstwashed with a suitable solvent. Rinse solvent is collected in clear plastic-covered glass bottlesmarked ‘CARCINOGENIC WASTE’ and ‘RADIOACTIVE WASTE’ and one of the followingtwo labels: "ORGANIC SOLUTION" or "AQUEOUS SOLUTION". The glassware is thenplaced in trays provided for the decontamination of radioactive glassware (Chapter 8).

Glassware which has contained a pure carcinogen should be decontaminated as indicatedabove, if possible, or placed in a bin for incineration.

Cleaning of bench tops and fume cupboards is carried out by an outside company under thesupervision of laboratory technicians, according to a schedule organised by the LaboratorySafety Officer and ASO. If necessary, laboratory bench tops and fume cupboards should bedecontaminated by laboratory personnel before such cleaning, and no containers in whichcarcinogenic or toxic compounds have been stored are to be handled by the cleaningpersonnel.

CARCINOGENIC WASTES

ORGANIC - ORGANIQUES


CARCINOGENIC WASTES

ACQUEUX - AQUEOUS


45


Agents and groups of agents

4-Aminobiphenyl

Aristolochic acid

Arsenic

Asbestos

Azathioprine

Benzene

Benzidine

Benzo[a]pyrene

Beryllium

N,N-Bis(2-chloroethyl)-2-naphthylamine(Chlornaphazine)

Bis(chloromethyl)ether and chloromethyl methylether

1,3-Butadiene

1,4-Butanediol dimethanesulfonate (Busulphan;Myleran)

Cadmium

Chlorambucil

1-(2-Chloroethyl)-3- (4-methylcyclohexyl)-1-nitrosourea (Methyl-CCNU; Semustine)Chromium[VI]

Cyclophosphamide

Cyclosporine Diethylstilboestrol

Dyes metabolized to benzidine

Epstein-Barr virus

Erionite

Estrogen-progestogen menopausal therapy (combined)

Estrogen-progestogen oral contraceptives (combined)

Estrogens, nonsteroidal

Estrogens, steroidal

Estrogen therapy, postmenopausal

Ethanol in alcoholic beverages

Ethylene oxide

Etoposide

Etoposide in combination with cisplatin and bleomycin

Formaldehyde

Gallium arsenide

[Gamma Radiation: see X- and Gamma (γ)-Radiation]

Helicobacter pylori (infection with

Hepatitis B virus (chronic infection with)

Hepatitis C virus (chronic infection with) 1

Human immunodeficiency virus type 1 (infection with)

Human papillomavirus types 16, 18, 31, 33, 35, 39,45, 51, 52, 56, 58, 59 and 66

Human T-cell lymphotropic virus type I

Melphalan

8-Methoxypsoralen (Methoxsalen) plus ultravioletA radiation

Methylenebis(chloroaniline) (MOCA)

MOPP and other combined chemotherapyincluding alkylating agents

Mustard gas (Sulfur mustard)

2-Naphthylamine

Neutrons

Nickel compounds (Vol. 49; 1990)

N'-Nitrosonornicotine (NNN) and 4-(N-Nitrosomethylamino)-1-(3-pyridyl)-1-butanone (NNK)

Oestrogen: see Estrogen

Opisthorchis viverrini (infection with)

Oral contraceptives, combined estrogen-progestogen: see Estrogen-progestogen oralcontraceptives (combined)

Oral contraceptives, sequential

Phenacetin

Phosphorus-32, as phosphate

Plutonium-239 and its decay products (may containplutonium-240 and other isotopes), as aerosols)

Radioiodines, short-lived isotopes, including iodine-131, from atomic reactor accidents and nuclearweapons detonation (exposure during childhood)

Radionuclides, α-particle-emitting, internally deposited

(NB: Specific radionuclides for which there issufficient evidence for carcinogenicity to humansare also listed individually as Group 1 agents)

Radionuclides, β-particle-emitting, internally deposited

(NB: Specific radionuclides for which there issufficient evidence for carcinogenicity to humansare also listed individually as Group 1 agents)

Annex : Group 1- Human carcinogens

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Radium-224 and its decay products



Radon-222 [10043-92-2] and its decay products

Schistosoma haematobium (infection with)

Silica [14808-60-7], crystalline (inhaled in the formof quartz or cristobalite from occupational sources)

Solar radiation

Talc containing asbestiform fibres

Tamoxifen

2,3,7,8-Tetrachlorodibenzo-para-dioxin

Thiotepa

Thorium-232 and its decay products, administeredintravenously as a colloidal dispersion of thorium-232 dioxide

ortho-Toluidine

Treosulfan

Vinyl chloride

X- and Gamma (γ)-Radiation

Mixtures

Aflatoxins (naturally occurring mixtures of)

Alcoholic beverages

Areca nut

Betel quid with tobacco

Betel quid without tobacco

Coal-tar pitches

Coal-tars

Household combustion of coal, indoor emissionsfrom

Mineral oils, untreated and mildly treated

Phenacetin, analgesic mixtures containing

Plants containing aristolochic acid

Salted fish (Chinese-style)

Shale-oils

Soots

Tobacco, smokeless

Wood dust

Exposure circumstances

Aluminium production

Arsenic in drinking-water

Auramine production

Boot and shoe manufacture and repair

Chimney sweeping

Coal gasification

Coal-tar distillation

Coke production

Furniture and cabinet making

Haematite mining

Involuntary smoking (exposure to secondhand or'environmental' tobacco smoke)

Iron and steel founding

Isopropyl alcohol manufacture

Magenta production

Painter (occupational exposure as a)

Paving and roofing with coal-tar pitch

Rubber industry

Strong-inorganic-acid mists containing sulfuric acid(occupational exposure to)

Tobacco smoking and tobacco smoke

SAFE USE OF RADIOACTIVE SUBSTANCES

Certain unstable isotopes spontaneously emit particles or rays as they decompose to a morestable state in a process called radioactivity. This process is called nuclear disintegration. Everyradioactive source is defined as follows:

• Its activity, which represents the number of spontaneous nuclear transformations that are produced by unit of time. This is expressed in bequerels.1Bq = 1 disintegration / second.

• Its radioactive period or half-life: time after which its activity has been reduced by half. After 10 periods, the activity will be very close to zero. The radioactive period is specific to each radioelement. It can vary from a few fractions of a second to several millions of years.

Radioisotopes are classified into different groups depending on their radiotoxicity.

The authorised radioisotopes which may be handled in IARC laboratories, as open sources, are:

Any person at IARC who works with such radioactive substances must follow the appropriateprocedures and security rules outlined in this chapter.

Additional information is provided in the annexes to this chapter and can also be obtained fromthe Radiation Safety Officer. She should also be contacted if the need arises to handle otherradioisotopes.

47

Radioisotope Radiotoxicity Group Radiation type Technical sheet in Annex

Tritium 3H Very slight 5 ß III

Carbon 14C Slight 4 ß IV

Phosphorus 32P High 2 ß V

Phosphorus 33P Very slight 5 ß VI

Sulphur 35S Very slight 5 ß VII

Iodine 125I Moderate 3 α VIII

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8 SAFE USE OF RADIOACTIVE SUBSTANCES

AUTHORIZATION TO USE RADIOISOTOPES AND MEDICAL SURVEILLANCE

Registration

Registration with the French Ionising Radiation Protection Service (Institut deRadioprotection 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 approvalto handle radioactive isotopes in IARC laboratories by contacting the Radioactive SafetyOfficer. The person must follow a course available on the Intranet, fill in a correspondingquestionnaire and have it validated by the Radioactive Safety Officer.

The person must also have medical clearance with the Staff Physician if they do not have arecent medical certificate.

The list of persons authorised to handle radioisotopes is updated every month, given to theIARC Staff Physician and posted on every laboratory floor.

All registered users must fill in a form each month noting the quantities of the differentradioisotopes they have manipulated during the past month. These forms are sent directly toeach user and should be returned to the Radioactive Safety Officer, even if no isotopes havebeen 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 becomeaware that they are pregnant, as pregnant women are not normally allowed tohandle radioisotopes. After discussion with the Staff Physician, exceptionalauthorization may be given on an individual basis.

Systematic surveillance carried out by the IRSN

Urinary bioassaysThis surveillance is carried out twice a year and is compulsory for all those registered tohandle radioisotopes. It requires the collection of a twenty-four hour urine sample (see AnnexIX).The user should complete the identification tag attached to the specimen bottle beforeleaving it at the collection point. Each radioisotope user is expected to comply with thedesignated urine collection dates, which are organised to allow all the samples to be dispatchedtogether to the IRSN.

