Biotechnology Process Engineering Center
CHEMICAL HYGIENE AND SAFETY PLAN
April 2, 2003
Revised By:
Jean-François P. Hamel
Chemical Hygiene Officer, 2003-2004
and
Daniel Bauer
Assistant Chemical Hygiene Officer, 2003-2004
TABLE OF CONTENTS
I. Policy and Purpose ..................................................................................................................... 1
I. A. POLICY 1
I. B. PURPOSE 1
I. C. PERSONNEL COVERED BY THIS PLAN 1
I. D. STANDARD OPERATING PROCEDURES FOR BPEC LABS IN BUILDINGS 16 AND 56 1
II. Responsibility, Authority, and Resources ................................................................................. 2
II. A. BPEC AND CHEMICAL ENGINEERING DEPARTMENT HYGIENE PLAN REPRESENTATIVES2 II. A. 1. Center Director .......................................................................................................................................2 II. A. 2. Chemical Hygiene Officer......................................................................................................................2 II. A. 3. Laboratory Supervisor ...........................................................................................................................3 II. A. 4. EHS coordinator responsibilities ...........................................................................................................5 II. A. 5. EHS representative responsibilities .......................................................................................................6 II. A. 6. DLC EHS Committee .............................................................................................................................6 II. A. 7. Employees, Staff, and Students.............................................................................................................11
II. B. VISITORS AND MIT SERVICE PERSONNEL 13
II. C. ENVIRONMENTAL HEALTH & SAFETY TEAM 13
II. D. MIT CHEMICAL, SAFETY AND EMERGENCY SERVICES 16
III. Preparation, Implementation, Annual Review, and Update ............................................. 16
III. A. PREPARATION AND APPROVAL 16
III. B. DISTRIBUTION AND IMPLEMENTATION 17
III. C. COMPLIANCE AND ENFORCEMENT 17
III. D. ANNUAL REVIEW AND UPDATE 18
IV. Basic Chemical Hygiene and Laboratory Safety Equipment ........................................... 19
IV. A. PERSONAL PROTECTIVE EQUIPMENT 19
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IV. A. 1. Eye Protection.....................................................................................................................................19 IV. A. 2. Protective Apparel ..............................................................................................................................21 IV. A. 3. Respirators..........................................................................................................................................22
IV. B. CONTAINMENT EQUIPMENT AND PROCEDURES 22 IV. B. 1. Chemical Purchase .............................................................................................................................22 IV. B. 2. Chemical Transportation ....................................................................................................................23 IV. B. 3. Chemical Storage................................................................................................................................24 IV. B. 4. Fume Hoods........................................................................................................................................25
IV. C. SAFETY EQUIPMENT 27 IV. C. 1. Fire Extinguishers and Blankets.........................................................................................................27 IV. C. 2. Bandages ............................................................................................................................................28 IV. C. 3. Safety Showers and Eyewashes...........................................................................................................28 IV. C. 4. Spill Kits .............................................................................................................................................28
V. Basic Chemical Hygiene and Laboratory Safety Procedures .................................................. 29
V. A. ACCIDENTAL RELEASE OF HAZARDOUS SUBSTANCES (SPILLS) 29
V. B. AUTOCLAVE AND STEAM USE 32
V. C. CHECKING OUT OF THE LABORATORY 33
V. D. CHILDREN AND PETS IN LABORATORIES 33
V. E. ELECTRICAL SAFETY 34
V. F. EMERGENCY ACTION PLAN 37 V. F. 1. Emergency Information Cards and Phone Lists...................................................................................38 V. F. 2. Emergency Evacuation.........................................................................................................................38 V. F. 3. Fires .....................................................................................................................................................39
V. G. EQUIPMENT MAINTENANCE AND MALFUNCTION 41
V. H. GAS CYLINDERS AND HIGH PRESSURE GAS SYSTEMS 42
V. I. GLASSWARE 46
V. J. HOUSEKEEPING 47
V. K. INCOMPATIBLE SUBSTANCES 48
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V. L. PERSONAL HYGIENE AND SANITATION 49
V. M. REU AND UROP REGULATIONS 50
V. N. SIGNS AND LABELS 50
V. O. HAZARDOUS WASTE DISPOSAL 53
V. P. WORKING AFTER HOURS 57
VI. Hazardous Chemical Evaluation....................................................................................... 57
VI. A. CLASSIFICATION OF SUBSTANCES 57 VI. A. 1. MSDS’s and Other Information Sources.............................................................................................57 VI. A. 2. Substance Evaluation..........................................................................................................................58
VI. B. INVENTORY CONTROL FOR RADIOACTIVE SUBSTANCES 59
VI. C. PARTICULARLY HAZARDOUS SUBSTANCES 59
VI. D. STANDARD OPERATION PROCEDURE FOR PARTICULARLY HAZARDOUS SUBSTANCES59
VI. E. ELIMINATION OR SUBSTITUTION 60
VII. Facilities for Particularly Hazardous Substance Use and Disposal ................................. 60
VII. A. BIOHAZARDOUS MATERIALS/ RECOMBINANT DNA RESEARCH 60 VII. A. 1. Control Measures ..............................................................................................................................62 VII. A. 2. Purchase and Transportation ............................................................................................................63 VII. A. 3. Storage...............................................................................................................................................64 VII. A. 4. Biological Safety Cabinets (or biosafety cabinets) ............................................................................65
VII. B. SPILLS AND DISPOSAL 67 VII. B. 1. Accidental Release of Biohazardous Substances (Spills)...................................................................67 VII. B. 2. Waste Disposal and Decontamination...............................................................................................68 VII. B. 3. Summary of Sterilization Methods and Procedures...........................................................................72
VII. C. LASERS 78
VII. D. RADIOACTIVE MATERIALS 79 VII. D. 1. Authorization for Use ........................................................................................................................80 VII. D. 2. Radiation Benches (Designated Areas).............................................................................................80 VII. D. 3. Purchase, Inventory Control and Transportation .............................................................................81
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VII. D. 4. Storage Facilities for Radioactive Materials ....................................................................................81 VII. D. 5. Working with Radioactive Materials.................................................................................................82 VII. D. 6. Accidental Release of Radioactive Substances (Spills) .....................................................................84 VII. D. 7. Waste Containers and Disposal ........................................................................................................87
VII. E. EQUIPMENT FOR RADIOISOTOPES 87
VIII. Enclosure, Isolation and Regulated Areas ........................................................................ 88
VIII. A. DESIGNATED AREAS - DEFINITION 88
VIII. B. FUME HOODS 89
VIII. C. RADIOACTIVE BENCHES 89
VIII. D. SHORT-TERM USE HAZARDOUS MATERIALS 89
IX. BPEC Laboratory Standard Operating Procedures (SOP’s)............................................. 89
IX. A. SOP’S FOR HAZARDOUS AND PARTICULARLY HAZARDOUS CHEMICAL CATEGORIES90 IX. A. 1. Carcinogens ........................................................................................................................................90 IX. A. 2. Reproductive Toxins............................................................................................................................91 IX. A. 3. Radioactive Materials .........................................................................................................................92 IX. A. 4. Biohazardous Materials......................................................................................................................92 IX. A. 5. Compounds with a High Degree of Acute Toxicity .............................................................................93 IX. A. 6.Organic Liquids; Combustible, Explosive, Flammable, and Volatile Materials; Liquid Inhalation Hazards that are Acutely or Chronically Toxic or Acute Contact Hazards.......................................................94 IX. A. 7.Solid Inhalation Hazards (Powders) that are Acutely or Chronically Toxic or Acute Contact Hazards...........................................................................................................................................................................94 IX. A. 8.Gases that are Flammable or that are Acutely or Chronically Toxic or Acute Contact Hazards ........94 IX. A. 9.Acids; Caustics; Liquid and Solid Eye, Oral and Skin Acute Contact Hazards....................................94 IX. A. 10. Highly Reactive Material ..................................................................................................................95 IX. A.11. SOP’s for Equipment ..........................................................................................................................95
X. Education and Training............................................................................................................ 95
X. A. INITIAL TRAINING 95
X. B. INSTITUTE-WIDE TRAINING FOR PARTICULARLY HAZARDOUS SUBSTANCES 96 X. B. 1. Biohazardous Material.........................................................................................................................96 X. B. 2. Lasers ...................................................................................................................................................97 X. B. 3. Radioactive Material............................................................................................................................97
X. C. CLEARANCE FORMS FOR OUTSIDE OR TEMPORARY PERSONNEL 97
X. D. CPR CERTIFICATION 97
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X. E. TRAINING UPDATES 97
XI. Monitoring, Medical Surveillance and Employee Exposure ............................................ 98
XI. A. MONITORING AND MEDICAL SURVEILLANCE 98
XI. B. ACCIDENTS AND EMPLOYEE EXPOSURE REPORTING 100
XII. Recordkeeping.................................................................................................................... 101
XII. A. AUTHORITY FOR CHEMICAL USE 101
Standard Operating Procedure For Autoclaves........................................................................... 113
Protective apparel .......................................................................................................................... 115
Preparation and Use of the BSC...................................................................................................... 115
SPECIALIZED EQUIPMENT 115
XII. C. CHEMICAL HYGIENE AND SAFETY CLEARANCE FORMS 116
XII. E. LARGE-SCALE BATCH FERMENTATION RUN LOGS 120
XII. F. MEDICAL SURVEILLANCE AND ACCIDENTS 120
XII. G. MSDS 120
XII. H. RADIOISOTOPE WORK 120
XII. I. STANDARD OPERATING PROCEDURES 120
XII. J. TRAINING 122
XIII. References......................................................................................................................... 123
XIV. Appendices........................................................................................................................ 124
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Acknowledgments Portions of this Biotechnology Process Engineering Center (BPEC) Chemical Hygiene and Safety Plan (CHASP) were drawn from the following resources: 1) The MIT template for Chemical Hygiene Plans, 1990. 2) The MIT Chemistry Department’s Chemical Hygiene Plan and Safety Manual, 1991.
Over one-third of the BPEC plan was modeled after this work. Without this resource, the BPEC plan would be far from complete.
3) The BPEC Safety Manual, 2001. 4) The MIT Chemical Engineering Department’s Chemical Hygiene Plan, 1991 and 2000,
after which many sections were modeled.
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I. Policy and Purpose
I. A. Policy It is the policy of the Massachusetts Institute of Technology (as is represented by the MIT Corporation and the Office of the President) to provide a safe and healthy workplace in compliance with the Occupational Safety and Health Act of 1970 and regulations of the Department of Labor including 29 CFR 1910.1450 "Occupational Exposure to Hazardous Chemicals in Laboratories".
I. B. Purpose Chemical Hygiene Plans are required of all departments maintaining laboratories that utilize toxic chemicals, as defined by law. This document is the Chemical Hygiene Plan (CHP) required by the above regulation and the Biotechnology Process Engineering Center (BPEC) Safety Manual, combined in one document to simplify the implementation of safe practices and to facilitate the transfer of information to employees. The purpose of this Chemical Hygiene and Safety Plan (CHASP) is to describe proper practices, procedures, equipment and facilities to be used by employees, students, research staff, visitors, and other personnel working in each laboratory of the Biotechnology Process Engineering Center in order to protect them from potential health hazards presented by chemicals used in the laboratory workplace, and to keep exposures below specified limits. It is the responsibility of faculty, administration, research and supervisory personnel to know and to follow the provisions of this Plan. J.-F. Hamel is the Laboratory Supervisor and the Chemical Hygiene Officer for the BPEC Laboratories. Professor Douglas Lauffenburger is the Center Director for the BPEC CHASP.
I. C. Personnel Covered by this Plan This Chemical Hygiene and Safety Plan applies to all work that is conducted in space assigned to the Biotechnology Process Engineering Center as defined in Section II. A. 5. of this plan. Individuals holding appointments in other MIT departments or outside investigators who plan to conduct research in BPEC laboratories must undergo the chemical hygiene training outlined in Section X. of this plan and file a Chemical Hygiene and Safety Clearance Form (Section XII. C.) with the BPEC CHO before beginning work.
I. D. Standard Operating Procedures for BPEC labs in Buildings 16 and 56 The BPEC laboratories (16-436, 16-476) and instrument rooms (16-459, 16-463, 16-479, 56-421) all follow the BPEC Chemical Hygiene and Safety Plan. All workers are subject to the BPEC regulations when using the BPEC facilities. STANDARD OPERATING PROCEDURES FOR BPEC LABS:
• Autoclave use • Long Experiments • Use of tissue culture rooms
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II. Responsibility, Authority, and Resources
II. A. BPEC and Chemical Engineering Department Hygiene Plan Representatives
II. A. 1. Center Director The Center Director for the Biotechnology Process Engineering Center is Professor Douglas Lauffenburger, rooms 16-429 or 56-341, telephone extension x2-1629.
• The Center Chairperson has the responsibility and the authority to see that the Chemical Hygiene Plan is written, updated and implemented.
• The Center Chairperson has appointed the Chemical Hygiene Officer in accordance with
the definition provided in paragraph (b) of the 29 CFR1910.1450 and requirements outlined in section B. of this document.
The Center Chairperson has the final responsibility for the safety and health of the employees (and students) conducting work in his/her department and visitors.
II. A. 2. Chemical Hygiene Officer For BPEC, the Chemical Hygiene Officer is Jean-François Hamel Room: _56-483____ Extension: __x8-6665______Email:[email protected]________
REQUIREMENTS:
The OSHA laboratory standard requires designation of a Chemical Hygiene Officer. Within MIT, this means that each department or other major administrative unit, which uses laboratory chemicals, must appoint its own Chemical Hygiene Officer. DEFINITION:
The Chemical Hygiene Officer is an employee designated by the employer (Dr. Douglas Lauffenburger), and who is qualified by training or experience, to provide technical guidance in the development and implementation of the Biotechnology Process Engineering Center written Chemical Hygiene Plan. Chemical Hygiene Officers at MIT will be technically competent and have appropriate authority to assist with development and administration of departmental plans. In most cases, Chemical Hygiene Officers will be tenured faculty, or similarly senior scientists, with supervisory responsibilities.
DUTIES:
The Chemical Hygiene Officer will assist responsible Principal Investigators and Departmental Chairs to accomplish the following:
1. Work with Administrators to develop Chemical Hygiene Plans for the department and to
implement plans at the level of individual laboratories.
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2. Work with Administrators and Principal Investigators to monitor safe procurement, use, and disposal of chemicals.
3. Assist responsible Principal Investigators with required safety audits and their documentation
(which includes documentation of training). 4. Advise Principal Investigators concerning adequate facilities and procedures under the
regulation. 5. Seek ways to improve the Chemical Hygiene Program. 6. Act as a liaison between the EHS Coordinator and Faculty. 7. Co-Chair the BPEC-EHS Committee
In addition, the Chemical Hygiene Officer will be responsible for knowing the contents of the relevant regulation (Occupational Exposures to Hazardous Chemicals in Laboratories, 29 CFR 1910.145) as well as the BPEC Chemical Hygiene Plan. RESOURCES:
The Chemical Hygiene Officer may call upon BPEC Administrative Officers for administrative support, upon the EHS Office, EHS Coordinator, and EHS Representative for assistance, as well as upon Principal Investigators who will provide specific information concerning their laboratories.
II. A. 3. Laboratory Supervisor The Supervisor’s duties, as defined in the OSHA Laboratory Standard and the MIT Chemical Hygiene Plan, are the responsibility of the Principal Investigator. For laboratories with no Principal Investigator, the Supervisor’s duties are assumed by the person with authority over all laboratory functions. That person shall be appointed by Dr. D. Lauffenburger for BPEC. The laboratory supervisor at BPEC is Jean-François Hamel, Room 56-483, ext. 8-6665.
The primary responsibility of the Supervisor is to institute the Chemical Hygiene Plan and ensure compliance with the OSHA Laboratory Standard. The duties include the following: 1. Ensure that all work is conducted in accordance with the BPEC Chemical Hygiene
Plan;
2. Define the location of work areas where toxic substances and potential carcinogens will be used, and ensure that the inventory of these substances is properly maintained;
3. Obtain, review, and approve standard operating procedures, detailing all aspects of proposed research activities that involve hazardous agents;
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4. Prepare a Standard Operating Procedure for use of test substances when this use involves alternate procedures not specified in these guidelines. The S.O.P. shall include a description of the alternate procedure and an assessment of the alternate controls that will be used;
5. Define hazardous operations, designating safe practices, and selecting protective equipment;
6. Ensure that program and support staff receive instructions and training in safe work practices, use of personal protective equipment, and in procedures for dealing with accidents involving toxic substances;
7. Ensure that employees understand the training received;
8. Ensure that all personnel obtain the medical examinations and protective equipment necessary for the safe performance of their job;
9. Monitor the safety performance of the staff to ensure that the required safety practices and techniques are being employed;
10. Arrange for workplace air samples, swipes or other tests to determine the amount and nature of airborne and/or surface contamination, inform employees of the results, and use data to aid in the evaluation and maintenance of appropriate laboratory conditions;
11. Assist the EHS Office when necessary.
12. Conduct formal laboratory inspections at least twice a year to ensure compliance with existing laboratory S.O.P’s;
13. Prepare procedures for dealing with accidents that may result in the unexpected exposure of personnel, or the environment, to a toxic substance.
14. Investigate accidents and report them to the Chemical Hygiene Officer. Include procedures that will minimize the repetition of that type of accident;
15. Report to the Chemical Hygiene Officer incidents that (1) cause personnel to be seriously exposed to hazardous chemicals or materials, such as through the inoculation of a chemical through cutaneous penetration, ingestion of a chemical, or probable inhalation of a chemical, or that (2) constitute a danger of environmental contamination;
16. Ensure that action is taken to correct work practices and conditions that may result in the release of toxic chemicals;
17. Properly dispose of unwanted and/or hazardous chemicals and materials;
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18. Document and maintain compliance with all local, state and federal regulatory requirements;
19. Make copies of the approved safety plan available to the program and support staff.
II. A. 4. EHS coordinator responsibilities
The EHS Coordinator oversees the day-to-day activities within BPEC to monitor compliance with environmental, health & safety (“EHS”) laws and regulations and MIT practices, and that BPEC and its Principal Investigators/Supervisors are implementing their responsibilities under MIT’s EHS Management System (“EHS Requirements”). The EHS Coordinator supports the BPEC head in his or her responsibility for BPEC EHS performance. The EHS Coordinator will work closely with the MIT EHS Office lead contact and other designated EHS team members to BPEC to assure that EHS Requirements are satisfied. A description of the EHS Coordinator’s responsibilities follows.
1. Meet regularly with EHS Office Lead Contact to discuss BPEC EHS service /
performance needs. 2. Report to the BPEC Head. Partner with EHS Office Lead Contact. 3. Coordinate EHS-MS (Environment Health and Safety Management System) activities
of EHS Representatives within BPEC. 4. Coordinate closely with BPEC and CHO and assist in developing EHS related SOP’s
such as hazardous waste, biosafety, chemical hygiene, laser safety, and emergency action plans.
5. Ensure BPEC personnel receive appropriate regulatory training. 6. Participate in periodic inspections and/or review inspection reports of BPEC
laboratories and facilities. Review and address EHS incidents/ performance issues that occur in BPEC.
7. Assist PIs/ EHS Representatives with laboratory or space registration. 8. Serve as chair or co-chair of the BPEC EHS Committee. 9. Serve as steward of BPEC EHS training, auditing & inventory record keeping
(including providing EHS Office with SARA Title III reporting information). 10. Meet with other EHS Coordinators regularly to share information, discuss EHS-MS,
new or changing EHS regulations, and best practices.
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11. Review significant EHS issues for proposed experiments with PI and EHS Lead Contact/Team Review especially changes that could impact compliance with EHS requirements.
12. Arrange for EHS decommissioning of lab spaces or special inspections as necessary.
13. Meet with regulators during inspections if requested by EHS to do so.
II. A. 5. EHS representative responsibilities The EHS Representative assists the principal investigator or supervisor in charge of the individual laboratory or facility space in complying with environmental health and safety (“EHS”) regulations and MIT practices under MIT’s EHS Management System (“EHS Requirements”). The EHS Representative will work closely with the principal investigator/supervisor and BPEC EHS Coordinator to ensure that EHS Requirements are satisfied.
EHS Representative assists the Principal Investigator or laboratory/facility supervisor with
monitoring that activities in the individual laboratory or facility space achieve EHS Requirements including:
1. Assisting with contacting BPEC EHS Coordinator and /or MIT EHS Office lead contact
person or other MIT EHS Office designated team members for assistance with potentially hazardous materials and potentially hazardous activities.
2. Assisting with providing or arranging for EHS training for personnel working in the
laboratory/facility (including employees, students, and visitors).
3. Assisting with disseminating EHS information to laboratory/facility personnel, including information on methods to reduce or avoid the use of potentially hazardous or toxic materials.
4. Assisting with conducting brief inspections of the laboratory or facility for compliance
with EHS Requirements. Assisting with evaluating and making recommendations on EHS issues that concern activities and materials in the laboratory/facility.
II. A. 6. BPEC EHS Committee
BPEC EHS Committee consists of senior faculty member as Chair (may be Chemical Hygiene Officer), EHS Coordinator as Co-Chair, EHS Lead Contact, CHO (if not Chair or EHS coordinator) and other faculty, staff, or students of BPEC. The committee is accountable to the BPEC Head. Committee responsibilities are described below.
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1. Discuss EHS-MS and new or changing EHS regulations and practices and how best to implement them in BPEC.
2. Review significant changes that could impact compliance with EHS requirements.
3. Participate in periodic inspections and/or review inspection reports of BPEC’s
laboratories and facilities.
4. Review EHS incidents/performance issues that occur in BPEC and recommend appropriate corrective action.
The BPEC EHS Committee for BPEC is comprised of the following members: Room # Phone Extension Chair: Jean-François Hamel 56-483 8-6665 Co-Chair: Daniel Bauer 16-436 3-2765 EHS Lead Contact: Richard Fink Administrative Safety Coordinator: Audrey Jones-Childs 16-429B 3-2504 Graduate Student Safety Coordinators: Kathryn Miller 16-436 3-2165 (lab) 3-7594 (off) Henry Lam 16-476 3-2226 The CHO and/or Laboratory supervisor are responsible for assignment of new personnel. Although personnel from the MIT Industrial Hygiene Program and Safety Program are not members of this Committee, they are consulted as necessary. The following people assist the Chemical Hygiene Officer in implementation and enforcement of all aspects of the Chemical Hygiene and Safety Plan, specific responsibilities of the safety and fire emergency personnel, as follows: Assistant Chemical Hygiene Officer
The Assistant Chemical Hygiene Officer is Daniel Bauer.
1. Ensure that 3 copies of the approved CHASP are available at all times for the Graduate Student Safety Coordinators in lab 16-436 and lab 16-476.
2. Verify that BPEC and guest researchers have received the proper training, and have
completed and signed all required documents before beginning work in the BPEC labs. Submit completed training documents to the Chemical Hygiene Officer.
3. Instruct all lab members that they must contact MIT’s EH&S Team when considering
conducting new experiments involving toxic substances.
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4. Instruct all personnel to conduct work in accordance with the Department’s/Center’s Chemical Hygiene and Safety Plan.
5. Develop and review Standard Operating Procedures for work involving hazardous
substances. This includes preparation of Standard Operating Procedures for use of test substances when this use involves alternate procedures not specified in a general SOP for the given class of chemical being tested. The SOP shall include a description of the alternate procedure and an assessment of alternate controls that will be used. Submit draft SOPs to the Chemical Hygiene Officer, for final review and approval.
6. When required by EH&S Team, by the Chemical Hygiene Officer or the Laboratory
Supervisor, arrange for workplace air samples, swipes, or other tests to determine the amount and nature of airborne and/or surface contamination, inform employees of the results, and use data to aid in the evaluation and maintenance of appropriate laboratory conditions. Provide written results to the EH&S Office, the Chemical Hygiene Officer and the Laboratory Supervisor.
7. Ensure that the sash of the chemical fume hood is kept at an appropriate level, and inform
the Laboratory Supervisor of frequent violators.
8. Work together with MIT’s EH&S Team to ensure that hazardous operations and safe practices are designated and that protective equipment is specified. Seek supervisory assistance from the Chemical Hygiene Officer and the Laboratory Supervisor, when needed. Provide written updates to the Chemical Hygiene Officer and the Laboratory Supervisor.
9. Suggest corrective actions for work practices and conditions that may result in the release
of toxic chemicals, to the Laboratory Supervisor and the Chemical Hygiene Officer. Seek supervisory assistance from the Chemical Hygiene Officer and the Laboratory Supervisor, whenever needed. Provide written updates to the Chemical Hygiene Officer and the Laboratory Supervisor.
10. Move empty gray sharp containers from the corridor to the labs, weekly. 11. Ensure that full and closed gray sharp containers, placed in the corridor, have a lid locked
with a cable tie. If not, place missing cable ties and inform appropriate lab members, and the Laboratory Supervisor.
12. If large empty boxes or crates are in the corridor, ask the appropriate lab member to
contact [email protected] to request removal, and inform the Laboratory Supervisor. If the follow-up does not happen, place the request by e-mail, with a cc. to the lab supervisor.
13. Investigate accidents and report them to the Chemical Hygiene Officer and the
Laboratory Supervisor. Assist in developing procedures that will minimize the repetition of that type of accident. Provide written updates to the Chemical Hygiene Officer and the Laboratory Supervisor.
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14. Report to the Chemical Hygiene Officer and the Laboratory Supervisor incidents that
cause (i) personnel to be seriously exposed to hazardous chemicals or materials, such as through the inoculation of a chemical through cutaneous penetration, ingestion of a chemical, or probable inhalation of a chemical, or that (ii) constitute a danger of environmental contamination.
15. Assist the Chemical Hygiene Officer and the Laboratory Supervisor in developing tools
to verify that employees understand the training received (e.g. the safety test). 16. Update the list of chemicals including the SARA Title III toxic chemicals, yearly. Inform
the Chemical Hygiene Officer and the Laboratory Supervisor when the lists are updated. 17. Help maintain lab safe, secure and fully operational by 1) keeping traffic and delivery
areas clear of boxes, 2) removing boxes or items blocking the emergency door between lab 16-476 and lab 16-436, 3) verifying that the lab doors of BL-2 areas are closed at all times, and locked when no one is around or after 5 PM.
18. Conduct formal and informal laboratory inspections at least twice per year to ensure
compliance with existing laboratory SOP's and regulations. Report results in writing to the Chemical Hygiene Officer and the Laboratory Supervisor.
19. Update green emergency information cards on all BPEC laboratory doors. Document
action with an email to the Laboratory Supervisor and when completing the monthly report.
20. Ensure that the chemical fume hood is kept safe and that waste chemicals are processed
and picked up regularly. Inform the Graduate Student Safety Officer of the current situation, at least monthly, with a cc. to the Chemical Hygiene Officer and the Lab Supervisor.
21. Provide a monthly activity report to the Chemical Hygiene Officer and to the Laboratory
Supervisor. Administrative Safety Coordinators a) Assist with scheduling and announcements of chemical hygiene and safety meetings for
all research groups in the BPEC. b) Assist with organizing special events such as CPR training, safety seminars as part of
seminars, and social gatherings to encourage social contacts between UROP students, REU students, graduate students, post-doctoral fellows, visiting scientists and other staff members.
c) Assist Safety Officers with updating of the laboratory safety and Emergency Action Plan
portions of the CHASP.
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d) Provide safety training to new administrative (non-lab) personnel (Emergency Action Plan, Section V. F., only, no lab hygiene or safety training).
e) Attend all BPEC chemical hygiene and safety meetings. These meetings operate on a
consensus basis and are scheduled at a time convenient for everyone. It is essential that Administrative Safety Coordinator participate.
Graduate Student Safety Coordinators
1) Update emergency telephone number lists next to each laboratory phone.
2) Ensure that all laboratories have the proper biohazardous, radioactive, designated area and safety labels and signs.
3) Ensure that wastes are correctly disposed of in a timely manner.
4) Assist with internal and external laboratory inspections.
5) Provide safety apparel for all lab personnel and ensure that these personnel are trained in
the use of this apparel and that they are using this apparel as required by the CHASP and particular SOP.
6) Ensure that all lab personnel have received safety training. Assist the Laboratory Supervisor with record keeping for safety training.
7) Bring safety procedural changes or concerns to the attention of the Chemical Hygiene Officer.
8) Be a listening ear for researchers and general laboratory personnel. Help organize
Chemical Hygiene and Safety Meetings, and contribute to meetings agenda.
9) Take appropriate steps to make the labs safer, such as removal of hazardous chemicals or objects, and making sure that safety equipment (eyewashes, fire blanket, fire extinguisher, safety shower, etc.) is fully accessible at all times.
10) Implement the chemical hygiene and safety measures discussed and agreed upon during the CHASP meetings, such as installing fire blankets, applying for respirators, ordering and installing safety cabinets, and correcting any problems.
11) Attend the BPEC chemical hygiene and safety meetings. These meetings operate on a
consensus basis and are scheduled at a time convenient for everyone. It is essential that the Graduate Student Safety Coordinators participate.
12) Check safety equipment (fire blankets, fire extinguishers, spill kits, showers, eyewashes,
respirators, etc.) for presence or proper operation. Document inspections or observations with an email to the Laboratory Supervisor.
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13) Notify the EH&S Office for maintenance, repair or replacement of safety equipment. Document the actions by sending an email to the Laboratory Supervisor.
14) Ensure that safety equipment is fully accessible and that emergency exits are not blocked.
15) Remove, replace or properly store hazardous chemicals or objects. Implement new safety
measures if necessary; such as the purchase new safety equipment.
16) Review safety procedures with safety regulation violators. Document actions with an email to the Laboratory Supervisor.
II. A. 7. Employees, Staff, and Students Current BPEC lab members and guests: Kathryn Miller, Steve Fox, Albert Hwa, Daniel Bauer, Dora Farkas, Karel Domansky, Artemis Kalesi, Peng Jiao, Zhiwei Song, Jin Yin,, Csanad Varga, Jane Roberts, Brent Schreiber, Maritza Rodriguez, Anand Sivaraman, Elise Bender, Henry Lam, Wendy Prudhomme, Kumar Shreyajit, Daryl StLaurent, Siddhi Shah, Jessica Kosa, Haiyan Wu, Cameron Haase-Pettingell, Katy Wack, Ji-Eun Kim, Pete Heinzelman, Judy Yeh, Daryl St. Laurent, Yas Hashimura, Henrik Lund, Alexandria Victoria Sams, Joseph Moritz, Nathan Tedford, Eugenio Usandizaga, Sing Fee Lim, Jennifer Clarke and Jean-Francois Hamel. These lab members comprise the BPEC and are thereby required to conduct research in compliance with the BPEC Chemical Hygiene and Safety Plan (CHASP). BPEC personnel lists are updated regularly using: [email protected] and [email protected].
Employees, as defined by the BPEC-CHASP, are those staff under the direction of the Supervisor, as defined by the Plan. Employees not under the direction of the Supervisor, but who are in an area under the direction of the Supervisor, are also subject to the BPEC-CHASP, Standard Operating Procedures in effect in that area. Non-employees, such as students, are equally subject to the plan, as described below. The primary responsibility of the employee is to follow the procedures outlined in the BPEC-CHASP and all Standard Operating Procedures developed under that plan. These would include the following: 1. Understand and follow all Standard Operating Procedures;
2. Understand all training received;
3. Understand the function and proper use of all personal protective equipment. Wear
personal protective equipment when mandated or necessary;
4. Report, in writing, to your supervisor of any significant problems arising from the implementation of the Standard Operating Procedures;
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5. Report to your supervisor all facts pertaining to every accident that results in the exposure to toxic chemicals, and any action or condition that may exist that could result in any accident.
Contact your supervisor, the Chemical Hygiene Officer, the EHS Coordinator, or the EHS Office if any of the above procedures are not clearly understood.
General safety tasks and related Chemical Hygiene and Safety Plan duties are assigned to all graduate students and researchers. These responsibilities are as follows: All Graduate Students and Researchers a) Follow emergency evacuation procedures (refer to the Emergency Action Plan in
Section V. F. 2. of this plan). i) If you see smoke or fire, call 100. ii) Ensure evacuation in your office and lab; assist elderly and handicapped persons. iii) ON THE WAY OUT FROM THE OFFICE AREAS, if possible, shut off air
conditioning, computers, radios and close windows and doors. iv) ON THE WAY OUT FROM THE LAB AREAS, if possible, stabilize or shut
down dangerous or critical equipment and experiments. Shut down fuel supplies such as gas. Shut down critical operations such as pressure vessels.
b) Provide chemical hygiene and safety training to all personnel working with or for
you, including visiting scientists, UROP and REU students. This includes: i) Supplying a copy of the BPEC CHASP to read and return. ii) Providing a walk-through of the lab and verbal communication of safe practices
outlined in the BPEC Safety Checklist, located in the Safety Checklist folder of the BPEC CHASP files and in Section XII. B. Record keeping.
iii) Administering a brief Safety Test containing questions based on the lab walk-
through. A copy of the test is located at the end of this manual. iv) Working with the BPEC CHO to ensure that the researcher has read, understood
and completed all aspects of the BPEC CHASP before beginning work in the lab. c) If applicable, complete institute-wide training on the use of specific hazardous
substances. Ensure that all personnel working with or for you, including visiting scientists, UROP and REU students also complete this training.
i) MIT training on the use and handling of biohazardous materials is not required
for researchers using biohazardous materials, but it is suggested to take the training as the class is very good. This class focuses on aseptic technique and
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basic biological procedures for research with bacterial and yeast cultures. Although not directly applicable to animal cell culture, the techniques are useful to learn. Refer to Section X. B. 1. for more information on this training.
ii) For researchers using radioisotopes, read appropriate sections of the BPEC
CHASP and the documents provided by Radiation Protection Office, and complete the institute-wide training as described in Section X. B. 3. of this plan. Submit all required forms as described.
iii) For researchers using lasers, complete the institute-wide training as described in
Section X. B. 2. of this plan, and submit all required forms as described.
II. B. Visitors and MIT Service Personnel Visitors shall have access to laboratories only when accompanied by a researcher in that laboratory who is knowledgeable about the required safety practices and hazards present. Visitors will follow the instructions of their host. Since 2000, visitors are asked to sign in a guest book located on the bench behind the main entrance of lab 16-436. Service personnel who regularly visit the laboratories in the course of their duties, such as mail persons, janitorial staff, and Facilities personnel are not subject to the restriction cited above. These personnel have been trained by the Institute to conduct their duties in a safe manner.
II. C. Environmental Health & Safety Team In 2000, MIT re-organized its safety and medical services programs, and created a new organization structure, called the Environmental Health & Safety Team (EH & S Team), headed by Jamie Lewis Keith. Under this new structure, several programs provide specific services to the MIT community. The EH & S Team is charged with responsibility for control, review, monitoring and advice with respect to exposure to chemical, radiological, and biological agents used in research and teaching. The EH & S Team includes the following programs: Safety, Industrial Hygiene, Radiation Protection, Biosafety and the Environmental Management Programs. The Safety Program is a component of the operations group charged with responsibility for oversight and control of physical hazards in the workplace, including fire protection, electrical and other safety hazards, and chemical waste disposal arising from work at the Institute. The Environmental Management provides oversight in the management of chemical waste disposal. The EH & S Team has authority to stop any activity that in their judgment is immediately hazardous to life or health. Radiation Protection and Reactor Radiation Protection have regulatory authority as part of MIT's license to use radioactive materials from the Nuclear Regulatory Agency. The EH & S Team has professional staff that can be called upon for advice and help on safety and environmental health problems. These staffs offer the following services to the Institute: The EH & S Team consists of professional staff, including several health physicists,
microbiologists, industrial hygienists, and industrial hygiene engineers, devote their skills to the protection of the Institute community from radiation, toxic chemical and biological
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hazards. All members of the Institute community should feel free to consult with the EH & S Team if they are concerned about the safety of operations involving a potential toxic chemical, microbiological or radiation exposure.
The duties of the programs are as follows: BioSafety ( http://web.mit.edu/environment/programs/biosafety.html)
provides technical guidance and training in biological safety. Principal services include:
i. Providing lectures on biological safety.
ii. Providing hands on training in aseptic technique.
iii. Inspecting recombinant DNA laboratories.
iv. Providing advice on biological spill cleanup procedures.
v. Investigating the accidental release of microorganisms.
vi. Providing advice on the sterilization, decontamination and disposal of biological waste. Analyze Bacillus spore vials, as part of autoclave validation tests.
vii. Providing assistance in the control of microbiological contamination.
viii. Sampling and analyzing air and water.
ix. Assisting in facility design.
x. Acting as administrative arm of the Institute Committee on Assessment of
Biohazards. Industrial Hygiene (http://web.mit.edu/environment/programs/hygiene.html) The Industrial Hygiene Program provides services for the recognition, assessment and
control of health hazards due to the use of chemical and physical agents. Principal services include:
1) Providing lectures and training sessions on industrial hygiene subjects. 2) Providing advice on the toxicity of materials and on substitution of less toxic substances. 3) Dispensing proper respiratory protective equipment for short-term use and providing
advice on the procurement of respirators for long-term use. 4) Evaluating existing laboratory ventilation and assisting in designing new ventilation.
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5) Providing information and personal protective equipment for spill clean-up procedures. 6) Evaluating potential health hazards by analysis of air in the work environment. 7) Analyzing air and biological specimens for toxic components. 8) Evaluating exposure to noise, recommending necessary controls and providing hearing
protectors. 9) Serving on the Institute Toxic Chemicals Committee.
Radiation Protection (http://web.mit.edu/environment/programs/radiation.html)
The Radiation Protection Program staff consists of professional health physicists and
radiation protection technicians who provide services required for radiation protection and compliance with federal, state and local regulations. Principal services include:
1) Providing lectures on radiation safety and other radiation related issues. 2) Registering and training radiation workers. 3) Reviewing proposed experiments using radioactive materials. 4) Providing hands-on review of radiation workers handling of radioactive materials. 5) Surveying radiation laboratories and environmental monitoring. 6) Supervising decontamination activities. 7) Ensuring proper use of radiation generating equipment. 8) Administering the personnel dosimeter program. 9) Calibrating radiation survey instruments.
10) Collecting and disposing of radioactive waste.
11) Providing consultation for laboratory design, shielding and radiation control devices and methods.
12) Acting as administrative arm of the Institute Radiation Protection Committee.
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Safety (http://web.mit.edu/environment/programs/office.html) The Safety Program has related responsibilities. The Safety Program evaluates and
implements safety policies and reviews new and existing equipment and operating practices to minimize hazards to the Institute community and visitors from fire, electricity, explosion, pressure, and machinery. The Safety Program conducts accident investigations and suggests remedial measures and procedures. It publishes the MIT Accident Prevention Guide. Training and assistance in conducting special accident prevention programs are available as required.
The services of this program are available both in emergency situations and in an advisory capacity to answer questions from anyone at the Institute. Both services provide 24-hour on-call personnel to respond to off-hours needs. They can be reached through the Operations Center, FIXIT (253-4948), Campus Police (253-1212), or the Medical Department (253-4481). Dial 100 for Assistance in any emergency. Addresses and emergency telephone numbers for these offices and other MIT emergency services are listed below.
II. D. MIT Chemical, Safety and Emergency Services SERVICE LOCATION TELEPHONE NUMBER MIT Police (includes fire and ambulance) W31-215 253-1212 or 100 MIT Medical Emergency Care E23-189 253-1311 (24 h emergency) 253-4481 (general information) Facilities Emergency Service E19-135 253-4948 (FIXIT) Nightline (Student Staffed) 253-8800 EHS Radiation Protection Program N52-496 452-3477 Industrial Hygiene Program N52-496 452-3477 Biosafety Program N52-496 452-3477 Safety Program N52-496 452-3477 Environmental Management Program N52-496 452-3477
III. Preparation, Implementation, Annual Review, and Update
III. A. Preparation and Approval The Chemical Hygiene Officer oversees the preparation of the Chemical Hygiene and Safety Plan for the Biotechnology Process Engineering Center. She/he is responsible for seeing that the
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plan meets requirements set forth in the 29 CFR 1910.1450. Assistance in creating the Chemical Hygiene and Safety Plan is provided by the Laboratory Supervisor and the MIT Environmental Health & Safety Team.
III. B. Distribution and Implementation The Chemical Hygiene Officer ensures that the Chemical Hygiene and Safety Plan is updated, as needed, and that the updates are distributed to or made available to those who are affected by it. Because of the length of the BPEC CHASP and the requirement of annual updates, individual copies are not distributed to each employee or researcher. Several copies of the plan are available in the BPEC laboratory, the CHO’s office and the Center Director’s Headquarters. One copy of the Chemical Hygiene and Safety Plan and all updates are provided to the Industrial Hygiene Program.
III. C. Compliance and Enforcement MIT Policy The Laboratory Supervisor is responsible for establishing and maintaining records for employee training, employee and environmental monitoring, and quantity of chemicals stored in the work place. In practice, the Chemical Hygiene Officer may assist with this work. The Laboratory Supervisor enforces the Chemical Hygiene Plan by making sure the Chemical Hygiene rules are known, and followed. The Chemical Hygiene Officer and Assistant Chemical Hygiene Officer advise and assist in this work and helps with documentation. The Chemical Hygiene Officer assists with chemical hygiene, laboratory and housekeeping inspections. When there are significant changes in existing policies or work practices, an inspection should be conducted soon after the new policy or process is implemented. The Industrial Hygiene Program is available to assist the Chemical Hygiene Officer in the inspection process and in all related matters. The Chemical Hygiene Officer reviews and updates the Chemical Hygiene Plan annually, with assistance from the laboratory supervisor. BPEC Regulation To assist the Laboratory Supervisor with compliance, the following routine training and documentation tasks have been established. Details on education and training are outlined in Section X. Required forms are presented in the Recordkeeping Section of this plan (Section XII.). .
1) Each person under the BPEC Chemical Hygiene and Safety Plan is required to read the plan and to sign a Chemical Hygiene and Safety Clearance Form (Section X.) certifying that they have read and understood the contents of the plan.
2) As required by this same Form, each researcher must take a general lab safety and
biohazardous materials handling orientation and test. These documents are kept as documentation of training.
3) To complete the Clearance Form, each person must attend specialty Institute Training
Courses on particularly hazardous substances (biohazards, lasers and radioisotopes) if relevant to their work. Personnel need attend these seminars only once during their time at MIT.
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4) Researchers can incorporate their chemicals into the common stock which is inventoried annually.
5) Individuals from outside labs or corporations who plan to conduct short-term research in
the BPEC laboratories must also undergo the above training, with the exception of the seminars in part 3), and file a Chemical Hygiene and Safety Clearance Form (Section XII) with the CHO before beginning work.
6) Researchers must attend any BPEC chemical hygiene and safety seminars. Attendance
sheets are collected.
7) Planned and impromptu laboratory inspections are performed and documented and new procedures developed as a result of the inspection are implemented as necessary.
8) The Chemical Inventory is updated annually by the Laboratory Supervisor and the CHO.
9) Additional records including, radioisotope and medical monitoring records, specialized
training forms (such as for Institute Radioisotope Training), accident reports, etc., as relevant to the researcher’s work, also serve to validate compliance.
III. D. Annual Review and Update The Chemical Hygiene Officer is responsible for seeing that the Chemical Hygiene and Safety Plan is reviewed on an annual basis and updated to accommodate changes in the 29 CFR 1910.1450, departmental procedures, personnel, MIT policy and other pertinent materials. The BPEC Chemical Hygiene Officer also see that the BPEC Chemical Hygiene and Safety Plan is updated to include procedures regarding new hazards and processes as they are introduced. The Chemical Hygiene and Safety Plan and updates are reviewed by staff of the EH & S Team for final approval. Updates are usually due annually in January. Components of the BPEC CHASP that require annual review include (but are not limited to): 1) Personnel List 2) CHASP Representatives: CHO, Laboratory Supervisor, and Chemical Hygiene and
Safety Committee (Section II. A.) 3) Education and training methods and documentation for new and existing personnel 4) Chemical Inventory and chemical evaluation procedures 5) Laboratory inspection forms 6) Changes in MIT safety and hygiene regulations 7) Evaluation of new processes and procedures
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8) Assessment of current hygiene and safety equipment and procedures 9) Evaluation of enforcement methods and records. Are they working? If not, redefine
methods or procedures so that they are more easily implemented and followed.
IV. Basic Chemical Hygiene and Laboratory Safety Equipment
IV. A. Personal Protective Equipment MIT policy and BPEC regulations for the use of eye protection, protective apparel and respirators are given below. If equipment other than the types listed below is required for the use of a given chemical or procedure, it will be identified, and its proper use outlined, in the relevant Standard Operating Procedure. The type and level of equipment designated in each SOP has been determined with the aid of MSDS’s and other information sources as well as through consultation with the MIT Environmental Health & Safety. Any use of personal protective equipment is considered only after the options of reducing the hazards.
IV. A. 1. Eye Protection To minimize the risk of eye injury, MIT policy requires that all personnel, including visitors, wear eye protection at all times in the laboratory. This policy includes the BPEC laboratories. This eye protection policy is necessary in order that the Institute comply with both Massachusetts and Federal Law (e.g. Code of Federal Regulations, Title 29, Section 1910.133). Eye protection is required at all times in the lab, not just when performing a chemical operation, unless the Standard Operating Procedure specifically excludes its use. The type of eye protection required is stated in each Standard Operating Procedure. Safety glasses may be removed while using microscopes, working at desks or when researchers wear adequate prescription glasses. Visitors and MIT Facilities personnel should not be allowed to enter a lab unless they wear appropriate eye protection. A supply of safety glasses is available in each BPEC lab. Researchers should keep their safety glasses in their lab coats or other easily accessible area. Non-prescription Eye Protection SAFETY GLASSES WITH SIDE SHIELDS provide the minimum protection acceptable for regular use. For non-prescription eyeglass users, they are the minimum required protection to be worn at all times while in the BPEC laboratories. Safety glasses must meet the American National Standards Institute standard Z87.1-1989 that specifies a minimum lens thickness (3 mm), certain impact resistance requirements, etc. Although these safety glasses can provide satisfactory protection against injury from flying particles, they do no fit tightly against the face and, therefore, offer little protection against splashes or sprays of chemicals or large, flying objects. GOGGLES should be worn whenever a significant chemical splash, vaporization or volatilization hazard exists. FULL FACE SHIELDS should be worn when conducting particularly hazardous laboratory operations where there is a hazard to the face and neck.
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UV PROTECTIVE GLASSES should be worn when the use of ultraviolet light sources or lasers is required. Only those types of UV protective glasses approved by the Radiation Protection Program may be used. All of these types of eye protection can be purchased through any lab safety supply company. Contact the MIT Safety Program for vendors of eye protection as well as ordering information. Prescription Eye Protection and Contact Lenses CONTACT LENSES offer no protection against eye injury. Under many laboratory circumstances they can be a significant threat to the eye. Contact lenses can: a) Absorb chemical fumes, b) Allow long and intimate contact of toxic chemicals with the eye, c) Trap chemicals and particles in the space between the contact lens and the eye, d) Be difficult to remove in an emergency due to muscle spasms of the eye, and e) Interfere with flushing procedures, Although wearing contact lenses is allowed in the BPEC laboratories, contact lenses should not be worn when performing operations in which there is a significant risk of a chemical splash, vaporization or volatilization. In the event of an accident, contact lenses can interfere with first aid and eye-flushing procedures. If for medical or personal reasons, a person must wear contact lenses, tight fitting goggles are recommended at all times, and are required when there is a significant risk of a chemical splash, vaporization or volatilization. The minimum requirement for contact lens users at all times while in the BPEC laboratories is to wear non-prescription safety glasses equipped with side shields. When working in an area where chemicals or other harmful materials are used, hands should be washed and rinsed thoroughly with soap and water before handling contact lenses. This procedure applies even if gloves are worn. Never leave or store contact lenses in any lab. PRESCRIPTION SAFETY GLASSES, instead of ordinary prescription glasses, are required when carrying out a chemical operation in which there is a significant risk of flying debris. Ordinary prescription glasses do not provide adequate protection against injury. Prescription safety glasses are the minimum required protection to be worn at all times by prescription eyeglass users while in the BPEC laboratories. Goggles worn over ordinary prescription glasses or prescription safety glasses are required when there is a significant risk of a chemical splash, vaporization or volatilization. Contact the MIT Safety Program for the purchasing procedure for prescription safety glasses approved by both the ANSI and the MIT Safety Program. The procedure is quite lengthy, requiring an eye exam, selection of eye-glass frames, departmental approval signature and purchase order, and placement of the order with the MIT Safety Program. Eye exams and
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prescription eyeglass frames are now available at MIT Optical, located in the basement of the Student Center. Delivery takes 6 to 8 weeks.
IV. A. 2. Protective Apparel The requirement of protective clothing, including gloves, is determined by the employee, Laboratory Supervisor and the Industrial Hygiene Program, based on the hazardous substance/process being used and on regulations (for example, as they exist in a BL-2 lab). Contact the Industrial Hygiene Program for selection of the type of clothing required for a given task. A LAB COAT and CLOSE-TOED SHOES constitute the minimum required protective clothing when conducting experiments that involve hazardous/biohazardous substances or chemicals in all BPEC laboratories. In addition, researchers working in the designated BL-2 areas of the lab must wear a lab coat at all times. The lab coat should be knee length or longer. To make sure that researchers have a cleaned lab coat at all times, it is recommended that all personnel wearing a lab coat have at least two lab coats. Close-toed shoes include sneakers, loafers and women’s pumps. Sandals and open toe versions of the above shoes are not allowed when conducting experiments that involve hazardous/biohazardous substances or chemicals. Lab coats should be hanged on hangers dedicated to the bench used by the researcher or on the coat rack in lab 16-436. BPEC lab members and guests should leave their lab coats in the BPEC laboratories (either 16-436 or 16-476) when their work in completed. The only exception is when they need to work in one of the instrumentation rooms, or the cold room, or the autoclave room. Do not bring or wear lab coats and/or gloves into offices, lounges and food areas. VINYL OR LATEX GLOVES are required for all manipulations of biohazardous materials, radioisotopes and toxic chemicals. When working in the BL-2 areas with biohazardous substances, with potentially infectious materials, contaminated surfaces or equipment, researchers must wear gloves. Gloves should be removed before handling common-use objects in the lab such as telephones, doorknobs, pens and log books. Gloves used for radioisotope work must be removed and disposed of before leaving the radiation bench. Gloves must be removed before exiting the lab to prevent transfer of these materials to doorknobs and areas outside the laboratory. Hands should also be washed before leaving the lab. Other types of chemical or heat resistant gloves may be specified by the SOP’s for very hazardous substances or operations. A glove selection program is available at VWR. LEG COVERING is required when handling particularly hazardous substances. Medical scrubs, long pants, leggings and long socks that cover all otherwise exposed portions of the leg are permissible types of leg covering. Shorts, skirts and dresses are acceptable for minimally to not hazardous chemical operations Long hair, loose clothing and dangling jewelry should be confined or avoided when working in lab.
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IV. A. 3. Respirators It is the Institute’s goal to control respiratory hazards at their point of generation by using engineering controls and good work practices. In keeping with this goal, the use of respirators as the primary means of protecting employees from airborne hazards is considered acceptable only in very specific situations. These situations include short-term temporary experiments where engineering controls are not feasible, and situations in which the use of respiratory protection is an added or supplemental control. At this time no BPEC researchers are using respirators. Contact the MIT Industrial Hygiene Program if you need one, and inform the Laboratory Supervisor and CHO.
IV. B. Containment Equipment and Procedures The following equipment and procedures are used to reduce the risk of chemical exposure, spills and/or accidents. Hazard control equipment and procedures for particularly hazardous substances are presented in Section VII.
IV. B. 1. Chemical Purchase Chemicals MIT has developed a policy to discourage the practice of bulk ordering of chemicals that reduces the chemical cost per unit volume. Although bulk orders save individual departments money, in the long run the cost of disposal of unused chemicals far outweighs the saving obtained from the bulk order. The following points should be addressed to determine the proper volume of any chemical to order. a) Properties that may preclude long-term storage should be evaluated before the quantity of
chemical to be ordered is decided. b) Chemicals should be purchased in quantities that ensure usage within the shelf life of the
chemical. Although many chemicals can be safely stored over long periods of time, decomposition can result in explosions, ruptured containers and the formation of hazardous by-products.
c) When chemicals are received, they should be marked with the owner’s initials and
dated with date received, and shelved in a manner that enables older chemicals to be consumed first. The date the chemical is opened should be marked if it is important to degradation.
d) The current Chemical Inventory should be consulted before ordering a chemical. It is in
the form of two binders, located in Rooms 16-436 and 16-463. If a chemical is listed in the inventory, go to the shelf to check the date of the chemical. If the chemical is past its expiration date, throw it out.
Hazardous or Dangerous Materials MIT has developed a policy for the purchase of hazardous or dangerous materials.
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Details of the policy are provided at: http://web.mit.edu/purchasing/pol&proced/4.2.pdf. Requisition for hazardous or dangerous materials.
IV. B. 2. Chemical Transportation MIT policy dictates that chemicals must be packaged, transported, and stored in a manner that minimizes the threat of release via container breakage. The requirement of break-resistant primary or secondary containers is based on the physical state of the chemical (liquid versus solid), the volume of the container and the hazard of the chemical (e.g. corrosive, flammable, infectious, nuisance, radioactive, reactive, toxic, etc.). Break resistant implies a container made of metal, plastic, plastic-coated glass, or metal overpack of glass. Secondary containers refers to a commercially available bottle carrier made of rubber, metal or plastic with carrying handle(s) and which is of large enough volume to hold the contents of the primary chemical container. Lids or covers are desirable but not necessary on secondary containers. Rubber or plastics should be used for acids and alkalies. Metal, rubber or plastic should be used for organic solvents. Compliance with the following regulations is mandatory: a) Requisitioners will acquire chemicals from the VWR stockroom (basement of Building
56) whenever possible. Chemicals should always be transported in a secondary container. When ordering chemicals from outside sources, requisitioners shall specify break-resistant containers whenever available.
b) Purchasing agents will monitor/issue purchase orders so as to be in compliance with this
policy. Present purchasing procedures may be revised to ensure compliance. c) Stockroom personnel will not dispense or sell chemicals in breakable containers of any
size unless the customer has an approved secondary container in which to place the chemical for transporting before leaving the stockroom.
d) Carts used to transport chemicals from one area to another should be stable and in good
condition. Carts should have edges to contain spills. Only easily manageable quantities of chemical should be transported. Routes should be planned so as to avoid steps or stairs.
e) Freight elevators, not passenger elevators should be used to transport hazardous
chemicals whenever possible. The closest freight elevator to BPEC headquarters is located in Building 26. The individual transporting the hazardous chemicals should operate the elevator alone if possible. Persons should avoid boarding an elevator being used for transport of hazardous chemicals.
f) It is unlawful to drive a motor vehicle containing hazardous materials in all highway
tunnels in Massachusetts. Anyone transporting hazardous chemicals in motor vehicles should check with the Safety Program for information of State and Federal regulations.
g) Users may be assessed a portion of the cleanup cost if breakage and spill results from a
total disregard of this chemical transportation policy.
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IV. B. 3. Chemical Storage MIT policy dictates that chemicals must be stored in a manner that minimizes the possibility of interaction between incompatible chemicals, the chance of contact with other substances that could elicit an adverse reaction (such as solvents with fire), and the threat of release via container breakage. In the BPEC laboratories, the following storage facilities and procedures are employed: a) Both solvents and corrosives are stored (separately) in specially designed steel storage
cabinets. Only small quantities of these chemicals should be kept at laboratory benches. b) Strong oxidizers and organic chemicals are stored separately. c) All toxic and hazardous substances are stored appropriately in ventilated areas. d) Chemical containers larger than one gallon, and all acids and bases are stored below eye
level. Hazardous chemicals are never stored on the floor. e) Breakable containers, one gallon or larger, that are not stored in specially designed
cabinets are stored within approved secondary containers when not in use. Very hazardous materials should be stored in trays or secondary containers to minimize distribution of material in the event of a container leak or break. The design of the solvents and corrosives cabinets satisfies this requirement.
f) Individual shelves at the bottom of a refrigerator or freezer, or an entire refrigerator or
freezer is dedicated to the storage of radioisotopes and other particularly hazardous chemicals in each BPEC lab. The bottom shelves are used to reduce contamination of other chemicals or users that may be reaching into the refrigerator or freezer after a spill or leak. A radioactive sign is located on the front of each of these refrigerators and freezers that store such materials.
g) Chemical containers must be labeled regardless of where they are stored (personal or
common spaces). The label must be the original manufacturer’s label or a label describing the chemical(s) contained in the substance or mixture, the owner’s initials, date received, date opened (if applicable to degradation), and special hazards (e.g. acid, flammable, poison, radioactive, etc.) when applicable. During laboratory inspections unmarked containers are brought to the attention of the lab worker for identification.
h) Chemical stock is periodically checked for signs of container or chemical degradation.
Deformation of container, discoloration of chemicals, or the formation of precipitates in liquids may indicate that a chemical has decomposed and should be disposed of. If the integrity of the chemical is not affected but the container is damaged, the chemical is transferred to a new container and properly marked.
i) Containers for waste chemicals should be break resistant or kept in an approved
secondary container, and in accordance with provisions of the MIT policy on waste chemicals.