In the case of an accidental internal exposure, the user should immediately contact theRadioactive Safety Officer who will organise, in consultation with the Staff Physician, theappropriate medical surveillance.

External dose measurementsPersonal dosimeters (chest and wrist film badges) are obligatory for all staff who work withthe radioisotopes 32P and 125I. They are provided and checked every 3 months by the IRSN.New users registered during the current term should contact B. Chapot, who will issuetemporary dosimeters until the delivery of nominative dosimeters.

49

SAFE USE OF RADIOACTIVE SUBSTANCES 8

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 bestored 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 thermalluminescent finger dosimeter must be worn. These dosimeters are provided and controlled bythe French Institute of Nuclear Physics (IPN). They are available from the Radioactive SafetyOfficer as needed.

IARC LABORATORIES AUTHORIZED FOR THE USE OFRADIOACTIVE MATERIALS

All areas within IARC where radioisotopes are handled must have prior authorization fromthe French Interministerial Commission.

It is therefore compulsory to handle radioisotopes only in these authorizedlaboratories. They are well defined according to the French legislation. Aprecise analysis of the experimental protocols and of the potential risk ofexposure has allowed the classification of IARC laboratories as zones undersurveillance. They are marked with a blue radioactive label.

The total amount of radioactivity (MBq) and the radioisotopes authorized to be stored andhandled in each IARC laboratory is strictly controlled. The present authorised limits are listedbelow. The nature of the radioisotopes that can be handled and stored in each laboratory islisted on the door and must be complied with.

Room 805 is also used for stocking and preparing carcinogenic solutions (see Chapter 7). Itis equipped with 2 extractors, a fume cupboard for the handling of radioactive products, anda laminar flow hood equipped with activated coal filters for the handling of carcinogenicproducts. These two extractors cannot function properly simultaneously (not enough air flowin the room). It is therefore necessary to make sure that enough time is allowed for use ofthe room in order to work in good conditions. This room has restricted access, andauthorisation for regular use should be requested from the Laboratory Safety Officer.

*A table for converting Ci to SI units (MBq) is given in Annex I, p. 61.

Laboratory Radioisotypes authorized Maximum quantity authorized602 32P, 33P 110 MBq

703/704 32P, 33P,35S,14C,3H 74 MBq805 32P, 33P,35S,14C,3H 555 MBq

E06/E07 125I, 3H 18 MBqS05 32P, 33P 740 MBq (storage rooms in basement)

SMH S3 125I, 35S,14C,3H 740 MBq (storage rooms in basement)

50


Laboratories 703/704 and 805 have absolute filters in the extraction system and should be usedwhen handling volatile radioactive products such as 35S-methionine.

Exceptionally, and with the authorization of B. Chapot, work may be carried out in non-authorized areas, but the following quantities must not be exceeded:

Handling of sealed sources

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 instrumentsare taken out of service) must respect special procedures. In both cases contact theLaboratory Safety Officer for information about the appropriate procedures to follow.

ORDERING RADIOACTIVE COMPOUNDS

Orders for radioactive compounds should be made on standard order forms. A specialnumbered form supplied by the IRSN is completed by the Supplies Office (SUP) for each neworder sent to the supplier.

STORING RADIOACTIVE COMPOUNDS

In order to comply with French law concerning the traceability ofradioactive sources in the building, every new compound is deliveredtogether with a Radioisotope Source Tracing Sheet. This sheet indicatesthe date of reception, radioisotope, quantity and name of the person whoordered the product. This sheet must be placed on the door of therefrigerator or freezer where the compound is stored. It should be filledin by each user and after discarding the source, the sheet should bereturned to the Radoactive Safety Officer.

Refrigerators and/or freezers containing radioactive compounds should be clearly identifiedwith the appropriate label and kept locked. If they are also used to store other products,radioactive sources should be placed in clearly identified, closed boxes intended for thispurpose.

All radioactive compounds having a long half-life (3H and 14C) must be stored in room 805. Thisroom has restricted access, therefore please contact the Radioactive Safety Officer for theprocedure to follow.

3H, 35S 5000 kBq

14C, 32P, 33P 500 kBq

125I 50 kBq

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A computer listing of all the long half-life radioactive compounds stored at IARC is maintainedand checked yearly. Each person is responsible for the products they order and must keep upto date the paper copy of the listing kept in room 805. It is also very important to indicateon the Radioisotope Source Tracing Sheet the quantity of radioisotope used so that the personresponsible for radioprotection can update the table of stocks of radioactive substances eitheras source or as waste. This is a requirement of the Nuclear Safety Authority.

WORKING SAFELY WITH RADIOACTIVE SUBSTANCES

Radioisotope users must understand the nature of the hazard and be aware of thecharacteristics of the particular isotope they are working with, especially the potential risksof internal or external contamination. Additional technical information is available in AnnexesIII to VIII.

Personal protection

You should never work with unprotected wounds or cuts in the skin, especially on the handsor forearms.

As in all laboratories, you are not allowed to eat, drink or apply cosmetics in an area whereradioactive material is handled. The use of paper handkerchiefs is recommended and of coursemouth pipetting of any solution is strictly forbidden.

A laboratory coat reserved for this kind of experiment and gloves must be worn when workingwith radioactive compounds, and where appropriate safety glasses and disposable arm coversshould also be used.

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 are alsoadvised to wear two pairs of gloves, particularly when handling sources.

You must always store isotopes, sources or probes in appropriate closed containers (inplexiglas for 32P), with clear labels indicating the isotope, activity, date and name of user. Thecontainer should be stored in a refrigerator or freezer dedicated to this purpose

Low energy ß radiation

For the radioisotopes 14C, 3H, 35S, 33P, the risk of external exposure is practically non-existent.Latex gloves, if changed frequently, and safety glasses give sufficient protection fromcontamination in most situations.

For tritium (3H), however, there is a potential risk of internal exposure either by inhalation orthrough the skin. Work with this radioisotope should preferably be carried out in a fumecupboard.

For 35S, and in particular when using 35S-methionine where volatile breakdown products areformed, there is an increased risk of internal exposure, mainly from inhalation or through theskin. Work with 35S-methionine must be carried out in a fume cupboard. It is also importantto store all the waste containing these volatile degradation products under the hood in aclosed bin.

52


High-energy ß radiation

For 32P, there is a risk of external exposure to the skin. Contact with the skin shouldbe avoided and adequate protection (gloves, safety glasses, tweezers, protective screens) shouldbe used. Gloves are useful for avoiding chemical contamination but they do not preventexternal exposure from 32P radiation. The wearing of two pairs of gloves, one over the other,is recommended.

For experiments using 37 MBq or more, it is also necessary to usethermo- luminescent finger dosimeters for hand exposure surveillance.These are available from the Radioactive Safety Officer. These moresensitive dosimeters worn on the fingers enable a better evaluation ofexposure. In the case of positive exposure, the Radioactive SafetyOfficer will be able to revise the experimental procedure with theperson concerned in order to try and reduce exposure.

X and γ radiation

These forms of radiation are emitted by 125I. The use of both chest and wristdosimeters is compulsory when handling this isotope.The main use of this isotope atIARC is as the labelling component in diagnostic kits, a technique that uses low amounts ofradioactivity. In this particular case the isotope may be handled on a bench with a minimal riskof internal contamination. The quantity of waste generated has led to a leaded glass screenbeing used in order to provide sufficient protection against this type of radiation.

Safe handling of radioactive substances

As a function of the radiotoxicity, activity and physical chemical nature of the radioisotopebeing used, work should be carried out.

• on the bench top for non-volatile compounds with low activity• under a ventilated fume cupboard, or• behind protective screens for 32P.

Plan your experiment to minimise the time you spend handling radioactivity. The shorter thetime, the smaller the dose to which you are exposed.

Distance yourself from the sources of radiation: doubling the distance from the source quartersthe radiation dose (inverse square law).

The use of both chest and wrist dosimeters (filmbadges) is compulsory when working with any

concentration of 32P.

53


Check the work area for contamination before starting your experiment, check the area andyour hands frequently during manipulation and check after completion of work. Check yourhands again and make sure you wash them.

The outer limits of the work zone on the bench should be delineated. You are strongly advisedto use a plastic tray which has been especially designed for this type of work and which canprovide a limited work zone. The surface of the work area should be protected by a protectiveplastic liner that is easy to clean or dispose of in the case of heavy contamination.

Do not use aluminium paper to protect the work area for this can easily create the formationof aerosols which are very contaminating.

Before beginning an experiment, you should have available:

• 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 pipettes and glassware • a container for organic and/or aqueous waste,• suitable containers for solid wastes (see next page)• vermiculite, if necessary• safety glasses• protective gloves • paper tissues• Benchkote absorbent paper

When working with a high-energy radioisotope such as 32P, you will need to use plexiglasscreens, syringe guards and tweezers.

Appropriate radiation detectors are available in each laboratory in which the handling ofradioisotopes is authorised. Any faulty material should be reported to the Radioactive SafetyOfficer, who can organise a temporary replacement and the repair of any defective equipment.

Several scintillation counters located in different research units are also available. Extremecare should be taken if it is necessary to transport radioactive samples between differentfloors, and in particular for 32P-labelled samples, which must be placed in an appropriate sealedplexiglas box to avoid any risk of external contamination.

54


RADIOACTIVE WASTE

All waste generated by radioisotope users must be separated from other, non-radioactivewaste and must be disposed of in accordance with strict regulations as detailed below inAnnex II.

The method of waste elimination depends on theradioisotope’s physical and chemical properties, decay half-life and activity. Thus, it is very important that radioactivewastes 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

These types of waste are stored in the building, awaiting sufficient radioactive decay (at least10 half lives) before elimination as non-radioactive waste.

Long radioactive decay half-life (>100 days)• 3H : 12.35 years• 14C : 5630 years

3H and 14C radioactive waste is stored until removal by the National Agency for the processingof radioactive waste (Agence Nationale pour la Gestion des Déchets Radioactifs - ANDRA).The removal of such waste is very expensive. It is therefore important to carefully sort outthese radioactive wastes.

Storage and disposal of radioactive waste

The storage and disposal of radioactive waste must be done strictly in accordance with Frenchregulations and is under the supervision of the ASO Safety Officer and the Radiation SafetyOfficer. Each waste container must have a special label (or adhesive tape) identifying theradioisotope it contains. When a waste container is full, it should be closed and left in anauthorised location for collection. Additional waste containers can also be obtained as required.

No radioactive waste may be disposed of in sinks orordinary bins by laboratory personnel.

Both solid and liquid radioactive waste are removed once aweek, or on special request, by the ASO Safety Officer.

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Under no circumstances may radioactive biological infectious waste be eliminatedwithout prior decontamination of the infectious agent.

Liquid radioactive wastes

Several categories of waste have been defined, depending on the physical and chemicalproperties (French two-letter designations are given in brackets):

Aqueous liquid waste (LA)

• 3H or 14C aqueous liquid waste is stored in 2-litre plastic bottles or in airtight 30-litre containers.

• 32P or 33P very radioactive aqueous liquid waste should be placed in appropriately labelled clear plastic-covered 1-litre glass bottles stored in cylindrical plastic containers (green cap).

• 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 by the ASO Safety Officer and replaced with an empty container.

• 35S aqueous liquid waste should be placed in appropriately labelled clear plastic-covered-1-litre glass bottles.

• 125I aqueous solutions should be placed in 20-litre plastic containers which are easily identifiable.

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Organic liquid waste (LS)

• 3H or 14C organic liquid waste or solvents are collected in 2 litre plastic bottles. Thecontents of these containers are subsequently combined for storage in the basement in 30-litre containers. Radioisotope concentrations are checked by the Radiation Safety Officerbefore final disposal by ANDRA.

• 32P, 33P, 35S and 125I organic liquid wastes should be collected in separate appropriatelylabelled clear plastic-covered 1-litre glass bottles. These containers are subsequently storedin the basement until the radioactivity has decreased sufficiently to permit disposal as non-radioactive waste.

Carcinogenic radioactive liquid waste

• 3H and 14C carcinogenic radioactive liquid wastes are collected in appropriatelylabelled plastic-covered 4-litre glass bottles which cannot be chemically attacked by the solventor the carcinogen. The waste solution bottles, which must be labelled as either organic oraqueous, must also carry a radioactive label as follows:

The contents of these bottles are subsequently combined into larger containers in thebasement and, after measurement of the radioactivity levels, they are discarded, depending onthe amount, as either carcinogenic or radioactive waste.Before carrying out any new experiment using a radiolabelled carcinogen, please contact theRadioactive Safety Officer before starting work to organise appropriate waste disposal facilities.

Mixed radioactive wastes

This usually consists of scintillation vials and their contents (radioisotopes and scintillationfluids). Plastic and glass vials should be discarded in separate containers.

• 3H or 14C waste are disposed of in appropriately labelled blue 30-litrebarrels. Separate containers should be used for low- (below 1000cpm/ml) or high-activity (above 1000 cpm/ml) waste. Indicate the totalvolume and total cpm or dpm of the vials disposed of on the label on thelid of the barrel.

• 32P or 33P mixed waste should be placed in the appropriatelylabelled blue barrels.

• 125I mixed waste is placed in yellow plastic containers reserved forthis purpose.

CARCINOGENIC WASTESORGANIC - ORGANIQUES


CARCINOGENIC WASTESAQUEUX - AQUEOUS


57


Solid radioactive waste

Solid compactable waste (SP)

Solid material contaminated with 3H, 14C, 35S or 125I such as disposable syringes, gloves, filterpaper, etc., should be placed in cardboard burn bins. Each container should be labelled clearlyto 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 binlabelled with radioactive tape.

Very low radioactive solid waste

Large metallic green bins have been placed in several laboratories whereradioactivity is handled. These are intended for items of laboratory waste (suchas gloves, filter paper…) with no or a very low level of radioactivecontamination, which must not be put in the ordinary rubbish bins. The ASOSafety Officer checks and collects the full bags.

Solid waste is sorted before being :

• disposed of as ordinary waste after radioactive decay, in the case of radioisotopes with a short half-life

• disposed of by ANDRA, in the case of long half-life radioisotopes

IMPORTANT

All containers (bottles, boxes, bags) which contain radioactive waste must, atthe end of each day, be clearly labelled and stored in one of the laboratories

authorised to avoid their inadvertent removal by the cleaning staff.

It is extremely important that all radioactivity warning labels are removedfrom any packaging, containers or other rubbish before they are discarded

as normal non-radioactive waste.

58


PROCEDURE IN CASE OF CONTAMINATION

In the event of a spill:

• Warn all people in the surrounding area.• Isolate the contaminated zone and restrict unnecessary movement into and through the area.• Call the Radiation Safety Officer or ASO Safety Officer or press the red emergency button.• Treat contaminated personnel first. In the case of skin contamination, the contaminated

part of the body should be rinsed abundantly. Use the eye wash or the shower located in the corridors (see Annex X).

• If contamination persists and/or in the case of 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.

DECONTAMINATION

Decontamination of equipment

At the end of each experiment, the equipment and workbench should be checked for possiblecontamination, using an appropriate detector for the radiation emitted (medium or high ß, Xor 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 5 to10% Gigapur in demineralized water or the spray version.Contaminated glassware or equipment should be rinsed immediatelywithout being left to dry and then soaked in two successivedecontamination baths for at least 12 hours of Gigapur 10%(decontamination bath and rinse liquids should be saved as LA wastes).

Decontamination of work areas

Any radioactive contamination should be eliminated without delay. If solutions have beenaccidentally spilt on the floor or on the benchtop, use an absorbent powder such as vermiculitewhich should then be disposed of as radioactive waste. Surfaces should then be washed witha 10% Gigapur solution. Check to see if decontamination is complete using either theappropriate radiation detector or a swab soaked in ethanol, which is then measured in ascintillation counter.

To prevent contamination by radioactive aerosols, which pose a particularly high risk of internalcontamination by inhalation, volatile radioactive substances should only be handled in the fumecupboards fitted with special absolute filters in the authorised laboratories.

59


Personal decontamination

The correct use of gloves, safety glasses, laboratory coats, cuffs or special clothing should besufficient to avoid bodily contamination. However, in the case of accidental contamination,the contaminated part of the body should be washed abundantly (see Annex X).

Staff are strongly advised to wear two pairs of gloves: the top pair can be disposed ofimmediately when the handling of the radioisotope source is finished or in the case ofaccidental contamination.

Care should be taken to remove gloves before touching any non-radioactive object or material(pens, telephones, doorknobs, note-books, detectors…) to prevent spreading contamination.

The person responsible for radioprotection should be contacted immediately whenevercontamination occurs, whether heavy or not.