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IV. B. 4. Fume Hoods Local exhaust ventilation is the primary method used to control inhalation exposures to hazardous substances. The laboratory fume hood is the most common local exhaust method used in laboratories. Fume hoods are used for manipulation of chemicals only, and they are unsuitable for using cells and viruses. Cells and viruses must be manipulated in biological safety cabinets only. Proper Use A laboratory fume hood should be used when working with all hazardous substances. Many BPEC Standard Operating Procedures for particularly hazardous substances dictate the use of a fume hood. A properly operating and correctly used fume hood controls the vapors released from volatile liquids as well as dust and mists. No modifications to hoods or duct pipes should be initiated without first contacting the Lab Supervisor and the Chemical Hygiene Officer who will discuss proposed changes with the Industrial Hygiene Program. Any changes made to local exhaust systems must be approved by the Industrial Hygiene Program. Biosafety cabinets and hoods must be re-inspected if changes of any kind are made to the laboratory hood system. If there is any question about a laboratory hood’s operation the Industrial Hygiene Program should be called immediately. Repairs can be made through Facilities, extension 3-4948 (FIXIT). When a new laboratory hood is installed, it is the responsibility of the Laboratory Supervisor to ensure that no hazardous substances are used in the hood until it is surveyed and labeled by the Industrial Hygiene Program. Do not use a fume hood for large pieces of equipment unless the hood is dedicated to this use. Large obstructions will change the airflow patterns and render the fume hood unsafe for other uses. It is generally more effective to install a specially designed enclosure for large equipment so the fume hood can be used for its intended purpose. The following general rules should be followed when using fume hoods:
a) Unless the hood is marked with a performance certification label (bright red label for toxic chemicals) that is less than one year old, it should not be used. This performance certification label should give the average air velocity when the hood sash is at the demarcated level. Ensure that the hood has an approval labeled for the type of chemicals being used (e.g. toxic materials, radioisotopes, etc.) and that this approval label is dated within this last year.
b) Always try to keep hazardous chemicals at least six inches behind the plane of the sash. c) Never put your head inside an operating laboratory hood to check an experiment. The
plane of the sash is the barrier between contaminated and uncontaminated air. d) Work with the hood sash in the lowest possible position. The sash will then act as a
physical barrier in the event of an accident in the hood. Effective hood containment is provided when on low fan mode. Do not work with the hood sash higher than the maximum safe operating sash height, demarcated on the hood with a performance certification label. Keep the sash closed when not conducting work in the hood.
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e) Do not use a fume hood for chemical storage. A hood cluttered with bottles may not
contain releases effectively. Store chemicals in a chemical storage cabinet. f) Do not clutter the hood with bottles or equipment. Keep the hood clean and clear. Only
materials actively in use should be in the fume hood. Operation under these conditions provides optimal containment and reduces the risk of extraneous chemical being involved in fire or explosions that may occur in the hood.
g) Regularly clean the grill along the bottom slot of the hood to prevent clogging from papers and dirt.
h) Promptly report any suspected hood malfunction to the Industrial Hygiene Program
(452-3477). Further information on fume hoods is available from the Industrial Hygiene Program, http://web.mit.edu/environment/programs/fumehoods.html, telephone: 452-3477. Surveillance The Industrial Hygiene Program inspects all laboratory hoods annually. This inspection consists of measuring the face velocity of the hood and using a smoke stick to visually check its containment effectiveness. If the hood passes both the face velocity and smoke containment tests, the hood performance certification label is updated. If the hood does not pass the survey and the problem is so severe that the fume hood is unsafe for use, then it is labeled with a "DO NOT USE" sign. If the problem if less severe so that the fume hood can still be used even though it does not pass the fume hood survey, it is not updated and a list of problem hoods is sent to the Chemical Hygiene Officer along with a description of the problem. If the problem is one that Facilities can correct (e.g. a slipping fan belt, cracked duct work, etc.) then the Industrial Hygiene Program submits a work order to Facilities to have it repaired. Facilities notifies the Industrial Hygiene Program when the repairs have been made and the fume hood is re-inspected. If the hood fails due to its incorrect use (e.g. a cluttered hood), then the Industrial Hygiene Program notifies the Chemical Hygiene Officer. It is the Chemical Hygiene Officers responsibility to have the problem fixed and contact the Industrial Hygiene Program to re-inspect the fume hood. It is the responsibility of the researchers and the Laboratory Supervisor to notify the Industrial Hygiene Program if a fume hood does not have an updated certification label (the inspection date must be less than 1 year old). No surveillance information is kept in the BPEC CHASP files. The performance certificate on each hood will suffice to verify proper operation. Location of Fume Hoods in BPEC Fume hoods are located in all laboratories where organic solvents or other flammable materials and inhalation hazardous chemicals are used. One fume hood is located in each of the following BPEC labs: 16-436, 16-476. All fume hoods are designated areas.
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IV. C. Safety Equipment All labs are equipped with fire extinguishers, safety showers, eyewash stations and spill kits. MIT Emergency and BPEC researcher’s telephone numbers are listed next to every lab phone. Each laboratory door is posted with an up-to-date green Emergency Information Card which lists the names and home and work telephone numbers of several emergency contact persons for the laboratory. These persons are familiar with equipment and chemicals in the laboratory that may pose a safety or even life-threatening problem in the event of an emergency. The safety equipment is checked during the lab inspections, twice a year.
IV. C. 1. Fire Extinguishers and Blankets It is MIT policy that personnel are not required to extinguish fires that occur in their work areas. However, under certain circumstances suitably trained personnel may attempt to extinguish fires. Extinguishing fires should only be attempted if you are confident that you can do so successfully and quickly, and from a position in which you are always between the fire and an exit from the laboratory. Refer to Section V. F. 3. on the BPEC Emergency Action Plan for additional information. All BPEC laboratories are equipped with dry chemical/powder. All office and laboratory personnel should be familiar with the location, use, and classification of the extinguishers in their area. Several types of extinguishers are described below, as well as their classification and suitability for use with different types of fires. Classes of Fires Class A Ordinary combustible solids such as paper, wood, coal, rubber, and textiles. Class B Petroleum hydrocarbons (diesel fuel, motor oil and grease) and volatile flammable
solvents. Class C Electrical equipment Class D Combustible or reactive metals (such as sodium and potassium), metal hydrides, or
organometallics (such as alkylaluminums). Types of Extinguishers a) Carbon dioxide extinguishers are effective against Class B fires, (involving burning
liquids such as solvents), and Class C (electrical) fires. They are not as effective against burning paper or trash fires. Do not use a carbon dioxide extinguisher against fires involving alkali and certain other metals (such as aluminum, magnesium, and zinc) and compounds such as lithium aluminum hydride.
b) Dry powder extinguishers can also be used against Class B (liquid) and C (electrical)
fires. These extinguishers contain sodium bicarbonate and are not recommended for fires involving delicate instruments or optical systems.
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c) Water extinguishers should only be used for Class A (paper and trash) fires. The use of water against electrical, liquid, and metal fires is not recommended and can intensify or spread the fire instead of extinguishing it.
d) Sand can be used on any type of fire, Class A, B, C or D and is especially useful against
small Class D (metal) fires. Location of Fire Extinguishers and Fire Blankets in the BPEC Laboratories The fire extinguishers in Building 16 are located just inside the door of each laboratory. Fire blankets are located in the hallway by the door. For room 56-421, the closest fire blanket is located by door 56-422, the one to the right of door 56-428. Inspection and Recharging Any time a fire extinguisher is used, no matter for how brief a period, it should be inspected and recharged. For recharging, replacement, inspection, or information regarding the type of extinguisher best suited for the laboratory, call the MIT Operations Center at 253-4948. It is the responsibility of the Laboratory Safety Officers to inspect the portable fire extinguishers in or near their laboratories twice per year. Fire extinguishers are also inspected and tagged annually by Department of Facilities. Student Safety Officers are required to maintain unobstructed access to fire extinguishers and fire blankets.
IV. C. 2. Bandages A box of bandages is located in BPEC laboratories 16-436 and 16-476. Student Safety Officers are responsible for inspecting the kits twice per year and informing the Laboratory Supervisor, if new bandages are needed.
IV. C. 3. Safety Showers and Eyewashes Safety showers are located just outside the main entrance of lab 16-436 and lab 16-476, and outside 56-422. The eye stations are located to the right of the sinks in labs 16-436 and 16-476. This equipment meets the requirements of the OSHA regulation 29 CFR 1910.151(c). All laboratory personnel should be familiar with the location and use of safety showers and eyewash stations. This safety equipment is inspected during the lab inspections to insure that the equipment is unobstructed. The Safety Program or Facilities also conducts inspections of this equipment regularly.
IV. C. 4. Spill Kits Spill kits are available in rooms 16-436 and 16-476. In room 16-436 in particular, there is a biological spill kit, located at the end of benches 13/14, near the area dedicated to the adenovirus projects. The spill kits are inspected during the lab inspections to insure that the equipment is unobstructed. Each of these laboratories has a designated safety shelf on which spill kits are located. This shelf is located near an exit for ready access. These shelves may be stocked with buckets, bags, mops, sponges, brooms, protective apparel, solvent pillows, and/or commercial spill absorbent kits. Each laboratory has supplies tailored to deal with the potential hazards of the materials being used in that laboratory. Typical spill control kits may include:
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a) Spill control pillows (commercially available from many laboratory safety vendors). These can generally be used for absorbing solvents, acids, and caustic alkalies, but not hydrofuran.
b) Inert absorbents such as vermiculite, clay and sand. c) Neutralizing agents for acid spills such as sodium carbonate and sodium bicarbonate. d) Neutralizing agents for alkali spills such as sodium bisulfate. e) Large plastic scoops and other equipment such as brooms, pails, bags, dust pans, etc. as
appropriate.
V. Basic Chemical Hygiene and Laboratory Safety Procedures The following standard chemical safety and control measures are enforced in all BPEC laboratories.
V. A. Accidental Release of Hazardous Substances (Spills) Minor hazardous materials or waste spills that present no immediate threat to personnel safety, health, or to the environment can be cleaned up by laboratory personnel that use the materials or generate the waste. A minor hazardous material spill is generally defined as a spill of material that is not highly toxic, is not spilled in large quantity, does not present a significant fire hazard, can be recovered before it is released to the environment, and is not in a public area such as a common hallway. Such a spill can usually be controlled and cleaned up by one or two personnel. Major hazardous material and waste spills should be reported to the MIT emergency number (x100) to receive immediate professional assistance and support in the control and clean up of the spilled material. Major hazardous materials or waste spills are generally defined as having a significant threat to safety, healthy, or the environment. These spills generally are a highly toxic material or is spilled in large quantity, may present a significant fire hazard, cannot be recovered before it is released to the environment, or is spilled in a public area such as a common hallway. Upon reporting such a spill personnel should stand by at a safe distance to guide responders and spill clean up experts to the spill area. Reporting personnel should also keep other personnel from entering into the spill area. In the case of a spill that presents a situation immediately dangerous to life or health, or a situation with significant risk of a fire, personnel should evacuate the area and summon emergency assistance by dialing the MIT emergency number (x100), activating a fire alarm station, or both. Hazardous material users and hazardous waste generators must be aware of the properties of the materials they use and the waste they generate. Properties of materials are most commonly found in material safety data sheets and many publications. A good guide to finding other sources of information is found in “Prudent Practices in the Laboratory.”
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Prevention Take precautions to reduce the risk of a spill and to facilitate a safe cleanup if an accident such as the following occurs: 1) Plan experiments so as to minimize the possibility of an accidental release of hazardous substances. Design experiments to use the minimal amounts of hazardous materials practical and always transport and store such materials properly using break-resistant bottles or secondary containers. 2) Be familiar with the properties (physical, chemical, and toxicological) of hazardous substances before working with them. Develop a contingency plan to handle spills when working with hazardous substances. 3) Make sure that the necessary safety equipment, protective apparel, and spill control materials are readily available. 4) Know the location of the nearest fire alarm and telephone and know what numbers to call in the event of an emergency. All emergency telephone numbers are next to each BPEC lab phone. In the Event of a Spill In the event of a spill, the following general guidelines for handling spills should be followed in the indicated order of priority. 1) Attend to any injured or contaminated personnel and, if necessary, request help by calling
the Campus Police emergency number ext. 100. 2) Notify other personnel of the accident and, if necessary, evacuate the area. 3) Take steps to confine and limit the spill if this can be done without risk of injury or
contamination. 4) Cleanup the spill. Dispose of contaminated materials properly (Section V. O.). Procedures for carrying out the above operations are discussed below. Treating Injured and Contaminated Personnel If an individual is injured or contaminated with a hazardous substance, then tending to them will generally take priority over the spill control measures outlined below. It is important to obtain medical attention as soon as possible. Dial 100, the Campus Police 24-hour line, in case of emergency. For spills covering small amounts of skin, immediately flush with flowing water for no less than fifteen minutes. If there is no visible burn, wash with warm water and soap, removing any jewelry to facilitate removal of any residual materials. Check the MSDS to see if any delayed effects should be expected. A MSDS Web site link is provided by MIT’s Safety Program at http://web.mit.edu/environment/programs/msds.html. Researchers should ensure that they know where to access a particular MSDS before there is an emergency. It is advisable to seek
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medical attention for even minor chemical burns. For spills on clothing, do not attempt to wipe the clothes. Quickly remove all contaminated clothing, shoes and jewelry while using a safety shower. Seconds count, and no time should be wasted because of modesty. Be careful not to spread the chemical on the skin, or especially in the eyes. Use caution when removing pullover shirts or sweaters to prevent contamination of the eye; it may be better to cut the garments off. Immediately flood the affected body area with warm water for at least fifteen minutes. Resume if pain returns. Do not use creams, lotions or salves. Get medical attention as soon as possible. Contaminated clothing should be discarded or laundered separately from other clothing. For splashes into the eye, immediately flush the eye with tempered potable water from a gently flowing source, such as an eyewash, for at least fifteen minutes. Hold the eyelids away from the eyeball, moving the eye up and down and sideways to wash thoroughly behind the eyelids. An eyewash station should be used, but if one is not available, injured persons should be placed on their backs and water gently poured into their eyes for at least fifteen minutes. First aid must be followed by prompt treatment by a member of a medical staff or an ophthalmologist especially alerted and acquainted with chemical injuries. Notify Personnel in the Area Alert other workers in the laboratory of the accident and the nature of the chemicals involved. Post personnel at entrances to prevent other workers from inadvertently entering the contaminated area. In the event of the release of a highly toxic gas or volatile material, evacuate the laboratory (or building) by activating the nearest fire alarm. Call 100 to obtain emergency assistance from the Cambridge Fire Department and the MIT Industrial Hygiene Program. Cleanup Spills Promptly Specific procedures for cleaning up spills will vary depending on the location of the accident (e.g. corridor, chemical store room, elevator, laboratory, or laboratory hood), the amount and physical properties of the spilled material (e.g. volatile liquid, solid, toxic gas, or biohazardous material), and the degree and type of toxicity. Outlined below are some general guidelines for handling several common spill situations. 1) Materials of low flammability which are not volatile or which have low toxicity.
This category of hazardous substances includes inorganic acids (e.g. sulfuric, nitric, etc.) and caustic bases (e.g. sodium and potassium hydroxide). For cleanup, wear appropriate protective apparel including gloves and, if necessary shoe coverings. Neutralize the spilled chemicals with materials such as sodium bisulfate for alkalies and sodium carbonate or bicarbonate for acids. Use acid and base neutralizing agents, spill control pillows, scoops and brooms (from the BPEC lab or from the Industrial Hygiene Program). Absorb the material with inert clay or vermiculite, scoop it up, and dispose of it according to the procedures detailed in Section V. O.
2) Flammable solvents. Fast action is crucial in the event that a flammable solvent of
relatively low toxicity is spilled. This category includes pet ether, pentane, diethyl ether, dimethoxyethane, and tetrahydrofuran. Immediately alert other workers in the laboratory, extinguish all flames using a carbon dioxide extinguisher, and turn off any spark-producing equipment. In some cases the power to the lab should be shut off with the circuit breaker. As quickly as possible, the spilled solvent should be soaked up using
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spill control pillows. These should be sealed in container and treat as a solvent for disposal; call the MIT Environmental Program for pickup.
3) Highly toxic substances. Do not attempt to cleanup a spill of a highly toxic substance by
yourself. Notify other personnel to stay out of the area of the spill and contact the Industrial Hygiene Program (452-3477) or the Cambridge Fire Department (dial 100) to obtain assistance in evaluating the hazards involved. The Industrial Hygiene Program and the Cambridge Fire Department have special protective equipment to permit safe entry into areas contaminated with highly toxic substances.
4) Mercury. Mercury can be released upon breakage of mercury thermometers. Personnel
should wear latex, rubber or vinyl gloves before cleaning up mercury spills as mercury readily absorbs through the skin. A plastic bottle should be used for storage of broken thermometer pieces. Due to its high surface tension, mercury will easily form into balls. A cloth or brush should be used to collect the mercury into large balls. Alternatively, a vacuum cleaner dedicated to mercury spills can be borrowed from the Industrial Hygiene Program. The surface onto which the mercury was spilled should then be sprinkled with sublimed sulfur. The sulfur will react with the mercury to form an inert, black powder. This powder can be placed in the trash. If mercury is spilled into water, add enough sublimed sulfur to completely react with all the mercury, turning it black. Evaporate the water and place the black powder in the regular trash.
Spills involving biohazardous and radioactive materials are covered in Section VII.
V. B. Autoclave and Steam Use Always wear protective clothing; thermal gloves, long sleeves, and goggles when working with an autoclave or steam. Procedures for how to use the BPEC autoclaves are posted next to each autoclave. When using the autoclaves: 1) Capped containers should be placed in the autoclave with the cap partially open. This will
allow pressure equilibration and ensure proper airflow for complete sterilization. 2) For autoclaving liquids, the autoclave should be placed on liquid cycle to enable slow
exhaust to minimize boiling. Containers with liquid should be placed in a pan to contain any spilled liquid from boiling.
3) To open an autoclave at the end of a sterilization cycle, stand back from the autoclave
and slightly crack the door to allow excess steam to escape. To prevent steam burns, allow the door to remain cracked for five minutes before attempting to retrieve items.
4) At the end of a cycle, autoclaved bottles of liquid should be left in the autoclave with the
door open until they are cool, about ten minutes. Removing a hot bottle may cause superheated liquid to boil over and cause injury.
5) Assume glassware in the autoclave is hot until proven otherwise. Do not tightly cap a hot
container. Allow the container to cool for fifteen minutes before tightly capping.
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Specific procedures on autoclave sterilization temperatures and times required for different materials are covered in Sections VII. B. 6. and 7. Autoclave Validation Biological indicator controls (ampoule tests) are used to certify all autoclaves for operating efficiency once per month. Validation tests and results are recorded on the Autoclave Validation Sheets, posted next to the autoclaves. It is the responsibility of the researcher assigned to autoclave maintenance to perform this validation. Currently, Daniel Bauer is the person in charge of the validations.
V. C. Checking Out of the Laboratory On termination of a research project or completion of a theses, graduate students, post-doctoral fellows and visiting scholars/students need to clear all of their supplies from the lab and complete and Exit Clearance Form. A copy of this form is given in Recordkeeping, Section XII. D. Research personnel are required to: 1) Account for all of their chemicals, by disposing of them, or reassigning them to another
researcher, or a combination of both. MIT chemical disposal regulations must be followed when disposing of chemicals (Section V. O.). All unused chemicals to be kept by the laboratory shall be labeled as common and placed in common chemical storage areas.
2) Discard or reassign any personal equipment. Commonplace equipment belonging to the
researcher such as pipettors, glassware, small reactors, timers, etc. can be put into the lab common supplies.
3) Remove personal files from the computers and any personal diskettes from the computer
room. Personal diskettes can be cleared of data and submitted to the common diskette box.
4) Clean out their desk and related office-space; file cabinets, bookshelves, storage closets,
refrigerators, etc. The Exit Clearance Form must be signed by two coworkers, the Laboratory Supervisor and the researcher’s advisor to obtain exit clearance. The researcher’s advisor should not sign off on the researcher’s thesis or related final document or forms until the Exit Clearance Form is completed and submitted to the CHO.
V. D. Children and Pets in Laboratories Children are only permitted in BPEC laboratories while under the direct supervision of their parent or other guardian. Pets such as dogs are not permitted in laboratories where hazardous substances are stored or are in use. Pets are especially excluded from biological containment facilities as they may breach containment or become contaminated themselves. Most of the BPEC laboratories operate under biological containment level 2 (BL2). Refer to Section V. N. for more information on biological containment levels.
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V. E. Electrical Safety Effects of Electricity on the Body There are inherent dangers involved when using any electrical equipment and therefore care must be exercised when operating and especially when installing, modifying, and/or repairing any electrical equipment. Electrical shock, the passage of current through the human body, is the major electrical hazard. Factors involved in electrical shock are: The quantity of current flowing through the body
Current (amperes) is the killing factor in electrical shock, not the voltage. The voltage only determines how much current will flow through a given body resistance. In general, the body’s resistance to electrical shock is minimal (150 to 600,000 Ohms). Even contact with standard 110-volt circuits can be lethal under certain conditions.
The current path through the body from entry to exit
Hand-to-hand, hand-to-foot, head-to-foot, and ear-to-ear current paths are the most dangerous because they may cause severe damage to the heart, lungs and brain. This is why it is important not to wear metal jewelry, and not to lean against or use both hands on electrical equipment so as not to become part of the circuit.
The length of time the body is in the circuit
The longer the body is in the circuit, the greater the damage. You may be unable to let go of a circuit that exceeds 15 to 20 milliamps. The body temperature may increase possibly damaging tissues, bones, and organs.
The table on the following page describes the effects of different levels of current on the body. ______________________________________________________________________________ Current passing through Effects of 60 Hz Current on the Body the body in milliamps ______________________________________________________________________________ 1 or less . . . . . . . . . . . . . . . . may not be felt 1 to 8 . . . . . . . . . . . . . . . . . . . sensation of mild shock; can let go at will 8 to 15 . . . . . . . . . . . . . . . . . . painful shock, muscles contract; may be able to let go 15 to 20 . . . . . . . . . . . . . . . painful shock; cannot let go 20 to 75 . . . . . . . . . . . . . . . intense pain; can not let go; breathing may be paralyzed 100 to 200 . . . . . . . . . . . . . . . ventricular fibrillation fatal if normal heartbeat not restored; holds unconscious
victim to circuit 200 or more . . . . . . . . . . . . . heart stops, muscles contract intensely possibly breaking bones; severe burns;
breathing stops 5 milliamps is the maximum harmless current intensity Do not touch a person in contact with a live electrical circuit. If the current is high enough you will not be able to let go of the victim. You will also become part of the circuit. Disconnect the power first!
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Types of Electrical Equipment and Safety Guidelines Types of electrical equipment in the BPEC laboratories include power supplies, electrophoresis apparatus, Coulter Counters, hot plates and variacs. Work involving any of these types of equipment can lead to serious injuries if prudent electrical practices are not followed. In order to assure the safe operation of electrical equipment, all electrical equipment must be installed and maintained in accordance with the provisions of the National Electric Code (NEC) of the National Fire Protection Agency. The MIT Safety Program should be contacted for any questions on NEC, and before attempting any minor repairs, modifications, or installations of electrical equipment. A copy of the National Electric Code NFPA Code #70 is available from the MIT Safety Program (N52-496). Modifications and repairs to the receptacle and wiring in the walls is the responsibility of Department o Facilities and should not be attempted or carried out by anyone else. Also, it is strongly advised that all work on electrical equipment be carried out by qualified personnel. Before carrying out any minor repairs, modifications, or installations of electrical equipment, it is required that the equipment be de-energized and all capacitors safely discharged. Furthermore, this de-energized and/or discharged condition must be verified before proceeding. A lockout device is required for disabling power from circuit breakers to prevent other workers from inadvertently reactivating the breaker. The following electrical safety guidelines were published in the National Safety Council’s R&D Newsletter, Jan-Feb., 1987. 1) Design for safety. Consider safety as an integral part of the experimental design process.
Use the above mentioned sources of electrical safety review. Protective devices, warning signs or labels, and administrative procedures only supplement the design. These can never take the place of engineering controls, such as ground fault interrupters, that should be built into the experimental design.
2) Resist rush pressure. Work pressures can cause even the most seasoned veteran to
momentarily forget safe habits. Resist the temptation to take unsafe shortcuts to save a few minutes.
3) Identify hazards and anticipate problems. Think through what might go wrong and the
possible consequences. 4) De-energize equipment before working on it. Always disconnect at the source and
work towards the load. Then ensure that the equipment remains safe after it has been de-energized by following the department’s shut down and notification procedures. If energized equipment must be worked on, the operation’s hazards should be carefully reviewed to develop specific procedures. It is also a good idea to test serviced equipment with a multimeter to assure that it is grounded before reconnecting power.
5) Prepare emergency procedures. All personnel working in areas with high hazard
electrical equipment should be trained in emergency response procedures, including cardiopulmonary resuscitation (CPR). A panic switch that disconnects the main power to an area in an emergency should be located near an exit and legibly marked.
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6) Practice proper housekeeping. An orderly work area minimizes the possibility of
tripping and falling into live equipment, fires, and many other kinds of accidents. OSHA requires 36 inches of space around electrical equipment with more than 600 volts.
7) Maintain equipment for safety. Good maintenance is essential to safe operations.
Experimental designs should provide for the safe maintenance of equipment, including procedures, schedules and records. Electrical equipment should be fused to protect the user from electrical shock.
8) Document your work. Always update documentation, such as prints, operational
procedures, maintenance manuals and logs. These should be available to anyone working on potentially hazardous electrical equipment.
9) Periodically attend safety seminars. To avoid complacency about electrical hazards, be
aware of the current safety procedures and of the accidents that have occurred at MIT. Both of these are covered during the department’s safety seminars.
Electrical Wiring Standards 1) Proper wiring. The installation, replacement, modification, repair or rehabilitation of
any part of any electrical installation must be in compliance with NEC standards, which specify the proper wiring. For any piece of electrical equipment, there must be a switch in a convenient and readily accessible location that will disconnect the main power source to the apparatus in the event of an emergency. Temporary wiring should only be used when absolutely necessary and must be replaced with permanent wiring as soon as possible. Temporary wiring must also comply with NEC codes. Extension cords must be used only as temporary wiring for portable equipment. For permanent equipment, permanent wiring should be installed.
2) Grounding. All equipment should be grounded and fused in accordance with NEC
codes. 3) Insulation. All electrical equipment should be properly insulated. Any power cords that
are frayed should be replaced and any exposed hot wires should be insulated to prevent the danger of electrical shock due to accidental contact. When working with high voltage equipment, properly rated gloves and matting for electrical protection should be used. When in doubt, consult with the MIT Safety Program.
4) Isolation. All electrical equipment or apparatus that may require frequent attention must
be capable of being completely isolated electrically. All power supplies must be enclosed in a manner that makes accidental contact with power circuits impossible. In every experimental setup, an enclosure should be provided to protect against accidental contact with electrical circuits. This applies to temporary arrangements as well.
Specific Electrical Equipment Hazards in the BPEC Laboratories
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Gel electrophoresis equipment is probably the most frequently encountered electrical hazard in the BPEC laboratories. Individuals who use this equipment should be intimately familiar with electrical safety procedures for this equipment. Uncovered gel electrophoresis units are no longer allowed at MIT. For any specialized questions on such equipment, consult with the MIT Safety Program.
V. F. Emergency Action Plan This section addresses guidelines on the safe and effective protection of all personnel in the event of different types of emergencies. Treatment of injured and contaminated personnel as the result of an accidental release of a hazardous substance is covered in Section V. A. The handling of leaky gas cylinders in addressed in Section V. H. Emergencies that can occur in a laboratory include chemical spill or release, explosion, fire, and medical or other health threatening accidents. General procedures to be followed in any emergency in the indicated order of priority are: 1) Render assistance to person(s) involved and remove them from exposure to further injury
if necessary and if this can be done safely. a) If a coworker has ingested a toxic substance, have the victim drink large amounts of
water while escorting them to the Medical Department. Attempt to learn exactly what substances were ingested and inform the medical staff as soon as possible.
b) If a person is found unconscious, dial 100 to get help. Perform CPR if you are trained
in this procedure. Never give anything by mouth to an unconscious person. c) If a coworker is bleeding severely, elevate the wound above the level of the heart and
apply firm pressure directly over the wound with a clean cloth or your hand. Call for medical assistance immediately; dial 100.
d) Do not touch a person in contact with a live electrical circuit. If the current is high
enough you will not be able to let go of the victim. You will become part of the circuit. Disconnect the power first!
2) Notify nearby persons who may be affected and call 100 to report the emergency and
seek assistance. Campus Police, the Cambridge Fire Department and the Cambridge Police Department can be reached by dialing 100. Note that Environmental Health & Safety Program is on-call 24 hours a day and can be reached by the Operations Center 3-1500.
3) Evacuate the area until help arrives. If necessary, pull the fire alarm to evacuate the entire
building. 4) Wait for emergency responders and assist them in handling the emergency. 5) Assist in the follow up investigation of the emergency.
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V. F. 1. Emergency Information Cards and Phone Lists Each laboratory door is posted with an up-to-date green Emergency Information Card which lists the names, home telephone numbers, work telephone numbers and work addresses of key emergency contact persons for the laboratory. These persons are familiar with equipment or chemicals in the laboratory that may pose a safety or life-threatening problem in the event of an emergency. It is the responsibility of the Safety Officer to ensure that these cards are updated regularly. Blank cards can be obtained from the Safety Program. General MIT emergency phone numbers are listed next to each lab phone. Those numbers should be checked as part of the lab inspections.