60


REFERENCES

In addition to the publications listed below, you can find links to relevant sites on this subjecton the committee’s website (http:intranet/labcom).

1. Décret n° 2007-1582 du 7 novembre 2007 relatif à la protection des personnes contre les dangers des rayonnements ionisants et portant modification du code de la santé publique (dispositions réglementaires).

2. Décret n°2007-1570 du 5 novembre 2007 relatif à la protection des travailleurs contre les dangers des rayonnements ionisants et modifiant le code du travail (dispositions réglementaires).

3. Arrêté du 15 mai 2006 relatif aux conditions de délimitation et de signalisation des zones surveillées et contrôlées et des zones spécialement réglementées ou interdites compte tenu de l'exposition aux rayonnements ionisants, ainsi qu'aux règles d'hygiène, de sécurité et d'entretien qui y sont imposées.

4. Arrêté du 26 octobre 2005 relatif aux modalités de formation de la personne compétente en radioprotection et de certification du formateur.

5. Arrêté du 26 octobre 2005 définissant les modalités de contrôle de radioprotection en application des articles R. 231-84 du code du travail et R. 1333-44 du code de la santé publique.

6. Arrêté du 30 décembre 2004 relatif à la carte individuelle de suivi médical et aux informations individuelles de dosimétrie des travailleurs exposés aux rayonnements ionisants.

7. Arrêté du 2 décembre 2003 fixant les seuils d'exemption d'autorisation.

8. Circulaire 2001-323 concernant les déchets radioactifs.

9. Arrêté du 1er septembre 2003 définissant les modalités de calcul des doses efficaces et des doses équivalentes résultant de l'exposition des personnes aux rayonnements ionisants.

10. Décret 2003-296 du 31 mars 2003 relatif à la protection des travailleurs contre les rayonnements ionisants.

11. 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

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Annex 1

INTERNATIONAL SYSTEM OF UNITS

The basic unit for measurement of radiation is the Becquerel (Bq). It is a very small unit. Theformer unit, which was much larger, was the Curie (Ci).

1 Ci = 37 109 Bq

Préfixes for SI units Prefix Symbol Factor

atto a 10-18

femto f 10-15

pico p 10-12

nano n 10-9

micro µ 10-6

milli m 10-3

kilo k 103

mega M 106

giga G 109

tera T 1012

penta P 1015

exa E 1018

Conversion Tables: Curies to Becquerels

µCi to kBqmCi to MBqCi to GBq

µCi to MBqmCi to GBqCi to TBq

1 372 743 1114 1485 1856 2227 2598 2969 333

10 37015 55520 74025 92530 1110

35 1.2940 1.4845 1.6650 1.8555 2.0360 2.2265 2.4070 2.5975 2.7780 2.9685 3.1490 3.3395 3.51

100 3.70

Converting SI units to non-SI units

From To Multiply by

becquerel (Bq)curie (Ci)gray (Gy)rad (rad)sievert (Sv)rem (rem)

curie becquerelradgrayremsievert

2.7 10-11

3.7 1010

1000.011000.01

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Annex II

DISPOSAL OF CARCINOGENIC AND/OR RADIOACTIVE WASTE

Pick up by an externalcompany for appropriate

treatment

RADIOISOTOPE

ACTIVITY

short half-life(<100 days)

long half-life(>100 days)

Treated as carcinogenic wastes

< 10Bq/lor

< 2 x backgroundnoise

(solids)

> 10Bq/lor

> 2 x backgroundnoise

(solids)

Treatment to getrid of the

associated risk

Storage in approvedcontainers as

radioactive waste

Request to ANDRAfor pick up

storage awaitingdecay during 10

periods

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Annex III

Working Safely with Tritium

Radioactive half-life (T1/2) 12.4 yearsPrincipal emission 8.6 keV beta (maximum)Radiotoxicity Group 5 (very low risk)Monitoring for contamination Swabs counted by liquid scintillationBiological monitoring Urine samplesDose per unit intake (DPUI)by ingestion or inhalation (tritiated water) 1.8 x 10-11 Sv/BqMaximum range in air 6 mmMaximum range in water 6 x 10-3 mmShielding required none

Special considerations

• Due 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 doses, it can lead to

hazardous internal contamination.• DPUI can vary considerably, e.g. DNA precursors such as tritiated thymidine are regarded

as more toxic than tritiated water partly because the activity is concentrated in cell nuclei.

3H

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Annex IV

Working Safely with Carbon-14

Radioactive half-life (T1/2) 5730 yearsPrincipal emission 0.156 MeV beta (maximum)Radiotoxicity Group 4 (low risk)Monitoring for contamination Thin end-window Geiger-Müller detectorBiological monitoring Urine samples

Breath measurements (CO2)Dose per unit intake (DPUI) by ingestionor inhalation 5,8 x 10-10 Sv/BqMaximum range in air 24 cmMaximum range in water 0.28 mmShielding required 1 cm Perspex/Plexiglas.

Thinner Perspex/Plexiglas down to 3 mm althoughadequate to reduce dose, does not have goodmechanical properties.


• Some organic compounds may be absorbed through surgical gloves.• Avoid the generation of CO2 which could be inhaled.

14C

65


Annex V

Working Safely with Phosphorus-32

Radioactive half-life (T½) 14.3 daysPrincipal emission 1710 keV beta (maximum)Radiotoxicity Group 2 (high-risk)Monitoring for contamination Geiger-Müller detectorBiological monitoring Urine samplesDose per unit intake (DPUI)by inhalation, 1.1 10-9 Sv/Bqby ingestion 2.4 10-9 Sv/BqMaximum range in air 790 cmMaximum range in water 0.76 cmDose rate from 1 MBq in 1 ml 210 mSv/h at surface

2.5 µSv/h at 1 mShielding required 1 cm Perspex (Plexiglas) stops betas and

minimizes production of Bremsstrahlung


• Avoid unnecessary exposure to this high energy radionuclide; use plexiglas racks or holders.• Wrist and finger dosimeters should be used when handling this isotope whatever the

quantity.• Finger dosimeters should also be be worn when handling quantities of 37 MBq and above.• Lead shielding is required when handling quantities above 370 MBq due to the production

of high energy Bremsstrahlung when the beta particles are absorbed.

32P

Proportion of a quantity of 32P remaining with timeHours

Days 0 12 24 36 48 60 72 840 1.000 0.976 0.953 0.930 0.908 0.886 0.865 0.8444 0.824 0.804 0.785 0.766 0.748 0.730 0.712 0.6958 0.679 0.662 0.646 0.631 0.616 0.601 0.587 0.57312 0.559 0.546 0.533 0.520 0.507 0495 0.483 0.47216 0.460 0.449 0.439 0.428 0.418 0.408 0.398 0.38920 0.379 0.370 0.361 0.353 0.344 0.336 0.328 0.32024 0.312 0.305 0.298 0.291 0.284 0.277 0.270 0.26428 0.257 0.251 0.245 0.239 0.234 0.228 0.223 0.21732 0.212 0.207 0.202 0.197 0.192 0.188 0.183 0.17936 0.175 0.170 0.166 0.162 0.159 0.155 0.151 0.14740 0.144 0.140 0.137 0.134 0.131 0.127 0.124 0.12144 0.119 0.116 0.113 0.110 0.108 0.105 0.102 0.10048 0.098 0.095 0.093 0.091 0.089 0.086 0.084 0.08252 0.080 0.078 0.077 0.075 0.073 0.071 0.070 0.068

66


Annex VI

Working Safely with Phosphorus-33

Radioactive half-life (T½) 25.4 daysPrincipal emission 0.249 MeV beta (maximum)Radiotoxicity 5 (very low risk)Monitoring for contamination Geiger-Müller detectorBiological monitoring Urine samplesDose per unit intake (DPUI)by inhalation 1.4 x 10-9 Sv/Bqor ingestion 2.4 x 10-10 Sv/BqMaximum range in air 49 cmMaximum range in water 0.6 mmShielding required 1 cm Perspex (Plexiglas).

Thinner Perspex/Plexiglas down to 3 mm, although adequate to reduce dose, does not have good mechanical properties.


• Phosphorus-33 does not require special precautions over and above those necessary for any beta-emitting radionuclide of this energy of emission.

33P

Proportion of a quantity of 33P remaining with timeHours

Days 0 1 2 3 4 5 6 7 8 90 1.000 0.973 0.947 0.921 0.897 0.872 0.849 0.826 0.804 0.78210 0.761 0.741 0.721 0.701 0.683 0.664 0.646 0.629 0.612 0.59520 0.579 0.564 0.549 0.534 0.520 0.506 0.492 0.479 0.466 0.45330 0.441 0.429 0.418 0.406 0.395 0.385 0.374 0.364 0.355 0.34540 0.336 0.327 0.318 0.309 0.301 0.293 0.285 0.277 0.270 0.26350 0.256 0.249 0.242 0.236 0.229 0.223 0.217 0.211 0.205 0.20060 0.195 0.189 0.184 0.179 0.174 0.170 0.165 0.161 0.156 0.15270 0.148 0.144 0.140 0.136 0.133 0.129 0.126 0.122 0.119 0.11680 0.113 0.110 0.107 0.104 0.101 0.098 0.096 0.093 0.091 0.08890 0.086 0.084 0.081 0.079 0.077 0.075 0.073 0.071 0.069 0.067100 0.065 0.064 0.062 0.060 0.059 0.057 0.055 0.054 0.053 0.051

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Annex VII

Working Safely With Sulfur-35

Radioactive half-life (T½) 87.4 daysPrincipal emissions 0.167 MeV beta (maximum)Radiotoxicity 5 (very low risk)Monitoring for contamination Thin end-window Geiger-Müller detectorBiological monitoring Urine samplesDose per unit intake (DPUI)by inhalation 1.3 x 10-9 Sv/Bq by ingestion 1.4 x 10-10 Sv/BqMaximum range in air 26 cmMaximum range in water 0.32 mmShielding required 1 cm Perspex/Plexiglas. Thinner

Perspex/Plexiglas down to 3 mm, although adequate to reduce dose does not have good mechanical properties.


• Vials should be opened and used in ventilated enclosures.• Avoid generation of sulphur dioxide or hydrogen sulphide, which could be inhaled.• Radiolysis of 35S-amino acids may lead to the production of labelled volatiles resulting in

an increased risk of internal exposure, mainly from inhalation or through the skin.

35S

Proportion of a quantity of 35S remaining with timeDays

Weeks 0 1 2 3 4 5 60 1.000 0.992 0.984 0.976 0.969 0.961 0.9541 0.946 0.939 0.931 0.924 0.916 0.909 0.9022 0.895 0.888 0.881 0.874 0.867 0.860 0.8533 0.847 0.840 0.833 0.827 0.820 0.814 0.8074 0.801 0.795 0.788 0.782 0.776 0.770 0.7640 0.758 0.752 0.746 0.740 0.734 0.728 0.7226 0.717 0.711 0.705 0.700 0.694 0.689 0.6837 0.678 0.673 0.667 0.662 0.657 0.652 0.6468 0.641 0.636 0.631 0.626 0.621 0.616 0.6129 0.607 0.602 0.597 0.592 0.588 0.583 0.57910 0.574 0569. 0.565 0.560 0.556 0.552 0.54711 0.543 0.539 0.534 0.530 0.526 0.522 0.51812 0.514 0.510 0.506 0.502 0.498 0.494 0.490

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Annex VIII

Working Safely with Iodine-125

Radioactive half-life (T½) 59.6 daysPrincipal emissions 35 keV gamma

(7% emitted, 93% internally converted)27-32 keV X-ray (140% Te K X-rays)

Radiotoxicity 3 (moderate risk)Monitoring for contamination Scintillation detectorBiological monitoring Thyroid scans (scintillation detector)Dose per unit intake (DPUI)by inhalation 1.1 x 10-9 Sv/Bqby ingestion 7.3 x 10-9 Sv/BqDose rate from 1 GBq point source at 1m 41 µSv/h First half value layer 0.02 mm lead


• Freezing 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 or polythene

alternatives 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 thiosulphate solution before

commencing decontamination.

125I

Proportion of a quantity of 125I remaining with timeHours

Days 0 2 4 6 8 10 12 14 16 180 1.000 0.977 0.955 0.933 0.911 0.890 0.870 0.850 0.830 0.81120 0.793 0.774 0.756 0.739 0.722 0.706 0.689 0.673 0.658 0.64340 0.628 0.614 0.600 0.586 0.572 0.559 0.546 0.534 0.521 0.50960 0.498 0.486 0.475 0.464 0.454 0.443 0.433 0.423 0.416 0.40480 0.394 0.385 0.377 0.368 0.359 0.351 0.343 0.335 0.328 0.320100 0.313 0.305 0.298 0.292 0.285 0.278 0.272 0.266 0.260 0.254120 0.248 0.242 0.236 0.231 0.226 0.221 0.215 0.211 0.206 0.201140 0.196 0.192 0.187 0.183 0.179 0.175 0.171 0.167 0.163 0.159160 0.156 0.152 0.149 0.145 0.142 0.139 0.135 0.132 0.129 0.126180 0.123 0.120 0.118 0.115 0.112 0.110 0.107 0.105 0.102 0.100200 0.098 0.095 0.093 0.091 0.089 0.087 0.085 0.083 0.081 0.079220 0.077 0.076 0.074 0.072 0.071 0.069 0.067 0.066 0.064 0.063240 0.061 0.060 0.059 0.057 0.056 0.055 0.053 0.052 0.051 0.050

69


Annex IX

Institut de Radioprotection et de Sureté NucléaireLaboratoire d’Analyses de Biologie Médicale31 rue de l’Ecluse/3578100 Le Vésinet CedexTel. 01.30.15.52.35 - Fax 01.30.15.52.64

Instructions for twenty-four hour urine specimens

Collection of all urine excreted over the 24 h period1. Empy the bladder in the toilettes2. Start the collection3. Empty the bladder in the specimen bottle4. Place the specimen bottle in the specified cold roomThe bottle must be securely closed (check both cap and package)

Fill out the identification form 478 and include with the sample.

Dispatch of the samples :The sample bottles must be dispatched to IRSN as soon as possible using the material supplied.

ATTENTION :

• Do not collect on working premises.• The sample bottle contains a preservative and must be protected from any sources of

radioactive contamination.• Expedition by normal postal service is not authorized.

The quality of the results of the tests depends on respecting the above rules ofurine sample collection and correct means of expedition.

1st Day 2nd Day

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Annex X

Decontamination techniques for external contamination

The following procedures can be carried out on the premises in case of external contaminationwith a radioisotope. There are emergency showers located on each laboratory floor and in thebasement of the tower building.

Shower using soap and water for extensive contamination

Take a warm shower and after extensive soaping, with light rubbing if necessary, thecontaminated area should be abundantly rinsed. Care should be taken not to spread thecontamination beyond its existing limits; in particular avoid the eyes and ears. Each shower should last a maximum of 5 minutes and if necessary, repeat this procedure upto three times before drying the washed skin, without rubbing, using a towel.

Washing with warm water and soap or detergent for a localisedcontamination.

Rinse and soap the contaminated area. The skin must never be rubbed so that it is broken, andcare should be taken that all folds of skin and the areas around and under finger-nails arethoroughly 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 threetimes; in the event of contamination persisting, medical help must be sought immediately.

Washing with water or physiological saline

This technique should be reserved for situations where the above procedures cannot be used,for instance where there is a cut in the skin. In this case, the skin should be abundantly rinsedfor at least three minutes with fast flowing water or physiological saline, so that any dirt orother particles are rinsed away.

In a situation where mucous membranes have been contaminated, the following proceduresshould be followed:

The mouthRinse the mouth repeatedly with water, taking extreme care not to swallow the washingsolution.

The noseBlow the nose gently using paper handkerchiefs, which should be kept to check the level ofcontamination by the medical services.

The eyesRinse eyes abundantly using water or physiological saline. Use eye washes. Physiological saline: 9g of sodium chloride (NaCl) per 1litre water: the solution must not bekept for longer than one month.

SAFE USE OF BIOLOGICAL SUBSTANCES

In a research institute such as IARC, where biological agents or compounds are stored andused, the risk of infection or contamination exists not only for laboratory personnel but alsofor other staff. In the interests of everyone, it is therefore essential that a certain number ofhealth and safety regulations be strictly followed when working with biological agents (bacteria,yeasts, viruses) or biological samples (cells, blood or blood-derived components, bodysecretions or organs of human or animal origin) in order to minimise these risks. All biologicalagents and samples must be handled in laboratories providing an appropriate level ofconfinement, and wherever possible special precautions must be taken to inactivate potentialdisease-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 otherbiological samples without having first been informed of the special procedures applicable fordecontamination or disposal of waste generated in the laboratory.