V. F. 2. Emergency Evacuation In the event of an emergency, the above general emergency procedures should be followed. The Institute rule for emergencies is complete evacuation. The Fire Marshals should perform their duties in the process of evacuation. Fire Marshal (currently Daniel Darling, room 16-429, Ph: 452-2464) a) Communicate an alarm: i) Dial 100 to report the emergency and, if time, send someone to pull the nearest fire
alarm. ii) Alert people in your assigned rooms that there is an emergency. Assigned rooms for
each Fire Marshal are given in the BPEC Chemical Hygiene and Safety Committee list (Section II. A. 4.).
Researchers a) If you see smoke or fire, report it to a Fire Marshal. b) Ensure evacuation in assigned room; assist elderly and handicapped persons. c) ON THE WAY OUT FROM THE OFFICE AREAS, if possible, shut off air
conditioning, computers, radios and close windows and doors. d) ON THE WAY OUT FROM THE LAB AREAS, if possible, stabilize or shut down
dangerous or critical equipment and experiments. Shut down fuel supplies such as gas. Shut down critical operations such as pressure vessels.
All personnel should study the floor plan for their building, posted in each laboratory and be familiar with evacuation routes and the locations of fire alarms and gas shut offs. All fire alarms and gas shut offs for BPEC laboratories are located in the corridors outside of the labs. Use stairwells and doors labeled “Exit” to evacuate. DO NOT use elevators. Report to the courtyard outside Building 16, by the concrete benches located next to Building 18 (the Chemistry Building). Wait for emergency responders and be prepared to explain the cause
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of the evacuation and to alert them of any hazardous substances in the labs that may hinder their investigation. Do not re-enter the building until given the “all clear” by emergency personnel.
V. F. 3. Fires MIT Policy states that personnel are not required to fight fires. If the fire is small and you have appropriate training, you may try to extinguish the fire yourself. Extinguishing fires should only be attempted if you are confident that you can do so successfully and quickly, and from a position in which you are always between the fire and an exit from the laboratory. Do not underestimate fires, and remember that toxic gases and smoke may present additional hazards. In most circumstances, immediate and complete evacuation is advised. In the Event of a Fire a) Pull a fire alarm (located in the corridor or near a stairwell). The alarm is activated by
pulling down on the handle. The alarm will make a loud whooping sound. OR b) Dial 100 on any Institute phone to alert authorities to secure help. Activating the fire
alarm should automatically alert the authorities. Give them your name, location, and the nature of the fire and other emergencies. Stay on the phone until the dispatcher hangs up.
c) Fire Marshals and Researchers should perform their duties (Section V. F. 2.) in the
process of evacuation. d) Assist the handicapped if you can. Use horizontal evacuation where available. If not
available, head for an enclosed stairwell and wait. If possible, dial 100 and let the Campus Police know where you are.
e) Drop and roll if your clothes catch on fire. Do not run! Running will only intensify the
burning. If you can safely get to a fire blanket, wrap it around yourself or the individual who is on fire. Make yourself or the individual drop to the ground and roll. Unfortunately fire blankets tend to hold in heat and increase the severity of burns. Remove the blanket as soon as possible. For all burn victims, remove contaminated clothing and if possible place wet, cold clothes on burned areas. Wrap the injured person in a blanket to avoid shock and get medical attention promptly.
f) Evacuate the building. Use the evacuation route designated by the department. This
should be posted in all rooms. If you have time, alert a BPEC Fire Marshal before evacuating. The Fire Marshal will ensure complete evacuation. Do not enter a stairwell if it is full of smoke. Do not use elevators. Find an alternative route.
g) Avoid Smoke. If you have trouble finding a clear exit, or if smoke intensifies. Drop to
you hands and knees and crawl to the nearest exit. Smoke will rise making air near the ground clearer.
h) Report to the Designated Meeting Area. Report to the courtyard outside Building 16,
by the concrete benches located next to Building 18 (the Chemistry Building). Wait for
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emergency responders and be prepared to explain the cause of the fire and to alert them of any hazardous substances in the labs that may react adversely with the fire.
i) Do not re-enter the building until given the “all clear” by emergency personnel. j) Inform the Safety Program and the BPEC Chemical Hygiene Officer or Laboratory
Supervisor of any fires or almost fires, no matter how small. Sharing this information may prevent someone else from encountering the same situation.
The following guidelines should be followed to prevent and minimize injury and damage from fires. General Guidelines for Fire Prevention and Rapid Evacuation a) Clutter causes accidents and impairs evacuation. Be neat! Do not leave items such as
bicycles, boxes, buckets and trash bags in the middle of the floor or blocking exits. b) In offices and labs, be alert to possible ignition sources such as extension cords, electrical
outlets, coffee pots, radiators, hot plates, soldering irons and Bunsen burners. Keep combustible materials away from these ignition sources. Turn off equipment after use. Turn off all non-essential electrical appliances and machinery at the end of the day.
c) Keep all doors to stairwells closed at all times. These are fire doors designed to block the
spread of a fire. Do not block or wedge these doors open. d) Store all flammable materials in solvent or other appropriate cabinets. Store flammable
materials upright. e) When using flammable materials, stay far away from ignition sources (e.g. hot plates,
Bunsen burners, soldering irons, etc.). f) When you have emptied a flammable materials container, uncap it, and place it in a fume
hood to let the last drops evaporate before discarding it. g) When lighting Bunsen burners, ensure that flammable material is not near the burner.
Always shut off natural gas outlets and Bunsen burners when you are done using them, or if they must be unattended, even for a short period.
h) Use of Bunsen burners in the biological safety cabinets is not recommended. If use is
desired, never use in conjunction with flammable materials and never leave unattended. The airflow pattern in the biological safety cabinets can easily spread the flame and even ignite the internal filters of the cabinet. If a fire breaks starts, shut off the airflow and close the glass sash. Shut down any other biological safety cabinets or fume hoods that operate off the same air supply duct and close their sashes. Air can still enter through the air supply duct, so the fire may not be extinguished until all internal filters have burned.
i) Be prepared! Know departmental evacuation routes and where the nearest exits, fire
alarm pull stations, fire extinguishers, gas shut offs, emergency numbers, safety showers,
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fire blankets, eyewashes, and other fire safety equipment are. Know how to use fire extinguishers and what types of fires they can be used for.
a) Know what the fire alarm sounds like in your building. The fire alarms vary from
building to building at MIT. b) If you hear a fire alarm, assume that it is genuine and evacuate the building immediately.
Specific Hazards that May Lead to Fires or Explosions The combination of certain compounds or classes of compounds can result in a violent chemical reaction leading to an explosion or fire. Other compounds pose explosion or fire hazards when exposed to heat, shock, or other conditions. Listed below are some of the specific compounds and combinations of compounds encountered in the BPEC laboratories that may pose explosion or fire hazards. This list is not intended to be complete. Researchers should always be familiar with the flammability and other properties of the chemicals involved in their research. a) Dimethyl sulfoxide decomposes violently on contact with a wide variety of active
halogen compounds. Explosions from contact with active metal hydrides have been reported.
b) Ethylene oxide has been known to explode when heated in a closed vessel. Experiments
using ethylene oxide under pressure should be carried out behind suitable barricades. Ethylene oxide sterilized materials should not be autoclaved.
c) Liquid nitrogen cooled equipment open to the atmosphere rapidly condense liquid air.
When coolant is removed, an explosive pressure buildup occurs, usually with enough force to shatter glass equipment. Hence, only sealed, evacuated or vented equipment should be cooled.
d) Oxygen cylinders. Serious explosions have resulted from contact between oil and high-
pressure oxygen. Oil should not be used on connections to an oxygen cylinder. Furthermore, only regulators bearing an oxygen seal can be used for oxygen cylinders.
e) Gas cylinders in general. Most cylinders are equipped with frangible discs, fusile plugs,
spring-loaded relief valves, or combinations of these devices to prevent the explosion of normally charged cylinders when involved in a fire. Cylinders containing toxic or poisonous gases, for which the risk of human exposure to fumes is more hazardous than the potential cylinder failure, do not have such relief devices.
V. G. Equipment Maintenance and Malfunction The responsibility for equipment maintenance is distributed among the groups sharing the space, i.e. the Griffith, Hamel, Lauffenburger and Wang groups. In the event of an equipment malfunction in a specific area, personnel can be contacted using the list of researchers located on the pigeon-hole mailboxes in 16-436 or 16-476. That list is organized by research area.
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In the event of a power failure, each office and laboratory door and equipment therein is marked with an Emergency Power Sticker that gives information on the length of time the equipment can sustain proper operation without power. If power is going to be off for longer periods of time than required for proper operation, notify Facilities, extension 3-1500, of the need for temporary power arrangements.
V. H. Gas Cylinders and High Pressure Gas Systems Ordering Gas and Liquid Nitrogen Cylinders Gas cylinders and Liquid Nitrogen should be ordered through B.O.C. Gases, which is located on campus (3-4761, fax 3-4968). To order from B.O.C., fill out a requisition and either call or fax in your order. Transportation and Storage of Cylinders 1) To transport gas cylinders within the BPEC labs, they must be capped with screw-on
safety caps and tightly strapped or chained to a cylinder cart. A maximum of two or three cylinders can be transported at a time, depending on the capacity of the cylinder cart.
2) Cylinders ordered from B.O.C. or other external vendors are to be delivered and picked
up by the vendor. 3) Upon receipt, all gas cylinders must be labeled and secured. Cylinders should already
have a label with the name of the gas in them. On the label, write the initials of the person responsible and the date the cylinder was received. Cylinders from most outside vendors are labeled with the order date. If the cylinders are for common use, such as carbon dioxide cylinders for the incubators, they need not be initialed. Check the condition of the valve by squiring a leak detector such as Snoop on it.
4) All gas cylinders must be secured to stationary supports with clamps, straps and/or
chains, or supported freestanding with a cylinder base unit. Gas cylinders are capped with screw-on caps when in storage.
5) The main storage location for compressed gas cylinders should be in a level, fire-resistant
area that is well ventilated and dry. Storage areas should be located away from sources of ignition or excessive heat, such as steam or hot water pipes, boilers, open flames, or sparking equipment, where leaking gas could ignite or explode. Cylinder temperature should not exceed 51°C (124°F), as the heat may activate the fusible metal relief device, releasing the contents of the cylinder.
6) Corrosive gases should be stored for the shortest possible time period; preferably under
three months. Using Cylinders and High Pressure Gas Systems and Moving Cylinders The procedures adopted for the safe handling of compressed gases are mainly centered on containment of the material, to prevent its escape to the atmosphere, and proper control of pressure and flow. The following procedures should be used when using or moving gas cylinders and when working with high pressure gas systems:
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1) All gas cylinders are sealed with two-stage pressure regulators when in use. Specially designed regulators must be used for certain gases, including oxygen that reacts explosively with oils that are used in many regulators. Check with the gas supplier for specifications on regulators for a given type of gas. In general, inert gases and carbon dioxide do not require special regulators, whereas oxygen, nitrogen, helium, hydrogen and many other gases require specially designed regulators. These special regulators denote the gases for which they are approved. These regulators are available through B.O.C. or other vendors. Regulators may be costly (approx. $800) but are essential to prevention of explosions and other adverse reactions.
2) Do not force the fittings of the regulator to the cylinder. If a regulator does not fit
properly, it may indicate that it is not intended for use with the gas cylinder, or the type of gas. Similarly, if the main valve cannot be opened, do not use a wrench to attempt to open it. This may result in breakage of the valve and leakage of the gas. Return the cylinder to the vendor or request a special valve wrench from the vendor.
3) When pressurizing a system from a gas cylinder or in-house compressed gas system wear
safety goggles. In-house compressed air contains particulate matter; do not place your hand or other body parts in front of a compressed gas jet.
4) When changing cylinders it is good practice to replace the plastic gasket. This will help
reduce leakage. Always test all connections for leaks after changing cylinders using Snoop™ or other leak detector.
5) It is good practice to check all connections in a compressed gas system once per month,
especially if an increase in gas usage is noticed. 6) To turn off the gas, turn off the cylinder valve first and then the regulator. Use the
cylinder valve, not the regulator valve to turn off the gas. 7) Whenever a gas cylinder is moved, the regulator must be removed and the cylinder must
be capped with a screw-on safety cap. To move the cylinder a short distance, slightly tilt the cylinder, no more than ten degrees and use a turning and swiveling motion to move the cylinder. This procedure will seem awkward and slow at first, but it is the only safe way to move a gas cylinder. Never tilt the cylinder more than about ten degrees. The cylinder can quickly become overbearing. If the cylinder is dropped, the pressurized gas may easily project the valve through walls and anything else in its path. The safety cap will help reduce this problem. In the event that a cylinder becomes overbearing or is dropped, try to get out of the way of the top of the cylinder in case the valve gives way. If gas cylinders need to be moved out of a room or further, use a cylinder cart for transportation. It is usually easier to pull than push a cylinder cart over doorway guards and other uneven surfaces.
8) If several different types of gases are manifold, one-way check valves should be placed
on the outlet line of each cylinder to prevent accidental gas mixture due to pressure differences.
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10) To prevent contamination, cylinders should not be completely emptied as back-flow of air, moisture or other gases into the cylinder may result in an explosive or corrosive mixture with the remaining gas. Always leave a least 25 psi of pressure in all cylinders.
Returning Empty Cylinders Cylinders ordered through B.O.C. Gases are generally picked up by B.O.C. when they deliver the replacement tanks. Used cylinders should be labeled “Empty” so that the B.O.C. personnel know to remove them from the lab. Specific procedures should be followed: 1) Empty cylinders are to be returned promptly, and it is the researcher’s duty to make sure
that B.O.C. picks-up the empties within a reasonable amount of time. Cylinders that are empty or no longer needed present a number of problems. They may corrode and the labels can become obscured or fall off. They may leak noxious or flammable gas that will contaminate the area and require special and expensive handling to prevent injury to personnel. They also take up lab space and are expensive to keep, costing $2.00 to $35.00 per month in cylinder rental charges.
2) When a gas cylinder is empty, ensure that the valve is tightly closed and the cylinder is
capped. If you need a cap, contact B.O.C. (3-4761). 3) When having the gas cylinders picked up from your lab, it is your responsibility to make
sure that: a) The cylinders remain secured until they are picked up. b) Proper instructions of the cylinder location are given to B.O.C. It is a good idea to
tag the cylinder(s) with a “for pickup” note. c) Someone is present in the lab for the pickup. Allow 1-2 working days for pickup. Handling Leaky Gas Cylinders Occasionally a cylinder or one of its component parts develops a leak. Most such leaks occur at the top of the cylinder in areas such as the valve threads, safety device, regulator, valve stem, and valve outlet. If a leak is suspected, do not use a flame for detection. Instead, use a flammable-gas leak detector or soap solution (Snoop is available in lab 16-436 at the end of benches 11 and 12). If the leak cannot be remedied by tightening a valve gland or a packing nut, emergency action procedures should be instigated and the supplier should be notified. Laboratory workers should never attempt to repair a leak at the valve threads or safety device. Instead, they should consult with the supplier for instructions. The following general procedures can be used for relatively minor leaks where the indicated action can be taken without the exposure of personnel to highly toxic substances. Note that if it is necessary to move a leaking cylinder through populated portions of the building, place a plastic bag, rubber shroud, or similar device over the top and tape it (duct tape preferred) to the cylinder to confine the leaking gas. Never transport a leaking cylinder containing highly toxic gas. 1) Flammable, inert, or oxidizing gases. Move the cylinder to an isolated area (away from
combustible material and ignition sources if the gas is flammable or an oxidizing agent) and post signs that describe the hazards and state warnings. If feasible, leaking cylinders
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should always be moved into laboratory hoods. While there are no hoods large enough for this practice in the BPEC labs, if the leaking cylinder can be moved safely in front of an operating laboratory fume hood that is suitable safety measure.
2) Corrosive gases may increase the size of the leak as they are released and some
corrosives are also oxidants or flammable. Move the cylinder to an isolated, well-ventilated area and use suitable means to direct the gas into an appropriate chemical neutralizer. Post signs that describe the hazards and state warnings. If the leak becomes extremely large, evacuate the area as the cylinder may eventually leak to the point of releasing an explosive force.
3) Toxic Gases. Follow the same procedure as for corrosive gases. Move the cylinder to an
isolated, well-ventilated area and use suitable means to direct the gas into an appropriate chemical neutralizer. Post signs that describe the hazards and state the warnings.
When the nature of the leaking gas or the size of the leak constitutes a more serious hazard, self-contained breathing apparatus and protective apparel may be required. Evacuate personnel from the affected area. Pull the fire alarm if the entire building or floor should be evacuated. Call Campus Police (dial 100) to obtain emergency assistance. Hazards and Safety Devices Associated with Gas Cylinders 1) Diffusion of leaking gases may cause rapid contamination of the atmosphere, giving rise
to toxicity, anesthetic effects, asphyxiation, and rapid formation of explosive concentrations of flammable gases. The ignition point of a flammable gas under pressure is always lower than ambient, or room, temperature. Leaking gas can therefore rapidly form an explosive mixture with air.
2) Low boiling point materials can cause frostbite on contact with living tissue. This is
common among the cryogenic liquids, such as nitrogen and oxygen, but it can also result from contact with the liquid phase of a liquefied gas, such as carbon dioxide, fluorocarbons, and propylene.
3) Other hazards of compressed gases include corrosion, irritancy and high reactivity. 4) Most cylinders are equipped with frangible discs, fusel plugs, spring-loaded relief valves,
or combinations of these devices to prevent the explosion of normally charged cylinders when involved in a fire. Cylinders containing toxic or poisonous gases, for which the risk of human exposure to fumes is more hazardous than the potential cylinder failure, do not have such relief devices.
5) All cylinders and piping containing flammable gases should be grounded to prevent
explosion caused by buildup of static electricity. 6) As noted above, specially designed regulators are required for many types of gases,
including oxygen, which reacts explosively with oil or greased contained in many types of regulators.
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7) As noted above, flammable, oxidizing, corrosive and toxic gases pose many problems. Order the minimum amount of these gases required for experimentation, and promptly return empty or unused cylinders.
8) Contact the Safety Program for information on threshold limit values, flammability limits
in air, and major hazards of a variety of commonly used gases.
V. I. Glassware The individual users are responsible for washing and decontamination of glassware in all BPEC laboratories. However, certain rules for handling glassware and preparing glassware for cleaning apply to all BPEC personnel. Rules for Handling Glassware 1) All glassware is assumed to be contaminated (chemically or biologically), unless it is
known otherwise. Always wear protective clothing (lab coat, safety eyewear and gloves) when separating or handling contaminated glassware.
2) To protect researchers from contacting unknown hazards, all glassware must be rinsed
before being placed on a dirty glassware cart or tray. Glassware requiring biological decontamination should be decontaminated with chlorine, Lysol™, Wescodyne™ or other effective disinfectant, before placing on the dirty glassware cart for cleaning. Any glassware that has come into contact with a culture should be considered biologically contaminated. The decontamination or soaking solution may remain in the glassware if the item needs to be soaked before cleaning as long as the solution does not pose a contact hazard to others (e.g. solutions containing acids, bases, or solvents). Soaking for a minimum of 20 min is usually done for biologically contaminated glassware to ensure decontamination.
3) Sinks are not to be used as storage areas. Glassware or other labware should not be left
in the sink overnight. Place all decontaminated glassware on dirty glassware carts or trays.
4) Glassware from 16-436 must be kept separate from laboratory glassware in 16-476.
Bacterial contamination from 16-476 would be highly deleterious to the animal cell culture work being performed in 16-436. Because there is only one dishwasher in Building 16, it is the responsibility of the person doing the dishes to ensure that glassware remains separated during washing and decontamination. Laboratory researchers are responsible for ensuring that glassware is not inadvertently transferred between these labs during experiments.
Rules for Disposing of Glassware All glassware should be rinsed before discarding and biologically contaminated glassware must be decontaminated with bleach before disposing. If biologically contaminated glassware is broken or otherwise too difficult to decontaminate, throw it in one of the dedicated gray Biohazard Sharps container for disposal.
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V. J. Housekeeping Laboratory 16-436 is an approved Bio-Level 2 facility and must be operated under the guidelines BL2 containment (Section VII. B.). The following practices are required under BL2 regulations: 1) Do not prepare or store food or drink in the lab. 2) Do not wear or store lab coats outside of the lab. 3) Biohazardous samples are not to be taken out of the lab except in approved transportation
containers. 4) All exterior doors should remain closed at all times to maintain positive pressure in the
BL2 facility. Researcher's Responsibilities It is the responsibility of the researchers to maintain the laboratory in a neat and ordered state and to keep equipment and chemicals labeled and properly stored. Specific responsibilities include:
1. Clutter causes accidents and impairs evacuation. Be neat! Do not leave items such as bicycles, boxes, buckets and trash bags in the middle of the floor or blocking exits. In fact it is not allowed to store bicycles in labs 16-436 and 16-476, or in the instrument rooms.
2. Clean work areas when work is completed using recommended procedures. 3. Decontaminate common work areas such as biosafety cabinets and incubators after
use. 4. Restock shelves, drawers and carts with depleted materials. 5. Store apparatus that are not currently being used. 6. Replenish chemicals that are exhausted. Fill water baths as necessary. 7. Ensure that all gas cylinders are secured to stationary supports with clamps, straps
and/or chains, or supported free standing with a cylinder base unit, and that the cylinders are capped with screw-on caps when in storage.
8. Ensure that all chemical storage regulations are followed (Section IV. B. 3.). 9. Rinse dirty glassware and place on glassware carts. 10. Place all biohazardous wastes in proper containers and empty containers as necessary.
Refer to Section VII. B. 6. for proper biohazardous waste disposal procedures. 11. Maintain neat and clean refrigerator and freezer spaces. Assist with routine defrosting
of freezers and consolidation of cell banks. 12. Immediately chemically disinfect and discard contaminated cultures. Disinfect any
contacted surfaces or equipment as well. 13. Report equipment malfunctions to the researcher assigned to the faulty apparatus. 14. Report any personnel, equipment safety or housekeeping violations to the Laboratory
Supervisor and Laboratory Safety Officers.
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V. K. Incompatible Substances When transporting, storing, using or disposing of any substance, utmost care must be exercised to ensure that the substance cannot accidentally come in contact with an incompatible one. Such contact could result in a serious explosion or the formation of substances that are highly toxic or flammable or both. The following table is a guide to avoiding accidents involving incompatible substances.
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____________________________________________________________________________ CHEMICAL OR CLASS IS INCOMPATIBLE WITH ___________________________________________________________________________________________ Acids and alkalis . . . . . . . . . . . . . . organics Acetic acid . . . . . . . . . . . . . . . . . . . . chromic acid, nitric acid, perchloric acid, peroxides, permanganates Acetone . . . . . . . . . . . . . . . . . . . . . . concentrated nitric and sulfuric acid Alkali . . . . . . . . . . . . . . . . . . . . . . . water, carbon tetrachloride or other chlorinated hydrocarbons, carbon
dioxide, halogens Carbon tetrachloride . . . . . . . . . . . . sodium Chlorine (including clorox) . . . . . . . . ammonia, acetylene, butadiene, butane, methane, propane (or other
petroleum gases), hydrogen, sodium carbide, benzene, finely divided metals, turpentine
Chromic acid & Chromium trioxide . acetic acid, naphthalene, camphor, glycerol, alcohol, flammable liquids Flammable liquids . . . . . . . . . . . . . . . ammonium nitrate, chromic acid, hydrogen peroxide, nitric acid,
sodium peroxide, halogens Hydrocarbons . . . . . . . . . . . . . . fluorine, chlorine, bromine, chromic acid, sodium peroxide Hydrogen peroxide . . . . . . . . . . . . . . . copper, chromium, iron, most metals or their salts, alcohols, acetone,
organic materials, aniline, nitromethane, combustible materials
Mercury . . . . . . . . . . . . . . . . . . . . . acetylene, fulminic acid, ammonia Nitrates . . . . . . . . . . . . . . . . . . . . . sulfuric acid Nitric Acid . . . . . . . . . . . . . . . . . . . . acetic acid, aniline, chromic acid, hydrocyanic acid, hydrogen sulfide,
flammable liquids or gases, copper, brass, any heavy metal Nitrites . . . . . . . . . . . . . . . . . . . . . . acids Oxalic acid . . . . . . . . . . . . . . . . . . . . silver, mercury Oxidizers . . . . . . . . . . . . . . . . . . . . . organics Oxygen . . . . . . . . . . . . . . . . . . . . . oils, grease, hydrogen, flammable liquids, solids, or gases Perchloric acid . . . . . . . . . . . . . . . . acetic anhydride, bismuth and alloys, alcohol, paper, wood, grease, oils Sodium nitrite . . . . . . . . . . . . . . . . . ammonium nitrate and other ammonium salts Sulfides . . . . . . . . . . . . . . . . . . . . . acids Sulfuric acid . . . . . . . . . . . . . . . . . . potassium chlorate, potassium perchlorate, potassium permanganate,
(similar compounds of light metals such as sodium, lithium)
V. L. Personal Hygiene and Sanitation Personal Hygiene Proper personal hygiene and sanitation is essential for maintenance of a safe and orderly laboratory environment. The guidelines for the BPEC labs are as follows:
• Lab coats and safety glasses are to be worn at all times by all personnel while conducting hazardous experiments in lab and in the BL-2 areas. Lab coats and safety glasses are not to be worn or stored outside of the lab.
• Lab coats should be submitted for cleaning as necessary. • Vinyl or latex gloves must be worn for all manipulations with biohazardous,
radioactive and toxic materials. Also, gloves must be worn when manipulating in the BL-2 areas.
• Lab personnel are asked to wash their hands before leaving the lab. • All waste must be placed in the correct receptacle. • No eating, drinking, smoking, no mouth pipetting or applying makeup in lab.
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V. M. REU and UROP Regulations All REU and UROP students must undergo BPEC Chemical Hygiene and Safety Training (Section X.) and submit all forms before beginning work in the laboratory. Entry-level REU and UROP students may not work in the laboratory unsupervised. The supervising graduate student, post-doctoral fellow, staff or visiting scholar/student must be in the lab with the student at all times. Entry-level indicates a student who is still receiving training on equipment and procedures. Once an REU or UROP student is fully trained on all equipment and procedures with which he/she works, he/she may work unsupervised in the lab. However, the supervisor must be in or near the lab (in of the labs in Buildings 16 or 56, or in their offices). If the supervisor must leave the area he/she must assign supervision of the student to another graduate student, post-doctoral fellow, staff or visiting scholar/student. Supervisors should check on students frequently, especially if the student is the only person in the lab. Senior-level UROP students, those who have been in the program for more than one semester, may work unsupervised, without the requirement that their supervisors be in buildings 16 or 56, as long as the student is working on non-hazardous procedures and equipment with which he/she is familiar. However, at least one graduate student, post-doctoral fellow, staff or visiting scholar/student must be working in or near the lab (as defined above) when a senior-level REU and UROP student is working unsupervised. This person must also be notified that the student is working. As soon as this person desires to leave, the student must also leave. REU and UROP students cannot be the only person(s) in the lab area at any time.
V. N. Signs and Labels Signs Posted at all Entries to Laboratories Emergency Information Cards and Phone Lists
Each laboratory door is posted with an up-to-date green Emergency Information Card which lists the names, home telephone numbers, work telephone numbers and work addresses of key emergency contact persons for the laboratory. These persons are familiar with equipment or chemicals in the laboratory that may pose a safety or even life-threatening problem in the event of an emergency. Blank cards can be obtained from the Safety Program.
Protective Clothing Requirements
Labs that require special procedures or protective clothing be used upon entry are posted with signs advising personnel of these requirements.
Biohazard Containment Level
Lab 16-436 is approved for Bio-Level 2 containment (BL2) with designated envelopes of BL1 within. This laboratory is posted with a BL2 sign on every external door. Lab 16-476 is a BL-1 laboratory. Biosafety-Level containment classes are defined by local, national or Institute codes based on the perceived risk of the biohazardous agent being used. There are four levels of physical containment:
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BL1 Minimal or no hazard to personnel or the environment. Most work at MIT falls into this category. Note that all recombinant DNA research is at least BL1.
BL2 Potentially pathogenic organisms are used. This includes plant or animal
pathogens. Three examples are polio, hepatitis and human immunodeficiency (AIDS) viruses.
BL3 Threat of serious or lethal disease. A typical example would be work with certain
viruses that cause encephalitis. BL4 Extremely hazardous infectious agents. BL4 work is neither authorized or planned
at MIT owing in part to local codes. At BPEC, the following labs are currently classified at the BL-2 level: • Lab 16-436 • Lab 16-479 (the cell bank) To make it convenient to those of you who work on non-BL2 projects, a BL-1 area was created within lab 16-436. This has been established with and approved by MIT's Biosafety Program. As a result, in practice, part of the inside of lab 16-436 (a BL-2 lab) is allowed to operate as a BL-1 lab, except for the following dedicated BL-2 areas:
• Tissue culture Room A (16-436 a) • Tissue culture Room B (16-436 b) • Space marked with red tape on the floor that includes BSC #5 and dedicated bench • Space marked with red tape on the floor that includes Bench #9
Radioactive Material and Equipment
Radioisotopes are stored in the cell bank, and appropriate signs have been placed outside the door and at the appropriate locations.