The purpose of this chapter is to draw your attention to the circumstances in which anaccidental infection may occur and to outline the measures to be taken to minimise risks.

MEDICAL SURVEILLANCE

In addition to adopting good working practices, which are essential for accident prevention andsafety, vaccination against certain infectious agents can provide an extra measure of protectionfor laboratory staff. Vaccination against hepatitis B virus is compulsory for all laboratorypersonnel (except in cases where there are medical contra-indications). A vaccinationcertificate (showing immunological evidence of seroconversion) must be provided by all newpersonnel 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 informthe Staff Physician of any potentially dangerous laboratory activities that are carried out on aregular basis.

Staff are also strongly recommended to inform the Staff Physician in confidence if their stateof health is incompatible with the handling of chemical or biological products or to exposureof given infectious agents.

It is compulsory to inform the Staff Physician or Laboratory Safety Officer of any incident orwork accident, however minor.

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72

9 SAFE USE OF BIOLOGICAL SUBSTANCES

CONFINEMENT LEVEL FOR MANIPULATING GENETICALLYMODIFIED ORGANISMS (GMO) AND BIOLOGICAL SAMPLES

Unfortunately, it is impossible to evaluate rigorously the risks linked to the genetic manipulationof DNA, often called genetic engineering or biotechnology. Measures and recommendationshave been formulated in almost all countries to try to limit potential dangers linked to thistechnology. Four different levels of laboratory confinement (L1-L4) for the handling ofbiological agents have been established:

• 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, and possesses specific equipment (class II laminar flow hoods, incubators, centrifuges) as well as adequate equipment for decontamination and disposal of biohazard waste. IARC laboratories 604, 708, 717, 809A, 914, 915, 920, 1006 and 1007 are L2 laboratories, most of them being cell culture laboratories.

One person is responsible for each L2 laboratory (their name and telephone number are shown on the laboratory door). This person ensures that the laboratory functions correctly and supervises the work carried out by the cleaning personnel. Anyone using an L2 laboratory must have a laboratory coat reserved for use in this type of laboratory.

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. Before beginning work in this laboratory, the form “Registration form for experiments using infectious vectors, viruses or other biological agents to be performed at IARC” should be filled in (available on Intranet under “Forms and templates – IARC Templates”) and an appointment should be made with the Laboratory Safety Officer who will check that the user is aware of all the specific handling procedures. The person’s badge can then be activated to allow access to this laboratory.

• Laboratory Level L3: used for handling of biological agents in group III: "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 terms of access and equipment apply in order to better protect the laboratory worker and to avoid the dispersal of micro-organisms in the environment. IARC possesses an L3 laboratory on the 9th floor, access to which is restricted to several authorised persons (see Chapter 12 fordetails about its use).

• Laboratory Level L4: used for handling of biological agents in group IV: “causes serious disease in humans and represents a major risk for working personnel”. High risk of propagation in the surrounding area. Only a few laboratories in the world have this confinement level.

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SAFE USE OF BIOLOGICAL SUBSTANCES 9

Any project involving the use of genetically modified organisms must be authorised by theCommission of Genetic Engineering which comes under the French Ministry of Education andResearch. IARC has a global authorization for the different laboratory groups. Thisauthorization is given for five years after which it must be renewed. The projects, dependingon the agents manipulated, are classified into different groups. This grouping will determine theconfinement level and thus the laboratory in which the experiments can be performed. Inaddition, biological samples which have not necessarily been genetically modified but which mayhave been contaminated with viruses, other infectious agents or carcinogens, must also behandled in laboratories with an appropriate level of confinement. For example, any biologicalsample contaminated with hepatitis B or C viruses, (HBV or HCV) or humanimmunodeficiency virus (HIV) must be handled in a L3 laboratory. Full details about the levelof confinement required for the handling of samples contaminated with different viruses andother infectious agents are available from the Laboratory Safety Officer.

The Laboratory Safety Officer must be informed about any work that may lead to the use ofnew GMOs. He/she will then be able to request a modified authorization if necessary.Laboratory workers should follow the same rules and take the same precautions whenworking with DNA as when working with infectious agents or materials for which the dangerto 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 statusof the samples is unknown.

It is therefore compulsory to wear a laboratory coat, gloves and safety glasses tohandle all blood samples. Working in a class II laminar flow hood is highlyrecommended. When this equipment is not available, a fume cupboard should be usedinstead, which allows the work area to be well defined. All work surfaces must be thoroughlycleaned with a detergent-disinfectant solution (Bacinyl) or a diluted bleach solution at the endof every experiment.

Any biological samples that might be contaminated with an infectious agent mustbe handled in a laboratory which provides the appropriate level of confinement(contact the Laboratorty Safety Officer for details).

In the basement of the tower there is a small room containing a laminar flow hood. This roomis reserved for the sorting of samples, particularly for epidemiological groups which do notpossess a laboratory, but it can also be used by laboratory groups that do not possess such ahood on their floor to work with biological samples. This room is kept locked. If you wish touse it, please contact the Biological Resources Manager.

74


It is important to note that laminar flow hoods have been designed to protect the user, thesamples (sterile air under the hood) and the environment (air filtered through HEPA filterswhich block particles bigger than 0.2µm). However, these hoods do not protect against vapourswhich pass through the HEPA filters. As the air in the room is recycled, volatile toxic productsshould not be handled and DNA extractions should not be carried out, for example in ordernot to be exposed to phenol vapours. For further details, see the slide show “Cell culture andL2 facility” on the Committee’s web page: http://intranet/labcom/index.htm.

If tubes appear be contaminated on the outside, they must be wiped with a tissue impregnatedwith diluted bleach before opening them. The dirty tissue must be thrown away in a burn-binfor incineration.It is recommended, whenever experimental procedure allows, that tubes containing bloodproducts be heated at 56°C for 45 min before any manipulation to inactivate HIV or hepatitisviruses.

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 mucousmembranes (eyes, nose, and mouth) or onto broken skin.

It is highly recommended that disposable plastic items be used for all such experimentalprocedures and these be disposed of in specific waste boxes. If the use of glassware cannotbe avoided, it must be decontaminated with a detergent-disinfectant solution or diluted bleachsolution before being given to the glassware cleaning service. Always take off your gloves and wash your hands after finishing your work.

Tissue sections

Various laboratory techniques are carried out using tissue sections where the tissues haveusually 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 highlyrecommended.Handling frozen tissue has a far higher risk of contamination, as viruses and other infectiousagents are not inactivated. A protocol specific to the experiment and the samples being handledshould be prepared before starting any manipulation, taking into account any potentialcontamination of the samples and thus the level of confinement necessary.

Cell lines

Cell lines from human and animal samples are routinely established and cultured in IARClaboratories. They must be handled under laminar flow hoods that meet European safetyguidelines and which are located in L2 laboratories. Disposable gloves and dedicated laboratorycoats should be worn at all times. Particular care should be taken when establishinglymphoblastoid cell lines by infecting lymphocytes with Epstein-Barr virus (EBV). Thisexperiment should be carried out in L2 laboratory 920, which is equipped with additionalsafety conditions compared to an ordinary L2 laboratory. It is highly recommended that anyoneroutinely establishing or working with such cell lines should be aware of their own EBVimmunological status (check with the IARC Staff Physician) and that all necessary precautionsare taken when manipulating such cultures.

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SOURCES OF CONTAMINATION

The principal causes of individual contamination are negligence and overconfidence. Rememberthat, whatever experience you may have, accidents can happen anytime.

The main sources of contamination are:

Aerosols

Inhalation of aerosols constitutes one of the principal causes of laboratory infection. Asmentioned previously, the handling of blood samples can generate very contaminating aerosols.

• Urine, blood 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.

• The sonication and maceration of infected tissue can also produce an infected aerosol. Only blenders or mixers designed to minimise aerosol formation should be used. These risks can be further reduced by working under a tissue culture hood.

• The inoculation of infectious agents or the collection of infectious material can also lead to the formation of aerosols, therefore to a risk of contamination. These procedures should be carried out in a fume cupboard.

Scalpels, needles and syringes

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. The use of disposable scalpelsis highly recommended: changing a scalpel blade can be dangerous and lead to deep cuts.

Accidents frequently involve the use of syringes and needles. Therefore, work very carefullywhen handling them and use them only when necessary. Try to avoid drops of liquid at the endof a syringe needle just before injecting: many accidents are due to accidental injection, and adeep injection could result in tissue contamination.