Signs Posted on Equipment and Materials Within the Laboratory Biohazardous Signs
Biohazard signs are placed on all equipment used to store, grow or manipulate biohazardous material. This includes, but is not limited to, refrigerators, freezers, cell banks, incubators, centrifuges, microscopes, microwaves, biological safety cabinets, and reactors. Individual samples and cultures are not marked with biohazard signs; instead they are marked with information defining their contents, e.g. cell or virus type, cell suspension or supernatant.
Hood and Biosafety Cabinet Surveillance Performance Certification Signs
Each fume hood and biological safety cabinet is labeled with a performance certification sign. This sign serves to verify that the hood is in proper working order. The working airflow velocity and date of last certification are noted on the signs. These certifications are updated annually by the Industrial Hygiene Program. Certification of the biological safety cabinets (BSC) is the responsibility of the lab. BPEC has 6 BSC in lab 16-436 and 1 BSC in
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lab 16-476. Currently the BSCs are certified annually by the B&V Testing Company, every March/April.
Radioactive Signs Radioactive signs are placed on all radioactive workbenches and equipment that makes contact with radioactive materials either through use or storage. This includes, but is not limited to, refrigerators, freezers, cell banks, centrifuges, incubators, biological safety cabinets, reactors, gamma counters, pipettors, storage containers, waste receptacles, etc. Currently no radioactive work is done in the BPEC laboratory; only radioactive substances are stored in the cell bank.
Designated Area Signs
Designated area signs are placed on all areas where particularly hazardous chemical operations are being carried out that could result in the accidental release of harmful quantities of a toxic substance. The exception is operations involving biohazardous materials, as these materials are used in all parts of biological BPEC laboratories. Their use is denoted by the BL2 signs on all laboratory external doors and by the biohazard signs on all contacted equipment. The two cell culture rooms, labeled 16-436 A and 16-436 B are BL-2 areas. Furthermore, red tape has been placed on the floor around BSC #5, the bench adjacent to it and bench #9 to indicate that these areas are BL-2. Currently, the red-marked areas are dedicated to manipulation with adenoviruses. Designated area signs are placed on all radioactive benches and fume hoods in the BPEC laboratories.
Chemical Labels 1) Chemical containers must be labeled regardless of where they are stored (personal or
common spaces). The label may be the original manufacturer’s label or a label describing the chemical(s) contained in the substance or mixture, the owner’s initials, date received, date opened (if applicable to degradation), and special hazards (e.g. acid, flammable, poison, radioactive, etc.) when applicable.
2) Physical and chemical data and toxicity warning signs or symbols should be prominently
visible on the manufacturer’s label. Labels for decanted chemicals, chemical mixtures made in the lab, or containers that have lost their label are available at the VWR stock room, located in the basement of Building 56.
3) All containers of chemical waste must be labeled with the type of waste (e.g. solvent,
biohazardous, etc), dated when the container is full, and equipped with an appropriate waste disposal tag (available from the BPEC laboratory safety shelves or the Environmental Management Program, EMP). All wastes from the BPEC labs, except sharps waste, are properly treated and labeled with one of the following MIT waste disposal tags:
a) Autoclaved Noninfectious Biomedical Waste tag for all non-sharp, autoclave-
sterilized, biological waste. b) Chemically Treated Noninfectious Biomedical Waste tag for all non-sharp,
chemically disinfected, biological waste.
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c) Red Hazardous Waste tag for all non-biohazardous, non-radioactive, hazardous
waste such as solvents, heavy metals and toxic chemicals. 4) Radioactive waste receptacles are labeled as such. A disposal log is also located on the
receptacle. All radioactive waste is placed in this receptacle. 5) Gas Cylinders should be delivered with the label intact, on which the type of gas is
written. On the label, write the initials of the person responsible and the date the cylinder was received. Cylinders from most outside vendors are already labeled with the order date. If the cylinders are for common use, such as carbon dioxide cylinders for the incubators, they need not be initialed.
Signs and labels are reviewed in every lab inspection. Missing or outdated signs are replaced. Unmarked containers are brought to the attention of the lab worker for identification.
V. O. Hazardous Waste Disposal POLICY HAZARDOUS WASTE MAY BE GENERATED FROM LABORATORY OPERATIONS, CONSTRUCTION AND RENOVATION ACTIVITIES, PHOTO PROCESSING, AND A VARIETY OF OTHER ACTIVITIES AT THE INSTITUTE. THE PROPER DISPOSAL OF WASTE CHEMICALS AT THE INSTITUTE IS OF SERIOUS CONCERN, AND EVERY EFFORT MUST BE MADE TO DO IT SAFELY AND EFFICIENTLY. THE RESPONSIBILITY FOR THE IDENTIFICATION AND PROPER MANAGEMENTOF WASTE CHEMICALS WITHIN THE INSTITUTE RESTS WITH THE INDIVIDUALS WHO HAVE GENERATED THE WASTE. PROCEDURES THE FOLLOWING SUMMARY PROVIDES A GENERAL OVERVIEW OF REGULATORY REQUIREMENTS APPLICABLE TO HAZARDOUS WASTE GENERATORS.
1. WASTE IDENTIFICATION:
HAZARDOUS WASTE (HW) INCLUDES MATERIALS THAT POSSESS HAZARDOUS CHARACTERISTICS (E.G. TOXIC, IGNITABLE, CORROSIVE OR REACTIVE), OR SUBSTANCES THAT ARE LISTED AS HAZARDOUS WASTE BY THE REGULATORY AGENCIES.
2. LABELING:
CONTAINERS, WHICH STORE HAZARDOUS WASTE, MUST BE PROPERLY AND CLEARLY LABELED. LABELS MUST INCLUDE: 1) THE WORDS "HAZARDOUS WASTE"; 2) THE CHEMICAL NAMES OF CONSTITUENTS (E.G. "ETHANOL"); 3) THE HAZARDS ASSOCIATED WITH THE WASTE IN WORDS (E.G. "TOXIC”). THE LABEL MUST BE DATED AS SOON AS A SATELLITE ACCUMULATION CONTAINER BECOMES FILLED. THE HW LABELS ARE AVAILABLE FROM THE ENVIRONMENTAL MANAGEMENT PROGRAM (X2-3666 OR http://web.mit.edu/environment/wastepickup/index.html)
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Full containers of hazardous waste may not remain in a laboratory for more than 3 days. The EMP will pick up waste and transport them to the Waste Chemical Storage Area. They will require that each bottle is tagged with a red Hazardous Waste Tag and its contents recorded on a disposal inventory sheet supplied upon pickup. The disposal inventory sheet lists the quantity of containers, chemical name, solid or liquid, and hazard associated with the waste, i.e. flammable, toxic, water reactive, etc. Ensure that all waste is correctly and completely labeled before calling for a pickup. Volatile or flammable waste must be stored in a fume hood or safety cabinet until pickup. All other hazardous waste must be stored below eye level, off the floor, in a secure place that will not promote decomposition. Non-hazardous waste must be stored off the floor as well.
3. ACCUMULATION & STORAGE: THE INSTITUTE ALLOWS TWO TYPES OF HAZARDOUS WASTE MANAGEMENT AREAS: LESS THAN 90-DAY STORAGE AREAS AND AREAS THAT ARE OFTEN REFERRED TO AS SATELLITE ACCUMULATION AREAS (SAA). SATELLITE ACCUMULATION: SAA MUST BE ESTABLISHED AT THE POINT OF GENERATION AND REMAIN UNDER THE CONTROL OF THE PERSON GENERATING THE WASTE. A MAXIMUM OF 55 GALLONS OF HAZARDOUS WASTE OR 1 QUART OF ACUTELY HAZARDOUS WASTE ARE PERMITED AT EACH SATELLITE ACCUMULATION AREA. ONLY ONE CONTAINER IS ALLOWED PER WASTE STREAM. SATELLITE ACCUMULATION CONTAINERS MUST BE CLOSED UNLESS WASTE IS BEING ADDED TO THE CONTAINER. ONCE A HW CONTAINER IS FILLED, THE LABEL MUST BE DATED AND THE CONTAINER REMOVED FROM THE SATELLITE ACCUMULATION AREA WITHIN THREE DAYS. ENVIRONMENTAL MANAGEMENT PROGRAM PROVIDES A PICK-UP SERVICE FOR THE WASTE READY FOR DISPOSAL, OR YOU CAN MOVE THOSE CONTAINERS TO A LESS THAN 90 DAY STORAGE AREA IF ONE IS AVAILABLE. HW PICK-UP CAN BE REQUESTED ONLINE AT http://web.mit.edu/environment/wastepickup/index.html OR BY CALLING THE ENVIRONMENTAL MANAGEMENT PROGRAM OF EHS (X 2-EHSS). STORAGE: THE STORAGE AREAS MUST MEET THE SAME REQUIREMENTS AS THE SATELLITE ACCUMULATION AREAS EXCEPT FOR THE AMOUNT AND TYPE OF HW STORED. HOWEVER, THE MAXIMUM STORAGE TIME CANNOT EXCEED 90 DAYS. ENVIRONMENTAL MANAGEMENT PROGRAM WILL SET UP AND MANAGE YOUR LESS THAN 90-DAY STORAGE AREA. 4. INSPECTIONS HAZARDOUS WASTE AREAS (SATELLITE ACCUMULATION AREAS AND STORAGE AREAS) MUST BE INSPECTED ON A WEEKLY BASIS. PERSONNEL MANAGING SATELLITE ACCUMULATION AREAS ARE RESPONSIBLE FOR CONDUCTING THEIR AREA’S INSPECTION. ENVIRONMENTAL MANAGEMENT PROGRAM PERSONNEL CONDUCT THE WEEKLY INSPECTION OF ALL HAZARDOUS WASTE STORAGE AREAS.
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GUIDELINES FOR WASTE REDUCTION PLAN A PROCEDURE FOR WASTE DISPOSAL BEFORE YOU START ON A PROJECT. LABEL WASTE PROPERLY. IT IS UP TO EACH DEPARTMENT, GROUP, OR RESEARCHER TO IDENTIFY WASTE MATERIALS PROPERLY BEFORE DISPOSAL; INADVERTENT MIXING OF INCOMPATIBLE MATERIALS COULD HAVE SERIOUS CONSEQUENCES. PROTECTION OF THE ENVIRONMENT MAKES THE DISPOSAL OF LARGE QUANTITIES OF CHEMICAL AND SOLID WASTES A DIFFICULT PROBLEM. IT IS IN EVERYONE'S BEST INTEREST TO KEEP QUANTITIES OF WASTE TO A MINIMUM. THE FOLLOWING SUGGESTIONS MAY HELP: L. ORDER ONLY THE AMOUNT OF MATERIAL YOU NEED FOR YOUR PROJECT OR
EXPERIMENT EVEN IF YOU CAN GET TWICE AS MUCH FOR THE SAME MONEY.
2. USE ONLY THE AMOUNT OF MATERIAL THAT IS NEEDED FOR CONCLUSIVE RESULTS.
3. AVOID STORING EXCESS MATERIAL, PARTICULARLY IF IT IS AN EXTREMELY TOXIC OR FLAMMABLE MATERIAL, JUST BECAUSE YOU MAY WANT IT IN THE FUTURE.
4. BEFORE DISPOSING OF UNWANTED, UNOPENED, UNCONTAMINATED CHEMICALS CHECK
WITH OTHERS IN YOUR DEPARTMENT WHO MAY BE ABLE TO USE THEM.
5. ON TERMINATION OF A RESEARCH PROJECT OR COMPLETION OF A THESIS, ALL UNUSED CHEMICALS TO BE KEPT BY THE LABORATORY SHALL BE LABELED.
6. MAKE SURE ALL SAMPLES AND PRODUCTS TO BE DISPOSED OF ARE PROPERLY IDENTIFIED, LABELED WITH ITS CHEMICAL NAME AND CONTAINERIZED. DO NOT LEAVE THEM FOR OTHERS TO CLEAN UP AFTER YOU.
SPECIAL PROCEDURES REQUIRED UNKNOWN WASTE CHEMICALS UNKNOWN WASTE CHEMICALS CANNOT BE ACCEPTED FOR DISPOSAL. IT IS THE RESPONSIBILITY OF THE DEPARTMENT INVOLVED TO IDENTIFY ALL CHEMICALS AND THIS MAY REQUIRE POLLING LABORATORY PERSONNEL, STUDENTS AND FACULTY MEMBERS TO ASCERTAIN THE OWNER OF SUCH UNKNOWN WASTE AND ITS IDENTITY. IF IDENTIFICATION IS NOT POSSIBLE, THE ENVIRONMENTAL MANAGEMENT PROGRAM CAN ARRANGE FOR ANALYSIS OF UNKNOWN MATERIALS AND THE PRINCIPAL INVESTIGATOR/LAB GROUP WILL BE RESPONSIBLE FOR THE COST OF ANALYSIS AND DISPOSAL. GAS CYLINDERS ARE TO BE RETURNED TO THE SUPPLIER. SOME SMALL LECTURE BOTTLES ARE NON-RETURNABLE WHICH BECOME A DISPOSAL PROBLEM WHEN EMPTY OR NEAR
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EMPTY WITH A RESIDUAL AMOUNT OF GAS. THE ENVIRONMENTAL MANAGEMENT PROGRAM WILL ARRANGE FOR DISPOSAL OF LECTURE BOTTLES. HOWEVER, THE PRINCIPAL INVESTIGATOR/LAB GROUP IS RESPONSIBLE FOR THE COST OF DISPOSAL. CONTROLLED DRUGS TO BE DISCARDED CAN NOT BE DISPOSED OF AS HAZARDOUS WASTE. THE HANDLING, RECORDS, AND DISPOSAL OF CONTROLLED DRUGS ARE THE RESPONSIBILITY OF THE DEPARTMENT INVOLVED OPERATING WITHIN THE DRUG ENFORCEMENT AGENCY (DEA) REGULATIONS. HOWEVER, THE ENVIRONMENTAL MANAGEMENT PROGRAM CAN PROVIDE ASSISTANCE DURING THE PROCESS. RADIOACTIVE WASTE DISPOSAL IS HANDLED IN ACCORDANCE WITH PROCEDURES ESTABLISHED BY THE RADIATION PROTECTION PROGRAM (EXT. 2-3477). WASTES MARKED AS RADIOACTIVE MUST NOT BE SENT TO THE WASTE CHEMICAL STORAGE AREA. BIOLOGICAL WASTE IS HANDLED IN ACCORDANCE WITH PROCEDURES SET FORTH BY THE BIOSAFETY PROGRAM. (EXT.2-3477). WASTES MARKED AS BIOHAZARDOUS MUST NOT BE SENT TO THE WASTE CHEMICAL STORAGE AREA. SHARPS WASTE - CHEMICALLY CONTAMINATED MUST BE PACKAGED IN PUNCTURE PROOF CONTAINERS AND MUST BE LABELED AS HAZARDOUS WASTE WITH THE CHEMICAL CONTAMINANTS LISTED. CONTAINERS MUST BEMANAGED IN ACCORDANCE WITH HAZARDOUS WASTE REGULATIONS. CHEMICALLY CONTAMINATED SHARPS WASTE MUST NOT BE PACKAGED IN BIOHAZARD CONTAINERS. SHARPS WASTE - BIOHAZARDOUS AND INFECTIOUS MUST BE PACKAGED IN PUNCTURE PROOF BIOHAZARD CONTAINERS AND IS HANDLED IN ACCORDANCE WITH PROCEDURES SET FORTH BY THE BIOSAFETY PROGRAM (EXT. 2-3477). ALL NON-CHEMICALLY CONTAMINATED SHARPS WASTE ORIGINATING FROM A BL RATED LABORATORY IS CONSIDERED TO BE BIOHAZARDOUS. SHARPS WASTE – CLEAN NEEDLES AND SYRINGES ARE CONSIDERED TO BE “MEDICAL WASTE” AND MUST BE PACKAGED IN PUNCTURE PROOF CONTAINERS. NO TAGS ARE NEEDED, BUT LAB NUMBER AND PI NAME MUST BE MARKED ON THE CONTAINER. IF THE LAB PRODUCES A LARGE VOLUME OF NEEDLES AND SYRINGES, A BIOSYSTEMS CONTAINER WILL BE PROVIDED AND A REGULAR PICK-UP SCHEDULE WILL BE ARRANGED. ERIC COOK (EXT. 8-5648) IS THE CONTACT FOR THE BIOSAFETY PROGRAM. SHARPS WASTE – CLEANED AND RINSED GLASS BOTTLES, GLASSWARE, BROKEN GLASS, WIRES, RAZOR BLADES, TOOTH PICKS AND OTHER SHARPS FROM NON-BL RATED LABORATORIES SHOULD BE COLLECTED IN A IN A VWR GLASS BOX OR OTHER STURDY PUNCTURE RESISTANT CARDBOARD OR PLASTIC CONTAINER. ANY CHEMICAL LABELS MUST BE DEFACED. MARK THE BOX “CLEAN BROKEN GLASS” WITH THE LAB # AND PERSON RESPONSIBLE. WHEN FULL, TAPE SHUT AND SECURE. PLACE CONTAINERS IN THE HALL. CUSTODIANS WILL PICK UP. IF THERE ARE ANY PROBLEMS OR QUESTIONS, CONTACT KEVIN HEALY, RECYCLING COORDINATOR, AT X3-6360 OR [email protected]. NO CHEMICAL, RADIOACTIVE, BIOLOGICAL OR HAZARDOUS WASTE RESIDUE.
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V. P. Working After Hours Although researchers are allowed to work alone after hours, they should avoid conducting work with hazardous substances when they are alone in the laboratory. When working alone, arrange with Campus Police or workers in other laboratories to check on you periodically. Hazardous experiments should not be conducted alone under any circumstances. It is the responsibility of the researcher to consult with their supervisor or CHO to assess potentially hazardous operations. Laboratory operations or experiments involving hazardous substances are sometimes carried out continuously or overnight. It is the responsibility of the researcher to design these experiments with provisions to prevent the release of hazardous substances in the event of interruptions in utility services such as electricity, cooling water, and inert gas. Laboratory lights should be left on and appropriate signs should be posted identifying the nature of the experiment and the hazardous substances in use. In some cases arrangements should be made for periodic inspection of the operation by other workers or yourself. Information should be left indicating how to contact you in the event of an emergency. If you must take samples or check on the equipment at a time when you will be alone, arrange for Campus Police or workers in other laboratories to call you if you have not called them by a certain time. A sign is posted next to continuous experiments with contact information, in case of emergency (see appendix).
VI. Hazardous Chemical Evaluation
VI. A. Classification of Substances
VI. A. 1. MSDS’s and Other Information Sources Chemical hazards are identified through use of Material Safety Data Sheets (MSDS’s), from the manufacturer of the chemical or from one of the following resources.
Chemical Hazards of the Workplace, Fourth Edition, N.H. Proctor, J.P. Hughes, and M.L. Fischman, 1996. 704 pages of fairly detailed discussions of the toxicology of many hazardous substances.
Dangerous Properties of Industrial Materials, Seventh Edition, N. Irving Sax, Van Nostrand Reinhold Company, New York, NY, 1989.
The Merck Index, Eleventh Edition, M. Windholz, Ed., published by Merck & Co. Inc., 1989.
The Sigma-Aldrich Library of Chemical Safety Data, Second Edition, R.E. Lenga, Ed., 2 volumes, 1988. This 4,098 page work provides information on the properties of over 14,000 compounds. Toxicity data, health hazards, and suggests methods of first aid, handling, storage, and waste disposal are also included.
Patty’s Industrial Hygiene and Toxicology, 4th Edition, G.D. and F.E. Clayton, Ed., 1991.
The Industrial Hygiene Program (IHP), room N52-496, extension 3-2596, for information on hazardous chemicals.
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MATERIAL SAFETY DATA SHEETS MATERIAL SAFETY DATA SHEETS, (MSDS) ARE BULLETINS PREPARED BY MANUFACTURERS TO SUMMARIZE THE HEALTH AND SAFETY INFORMATION ABOUT THEIR CHEMICAL PRODUCTS. AN MSDS SHOULD BE REVIEWED BEFOR BEGINNING WORK WITH A CHEMICAL IN ORDER TO DETERMINE PROPER USE AND SAFETY PRECAUTIONS. ONCE A CHEMICAL IS PRESENT IN THE LAB, THE MSDS SHOULD BE EITHER BOOK MARKED ELECTRONICALLY OR A HARD COPY KEPT ON HAND FOR REFERENCE IN CASE OF EMERGENCIES. INFORMATION THAT IS REQUIRED BY OSHA TO BE ON AN MSDS INCLUDES:
PRODUCT IDENTITY HAZARDOUS INGREDIENTS PHYSICAL/CHEMICAL PROPERTIES SPECIAL PRECAUTIONS PROTECTIVE EQUIPMENT HEALTH HAZARDS REACTIVITY HAZARDS FIRE AND EXPLOSION HAZARDS SPILL CLEANUP
MSDS ARE AVAILABLE THROUGH A NUMBER OF SOURCES: 1) THE INTERNET IS A GOOD SOURCE OF MSDS’S. SEE THE EH&S TEAM HOMEPAGE FOR
LINKS TO THE MOST HELPFUL SITES (http://web.mit.edu/environment/programs/msds/html)
2) LAB CHEMICAL SAFETY SUMMARIES HAVE BEEN WRITTEN FOR A LIMITED NUMBER OF RELATIVELY COMMON HAZARDOUS LABORATORY CHEMICALS. THESE ARE PARTICULARLY USEFUL BECAUSE THEY ADDRESS LABORATORY USE OF CHEMICALS. THEY ARE AVAILABLE FROM THE HOWARD HUGHES MEDICAL INSTITUTE WEB SITE (HTTP://HHMI.ORG/RESEARCH/LABSAFE/OVERVIEW.HTML).
3) A REQUEST MAY BE MADE DIRECTLY TO THE CHEMICAL MANUFACTURER OR SUPPLIER. IN FACT, THIS IS BEST FOR “PRODUCTS” OR “MIXTURES” TO DETERMINE WHAT HAZARDOUS INGREDIENTS ARE CONTAINED IN THE FORMULATION.
4) A REASONABLY COMPLETE FILE OF MSDS’S FOR COMMON CHEMICALS WHICH ARE IN USE AT MIT OR HAVE BEEN USED AT MIT IS AVAILABLE THROUGH THE EH&S TEAM ON THE FOURTH FLOOR OF BUILDING N52 (EXT. 2-EHSS). THAT PROGRAM SHOULD BE CONTACTED IF ANYONE NEEDS HELP IN INTERPRETING MSDS INFORMATION.
VI. A. 2. Substance Evaluation Classes of Chemicals Based on information available in MSDS’s and other chemical evaluation resources, all substances are classified into one of the following classes of chemicals:
Carcinogen Reproductive Toxin Radioactive Biohazard Acutely Toxic
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Chronically Toxic Acute Contact Hazard: eye, inhalation, oral, skin Highly Flammable or Explosive Highly Reactive Moderate to Low Contact Hazard: eye, inhalation, oral, skin Minimally or Not Hazardous
Substances are classified by their most deleterious effect in a "normal and appropriate use" situation; i.e. a substance that is acutely toxic if swallowed and also an acute eye contact hazard is listed under 'Acute Contact Hazard: eye' since it is not likely to be swallowed under normal and appropriate use, but it may contact the eye in the event of a spill or splash. This classification is recorded on the Chemical Evaluation Form for each chemical.
Although there are hundreds of chemicals in the BPEC laboratories, most of them are used in small quantities. Most chemicals are ordered in quantities less than one gallon or one kilogram, not more than three times per year and are shared by up to 30 researchers. For this reason, hazard classification and control methods are based only on hazard type and not on chemical quantity.
VI. B. Inventory Control for Radioactive Substances Separate ordering and inventory control procedures exist for radioactive substances. These inventory/ordering control measures are established by the MIT Procurement Office and MIT Radiation Protection Program. The Radiation Protection Program also oversees the disposal of radioactive wastes. Inventory/ordering control and waste disposal procedures for radioactive substances used in the BPEC laboratories are given in Section VII. D. and the SOP for radioactive substances.
VI. C. Particularly Hazardous Substances The OSHA Laboratory Standard (29 CFR 1910.1450) defines a hazardous substance as “a chemical for which there is statistically significant evidence based on at least one study conducted in accordance with established scientific principles that acute or chronic health effects may occur in exposed employees. The term ‘health hazard’ includes chemicals which are carcinogens, toxic or highly toxic agents, reproductive toxins, irritants, corrosives, sensitizers, hepatotoxins, nephrotoxins, neurotoxins, agents which act on the hematopoietic systems, and agents which damage the lungs, skin, eyes, or mucous membranes.” Highly flammable and explosive substances comprise another class of hazardous compounds. In addition, certain carcinogens termed ‘select carcinogens’, all reproductive toxins and all compounds with a high degree of acute toxicity pose significant threats to human health and are classified as particularly hazardous substances. These major classes of hazardous and particularly hazardous substances are discussed in further detail in Section IX.
VI. D. Standard Operation Procedure for Particularly Hazardous Substances If a Standard Operation Procedure does not exist for a particularly hazardous chemical, it is the responsibility of the researcher who is using the chemical to write up an SOP, following the BPEC format established for SOP’s (Section XII. I.), and submit the SOP to the Laboratory Supervisor or CHO. In addition to the information required by the standard format for SOP’s, the researcher must also list the names of other researchers who will be
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working with the restricted substances and the approximate amounts that will be employed in the proposed research.
VI. E. Elimination or Substitution The first step in evaluating a new experiment, process or operation is to investigate the possibility of eliminating the use of hazardous materials or substituting a less hazardous material. For example, instead of using an organic solvent or chromic acid based material for washing glassware, one should substitute an aqueous based detergent. Aromatic compounds (e.g., benzene) and chlorinated hydrocarbons (e.g., methylene chloride) in some experiments should be replaced with aliphatic compounds or non-chlorinated hydrocarbons, if possible.
The particular process, experiment or operation may also be modified to reduce the quantity of the hazardous material(s) necessary or limit the potential emission release rate or exposure time. For example, the use of microscale techniques may be applicable in measuring boiling points of a material. Another example is the substitution of closed systems for open vessels. The use of a secondary containment device such as a pan can be helpful in preventing or minimizing the effects of chemical spills. The Institute has a inventory-control program (Section IV. B. 1.) that is administered by the Safety Program to reduce the purchase of large quantities of chemicals.
In many of the procedures used in the BPEC laboratories hazardous chemical reduction, substitution, or elimination has been implemented. Chromic acid is no longer used to wash glassware. Most all experiments are carried out on the smallest possible scale to reduce waste. All biological experiments are carried out in closed vessels to ensure asepsis. Radioactive experiments are performed with the lowest energetic level isotopes available.
VII. Facilities for Particularly Hazardous Substance Use and Disposal
VII. A. Biohazardous Materials/ Recombinant DNA Research Biological hazards are agents of biological origin that present a possible threat to the environment or the health of laboratory personnel. Such materials include, but are not limited to, viruses, spent media used for their propagation, animal or human tissues, genetic material of pathogenic organisms and any nucleic acid (when in cells) that is categorized as recombinant DNA. Principle Investigators/Center Directors are responsible for registering all laboratories in which biohazardous materials are used with the EH & S prior to beginning work in the labs. If live animals are to be used, the laboratory must also be registered with the MIT Division of Comparative Medicine (DCM). The specific registration procedures typically require a written application. Since the procedures vary greatly with the nature of the work being conducted, it is best to contact EH & S or DCM directly to obtain the appropriate registration forms. For some proposed work, detailed review by an Institute Committee (Committee on Assessment of Biohazards, Committee on Animal Care, etc.) must precede formal authorization. Registration procedures for projects involving recombinant DNA research and other work formally defined as biohazardous are given in the Registration and Approval Process for rDNA Protocols
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(http://web.mit.edu/environment/programs/bio-reg.html). Procedures for work with animals can be obtained from the Division of Comparative Medicine (http://web.mit.edu/comp-med/). Lab 16-436 is a Bio-Level 2 facility (BL2). BL2 containment encompasses work with recombinant DNA as well as potentially pathogenic organisms. This includes plant or animal pathogens. Three examples are polio, hepatitis and human immunodeficiency (AIDS) viruses. Refer to Section V. N. for a description of Bio-Level containment levels, BL1 through BL4. Current Recombinant DNA Project Registration Numbers: Daniel Wang - 107 and 413 Jean-Francois Hamel - 436 Douglas Lauffenburger - 378 and 405 New Project Applications In the event that one or more BPEC laboratories needs to be recertified for a different type of, or higher level of biohazardous containment, information on the MIT programs that register biohazardous laboratories and the types of biohazardous work they certify is given below. 1) Projects that must be registered with the Committee on Assessment of Biohazards are
given below. The contact program within MIT is the Biosafety Program (BSP). a) All recombinant DNA projects, including studies considered exempt under the NIH
guidelines. b) All studies with potentially pathogenic agents, such as hepatitis virus, human T-cell
leukemia virus (HTLV), and material from AIDS patients. 2) Projects that must be registered with the Committee on Animal Care. All projects involving live animals require the authorization of the Committee on Animal
Care. An example of a project needing this authorization would be one in which pathogenic viruses are to be isolated from the tissues of experimental animals. This project would require the authorization of the Committee on Animal Care (contact the DCM), as well as the approval of the Committee on Assessment of Biohazards (contact the BSP).