Never try to put needles back in their initial protective packaging, but dispose ofthem in appropriate containers: burn-bins or boxes labelled 'Jet Aiguilles'.

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Evaporation/Lyophilisation

Before handling a lyophiliser, you must know how to operate the machine correctly (if in doubt,ask the staff member responsible for the equipment before starting). Contamination can occurboth during the evaporation phase and when the machine is being stopped. When carryingout a lyophilisation, always verify that the vacuum tubes are correctly connected and alwayswork carefully with infectious, lyophilised material. Containers of lyophilised, infectious materialwhich are broken upon reception are often sources of laboratory contamination. Receptionand unpacking of tubes containing infectious material should be carried out under a laminarflow hood.

Centrifugation

Make sure you know how to operate the centrifuge before use. Keep centrifuges and bucketsclean and decontaminate them regularly. Contamination often occurs when buckets are filled,when bottle caps or liquid layers are removed or when tubes break during centrifugation, etc.The use of caps fitted to the buckets is recommended and compulsory when centrifugingblood samples; this will ensure that the sample remains confined in case of a problem andminimises the risk of contamination.

Transport of samples should be avoided whenever possible. All L2 cell culture rooms shouldbe equipped with a centrifuge.

Transport of biological samples between floors must only be done using the redlift, which is specifically for the use of transporting laboratory products.

Pipetting

Disposable plastic pipettes should be used whenever possible.

Mouth pipetting is absolutely forbidden. Use pipette bulbs or automatic aspirators. Never leavea 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 whenputting on or taking off the pipette bulb.

NEEDLES MUST NEVER BE DISPOSED OF IN ANORDINARY WASTE PAPER BASKET

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DISPOSAL OF CONTAMINATED SOLID WASTE

All biological waste should be incinerated. Solid waste must be disposed of in the “burn-bin"boxes or the plastic-lined cardboard boxes which are clearly labelled with a BIOHAZARD label.

A green label should be stuck on all boxes containing waste presenting abiological risk (cell cultures, biological samples) (see label opposite). This willindicate to the persons responsible for these boxes that they should be dealtwith in priority.The waste bins are collected every day and stocked prior to being collectedby an outside company.

DECONTAMINATION

Decontamination of biologically contaminated cell cultures and other material is the bestmeans of protection against infection for laboratory personnel. Decontamination is alsonecessary to protect the environment, i.e. to avoid spreading environmentally dangeroussubstances. In general:

• Work benches should be disinfected regularly with Bactinyl (available from the ASO Safety Officer) or ethanol at 70%. Do not ask cleaning staff to carry out this task.

• Glassware, plastic containers, solutions and other materials that 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.

Media which has served for cell culture, whether contaminated or not by viruses, should bedecontaminated with a bleach solution. One part of bleach per twenty parts of water (onedose for one litre of bleach) should be sufficient for decontamination. A sufficient proportionof bleach solution will make the medium change colour to yellow. It can then be discardeddown the sink (see Chapter 12 for waste generated in the L3 laboratory).

Blood samples should be decontaminated in a fume cupboard in a recipient containing dilutedbleach. Never add a concentrated solution of bleach to blood samples; it produces a vigorouschemical reaction that generates chlorine vapours.

Contaminated Pasteur pipettes, disposable tips and other glass- or plastic-ware should bedisposed of in 'burn-bins' for incineration. Never autoclave glassware covered in chlorinebleach, as this can produce toxic chlorine vapours which can damage the autoclave. Thoroughlyrinse all bleach-decontaminated material before autoclaving.

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Whenever possible all culturing of bacteria should be carried out in laboratory 919 on the 9thfloor, which has been set up for this purpose. Media and flasks which have been used forbacterial cultures must be autoclaved or decontaminated with bleach before being discarded.Contaminated petri dishes should be placed in bags before autoclaving or in the incinerationboxes.

Glassware

Erlenmeyer flasks, pipettes, tubes, test tubes and glass bottles are soaked after use, completelyimmersed, in a detergent bath (Bactinyl). Glassware that has been used for cell cultures shouldbe decontaminated by total immersion in Bactinyl solution for a minimum of three hours.

Disposable "Nalgene" filters

Disposable filters that have been used for filtering non-contaminated liquids can be discardedinto the normal waste. Those used for filtering viral solutions or contaminated liquids must bethrown away in boxes destined for incineration.

Scissors, tweezers and other instruments

Dissection instruments must be soaked in a concentrated detergent solution immediatelyafter use. 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 inordinary waste baskets meant for waste paper or plastic, but disposed of in "burn-bins" forincineration.

Inactivation of HIV (Human Immunodeficiency Virus)

HIV, the etiological agent of acquired immune deficiency syndrome (AIDS) belongs to a groupof RNA-containing viruses, called retroviruses. Transmission of HIV occurs either by sexualintercourse (semen, vaginal secretion) or by blood or blood products. However, the virus hasalso been isolated from saliva, tears, breast milk, cerebrospinal fluid, amniotic fluid, urine andother body fluids and secretions. These biological components should be considered aspotential sources of contamination, even though epidemiological evidence indicates that blood,semen, vaginal secretions, 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 clinicallaboratory workers will be exposed to blood and other bodily fluids and secretions frompersons infected with HIV is also increasing. It is important for these workers to be aware ofand to take all of the necessary precautions to avoid contamination with HIV or with otherhuman retroviruses.

The recommendations described in this chapter, if followed, are largely sufficient to reduce therisk of contamination by many infectious agents, including HIV. In addition, HIV is an envelopedvirus and therefore very sensitive to most of the usual detergents used in the laboratory.

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HIV is inactivated rapidly if exposed to commonly used chemical disinfectants atconcentrations much lower than those normally used. A 1 to 10% solution of sodiumhypochlorite (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 followingdisinfectants have also been proven effective in inactivating HIV:

• 10% RBS HBS 10 at 10% at 37ºC• Bactinyl• 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

Other precautions

Bench surface contamination can be avoided by working on sheets of absorbent plastic("Benchkote"). After use, these sheets should be thrown away in boxes destined forincineration.The use of trays inside the hood or on the bench to contain the products or the samplesreduces the spread of contamination in case of an incident and facilitates cleaning.

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.

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ACCIDENTS INVOLVING POTENTIALLY CONTAMINATED BIOLOGICAL FLUIDS

(See Annex I)

1. In case of an injury while working with potentially contaminated biological fluids, immediately clean the cut. Do not make it bleed.

2. Wash with soap and water, and rinse.

3. Disinfect by immersing the cut for at least 5 minutes 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.

If a biological sample was involved in the accident, evaluate rapidly the risk of potential infectionby consulting immediately Dr Cuche, 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 redalarm button, will ensure that the necessary action is taken.

The HIV and HCV status of blood samples can be assessed rapidly by the “Laboratoired’analyses medicales” located opposite IARC, at 184, avenue des Frères Lumiere (tel. 04 78 7403 58). 0.5ml of the blood sample involved in the accident is necessary and should be takendirectly to the laboratory.

It is important to note that if a treatment against HIV is to be considered, its efficiency will bemuch greater if started within 4 hours. But as it is a complicated therapy, with a lot of potentialside-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 helpcan be obtained by contacting:

- Hôpital E. Herriot - Pavillon P - Professor Touraine’s service - Tel: 04 72 11 02 29 - Hôpital de l'Hôtel Dieu - Professor Trepo’s service - Tel: 04 72 41 30 68, - Hôpital de la Croix Rousse - Professor Peyramond’s service - Tel: 04 72 07 17 54

All accidents, however minor, should be reported to Dr Cuche or B. Chapot.

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Annex

PROCEDURE IN CASE OF AN ACCIDENT INVOLVINGPOTENTIALLY CONTAMINATED LIQUIDS

WORKING WITH ANIMALS

The animal house, which was situated on the 13th floor of the IARC tower building, was closedat the beginning of 2007, but nevertheless it is still possible to continue experiments on mice:

• If you have a level 2 diploma in carrying out animal experiments, you can continue to work on animals in another animal house.

• It is also possible to subcontract all activities on animals.

You can contact the following animal houses in order to define the conditions forsubcontracting:

• Ulysse at the Laennec University• The PBES at the Ecole Normale Supérieure• Charles River at L’Arbresle

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COLLECTIONS OF BIOLOGICALSPECIMENS

DESCRIPTION OF THE COLLECTIONS AND INFRASTRUCTURE

All biological specimens (including human tissue, animal tissue, cell lines) kept at IARC mustbe registered in a centralised database. Each specimen must be listed by an identificationnumber unique to IARC.