3) Projects that must be registered with the Committee on the Use of Humans as
Experimental Subjects. An example of such a study would be one in which enzymes are isolated from a human
tissue, such as placenta. Owing to the significant incidence of transmissible diseases in the population, all human tissue should be considered as having infectious potential.
4) Projects that must be registered with the Radiation Protection Program (RPP). All projects involving radioactive substances require the authorization of the RPP,
regardless of whether or not a biohazard is generated. There are many experiments in
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which a radioactive biohazardous agent is generated (e.g. a radioactive AIDS virus or a pathogenic bacterial cell). The use and disposal of these agents requires special procedures, and investigators are required to contact the RPP for guidelines prior to the start of the experiment (Section VII. D.).
VII. A. 1. Control Measures Containment is the minimization of risk to personnel and the environment from biohazardous material. There are two levels of containment. Primary containment includes safety techniques and devices used to separate personnel physically from the biohazard. Specific examples include the use of protective laboratory clothing, the use of aseptic technique in a manner that avoids environmental spread of the rDNA or pathogenic material in use, and the use of biological safety cabinets to minimize the spread of aerosols. Secondary containment pertains to laboratory design and operation features used to protect the environment by physically containing the biohazardous agent within the laboratory. Specific examples of secondary containment considerations are the use of non-porous surfaces for bench tops and floors in the laboratory, the type of door used to separate the laboratory from public areas, the type of ventilation system in use (positive or negative pressure and filtering), and the location of autoclaves proximal to the site of biohazard research. Further information on the levels of containment can be obtained from the Biosafety Program (http://web.mit.edu/environment/). The required level of primary or secondary containment is in direct relation to the perceived risk of the biohazardous agent being used, defined by the containment level. Refer to Section V. N. for a description of Bio-Level containment levels. The following practices and procedures are used in the BPEC laboratories to satisfy containment requirements of BL2 control. a) Laboratory personnel are required to wear a lab coat and vinyl gloves when
working with biohazardous materials in the BL-2 areas. Gloves are removed before handling common laboratory equipment such as telephones, notebooks, and storage equipment. Gloves are removed before handling doorknobs or exiting the lab. Hands must be washed after working with biohazardous materials and before leaving the lab. Lab coats are removed before entering computer rooms or offices.
b) Biological safety cabinets are used to prevent contamination of cultures and to prevent
exposure of personnel to aerosolized biohazardous materials. c) Aseptic technique is practiced by all laboratory personnel to reduce the chance of culture
contamination, and the spread of biohazardous materials to the surrounding environment. e) Non-porous surfaces are used for bench tops and floors in the laboratory to prevent the
spread of biohazardous materials and to facilitate cleaning and decontamination.
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f) Laboratory doors remain closed at all times to maintain positive pressure in the laboratory.
g) Contaminated cultures are immediately, chemically disinfected and discarded. Any
contacted surfaces or equipment are disinfected as well. h) Glassware is rinsed with a decontaminating solution before submitting for cleaning and
sterilization. i) Waste disposal receptacles designed to contain and separate solid, solid sharps, and liquid
biohazardous material and to prevent subsequent spills are used. j) Bleach, ethanol, iodophor disinfectants and autoclaving are used for decontamination of
biohazardous material. k) Floors are vacuumed and washed biweekly with a disinfectant solution. Vacuum bags
are put into biohazardous waste receptacles. Floors are sealed with a wax coat approximately once every 18 weeks.
l) Biohazardous samples are not to be taken out of the lab except in approved transportation
containers. Details of each of these procedures is contained in the following five sections on the use of biohazardous materials. Biohazard Signs In addition to BL2 signs posted on the door of every laboratory using biohazardous materials, biohazard signs are placed on all equipment used to store, grow or manipulate biohazardous material. This includes, but is not limited to, refrigerators, freezers, cell banks, incubators, centrifuges, microscopes, microwaves, biological safety cabinets, and reactors. This is done to inform users of contact hazards and to ensure that researchers disinfect any biohazard-labeled equipment before submitting for repair. Individual samples and cultures are not marked with biohazard signs; instead they are marked with information defining their contents, e.g. cell or virus type, cell suspension or supernatant, etc.
VII. A. 2. Purchase and Transportation Rules for receiving biological specimens
a) Researchers receiving cultures or cells from other laboratories are requested to inquire that they are free of mycoplasma and other infectious agents.
b) Cell lines found to contain mycoplasma or other contamination are to be immediately
chemically disinfected and disposed of. All chemicals in contact with the culture are to be similarly disinfected and discarded.
Rules for shipment of biohazardous material a) Growing cell cultures - shipped at ambient temperature. Fill a polypropylene
centrifuge tube or cryo-ampoule vial (suspension cultures) or T-25 flask (monolayer culture) with growth medium, cap tightly and secure the cap with tape. Insert the tube or
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flask into a mailing tube (Fischer, Scientific #03-520E) filled with enough absorbent cotton to contain the contents should the centrifuge tube, vial or flask break or leak, and secure the cap tightly. If the material is infectious to humans, the mailing tube must be labeled with an Infectious Biohazardous Material label. Mail via overnight air.
b) Iced cultures using blue ice. The culture to be shipped is placed in a polypropylene
centrifuge tube or cryovial which is capped tightly and secured with tape. The tube is then placed in a leak-proof polyethylene envelope (Britton Liquid Bag #101) containing enough cotton to absorb the liquid, should the initial container break. Total liquid volume must be less than 50 mL. The envelope is then placed in a shipping container (cardboard with styrofoam liner) containing a frozen blue ice pack. If the material is infectious to humans, the shipping container must be labeled with an Infectious Biohazardous Material label. Mail via overnight air.
c) Iced cultures using dry ice. The frozen specimen is placed in a polypropylene centrifuge
tube or cryovial, which is capped tightly and secured with tape. The tube is then placed in a leak-proof polyethylene envelope (Britton Liquid Bag #101) containing enough cotton to absorb the liquid, should the initial container break. Total liquid volume must be less than 50 mL. The envelope is then placed in a shipping container (cardboard with styrofoam liner) containing about 10 pounds of dry ice. For shipments containing dry ice, a Shipper’s Certification for Restricted Articles form must be filed in compliance with DOT Hazardous Materials Regulations (49 CFR, CAB 82). If the material is infectious to humans, the shipping container must be labeled with an Infectious Biohazardous Material label. Mail via overnight air.
c) Etiologic agents under ice. If the material to be shipped is an etiological agent (disease-
causing, or suspect) the container should be placed in a leak-proof polyethylene envelope (Britton Liquid Bag #101) containing enough cotton to absorb the liquid, should the initial container break Total liquid volume must be less than 50 mL. The envelope is then placed in a shipping container (cardboard with styrofoam liner) containing about 10 pounds of dry ice or a frozen blue ice pack. For shipments containing dry ice, a Shipper’s Certification for Restricted Articles form must be filed in compliance with DOT Hazardous Materials Regulations (49 CFR, CAB 82). The package must be labeled with a special “Etiological Agents” label (see Methods in Enzymology, v. 58 p. 35). These procedures are necessary to comply with U.S. Public Health Service Interstate Quarantine Regulations (42 CFR, part 72). Mail via overnight air.
VII. A. 3. Storage All frozen cell stocks are stored in the liquid nitrogen cell bank or in the walk-in deep freeze in 16-479. All cells stored in these facilities are BL2 control or below. A lab coat, goggles and thermal gloves are required for filling the cell bank with liquid nitrogen and for retrieving samples. All other biohazardous materials, including culture samples are stored in refrigerators (+4°C), freezers (-20°C) or deep freezers (-70 to -125°C) located in each of the BPEC labs. Growing cultures are kept in temperature, humidity and carbon dioxide controlled incubators. These
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incubators are designed to reduce the possibility of contamination and are regularly decontaminated using built-in decontamination cycles. All biohazardous material is either too unstable or unable to survive at room temperatures. Contact the Biosafety Program for more information of storage regulations (http://web.mit.edu/environment/)
VII. A. 4. Biological Safety Cabinets (or biosafety cabinets) Biological safety cabinets, commonly referred to as biosafety cabinets, are laminar flow hoods equipped with germicidal (UV) lights and HEPA (0.2 µm) filters which, in combination provide an aseptic environments for biological manipulation. A properly operating and correctly used biosafety cabinets provides an aseptic work environment as well as protects the researcher from contact with biosafety cabinets vapors, dusts, aerosols and liquids. As far as possible, all cell, virus and other biohazardous material manipulations are conducted inside a laminar flow biosafety cabinets. Proper Use The following general rules should be followed when using a biosafety cabinet:
1. The air circulating fan should remain on at all times to maintain asepsis in the hood. The biosafety cabinet is designed to pull air in through the front face of the biosafety cabinet (where the user sits) and down into a filter that spans the length of the front of the biosafety cabinet, on the level of the working surface. This air is directed up through a set of 0.2 µm HEPA filters about three feet above and directly over the working area, which remove particles as small as viruses. This aseptic air is then directed down onto the working area where it is pulled both toward the front and back filters that span the length of the front and back of the biosafety cabinet on the level of the working surface. This downward flow of aseptic air provides a curtain through which air from outside the biosafety cabinet cannot penetrate, thus achieving an aseptic seal. A portion of the air is filtered and directed out the exhaust or duct work, which leads to the roof, to provide the driving force for pulling air in through the front face of the biosafety cabinet.
2. The user can reach through the curtain and place items in the aseptic environment of the
biosafety cabinet. Once items are inside, contaminating substances such as bacteria, yeast, fungus and viral particles that may have been transported into the hood are quickly swept away by the air curtain. The only mechanism of transport remaining is by touching a contaminated item to another item. This is prevented by aseptic technique.
3. The biosafety cabinet sash should remain at the prescribed level to ensure a proper air
flow pattern. This level is demarcated on the hood with a yellow sticker, indicating the air-flow rate at the indicated sash height. If the sash is raised or lowered from this height, a buzzing alarm will sound and will not turn off until the sash is returned to the proper height. The sash may be lifted to transport large items into the hood. Users should wait for about 15 seconds after returning the sash to its designated height, to allow the air flow pattern to be reestablished.
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4. The germicidal (UV) light should remain on when the biosafety cabinet is not in use, but should be turned off before working in the biosafety cabinet. The UV light from the germicidal light nicks DNA of living organisms, resulting in death of the organism. The light is one of the key features in maintaining asepsis in the biosafety cabinet. The germicidal light is damaging to both humans and microorganisms. Safe exposure time for the unprotected skin or eyes according to ACGIH TLV's is 23 seconds each 8 hour period inside the hood, and 3.16 hours at the hood sash (hand level) due to the reflected UV light. For these reasons, users should not sit at or use a biosafety cabinet with the germicidal light on. There is no UV hazard in the room at locations normally occupied while the UV lights are on. Refer to the biosafety cabinet SOP for recent UV measurements on these biosafety cabinets and additional operating guidelines.
5. For the best guarantee of asepsis, work should be performed as far as possible into the
biosafety cabinet, preferably 6 or more inches beyond the glass sash. Touching items together during transfers (such as pipettes to containers) should be avoided as this increases the probability of contamination.
6. Use of bunsen burners in the biological safety cabinets is not recommended. If use is
desired, never use in conjunction with flammable materials and never leave unattended. The airflow pattern in the biosafety cabinets can easily spread the flame and even ignite the internal filters of the cabinet. If a fire breaks out, shut off the air flow and close the glass sash. Shut down any other biosafety cabinets or fume hoods which operate off the same air supply duct and close their sashes. Air can still enter through the air supply duct, so the fire may not be extinguished until all internal filters have burned.
7. All biosafety cabinets in the BPEC laboratories are connected to house vacuum for
filtration or suction operations. At least one vacuum flask with approximately one inch of bleach is placed in line between the vacuum outlet and the point of application to collect and disinfect liquids drawn in by the vacuum.
8. After use, a biosafety cabinet (BSC) is to be cleared of equipment and its interior working
surface should be decontaminated with water followed by ethanol. A pre-rinse of the interior stainless steel working surface with water prevents staining caused by an ethanol-biological media combination. In addition, it is highly recommended that the main users of the BSCs clean them thoroughly with a solution of iodophor disinfectant, either Wescodyne or Roccal, on a regular basis, perhaps once every other week. After you are done using the BSC, the UV light should be turned ON. Please note that moving the sash completely down before turning the UV light ON is neither required nor recommended. Just leave the sash at the prescribed level for proper operation.
Surveillance All biosafety cabinets (BSCs) are decontaminated and operationally certified by B & V Testing (Waltham, MA) annually, presently in March. Decontamination certification involves an aerosol challenge of the HEPA filters. Operational inspection consists of measuring the face velocity of the BSC and using a smoke stick to visually check its containment effectiveness. If the BSC passes the aerosol challenge, face velocity and smoke containment tests, the hood performance certification label is updated. If the biosafety cabinet does not pass the survey, the
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words “DO NOT USE” or “VOID “ are written on the biosafety cabinet certification label and the biosafety cabinet is shut down until it can be recertified. Recertification involves evacuation of the room in which the biosafety cabinet is located for half a day, flooding the room with a gaseous disinfectant in order to kill any biohazardous material trapped in the biosafety cabinet filters and duct work (so as not to expose the certification team to biohazardous material), dismantling the hood and replacing filters and any air flow motors or devices that are worn, and reinspecting the face velocity and flow pattern. BSCs are also surveyed on an annual basis by the MIT Industrial Hygiene Program (IHP) if they are vented to the outside. All BPEC BSCs are vented to the outside. This survey also consists of a face velocity measurement and a smoke test, followed by recertification of the BSC with an IHP biosafety cabinet label if the cabinet passes inspection. This inspection, performed to satisfy the NIH Biocabinet Certification, does not require an aerosol challenge of the HEPA filters. The Industrial Hygiene Program recommends that biocabinets be certified on an annual basis and whenever moved. It is the responsibility of the Chemical Hygiene Officer to see that these inspection requirements are satisfied. \ Location of Biological Safety Cabinets (Biosafety cabinets) in BPEC Biosafety cabinets (BSCs) are located in all BPEC laboratories where biohazardous materials are used (16-436, 16-476). Six are located in lab 16-436, including two in each of cell culture rooms A and B, and one in lab 16-476. BSCs # 1 to 5 are for use with BL-2 level projects, BSC#3 is used for human-cell lines and BSC#5 is dedicated to work with adenovirus.
VII. B. Spills and Disposal
VII. B. 1. Accidental Release of Biohazardous Substances (Spills) Biohazardous Spill in or outside a Biological Safety Cabinet DO NOT turn off the Biological Safety Cabinet. Decontamination procedures should be initiated at once while the cabinet continues to operate to prevent escape of contaminants from the cabinet. a) Wear a lab coat, safety eyewear and disposable vinyl, latex or rubber gloves for this
procedure. b) Spray or wipe walls, work surfaces and equipment within the cabinet with the diluted
bleach stored in the Biological Spill Kit, located at the end of benches 9/10. c) Flood the affected surface area, and if a Class II cabinet, the drain pans and catch basins
below the working surface with the decontaminant and allow it to stand for ten to fifteen minutes.
d) Remove excess decontaminant from the tray by wiping with a sponge or cloth soaked in
the decontaminant. For Class II cabinets, drain the tray into the cabinet base, lift out the
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tray and removable exhaust grille work, and wipe off the top and underside surfaces with a sponge or cloth soaked in the decontaminant. Replace the cabinet pieces and drain the decontaminant from the cabinet base into a bleach bucket or other appropriate container. Gloves, wipe cloths and sponges should be discarded into non-sharp biohazardous trash receptacles.
Spills Requiring Emergency Action and Accidental Exposure of Personnel The Biosafety Program (BSP) should be immediately notified in the event of an accidental release of biohazardous material to laboratory drains or ventilation systems. People are exposed to biohazardous agents by four routes. These include ingestion (for example, by mouth pipetting - which is a violation of MIT safety procedures!), inhalation of aerosols, absorption of biohazardous material through membranes or through cuts in membranes, and accidental inoculation by needles or other sharp objects. Eye exposure to a biohazard should be treated immediately by flushing with water from an eyewash. Skin exposed to a biohazard material should be treated by showering it with water or, in selected cases, by alcohol wipes. Persons who have inhaled or inoculated a biohazard should be taken to the Medical Department for treatment. Workers exposed to biohazardous agents should immediately report the incident to the Principal Investigator responsible for the research project. The Principal Investigator should use his or her judgment in determining the need to report the accident to the BSP. Examples of accidents that need to be reported and those that do not are given in the Guidelines for Biological Research at MIT.
VII. B. 2. Waste Disposal and Decontamination The disposal of biological wastes is handled in accordance with procedures for deactivation that have been established by the department involved and the EH & S. Waste Disposal Containers for Use Prior to Decontamination Institute regulations dictate that solid biohazardous waste containers consist of a terminal (non-reusable) biohazard bag in a labeled, covered, leak-proof container located in an appropriately marked and confined area. Furthermore, BL1 liquid waste should be stored in a covered container in the laboratory prior to decontamination. In contrast, BL2 and BL3 waste must be chemically decontaminated immediately after its generation, and subsequently autoclaved. A diagram that is posted throughout the BPEC laboratories depicts the waste disposal containers employed in all BPEC laboratories.
a) Regular Trash (black trash bags). All non-sharp, non-biohazardous waste is discarded into regular trash bags. Included are pipette wrappers and other packaging for disposable plastic wear, centrifuge tube racks, non-contaminated paper towels and Kimwipe paper, and notebook paper.
b) Non-sharp Biohazardous Trash (clear biohazardous bags). All non-sharp
biohazardous waste, including plastic petri dishes, T-Flasks, centrifuge tubes, roller 68
bottles, filter flasks and any kimwipes, paper towels and other non-sharp items that have contacted biohazardous material are discarded into circular biohazardous trash bins. Double biohazard autoclavable bags are used in the bag holders to ensure containment in the event of a puncture of the inner bag. All containers must be emptied of spent medium and other biohazardous liquids before discarding the container into the clear biohazardous trash bags. The biohazardous liquids must be dumped into bleach buckets. Significant quantities of fluid in the biohazard bags could promote microbial growth and complicate autoclaving and disposal.
Do not dispose of “soft” biohazardous trash in the gray/beige sharps containers.
All non-sharp, disposable plastic tissue culture items must be discarded into non-sharp
biohazardous bags regardless of whether they have contacted biohazardous materials. The reason for this is that the BPEC laboratories have a general rule that all objects that are designed for contacting biohazardous materials should be disposed of as biohazardous waste, even if they did not contact biohazardous material in their use. Under this procedure, there is never a question as to whether an object is discarded in the correct receptacle.
c) Sharps Disposal Sharps disposal procedure is as described in a November 10, 1999
memo from Eric Cook of the MIT Biosafety Program. “All sharps (except radioactive or chemically contaminated sharps) must be disposed of in the gray or beige containers supplied by the Biosystems Company. Sharps placed in these containers are no longer required to be autoclaved prior to disposal. If you have any questions, please contact Eric Cook at 8-5648 or [email protected].”
All sharp waste, except non-biologically contaminated broken or disposable glassware, including razor blades, needles, and other sharp metal and glass items that can easily puncture the epidermis should be discarded in a biohazardous sharps disposal container, regardless of whether they have contacted biohazardous materials. The reason for this is that the BPEC laboratories have a general rule that all objects that are designed for contacting biohazardous materials should be disposed of as biohazardous waste, even if they did not contact biohazardous material in their use. Under this procedure, there is never a question as to whether an object is discarded in the correct receptacle. All needle/syringe units are to be disposed of intact. This means that needles are not recapped, bent, or broken. The use of needle chopping devices (guillotines) is not recommended due to the release of aerosols, which can contaminate both personnel and surfaces. These practices and devices also require increased needle handling which increases the risk of needle-stick injuries.
d) Biohazardous Liquid Waste (bleach buckets). All spent culture medium and other
liquids that have contacted biohazardous materials are to be dumped into bleach buckets. Bleach buckets in the BPEC labs are filled with at least one inch of bleach to prevent microbial growth in these containers and are fitted with a step-opening lid to prevent the awful smell due to evaporation of bleach and to confine aerosolized biohazardous
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material. Users are required to add an amount of bleach equal to the volume of liquid discarded into the bleach bucket to aid in decontamination.
Chemically contaminated glass, toxic chemical wastes, or metallic wastes (sharps) should never be placed in the biohazardous trash bags or the Sharps Collectors described above. If they have contacted biohazardous materials, they should be separately sterilized with a disinfecting agent, not by autoclaving since the heat may cause an adverse chemical reaction. The respective special disposal procedures should then be followed for these items. (Toxic materials are never mixed with biohazardous material under normal operation; it would kill cells.) Decontamination Procedures - MIT Policy There are many different methods of sterilizing biohazardous wastes, including wet heat sterilization (autoclaving), gaseous chemical sterilization (ethylene oxide), and liquid chemical sterilization (bleach or iodophor decontaminant such as Wescodyne or Roccal II). Once biohazardous liquid waste such as spent culture medium has been rendered non-infectious by autoclaving or chemical treatment, it can be disposed of by dumping down the drain. Please note that contaminated solid wastes and animal carcasses from laboratories that conduct research with animals should be collected in impermeable containers which are closed before removal from the work area for disposal by incineration. Jane Roberts does such collections in our lab, and she brings the material in closed plastic bags to the 8th floor of the Division of Comparative Medicine. There recently has been a new state regulation [105 CMR 480.000] requiring MIT and other institutions to document the storage and disposal of medical and biological waste. a) It is now necessary for all autoclave facilities to record the following information for each
autoclave run: the time and temperature setting of the autoclave, the volume and type of waste generated, and the name of the department and person responsible for the waste. As part of the logging process, an Autoclaved Noninfectious Biomedical Waste tag is affixed to the autoclaved biohazardous waste. Included on this tag is an ID number, the date, the Principal Investigator (BPEC) and the initials of the person who performed the sterilization.
b) Disposal of all hazardous solid waste must also be documented. There are three types of
solid waste: autoclaved solids (part a), chemically treated solids (treated with bleach or an iodophor decontaminant), and physically hazardous solids (sharps). Once these materials have been rendered noninfectious, all biohazard signs and symbols must be removed from sight.
Tags are available from the BSP. Decontamination Procedures - BPEC Regulation In the BPEC laboratories, all materials that contact cell cultures, viruses and other biohazardous materials are decontaminated by either chemical means (e.g. dilute bleach), or by autoclaving. The researchers generating soft biohazard wastes are responsible for making sure it is
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decontaminating and properly disposed of. However they are not responsible for decontaminating the gray sharp containers that are being handled by the Biosystems Company. .
a) Regular Trash (black trash bags). Regular trash is collected by MIT’s janitorial staff every other night. Since it contains no sharp or biohazardous materials, it does not require special processing.
b) Non-sharp Biohazardous Trash (biohazardous bags). It is the responsibility of the
researcher to replace these bags when they are more than 3/4 full. Unless the inner bag is punctured, only the inner bag should be removed from the bag holder. This bag should be replaced with a fresh bag. The full biohazard bag should then be transported to the autoclave room. A cup of water is also added to the bag to produce steam in the bag, ensuring proper sterilization of the entire contents of the bag. The bag should be left open (to allow steam to enter the bag) and autoclaved for at least 90 minutes at 126°C (250°F) .
c) An Autoclaved Noninfectious Biomedical Waste tag is attached to one of the autoclave
bags in each load. The blue line at the top of the tag will turn black if the autoclave reaches and maintains the proper sterilization temperature, indicating successful decontamination. The bags are placed in autoclave trays to contain any liquids that may escape during autoclaving. A “solids” autoclave cycle is used. The length of the autoclave cycle depends on the number of bags to be autoclaved. Generally, an autoclave cycle of 90 minutes at 126°C (250°F), followed by a drying cycle of 15 minutes is sufficient for three bags. Four or five bags need to be autoclaved for 105-120 minutes at the same temperature and dried for 15 minutes. The drying cycle basically ensures that the autoclave is fully depressurized at the end of the cycle.
d) Once the bags have been autoclaved, they must be placed in a black, regular trash bag to
comply with the MIT regulation that once biohazardous materials have been rendered noninfectious, all biohazard signs or symbols must be removed from sight. The bag is sealed shut with the Autoclaved Noninfectious Biomedical Waste tag. The disposal is logged in a Biohazardous Waste Disposal Notebook, located next to the large autoclave. The log includes the type of waste, (x-number of bags of solids), the waste generator, (BPEC), the tag number, the date, and the sterilization temperature and length. The black trash bag is then placed in the hall for pickup by the janitorial staff.
e) Sharps, Non-Biohazardous Waste AND Biohazardous. ALL sharps are to be disposed
of in the designated gray bins in the lab. These bins/containers can be lined with an orange/red biohazard bag. “All sharps” means broken glass, syringes (with or without needles), razor blades, plastic pipettes, plastic pipette tips, and anything that could cut or puncture the skin. It does not matter if it is contaminated or not, if it is sharp it goes in the bin. When about full, those gray sharp containers must be closed with the dedicated lid and secured with a tie cable. Next they are moved in the corridor, just outside the lab, on thursdays evenings. The Biosystems Company picks up the bins every Friday morning and replaces the full containers by empty ones.
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f) Biohazardous Liquid Waste (bleach buckets). When cell culture media is mixed with bleach, a pH change occurs, turning the solution from yellow to purple, and back to yellow again if sufficient quantities of bleach are present. Although this color change does not directly correlate with the microorganism killing ability of bleach, it is a reasonable indicator. When a bleach bucket is over half full, enough bleach is added to turn the solution yellow and then another volume of bleach equal to half the volume initially added is used to guarantee a complete kill. The solution is allowed to stand for one hour to ensure a complete kill. The solution is then poured down the drain with copious amounts of cold water. Warm or hot water makes the bleach vaporize which can be hazardous to the lungs and eyes as well as being hideously smelly.
g) Cultures found to contain any bacterial, yeast or fungal contaminations are to be
immediately chemically disinfected (with bleach or Wescodyne or Roccal II) and discarded. The container carrying the infected culture should be opened carefully to avoid aerosol formation, and chemical disinfectant added. The disinfectant should contact all surfaces of the contaminated vessel. Any chemicals or solutions that have come into contact with a contaminated culture are discarded. In the event that contamination is discovered, but the owner of the culture is unavailable, the container should be isolated from other cultures and sealed. Notice should be left for the owner indicating the time and date, the nature of the action, and the name of the person who isolated the contamination. Do not discard someone else’s culture without permission from the owner. The only exception is when the owner of the cultures is no longer at MIT and no one wants to claim the cultures. However, before disposing of someone else’s culture, the Laboratory Supervisor should be notified.
VII. B. 3. Summary of Sterilization Methods and Procedures Other General Procedures for Decontamination and Sterilization a) To minimize hazard to firemen or disaster crews, all biohazardous materials should be
placed in an appropriately marked refrigerator or incubator, sterilized, or otherwise confined at the close of each workday.
b) All materials, equipment, or apparatus contaminated with or containing potentially
infectious organisms should be sterilized before being washed and stored, or discarded. Autoclaving is the preferred method. Each individual working with biohazardous materials should be responsible for its sterilization before disposal.
c) The work area and the surrounding areas should be disinfected after completion of
operations involving plating, pipetting, centrifuging, and similar procedures with biohazardous materials. It is the responsibility of the Lab Supervisor to determine that the disinfectant, time and method of exposure is effective against the biological agents used in the facility.
d) All laboratory rooms containing biohazardous materials should distinguish two separate
areas or containers labeled: biohazardous - to be autoclaved, or non-infectious - to be cleaned.
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e) Special precautions should be taken to prevent accidental removal of materials from an autoclave before it has been sterilized or the simultaneous opening of both doors on a double door autoclave.
f) Dry hypochlorites, or any other strong oxidizing material, must not be autoclaved with
organic materials such as a paper, cloth or oil: oxidizer + organic material + heat, may produce an explosion
g) All floors, laboratory benches, and other surfaces in building where biohazardous
materials are handled should be disinfected as often as deemed necessary by the Lab Supervisor. Refer to the Housekeeping section, Section V. J. and the Control Measures section, Section VII. B. 1.
h) Floor cleaning procedures that minimize the generation of environmental aerosols should
be used. Wet mopping or wet vacuum pickup is recommended. Water used to mop floors should contain a disinfectant or disinfectant-detergent. Dry mopping or dusting should be avoided. Where wet procedures are not practical, dry vacuum cleaning with a HEPA filter on the exhaust, sweeping compound used with push brooms, or dry dust mop heads treated to suppress aerosolization may be used.
i) Floor drains should be flooded with water or disinfectant at least once each week in order
to fill traps and thus prevent the back flow of sewer gases. j) Stock solutions of suitable disinfectants are maintained in each laboratory for disinfecting
purposes. Specific methods of sterilization The following is a listing of many sterilization methods that may be used in addition to, or as a substitute for those procedures already being used in the BPEC laboratories if these procedures are found to be ineffective for a new or existing biohazardous material. a) Wet heat (the only method in use at BPEC). Wet heat is the most dependable
procedure for the destruction of all forms of microbial life. Steam sterilization generally denotes heating in an autoclave employing saturated steam under a pressure of approximately 15 psi to achieve a chamber temperature of at least 121°C (250°F). The critical factors in insuring the reliability of this sterilization method are proper temperature and time, and the complete replacement of the air with steam; i.e. no entrapped air.
Some autoclaves utilize a steam activated exhaust valve that remains open during the
replacement of air by live steam until the steam triggers the valve to close. Others utilize a pre-cycle vacuum to remove air prior to steam introduction. Physical controls such as pressure gauges and thermometers are widely used but are considered secondary method of insuring sterilization. The use of appropriate biological indicators at locations throughout the autoclave is considered the best indicator of sterilization. The biological indicator most widely used for wet heat sterilization is Bacillus sterothermophilus spores.
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b) Dry Heat (not in use at BPEC). Dry heat sterilization is less efficient than wet heat
sterilization and requires longer times and/or higher temperatures. The specific times and temperatures must be determined for each type of material being sterilized. Generous safety factors are usually added to allow for the variables that can influence the efficiency of this method of sterilization. Environment as well as the moisture history of organisms prior to heat exposure appear to affect the efficiency of dry heat sterilization.
Sterilization by dry heat can usually be accomplished at 160-170°C (320-338°C) for
periods of 2-4 hours. Higher temperatures and shorter times may be used for heat resistant materials. The heat transfer properties and the spatial relation or arrangement of articles in the load are critical in insuring effective sterilization.
c) Ethylene oxide sterilization (not in use at BPEC). Ethylene oxide (EtO) gas is lethal
for microorganisms including spores, viruses, molds, pathogenic fungi, and highly resistant thermophilic bacteria. Some of the principal variable which determine the rate that microorganisms are destroyed by ethylene oxide are as follows:
i) Temperature affects the penetration of EtO through microbial cell walls and
wrapping and/or packaging materials. The activity of ethylene oxide increases approximately 2.7 times for each 10°C (18°F) rise in temperature (between ranges of 5°C and 37°C, or 41°C and 98.6°C using a concentration of 884 mg/liter). Normally, EtO sterilization is conducted at temperature between 49°C and 60°C (120 - 140°F).
ii) Ethylene oxide concentration. Sterilization with EtO can be achieved in shorter
period when the concentration is increased. For practical sterilization, gas concentrations of 500 to 1000 mg/liter at approximately 49°C to 60°C (120-140°F) are recommended.
iii) Humidity. It is generally accepted that moisture is an essential condition in achieving
sterilization with EtO gas. The effect of moisture on the sterilizing action of ethylene oxide appears to be related to the moisture content of the exposed bacterial cell. A relative humidity of 30 to 60 % is frequently employed in EtO chambers during such exposure conditions.
iv) Exposure time. In most cases, the appropriate exposure time for attaining sterility is
determined experimentally using accepted biological indicators. Frequently, these controls are Bacillus subtilis var. niger spores placed on suitable carrier materials.
v) Precautions in using ethylene oxide. In contrast to steam sterilization where the
only residual of post-sterilization may be water vapor or moisture, EtO sterilization does result in post-sterilization residues of EtO, ethylene glycol, and depending on the chemical nature of the article sterilized, ethylene chlorohydrin. Sufficient time must be allowed for these toxic residues to escape. Current practices indicate that 5 to 7 days of ambient aeration are required for the removal of absorbed EtO in some plastics and rubber. The removal of residual EtO can be accelerated by mechanical aeration at elevated temperatures (50°C).
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Mixtures of 3 to 10% EtO in air are explosive. Commercially available mixtures of
EtO in Freon or carbon dioxide are not explosive and can be safely utilized. Ethylene oxide sterilization is not used in the BPEC laboratories because of the
danger associated with its use and its residues. Sterilization Procedures General criteria for sterilization of typical materials are presented below. Supervisors are encouraged to review the type of materials being handled and to establish standard conditions for sterilization. Treatment conditions to achieve sterility vary in relation to the volume of material treated, its contamination level, the moisture content, and other factors. a) Steam autoclave i) Laundry: 121°C (250°F) for a minimum of 30 minutes. ii) Trash: 121°C (250°F) for at least 1.5 hours. iii) Glassware: 121°C (250°F) for a minimum of 25 minutes. iv) Liquids: 121°C (250°F) for 1 hour for each gallon. v) Animals and bedding. Steam autoclaving is not recommended. The sterilization time
required would be at least 8 hours. Incineration in an approved facility is the recommended treatment of these wastes.
b) Gas sterilants i) Ethylene oxide gas: Sixteen hours exposure to a concentration of 750 mg/L (∼5%) at
30 - 60% relative humidity and at ambient temperature (70°F).
ii) Paraformaldehyde: Sixteen hours exposure to a concentration of 1.0 mg/L at 60% or greater relative humidity and at ambient temperature (70°F).
Disinfectants a) Phenolic compounds. These are recommended for the killing of vegetative bacteria,
including mycobacterium tuberculosis, fungi and lipid containing viruses (0.5 -2.0%). They are less effective against spores and non-lipid-containing viruses. Other properties are:
i) Low solubility in water unless combined with detergent ii) Stable in storage iii) Germicidal against gram-negative and Gram-positive organisms and tubercula bacilli
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iv) Less adversely affected by organic matter than other common germicides v) Effective over relatively large pH range vi) Limited sporicidal activity vii) Prolonged contact deteriorates rubber viii) Can cause skin and eye irritation ix) Not for use on food contact surfaces x) Many active against lipophilic viruses b) Quaternary ammonium compounds. These are acceptable as general use disinfectants
to control vegetative bacteria and non-lipid containing viruses. However, they are not active against bacterial spores at the usual use concentrations (1:750). Other properties are:
i) Stable in storage ii) No odor but act as deodorizers iii) Use dilution usually non-irritating to skin but prolonged skin or eye contact should be
avoided. iv) Effective at temperatures up to 212°F v) Effective against Gram-positive organisms, bacteriostatic in high dilutions vi) Generally ineffective against tubercula bacilli, spores, and viruses. vii) More effective in alkaline than acidic solutions viii) Neutralized by soap and anionic detergents ix) Effectiveness reduced by organic material x) Have built-in detergency properties xi) Some active against lipophilic viruses c) Iodophors. Although these show poor activity against bacterial spores, they are
recommended for general use (70 to 150 ppm). They are effective against vegetative bacteria, fungi and viruses. Other properties are:
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i) Combine iodine with non-ionic detergent ii) Rapid biocidal action iii) Effective against Gram-positive and Gram-negative organisms and tubercula bacilli iv) Most effective in acid solutions v) Vaporized at 120°F to 125°F should not be used in hot water vi) Effectiveness reduced by organic matter (but not as much as with hypochlorites). vii) Stable in storage if kept cool and tightly covered. viii) Iodophors are relatively harmless to man ix) Iodophors have a built-in indicator. If the solution is brown or yellow, it is still
active. x) Iodophors can be readily inactivated and iodophor stains can be readily removed with
solutions of sodium thiosulfate xi) Tarnish silver, silverplate, and copper d) Alcohols. In concentrations of 70 to 95%, alcoholic solutions are good general use
disinfectants, but they exhibit no activity against bacterial spores. Other properties are: i) Germicidal against a broad spectrum of bacterial species and many viruses ii) Fast acting iii) Leaves no residues iv) Compatibly combines with other disinfectants (quarternaries, phenolics, and iodine)
to form tinctures, extending alcohol’s biocidal action. v) Results from experiments conducted at NIH indicate that a combination of 60%
ethanol with 0.01 N HCL (pH 4) remarkably improved biocidal action against poliovirus and adenovirus.
e) Aldehydes. They are effective against a wide spectrum of bacteria, fungi and viruses, and
are also sporicidal when used properly. i) Formaldehyde solutions. At concentrations of 8% formalin exhibits good activity
against vegetative bacteria, spores, and viruses.
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ii) Formaldehyde-alcohol. Solutions of 8% formalin in 70% alcohol are considered very good for disinfection purposes because of their effectiveness against vegetative bacteria, spores and viruses. For many applications, this is the disinfectant of choice.
iii) Activated glutaraldehyde. Two percent solutions exhibit good activity against
vegetative bacteria, spores, and viruses. Its use, however, must be limited and controlled because of its toxic properties and the damage to eyes. They have limited stability after activation (for alkaline glutaraldehyde).
f) Chlorine compounds. These are recommended for certain disinfecting procedures
provided the available chlorine needed is considered. Low concentrations of available chlorine (50 to 500 ppm) are active against vegetative bacteria, fungi and most viruses. For bacterial spores, concentrations of approximately 2500 ppm are needed. The corrosive nature of these compounds, their decay rates and lack of residuals is such that they are recommended only in special situations. Other properties are:
i) Very active ii) Provides biocidal action against a wide variety of gram-negative and gram-positive
bacteria and against many viruses. iii) Solution of 2000 ppm available chlorine commonly used in laboratory as a soak for
contaminated equipment. iv) For optimal biocidal activity, dilute with low temperature water after soaked items are
autoclaved. v) Neutralized rapidly in the presence of organic matter. vi) A 1% hypochlorite solution is recommended for destruction of the Hepatitis B
antigen. vii) Acidic solutions are less stable but have a higher biocidal activity. g) Mercurials. Not recommended for general use; they have poor activity against vegetative
bacteria and are useless as sporicides. Although the mercurials exhibit good activity against viruses (1:500 to 1:1000 concentration), they are toxic and therefore are not recommended.
VII. C. Lasers All laser systems at MIT must be registered with the MIT Radiation Protection Program, Bldg. N52-496, telephone extension 3-2180. All researchers who will be using lasers must attend the MIT Institute-wide training program on laser safety once during their time at MIT. The following guidelines must be followed when using lasers:
1) The most common hazard encountered in work with lasers at MIT has been electric shock. Other hazards include eye damage from laser light, skin burns, and fire. Take
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precautions to minimize these hazards, such as equipment design and placement, wearing UV eyeglasses, and never crossing a laser beam with any portion of the body.
2) No person is allowed to use a laser unless authorized to do so by the Laboratory
Supervisor and until they have completed the Institute-wide training program on laser safety.
3) No person is allowed to work alone with an energized laser without specific instructions
and permission from their Laboratory Supervisor. 4) Laser laboratories should be arranged so that they are safe for individuals with
unprotected eyes. Laser beams should be confined to laser tables or designated beam paths. Flight tubes are recommended by are not required. Specular reflections from optics should be blocked. Special exceptions, such as alignment or use of a 1.06 µm light from the Nd:YAG laser, require an explicit warning sign to be posted on the door. A laser beam should never be aimed at a door, whether opened or closed. Using laser beams at eye level must be avoided if at all possible.
5) Safety glasses (designed for the laser wavelengths in use) should be worn when working
with any laser having an average power exceeding 50 mW or an energy/pulse exceeding 10 mJ (and below if appropriate). This is especially important for IR and UV lasers, and when aligning beams. The most dangerous time is during alignment of lasers and other optical elements during which everyone present in the laboratory must take maximal precautions against direct or specular reflections hitting the eye. Only those types of UV protective glasses approved by the Radiation Protection Program may be used.
6) Before turning on a laser, be sure that the safety interlocks and shields are in place and
(where appropriate) that the exhaust ventilation system is on. Also remember to turn on the warning light outside the laboratory, if appropriate.
7) Researchers should be aware of the “blue-light” hazard: the threshold for eye damage is
100 times lower for blue light than for red or near-ultraviolet. For example, the diffuse reflection of a multiwatt-argon ion laser beam falling on a black beam stop can be an eye hazard.
VII. D. Radioactive Materials Work with radioactive materials and with equipment that produces radiation (shorter than UV wavelength) is regulated by MIT’s Radiation Protection Program (RPP), telephone extension 3-2180. In addition to the instructions outlined in a SOP for radioactive substances, special authorization, ordering and inventory control procedures have been established by MIT’s Procurement and the Radiation Protection Offices. Because establishment and approval of these provisions must occur well in advance of using these substances, they are listed here rather than in the Radioactive Materials SOP. The responsibilities of the RPP are given in Section II. D.
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The MIT Required Procedures for Radiation Protection gives the Commonwealth of Massachusetts Department of Public Health Radiation Control Program and the MIT Radiation Protection Program guidelines for safe transportation, use, storage and disposal of radioactive
materials. A copy of this guidebook is available at the Radiation Protection Program in Bldg. N52-496.
VII. D. 1. Authorization for Use The Center Director must approve the use of radioactive materials in a particular laboratory. The Radiation Safety Officer for the lab or an individual researcher must submit an Application for Authorization form to the RPP. This application must include a protocol for the proposed work, identification of the types and amounts of radioisotope(s) to be used, and the amounts to be disposed of. The RPP then assigns an authorization number and returns the form to the Laboratory Supervisor or Center Director with a description of the amounts and types of materials that can be handled in the labs. Any changes to the type of radioactive work in the lab must be approved via an amendment request to the RPP. Following this authorization, individual workers must be trained and authorized by the RPP to work with materials and protocols previously approved by the RPP. This training includes a two- hour, Institute-wide training seminar held by RPP. Researchers are required to pass an exam on the material presented in the training seminar. The RPP has regular training sessions so that new personnel can generally receive instruction and authorization with two weeks of a request. Because BPEC also has its own set of rules and guidelines for handling radioactive materials, especially biohazardous radioactive materials, all new personnel should also receive training by a senior level BPEC graduate student or post-doctoral fellow who is familiar with BPEC rules. BPEC-Specific guidelines for handling radioactive material include rectifying leaky storage apparatus, even if the exposure complies to OSHA rules; if you can detect it with the Geiger counter in the general work areas, then it is not well shielded. Other training is associated with storage locations, sign-out cards, work-area sign-up, use of dedicated radioactive equipment such as pipettors, use of the gamma counter, etc. If the worker wants to use a new material or protocol, he/she must obtain approval. If the new protocol falls outside of the lab authorization, the latter must be amended as described above. Authorized personnel must be periodically retrained by the RPP. BPEC laboratories Approved for Radioactive Material BPEC labs are not currently approved for radioactive material use. The last approval for 14C, 35S, 125I, 3H and 32P under authorization #10-G-7 expired on 8/31/2002. BPEC lab 16-479 is still currently approved for storage of radioactive material.
VII. D. 2. Radiation Benches (Designated Areas) A key component of the OSHA Laboratory Standard is that all work with particularly hazardous substances be confined to designated areas. Particularly hazardous substances as defined by the OSHA Laboratory Standard (29 CFR 1910.1450) are certain carcinogens termed ‘select carcinogens’, all reproductive toxins and all compounds with a high degree of acute toxicity that pose significant threats to human health. Particularly hazardous substances require Standard Operating Procedures, Authority for Chemical Use Forms, and special equipment provisions, usually including Designated Areas. Because radioactive materials are acutely toxic, they are
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classified as particularly hazardous substances. When it applies (but not this year) designated area signs are placed at all radiation benches in the BPEC laboratories. In such a case, equipment required for the manipulation of radioactive samples are dedicated exclusively to these benches as is evident by their radioactive material tags and stickers. Any equipment with radioactive material labels cannot be removed from the radioactive benches for use in other areas of the lab. In addition, only RPP authorized and trained personnel may work at these benches and with any radioactive equipment in the laboratories.
VII. D. 3. Purchase, Inventory Control and Transportation For procurement of radioactive substances, it is necessary for the worker to include the radiation authorization number [10-G-2] and the amount and type of isotope to be procured on the requisition form. The Procurement Office has a special procedure for allocating P.O. numbers to radioactive requisitions. The Application for Authorization form identifies the types and amounts of radioisotopes to be used. It essentially acts as an inventory control device. Upon receipt of a radioisotope, the box is opened in a laminar flow hood; the package and packing material are thoroughly inspected for leakage before allowing the container into the general laboratory area. Radioactive materials must be transported with appropriate shielding, to minimize exposure to others and in a properly labeled, shater-proof container. Shielding must reduce exposure to less than 50 mrem/hour at the container surface, or 2 mrem/hour at 3 feet from the container. There should be no detectable contamination on the surface of the container as measured by a wipe test. Only authorized persons may transport the material. Transport of radioactive materials outside of the MIT complex without the permission and approval of the RPP is not allowed.
VII. D. 4. Storage Facilities for Radioactive Materials All radioactive materials must be kept in areas designated for radioactive materials storage. This includes the designed “radiation” work bench and any refrigerators or freezers approved for radioisotope storage, indicated by a radioactive materials label on the equipment. Radioactive material used in the BPEC laboratories must be stored in a locked refrigerator or freezer. All storage containers/facilities should provide adequate protection against, fire, explosion, or flooding, against accidental breakage, and against unauthorized removal. Several refrigerators and freezers have been approved for storage of radioactive materials. These designated storage devices must be kept locked when not in use, and laboratory personnel that use the refrigerator or freezer will have access to the key. The radioactive material may also be stored in a locked box secured to the refrigerator or freezer. At any given time, the researcher may have on the bench only the amount of radioactive material required for the experiment. The following limits indicate the quantities of radioactive material that may be removed from the stock vials and used at the designated areas within the lab while the material is not under continual active surveillance. To secure radioactive material that are in use, the laboratory doors must be locked when the authorized user is not present.
81 Radionuclide Appendix C Value (from 10 CFR 20)
3H 1000 µCi 14C 100 µCi 33P 100 µCi 32P 10 µCi 35S 100 µCi 51Cr 1000 µCi 125I 1 µCi Radioisotopes must be stored properly in the laboratory; this includes appropriate labeling and proper shielding to maintain a safe radiation level and prevent release of the material. All materials should be labeled with a radioactive substance label (yellow/orange in color with the radiation symbol and the words “CAUTION RADIOACTIVE MATERIAL”). The label should contain the name of the worker, type and amount of isotope, form of the isotope, date of receipt, and date of expiration. The materials must also be adequately shielded to minimize exposure to persons/chemicals in the laboratory. The BPEC criterion is that the Geiger counter should not be able to detect excess radiation within two feet of the container.
VII. D. 5. Working with Radioactive Materials Protective Apparel and Shielding a) All radioisotopes are treated as poisonous, hazardous materials. A lab coat, eye protection
and double gloves should be worn to protect against skin exposure when working with radioactive materials. Hands should be washed after handling any radioactive compound.
b) Shield larger quantities of radioactive material in appropriate containers, e.g. lead. Use
shielding barriers such as lead sheets or plexiglass face shields to protect oneself from exposure to materials that are not well contained.
c) Periodically monitor the effectiveness of shielding and check for contaminations. d) Wherever significant exposure is a possibility, film badges should be worn by all
personnel involved. Precautions a) Stainless steel, porcelain, or glass surfaces lined with absorbent paper should be used
when working with radiochemicals to prevent the spread of contamination. b) Avoid direct contact or ingestion with any radioactive materials. c) Minimize exposure by minimizing exposure time, maximizing distance from the body (or
body part) to the source, and maximizing shielding.
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d) Avoid and minimize spill danger by minimizing clutter and aliquoting radioactive
materials to several smaller amounts. e) Whenever possible, perform a trial run with low activity materials to ensure the adequacy
of procedures and equipment. Handling a) Radioactive work areas are complete with plexiglass and lead shielding, liquid and solid
waste containers, geiger counters, designated pipettors, and count-off for spill cleanup. b) Sign-up sheets and sign-out cards are attached to the bench area, the gamma counter and
the waste containers. c) A Geiger counter is available to monitor all laboratory objects. Before beginning work,
check the work area for previous contamination with the Geiger counter. d) Specific BPEC guidelines for handling radioactive material include rectifying leaky
storage apparatus, even if the exposure complies to OSHA rules; if you can detect it with the Geiger counter in the general work areas, then it is not well shielded.
e) Use pipettes dedicated for the use of radioactive materials. Eject pipette tips into a
disposable container such as a plastic bottle (a T-flask works well) to consolidate solid waste.
f) Work with small amounts of isotope at a time if possible to minimize exposure and spill
potential. Minimize clutter near and on the work area. This will assist in avoiding spills and contamination of apparatus carelessly placed in the way. Replace blotting paper with multiple layers if necessary to contain a possible spill.
g) Clean and screen the area after use of radioactive materials. Store equipment and waste
containers neatly. Replace table soakers if torn or soiled. Complete waste container sign-outs. Fill out the counter log indicating background counts before and after each use, as well as the number of samples measured.
h) Dispose of all items that have contacted radioactive materials in the solid radioactive
waste container. This includes gloves, soiled table soakers, pipette tips and transfer containers. Decontaminate any non-disposable equipment used to manipulate the biohazardous material. Screen the equipment to ensure proper decontamination.
i) Wash hands thoroughly after using radioactive materials. Survey exposed skin, hair and
clothing for contamination. The RPP should be notified immediately if residual contamination is detected. Store radiation badges in an area that shows only background levels of radioactivity. Return the badges to RPP at regular intervals for analysis.
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Exposure Limits Each worker who may receive in excess of 25% of the following amounts should be provided with a radiation badge by the RPP: Body Contact Weekly Quarterly Annually Whole body 100 1250 5000 Skin (beta) 575 7500 30,000 Extremities 1450 18,750 75,000 Note: Beta particles with less than 70 keV will not penetrate the skin. Since the maximum energy from tritium is 18.6 keV, exposure is not a problem (although ingestion is still dangerous). Other common isotopes are of more concern. 14C and 35S put out beta particles at up to 167 keV and 32P up to 1700 keV. 125I emits gamma rays at only 35 keV, but gamma rays penetrate light barriers such as skin. Fortunately, 125I at 1 microcurie can be shielded with a few millimeters of lead or with lead-lined plexiglass.
VII. D. 6. Accidental Release of Radioactive Substances (Spills) Minor hazardous materials or waste spills that present no immediate threat to personnel safety, health, or to the environment can be cleaned up by laboratory personnel that use the materials or generate the waste. A minor hazardous material spill is generally defined as a spill of material that is not highly toxic, is not spilled in large quantity, does not present a significant fire hazard, can be recovered before it is released to the environment, and is not in a public area such as a common hallway. Such a spill can usually be controlled and cleaned up by one or two personnel. Major hazardous material and waste spills should be reported to the MIT emergency number (x100) to receive immediate professional assistance and support in the control and clean up of the spilled material. Major hazardous materials or waste spills are generally defined as having a significant threat to safety, healthy, or the environment. These spills generally are a highly toxic material or is spilled in large quantity, may present a significant fire hazard, cannot be recovered before it is released to the environment, or is spilled in a public area such as a common hallway. Upon reporting such a spill personnel should stand by at a safe distance to guide responders and spill clean up experts to the spill area. Reporting personnel should also keep other personnel from entering into the spill area. In the case of a spill that presents a situation immediately dangerous to life or health, or a situation with significant risk of a fire, personnel should evacuate the area and summon emergency assistance by dialing the MIT emergency number (x100), activating a fire alarm station, or both. Hazardous material users and hazardous waste generators must be aware of the properties of the materials they use and the waste they generate. Properties of materials are most commonly found
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in material safety data sheets and many publications. A good guide to finding other sources of information is found in “Prudent Practices in the Laboratory.” Because of the diversity of types and energetics of radioactive materials, do not attempt to cleanup a radioactive material spill unless you have been specifically trained in cleanup procedures for the particular type of radioisotope used in your work. Notify other personnel to stay out of the area of the spill and contact the RPP (253-2180) to obtain assistance. The RPP has special protective equipment and materials to permit safe entry into areas contaminated with radioactive spills. The RPP is on call 24-hours a day through Facilities, extension 3-1500, or the MIT emergency number - 100. If you feel you are qualified to cleanup the radioactive spill, use the following procedures for radioactive or radioactive, biohazardous spills. Radioactive Spills a) Warn others not to enter the contaminated area. Rope off or guard the spill area against
re-entry. If the spill was a powdered, volatile, or gaseous material, evacuate all personnel and, if necessary, turn off critical equipment.
b) Assemble potentially contaminated persons in one location in or near the laboratory,
monitor for contamination, and detain them to prevent spread. If your garments were soiled during the spill, remove and place contaminated garments into a trash bag. Have the garments checked for radiation before submitting for cleaning. Thoroughly wash hands and face.
c) Put on a lab coat, safety eye wear and double disposable vinyl, latex or rubber gloves. d) Radioactive benches should always be fitted with multiple layers of absorbent, plastic
backed table soakers. If there is a spill on the bench, cut out the contaminated area of the soaker and dispose of in the solid radioactive waste container. Be cautious to avoid cross contamination of other workers or equipment.
e) If there is a spill on a non-disposable surface such as equipment and floors, surround the
spill with spill control pillows or other absorbent material f) In most cases, the spill will involve 14C or 3H that present no external hazard. However,
if more energetic beta or gamma emitters are involved, care must be taken to prevent hand and body radiation exposure. The Radiation Protection Officer must make this determination before the cleanup operation begins.
g) Once the Radiation Protection Officer approves, absorb the spill with the pillows. Wipe
the area clean with paper towels or cloths soaked in the count-off decontaminant. Discard the pillows and wipe cloths into the solid radioactive waste container, or give to the Radiation Protection Officer for disposal.
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h) A final radioactive survey should be made of the spill area and any non-disposable equipment used to cleanup the spill. This can be done using a Geiger counter, or a smear can be taken and counted in a liquid scintillation counter.
Radioactive, Biohazardous Spill Outside a Biological Safety Cabinet a) Hold your breath, leave the room immediately and close the door. b) Warn others not to enter the contaminated area. Rope off or guard the spill area against
re-entry. If the spill was a powdered, volatile, or gaseous material, evacuate all personnel and, if necessary, turn off critical equipment.
c) Assemble potentially contaminated persons in one location in or near the laboratory,
monitor for contamination, and detain them to prevent spread. If your garments were soiled during the spill, remove and place contaminated garments into a biohazard trash bag. Have the garments checked for radiation before submitting for cleaning. Thoroughly wash hands and face.
d) Wait 30 minutes to allow dissipation of aerosols created by the spill. Before cleanup
begins, a Radiation Protection Officer from RPP should survey the spill to determine the degree of risk.
e) Put on a lab coat, safety eye wear and disposable vinyl, latex or rubber gloves. For high
risk spills (such as viruses) a jumpsuit with tight-fitting wrists and use of a respirator should be considered.
f) Surround the spill with spill control pillows or other absorbent material. Pour an iodophor
decontaminant such as Wescodyne or Roccal II around the spill, within the pillows so that none of the spill contacts the pillows before contacting the decontaminant. Allow the decontaminant to flow into the spill. To minimize aerosolization of the spill, avoid pouring the decontaminant onto the spill.
g) Let the mixture stand for 20 minutes to allow adequate contact time. h) In most cases, the spill will involve 14C or 3H that present no external hazard. However,
if more energetic beta or gamma emitters are involved, care must be taken to prevent hand and body radiation exposure. The Radiation Protection Officer must make this determination before the cleanup operation begins.
i) Once the Radiation Protection Officer approves, absorb the spill with the pillows. Wipe
the area clean with paper towels or cloths soaked in the decontaminant. Discard the pillows and wipe cloths into a biohazardous bag or break-resistant container.
j) Before biological decontamination of the waste begins, a Radiation Protection Officer
must determine whether radioactive vapors may be released upon autoclaving (which would contaminate the autoclave). As a general rule, autoclaving should be avoided. If there is no vapor hazard, the material may be autoclaved. If a vapor hazard exists, place
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the waste in a break-resistant container and add sufficient iodophor decontaminant (Wescodyne or Roccal II) to submerge the waste. Cover the container and seal with tape. Affix radioactive and biohazard labels on the container or autoclave bag.
k) Place the autoclaved or iodophor-decontaminated waste in the radioactive waste disposal
receptacle or give to the Radiation Protection Officer for disposal. l) A final radioactive survey should be made of the spill area and any non-disposable
equipment used to cleanup the spill. This can be done using a Geiger counter, or a smear can be taken and counted in a liquid scintillation counter.
Spills Requiring Emergency Action The RPP should be immediately notified in the event of: a) Accidental release of radioactive substances to laboratory drains or ventilation systems. b) Ingestion of radioactive materials. In the event of ingestion, the RPP will perform
bioassay tests. c) Exposure to levels of radioactivity which exceed the quarterly limits described above
(Section VII. D. 5). The RPP can be reached at extension 3-2180 or 3-2360 or by dialing 100 after hours. When calling, report the type and amount of radiation contamination, the room number, injuries, your name and extension.