Biological specimens (frozen or in paraffin blocks) are mainly kept in a common storage roomin the basement of the Sasakawa building. This area contains small tanks of liquid nitrogen aswell as -80°C, -40°C and -20°C freezers including emergency freezers.

This room also contains a small laboratory dedicated to the handling of specimens which havebeen sent to IARC or are being sent out.

Another room is reserved for storing biological specimens at room temperature such as bloodsamples on blotting paper and also histological slides and paraffin blocks.

Both small and large tanks of liquid nitrogen are kept in the BRC building (rooms E01, E02, E03).

The rules and regulations concerning access to the common storage room are laid out in theEnglish manual “Guidelines for IARC Archives of Biological Material” (May 2003), which can beobtained from every Group Secretary.

Management of these resources is currently supervised by Dr Pierre Hainaut, Head of theMolecular Carcinogenesis Group (MOC) with the assistance of the Biological ResourcesManager, Elodie Caboux.

It should be noted that biological specimens stocked at IARC can represent a serious biologicalrisk. Rules and regulations should be followed scrupulously and when handling biologicalspecimens, it is compulsory to wear protective clothing (laboratory coat, gloves and safetyglasses).

HANDLING BIOLOGICAL SPECIMENS

Database

An IARC database, which is currently being developed, has been created to list all the biologicalspecimens that have been sent to the Agency as part of studies carried out in collaborationwith other institutes and those that have been produced at the Agency, such as cell lines andderived products such as DNA.

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11 COLLECTION OF BIOLOGICAL SAMPLES

Basement laboratory

A room has been fitted out in the Sasakawa basement to be used to receive, sort, aliquot andsend out biological samples. This room is equipped with a laboratory workbench and a laminarflow hood. All handling of biological samples should be carried out under this hood. Thislaboratory is kept locked at all times and a key is available at the switchboard and from theBiological Resources Manager. Every entry to and exit from this laboratory should be notedin a user’s register.

Procedure for the transport of biological samples

Sending biological samples by air is subject to the International Air Transport Association (IATA)regulations. An international classification of dangerous products exists namely for infectiousproducts. This classification takes into account the potential infectious risk (blood andderivatives) but also other more specific risks (dry ice, inflammable liquids, paraffin).

Before shipping specimens, please check whether or not they are subject to the DangerousGoods Reglementation (DGR) and, in any case, please follow the appropriate procedure inplace at IARC. Information related to shipment of biological material is available on the Intranetunder IARC Services and Resources/Labs/Biological Resources.

WORKING IN THE L3 LABORATORY

The L3 confinement level authorises the handling of Group III biological agents (could causea serious disease in humans and represents a major risk for laboratory staff) such as thehepatitis B virus or the AIDS virus. This level of confinement is governed by a very strictprotocol in order to protect laboratory staff and to prevent the leak of micro-organisms intothe environment.

ACCESS TO THE L3 LABORATORY

Access to the L3 laboratory is by magnetic badge only. The laboratory is situated in room 901on the 9th floor and is currently supervised by Dr Massimo Tommasino, Head of the Infectionsand Cancer Biology Group (ICB).

All rules and regulations concerning access to the L3 laboratory can be found in the manual“Instructions for handling infectious material and genetically modified organisms (GMO) inthe L3 laboratory”.

Anyone wishing to carry out experiments demanding a level L3 confinement must fill in theapplication form “Entry into L3 laboratory” which is available on the Intranet in the section:“Forms and templates/IARC template” and forward to the ICB Group Head after clearancefrom their own Group Head. Work in this type of laboratory can only go ahead with theauthorisation of the Staff Physician.

Before beginning the experiment, the person should have been trained in safety measures,experiment techniques and good practice. All this theoretical and practical training is set outin the document “Approval to work in the IARC L3 laboratory” and it is only once thisdocument has been completed that access to the laboratory can be granted.

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INDEX

AAccident, 6, 10, 11, 17, 18, 40, 71, 80Acrylamide, 42Aerosols, 53, 58, 68, 75Alarm, 9, 10Animal house, 83ASO Safety Officer, 6, 25, 33, 34, 35, 38, 54, 55,

58Assembly point, 9

BBack-up freezers, 26, 85Bacteria, 71, 78Bacteriology laboratory, 13Becquerel, 61Benzene, 41Biological Resources Centre (BRC), 7, 8, 9, 10,

11, 13, 14, 20, 25, 26, 27, 85Biological samples, 5, 6, 71, 73, 76, 77, 86Blood, 18, 71, 73, 74, 75, 76, 77, 78, 80, 85, 86

CCarbon-14, 47, 49, 50, 51, 54, 55, 56, 57, 64Carcinogen, 6, 13, 20, 22, 23, 28, 37, 38, 39, 41,

42, 43, 44, 56Cell lines, 74, 85Centrifugation, 76Children, 14Chloroform, 23, 41Cleaning personnel, 20, 27, 72Cold rooms, 21, 24Confinement level, 72, 73, 87Contact lenses, 23Controlled access, 13

DDefibrillator, 11Defrosting and decontamination of freezers

and refrigerators, 25Dichloromethane, 23, 41Domestic animals, 14Dosimeters, 48, 49, 52, 65Drinks, 21

EElectrocution, 10Emergency wardens, 9

Ethidium bromide, 41, 42Evacuation of the building, 9Explosion, 10, 26

FFire, 10, 12Fire extinguishers, 9, 12First aid supplies, 11First-aiders, 11Food, 21, 22, 79Formaldehyde, 41Fume cupboard, 24, 39, 40, 41, 42, 49, 51, 52,

73, 75, 77

GGas cylinders, 25Genetically Modified Organisms - GMOs, 73Glassware, 26, 27, 44, 53, 58, 74, 77Glassware washing service, 26Gloves, 22, 39, 53, 73, 85

HHBV (Hepatitis B virus), 45, 71, 73, 87HCV (Hepatitis C virus), 45, 73HEPA filters, 39, 74HIV (AIDS virus), 45, 73, 80

IIARC Monographs, 37Inflammable gases, 25, 35Inflammable substances, 28, 33, 34Iodin-125, 47, 48, 49, 50, 52, 54, 55, 56, 57, 68

KKitchen, 21, 22, 26

LL3 laboratory, 13, 72, 73, 77, 87Laboratory cleaning, 21Laboratory coats, 22Laboratory equipment, 29Laboratory Safety Officer, 5, 6, 7, 11, 13, 25,

28, 34, 38, 39, 43, 49, 50, 71, 72, 73Laminar flow hood, 39, 49, 73Laminar flow hoods, 39, 49, 72, 73, 74

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INDEX

Lifts, 22, 24Liquid nitrogen, 25, 29, 85Lyophiliser, 76

MMagnetic card, 13Medical surveillance, 17Microwave ovens, 26, 30

N‘Nalgene’ filters, 78Needles, 28, 75, 78Noise, 23Normal working hours, 19, 26, 31

OOccupational health and Safety Committee(OHSC), 5, 6, 8

PPeriodic medical examinations, 17Phosphorus-32, 47, 48, 49, 50, 51, 52, 53, 54,

55, 56, 57, 60, 65Phosphorus-33, 47, 49, 50, 51, 54, 55, 56, 57,

66Poison Control Centre, 10Pipetting, 76Pregnancy, 18, 48

RRadioactive contamination detector, 58Radioactive substances, 47Red button, 10, 40Red lift, 24, 35, 40Red telephone, 10Room 805, 22, 37, 38, 39, 40, 50, 51

SSafety glasses, 22, 23, 51, 73Safety procedures, 9Safety showers, 11, 12Scalpels, 75Security team, 6, 10, 80Smoking, 12Solvents, 12, 23, 24, 26, 33, 34, 35, 40, 41, 43, 51,

56Staff Physician, 6, 11, 17, 48, 71, 80, 87Sulphur-35, 47, 49, 50, 51, 54, 55, 56, 57, 67Syringes, 75, 78

TTheft, 12Tissue sections, 74Tritium (3H), 47, 49, 50, 51, 54, 55, 56, 57, 63

UUnattended equipment, 31Unattended laboratory experiments, 19UV radiation, 23

VVaccination, 18, 71

WWarning, 9, 57Waste - Biological, 28, 77Waste - carcinogenic, 28, 42, 43, 56Waste - radioactive, 28, 43, 54Waste - solid & non-contaminated, 27

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