VII. D. 7. Waste Containers and Disposal Radioactive material disposal is handled in accordance with procedures established by the RPP. Whenever possible, radioactive biohazardous waste should be chemically decontaminated to render it non-infectious, and then disposed of per RPP specifications. Expired materials should be disposed of promptly, and not accumulated in the storage area. Paper refuse or disposable laboratory equipment contaminated with radioactive materials should be disposed of only in appropriately marked yellow radioactive waste containers. Call RPP for disposal pickup when necessary.
VII. E. Equipment for Radioisotopes This section applies to the use of gamma counters, x-ray diffraction equipment, fluorescence analysis equipment, and any other equipment that is used to handle or produces ionizing radiation. Laboratory Supervisors must fill out an RPO-81 form describing the proposed use of any radiation equipment. Approval from the Radiation Protection Program (RPP) must be obtained before the equipment is operated. All personnel using the equipment must register in writing with the RPP and receive specific training from them for the equipment used.
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Use of radiation-producing equipment must be carried out in accordance with the instructions received by the RPP. A gamma counter is the only major radioisotope handling equipment in the BPEC laboratories. There is no radiation producing equipment in the labs. All other equipment for radioisotope use is typical material handling equipment such as shields, pipettors, and Geiger counters.
VIII. Enclosure, Isolation and Regulated Areas
VIII. A. Designated Areas - Definition A key requirement of the OSHA Laboratory Standard is that all work with particularly hazardous substances that could result in the accidental release of harmful quantities of a toxic substance be confined to designated areas. Particularly hazardous substances as defined by the OSHA Laboratory Standard (29 CFR 1910.1450) are certain carcinogens termed ‘select carcinogens’, all reproductive toxins and all compounds with a high degree of acute toxicity that pose significant threats to human health. A designated area is defined as a laboratory, an area of a laboratory, or a device such as a laboratory hood which is posted with warning signs that ensure that all employees working in the area are informed of the hazardous substances in use there. Particularly hazardous substances for which a designated area or areas is established are stored, used, and prepared for disposal only in these designated areas. The boundaries of a designated area are defined in the Standard Operating Procedure written for the specific particularly hazardous substance. In addition to establishing the physical boundaries that define the designated area, the procedures used in a designated area need to be outlined. Particularly hazardous substances require Standard Operating Procedures. For designated areas in the BPEC laboratories, the Standard Operating Procedures are described in the Standard Operating Procedure, Section IX. It is the responsibility of Laboratory Supervisors to define the designated areas in their laboratories and to post these areas with conspicuous (yellow) signs reading “DESIGNATED AREA FOR USE OF PARTICULARLY HAZARDOUS SUBSTANCES - AUTHORIZED PERSONNEL ONLY”. Printed designated area signs are available from the IHP. In some cases it may be appropriate to post additional types of signs describing unusual hazards present and/or identifying the specific hazardous substances in use. An example would be a sign reading “CARCINOGENIC MATERIAL ALIQUOTED IN THIS FUME HOOD” for hoods that are usually used only for volatile and flammable materials. Laboratory fume hoods and radioactive benches in all BPEC laboratories are classified as designated areas. Laboratory Supervisors are required to notify the CHO of the specific location of any designated areas established in their research area that are not laboratory fume hoods or radioactive benches. The exception is operations involving biohazardous materials, as these materials are used in all parts of biological BPEC laboratories. Their use is denoted by the BL2 signs on all laboratory external doors and by the biohazard signs on all contacted equipment and storage facilities.
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VIII. B. Fume Hoods Because fume hoods are required for the manipulation of many types of particularly hazardous substances, such as volatile, flammable, inhalation hazardous, carcinogenic and teratogenic substances, all fume hoods in the BPEC laboratories are classified as designated areas. Fume hoods are located in all laboratories where organic solvents or other flammable materials and inhalation hazardous chemicals are used. One fume hood is located in each of the following BPEC labs: 16-436, 16-476.
VIII. C. Radioactive Benches Because radioactive materials are acutely toxic, they are classified as particularly hazardous substances. Designated area signs have been placed at all radiation benches in the BPEC laboratories. Equipment required for the manipulation of radioactive samples has been dedicated exclusively to these benches as is evident by their radioactive material tags and stickers. Any equipment with radioactive material labels cannot be removed from the radioactive benches for use in other areas of the lab. In addition, only RPO authorized and trained personnel may work at these benches and with any radioactive equipment in the laboratories. No BPEC lab space is currently approved for radioactive material use.
VIII. D. Short-Term Use Hazardous Materials As new particularly hazardous chemicals are introduced into the BPEC laboratories for either short-term or permanent use, the need to assign a designated area for their exclusive use will be evaluated. For short-term use of particularly hazardous materials, users must: 1) Select a confined area for the work, and acquiring proper control equipment. 2) Demarcate the area to be used, and place a designated area sign and a sign identifying the
hazardous substance being used at the location. These signs should also indicate any contact hazards that other workers should be aware of when working near the area.
3) Submit an SOP for the procedures used to the Laboratory Supervisor for review and
acceptance. Laboratory Supervisors are required to notify the CHO of the specific location of any designated areas established in their research area that are not laboratory fume hoods or radioactive benches.
IX. BPEC Laboratory Standard Operating Procedures (SOP’s) Standard Operating Procedures (SOP’s) have been written for ten categories of hazardous chemicals and for equipment that poses some sort of hazard. All chemicals and gases in the BPEC laboratories are classified into one of seventeen classes (Section VI. B. 2.). All chemicals except those classified under 'Moderate to Low Contact Hazard' or 'Minimally or Not Hazardous' are considered hazardous or particularly hazardous
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chemicals and require Standard Operating Procedures with which all researchers must be familiar. Because of the large number of chemicals in the BPEC laboratories, and since all of the hazardous and particularly hazardous substances in the BPEC laboratories are used in very small quantities and often by only a few researchers, individual procedures for the handling of each chemical are not written. Instead, chemicals are grouped into categories and SOP's are written for each category of chemical. All substances are grouped into one of the following SOP categories: Category 1 Carcinogens Category 2 Reproductive Toxins Category 3 Radioactive Materials Category 4 Biohazardous Materials Category 5 Compounds with a High Degree of Acute Toxicity Category 6 Organic Liquids; Combustible, Explosive, Flammable, and Volatile Materials;
Liquid Inhalation Hazards that are Acutely or Chronically Toxic or Acute Contact Hazards
Category 7 Solid Inhalation Hazards (Powders) that are Acutely or Chronically Toxic or
Acute Contact Hazards Category 8 Gases that are Combustible or that are Acutely or Chronically Toxic or Acute
Contact Hazards Category 9 Acids; Caustics; Liquid and Solid Eye, Oral and Skin Contact Hazards Category 10 Highly Reactive Materials SOPs are written by the Laboratory Supervisor and the researchers using them. Existing SOP's are located in an appendix to this plan. The SOP under which each hazardous and particularly hazardous chemical is to be used is specified in the BPEC Chemical Inventory database as well. An example BPEC SOP format is given in the Recordkeeping section (Section XII. I.). New researchers in the BPEC laboratories are introduced to these SOP's at the Chemical Hygiene and Safety Plan Training Seminar at the beginning of each academic year. New chemicals that are classified as particularly hazardous are assigned an SOP at the time they are classified, or a new SOP is written if necessary.
IX. A. SOP’s for Hazardous and Particularly Hazardous Chemical Categories
IX. A. 1. Carcinogens Carcinogens are chemical or physical agents that cause cancer. Generally they are chronically toxic substances; that is, they cause damage after repeated or long-duration exposure, and their
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effects may only become evident after a long latency period. Chronic toxins are particularly insidious because they may have not immediate apparent harmful effects. Certain select carcinogens are classified as particularly hazardous substances and must be handled using the special precautions described in this SOP. Select carcinogens include compounds for which there is evidence from human studies that exposure can cause cancer. A select carcinogen is defined in the OSHA Laboratory Standard as a substance that meets one of the following criteria: a) The substance is regulated by OSHA as a carcinogen. b) The substance is listed as “known to be a carcinogen” in the latest Annual Report on
Carcinogens, published by the National Toxicology Program (NTP). c) The substance is listed under Group 1, “carcinogenic to humans”, by the International
Agency for Research on Cancer (IARC). OR d) The substances is listed under IARC Group 2A or 2B, “probably carcinogenic to
humans”, or under the category “reasonably anticipated to be a carcinogen” by the NTP, and causes statistically significant tumor incidence in experimental animals in accordance with any of the following criteria:
i) after inhalation exposure of 6-7 hours per day, 5 days per week, for a significant
portion of a lifetime to dosages of less than 10 mg/m3, ii) after repeated skin application of less than 300 mg/kg of body weight per week, or iii) after oral dosages of less than 50 mg/kg of body weight per day. The SOP for carcinogens lists the substances meeting criteria a), b), or c). For information on compounds meeting criteria d), see copies of the IARC Group 2A and 2B lists and the NTP lists, available in the Chemical Hazard Classification Information folder of the BPEC CHASP files. For a large number of other compounds there is limited evidence of carcinogenicity from studies involving experimental animals. These compounds should be handled using the general procedures for work with hazardous substances. A list of compounds that are reasonably anticipated to be carcinogens based on animal tests are given in this SOP.
IX. A. 2. Reproductive Toxins Reproductive toxins, are defined by the OSHA Laboratory Standard as including substances which cause chromosomal damage (mutagens) and substances with embryolethality (death of the fertilized egg, embryo or fetus) or teratogenic (malformations) effects on fetuses. Examples of embryotoxins include thalidomide and certain antibiotics such as tetracycline. Many reproductive toxins are chronic toxins which cause damage after repeated or long-duration exposures with effects that become evident only after long latency periods. The period of 91
greatest susceptibility to embryotoxins is the first 8-12 weeks of pregnancy, a period which includes time when a woman may not know she is pregnant. Consequently, women of child-bearing potential should take care to avoid all skin contact with chemicals; especially with chemicals that are rapidly absorbed through the skin such as formamide and trypan blue. Pregnant women and women intending to become pregnant should consult with their Laboratory Supervisor and the EH & S with regard to the type of work they may safely perform and the special precautions they should take. Information on reproductive toxins can be obtained from MSDS’s, by contacting the IHP, and by consulting the Catalog of Teratogenic Agents, Sixth Edition, T.H. Shepard, John Hopkins University Press, Baltimore, 1989. A discussion of “What Every Chemist Should Know About Teratogens,” is found in R.E. Beyler and V.K. Meyers, J. Chem. Ed., 59, 759-763, 1982. A list of known reproductive toxins is included in this SOP.
IX. A. 3. Radioactive Materials All work with radioactive materials and equipment producing radiation (shorter than UV wavelength) is regulated by the MIT Radiation Protection Program. In addition to the instructions outlined in this SOP, special authorization, ordering and inventory control procedures have been established for radioactive substances by the MIT Purchasing Office and Radiation Protection Program (RPP). Because establishment and approval of these provisions must occur well in advance of using these substances, they are listed in Section VII. D. 1. rather than in the radioactive materials SOP. The Center Director must approve the use of radioactive materials in a particular laboratory. The Radiation Safety Officer for the lab must submit an Application for Authorization Form to the RPP. The RPP then assigns an authorization number and returns the form to the Laboratory Supervisor with a description of the amounts and types of materials that can be handled in the labs. Any changes to the type of radioactive work in the lab must be approved via an amendment request to the RPP. Following this authorization, individual workers must be trained and authorized by the RPP to work with materials and protocols previously approved by the RPP. If the worker wants to use a new material or protocol, he/she must obtain approval. If the new protocol falls outside of the lab authorization, the latter must be amended as described above, or the lab must be recertified for handling higher level radioactive material. All radioactive materials must be used in Designated Areas (usually radioactive benches) in approved laboratories.
IX. A. 4. Biohazardous Materials Experiments involving biohazardous materials constitute the bulk of the research performed in the BPEC laboratories. Researchers should be intimately familiar with all procedures related to biohazardous materials. This information is given in Section VII. B. and in this SOP. Biohazardous material can only be used in laboratories labeled with Bio-Level signs.
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IX. A. 5. Compounds with a High Degree of Acute Toxicity A substance is acutely toxic if its toxic effects are manifested after a single or short-duration exposure. OSHA regulations (29 CFR 1910.1200 Appendix A) define toxic and highly toxic agents as substances with median lethal dose (LD50) values in the following ranges: Type of Lethal Dose Toxic Highly Toxic Oral LD50 (albino rats) 50-500 mg/kg < 50 mg/kg Skin Contact LD50 (albino rats) 200-1000 mg/kg < 200 mg/kg Inhalation LD50 (albino rats) 200-2000 ppm/air < 200 ppm/air
Substances which have a high degree of acute toxicity are interpreted by OSHA as being substances defined as “toxic” and “highly toxic” according to the above lethal dose (LD50) values, and substances which “may be fatal or cause damage to target organs as the result of a single exposure or exposures of short duration.” Substances with a high degree of acute toxicity include: a) Corrosive substances. Corrosive substances cause visible destruction of, or visible
alterations in living tissue by chemical action at the site of contact. Major classes of corrosive substances include strong acids (e.g. sulfuric, nitric, hydrochloric, and hydrofluoric acids), strong bases (e.g. sodium hydroxide, potassium hydroxide, and ammonium hydroxide), dehydrating agents (e.g. sulfuric acid, sodium hydroxide, phosphorus pentoxide, and calcium oxide), and oxidizing agents (e.g. hydrogen peroxide, chlorine, and bromine).
b) Irritants. Irritants are defined as non-corrosive chemicals that cause reversible
inflammatory effects on living tissue by chemical action at the site of contact. A wide variety of organic and inorganic compounds are irritants and consequently skin contact with all laboratory chemicals should always be avoided.
c) Sensitizers (allergens). A sensitizer (allergen) is a substance that causes exposed people
to develop an allergic reaction in normal tissue after repeated exposure to the substance. Examples of allergens include diazomethane, chromium, nickel, formaldehyde, isocyanates, arylhydrazines, benzylic and allylic halides, and many phenol derivatives.
Substances with a high degree of acute toxicity also include hazardous substances with toxic effects on specific organs such as: a) hepatotoxins - substances that produce liver damage such as nitrosamines and carbon
tetrachloride,
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b) nephrotoxins - agents causing damage to the kidneys such as certain halogenated hydrocarbons,
c) neurotoxins - substances which produce their primary toxic effects on the nervous
system such as mercury, acrylamide, and carbon disulfide, d) agents which act on the hematopoietic system - such as carbon monoxide and cyanides
which decrease hemoglobin function and deprive the body tissues of oxygen, and e) agents which damage lung tissue - such as asbestos and silica, and which damage skin,
eyes or mucous membranes. Use of these compounds requires full protective apparel, lab coats, leg covering, close toed shoes, safety eye wear, and appropriate gloves, and possibly hazard control equipment such as a fume hood.
IX. A. 6.Organic Liquids; Combustible, Explosive, Flammable, and Volatile Materials; Liquid Inhalation Hazards that are Acutely or Chronically Toxic or Acute Contact Hazards Explosive substances are materials that decompose under conditions of mechanical shock, elevated temperature, or chemical action, with release of large volumes of gases and heat. Flammable substances are substances that ignite when exposed to an ignition source that can be electrical or chemical, or static electricity. This category of chemicals basically covers all chemicals that should be used in a fume hood.
IX. A. 7.Solid Inhalation Hazards (Powders) that are Acutely or Chronically Toxic or Acute Contact Hazards This category of chemicals basically covers all solids that should be used in a fume hood. Refer to Section IX. A. 5. for examples of these types of chemicals, as many are acutely toxic. A substance is acutely toxic if its toxic effects are manifested after a single or short-duration exposure. Chronically toxic substances show their toxic effects after repeated or long-duration exposure and their effects usually become evident only after a long latency period. One example that is commonly used in the BPEC laboratories is solid trichloroacetic acid (TCA).
IX. A. 8.Gases that are Flammable or that are Acutely or Chronically Toxic or Acute Contact Hazards The only commonly used gas in the BPEC laboratories that falls into this category is oxygen. These cylinders should be stored and used away from all ignition sources and other flammable or explosive gases. Regulators designed specifically for oxygen only are used.
IX. A. 9.Acids; Caustics; Liquid and Solid Eye, Oral and Skin Acute Contact Hazards This category of chemicals basically covers all corrosive liquids that require full protective apparel, lab coats, leg covering, closed- toed shoes, safety eyewear, and especially appropriate gloves. Corrosive substances cause visible destruction of, or visible alterations in living tissue by chemical action at the site of contact. Major classes of corrosive substances include strong acids
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(e.g. sulfuric, nitric, hydrochloric, and hydrofluoric acids), strong bases (e.g. sodium hydroxide, potassium hydroxide, and ammonium hydroxide), dehydrating agents (e.g. sulfuric acid, sodium hydroxide, phosphorus pentoxide, and calcium oxide), and oxidizing agents (e.g. hydrogen peroxide, chlorine, and bromine).
IX. A. 10. Highly Reactive Material Highly reactive material encompasses many types of chemicals such as oxygen and hydrogen gas, as well as chemical combinations such as acids and solvents. This SOP describes procedures to prevent contact of highly reactive material with its respective reactant.
IX. A.11. SOP’s for Equipment The following pieces of equipment are considered hazardous to operate and require SOP's for use: Apparatus #1 Autoclave and Steam Use.
X. Education and Training MIT Policy The Chemical Hygiene Officer (CHO) or appointed individual(s) shall provide information and training concerning handling of hazardous chemicals in the laboratory. A training outline shall be prepared and used as the basis for lectures and demonstrations. The Industrial Hygiene and Safety Programs are available to assist the Chemical Hygiene Officer in developing and implementing training procedures and policies. The Industrial Hygiene Program also conducts training sessions for the Chemical Hygiene Officers at the Institute. Employees shall be informed of the presence of hazardous chemicals when assigned to a work area and prior to new exposure situations. Information available must include: 1) contents of the OSHA Laboratory Standard, 2) the applicable details and location of the Chemical Hygiene and Safety Plan, 3) emergency and personal protective equipment training, 4) physical and chemical properties of hazards used in the work place along with proper handling to minimize exposure, signs and symptoms of exposure associated with appropriate chemicals, and4) location and availability of reference material. This training should be provided immediately for new employees in the workspace and annually for other personnel.
X. A. Initial Training All laboratory members of the BPEC laboratories should be familiar with the hazards associated with the chemicals present in their work area. Researchers whose work involves the use of hazardous substances must be informed as to the proper handling procedures and measures they must take to protect themselves from these hazards. With a few exceptions, all new personnel in the BPEC laboratories must complete the following steps before beginning work in the areas where hazardous chemicals are in use. The exceptions are office staff, who are required to be familiar with the Emergency Action Plan only, Section V. F., researchers who hold appointments in other departments but occasionally perform work in the BPEC laboratories, and outside investigators that are performing short-term work in the BPEC laboratories. Individuals holding appointments in other MIT departments who plan to conduct research involving hazardous substances in the BPEC laboratories will be fully trained and legally bound 95
under their respective department’s CHP. However, these individuals will also be required to undergo the same BPEC CHASP training required of temporary personnel. Training for temporary laboratory personnel is outlined below. All Researchers Who Hold Appointments in the BPEC Laboratories and Guests The first group of personnel includes graduate students, post-doctoral fellows, visiting scientists, visiting students and scholars and undergraduate UROP and REU students, and laboratory technical staff. The second group of personnel includes temporary or visiting researchers from within the Institute or from outside institutions or corporations. Responsibility for the training of these individuals is assigned to an individual from the first group, except for undergraduate UROP and REU students. These appointed researchers are also responsible for retraining the new worker should that worker violate BPEC laboratory hygiene and safety policies and procedures.
• All new laboratory personnel must read and understand the BPEC Chemical Hygiene and Safety Plan (CHASP) and must study any other materials provided by their Laboratory Supervisor concerning the hazardous substances in use in their laboratory.
• All new laboratory personnel are required to walk through the lab and review the BPEC
Lab Safety Checklist with the Student Safety Coordinator (or a permanent member of BPEC) to introduce new personnel to basic laboratory hygiene and safety procedures required in the BPEC laboratories.
• Researchers who will be using biohazardous or radioactive materials or lasers must also
attend an Institute-wide training program on the specific hazard(s) and file the appropriate forms verifying that training was successfully completed with the CHO before beginning work in the laboratory. Information on these programs is given in Section X. B.
• New laboratory workers must submit a signed copy of the Chemical Hygiene and Safety
Clearance Form after they have completed parts 1,2, 3, and 4. This form certifies that the researcher has read and understood the BPEC CHASP and all BPEC training and has successfully completed applicable Institute-wide training programs for the hazardous materials or equipment they will use. A copy of this clearance form is given in the Section XII. C. of Recordkeeping.
X. B. Institute-Wide Training for Particularly Hazardous Substances Institute-wide training programs on biohazardous materials, radioactive materials and lasers must be successfully completed by all researchers involved with the use of these materials or equipment, before they are approved to begin working with these materials or equipment. These training sessions are held very frequently, as often as is required by the researcher. In most cases, researchers can receive training within two weeks of a request.
X. B. 1. Biohazardous Material MIT Institute-wide training on the use and handling of biohazardous materials is not required for researchers using biohazardous materials. The training course is recommended as the class is 96
very good. This class focuses on aseptic technique and basic biological procedures for research with bacterial and yeast cultures. Although not directly applicable to animal cell culture, the techniques are useful to learn.
X. B. 2. Lasers All researchers who will be using lasers must attend the MIT Institute-wide training program on laser safety once during their time at MIT. This training covers protective apparel, electrical dangers, eye and skin hazards, and specific requirements dependent on the power of the laser. If tests or forms are required for completion of the class, a copy must be submitted to the BPEC CHO to verify successful completion of the class.
X. B. 3. Radioactive Material For researchers using radioisotopes, read appropriate sections of the BPEC CHASP and all of the MIT Report on Radioisotopes, (provided by RPP), and complete the institute-wide training on radioactive materials. The two-hour training seminar covers purchasing, transportation, storage, use, permissible exposure dosages and disposal of radioactive material as well as required precautions based on the amount and energetics of the radioisotopes used. Radiation badges are assigned if the amount and energetics of the radioisotopes employed dictate their use. Procedures for turning in badges for analysis are reviewed. Researchers are required to pass an exam on the material presented in the training seminar. Submit a copy of this test to the BPEC CHO to verify successful completion of the class.
X. C. Clearance Forms for Outside or Temporary Personnel A Chemical Hygiene and Safety Clearance Form must be submitted by all temporary laboratory personnel before beginning work in the laboratory. This form certifies that the researcher has completed the BPEC Safety Checklist and Test, understands all additional BPEC training on the limited set of materials and equipment with which they will be working, and has successfully completed applicable Institute-wide training programs for the hazardous materials or equipment they will use. This Clearance Form must be signed by the researcher, his supervisor and by the Laboratory Supervisor. A copy of this Clearance Form is given in Section XII. C. of Recordkeeping.
X. D. CPR Certification Cardiopulmonary resuscitation (CPR) training is provided by the Safety Program. It is advised that at least one person in each laboratory and office in the BPEC be CPR certified.
X. E. Training Updates All personnel, temporary or permanent receive Chemical Hygiene and Safety updates following the Chemical Hygiene Officers (CHO) Meeting, as this is most likely when new procedures will be introduced to CHO. These information provided reviews the contents of the BPEC CHASP and expand upon new procedures or changes in procedures due to difficulty in implementation.
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XI. Monitoring, Medical Surveillance and Employee Exposure
XI. A. Monitoring and Medical Surveillance OSHA Laboratory Standard Requirements Employee exposure determination shall be done in accordance with paragraph (d) of the 29 CFR 1910.1450. Initial monitoring will be performed if there is reason to believe that exposure levels for a substance routinely exceed the action level (or in the absence of an action level, the PEL). If the initial monitoring performed discloses employee exposure over the action level (or in the absence of an action level, the PEL), the employer shall immediately comply with the exposure monitoring provisions of the relevant standard. Monitoring may be terminated in accordance with the relevant standard. Within 15 working days after the receipt of any monitoring results, the employee will be notified of these results in writing either individually or by posting results in an appropriate location that is accessible to employees. Anyone with a "reason to believe that exposure levels for a substance routinely exceed the action level, or in the absence of an action level, the PEL" may initiate the monitoring process. Requests for monitoring can be made to EH & S, the Chemical Hygiene Officer, a supervisor, etc. The Chemical Hygiene Officer must be notified of monitoring requests. Monitoring may be requested at any time. It is the responsibility of the Chemical Hygiene Officer to insure that periodic monitoring requirements are satisfied when necessary. In general, the EH & S is called upon to perform the monitoring in accordance with paragraph (d) of 29 CFR 1910.1450, however other qualified services may be employed, but results must be sent to EHS. The EH & S and the Chemical Hygiene Officer must both maintain records. MIT Policy Researchers requiring advice on the health effects of chemicals involved in their work should consult the MSDS file or other chemical evaluation literature (Section VI. B. 1.), available in the Industrial Hygiene Program (IHP), room N52-4th floor, extension 3-2596. Any individual who believes they may have been exposed to a hazardous substance should notify their Laboratory Supervisor immediately and contact the IHP. Individuals who experience any adverse health effects from exposure to hazardous substances should immediately report to the Medical Department. Emergency assistance can be obtained by calling extension 100. All accidents involving exposure to hazardous substances that require medical attention must be reported in writing to the Laboratory Supervisor and the CHO. It is the responsibility of the Laboratory Supervisor to identify and notify workers who will be engaged in research that may involve hazardous exposures requiring medical surveillance. The Laboratory Supervisor must provide names, work addresses, and social security numbers of these workers to the EH&S Office. The EH&S Office may also identify individuals or populations of individuals at risk and invite their participation. The purpose of medical surveillance is, as a secondary means of prevention, to detect early failures of primary means of work place protection that may result in work place illness. Medical surveillance is offered, to researchers exposed routinely to the following hazards:
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* Asbestos + Arsenic * Beryllium + Cadmium + Carcinogen * Lasers (Class III or IV) + Lead + Mercury * Noise (Hearing Loss) Respirator Use (See Respirator Policy, Section IV. A. 3.) + Thallium * Tower Climbing and the following materials: + Recombinant DNA +,* Potential Sources of Infectious Diseases (e.g. animal handlers, etc.) +,* Radioactive Material/Ionizing Radiation *Physician Visit +Laboratory Values and Report Only Researchers may enroll themselves in medical surveillance programs by calling the Medical Department, extension 3-9398. Workers can arrange consultations with physicians concerning work place risks, illnesses, and allergies by calling the Medical Department, extension 3-4481. There are no charges for employees or students who need these services. It is the responsibility of individuals to appear promptly for scheduled appointments. The departments or other administrative entities may, as a matter of written policy, require enrollment and participation in medical surveillance provided that such requirements apply equally to all affected individuals. Once individuals are enrolled, introductory and periodic invitations are automatically mailed to them. Those invitations are accompanied by lab slips and directions. On receipt, individuals are still responsible for calling the Medical Department, extension 3-4481, to set up a time of appointment. Laboratory Supervisors who believe that individuals have been inadvertently omitted from medical surveillance may enroll or re-enroll exposed individuals. In addition, individuals who believe that they have been inadvertently omitted may enroll or re-enroll. To receive invitations, call extension 3-9398. Right To Privacy: Your medical surveillance results are private. They are part of a medical record that is kept within the MIT Medical Department. The only individual who can initiate the dissemination of this information outside of the Medical Department is you. Right To Information:
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You have the right to be informed of the results of your medical surveillance, and this will happen normally as a matter of course. If your surveillance involves a physician or other
provider visit, you will be informed of findings at the time of your visit. If complete surveillance information is not given to you at the time of provider visits, because surveillance involved lab work only or because the full information was not yet available at the time of the visit, you will receive a letter informing you of results. If you have not received complete information about an encounter within one month from the time of the visit, or if you have questions about the information received, please call 253-5360 to obtain additional information. Rights of Supervisors: Supervisors have a right, and sometimes a legal duty, to know if individuals are medically fit to perform certain kinds of work (such as wearing respirators on the job). Supervisors are not entitled to know diagnoses, medical findings, or other personal information in the medical record. When supervisors need information, the information given to supervisors is in one of three categories: 1. Fit to perform the operation 2. Not fit to perform the operation 3. Fit to perform the operation with the following specific restrictions (weight limits, time
limits, equipment limits, etc.). Additional information can be given to supervisors only at the patient's written request. BPEC Regulation The only environmental monitoring currently required in the BPEC labs is for radiation exposure (radiation badges) for those researchers using radioactive materials. The results of the badge monitoring are kept on file at the Radiation Protection Program. The Institute offers a voluntary health-screening program for personnel doing recombinant DNA research. Details of this program can be obtained by contacting the Medical Department, extension 3-9398, http://web.mit.edu/envms/www/.
XI. B. Accidents and Employee Exposure Reporting Individuals who experience any adverse health effects from exposure to hazardous substances should immediately report to the Medical Department. Emergency assistance can be obtained by calling extension 100. Any individual who believes they may have been exposed to a hazardous substance should notify their Laboratory Supervisor immediately and contact the Industrial Hygiene Program (IHP), room N52-4th floor, extension 3-2596. All accidents involving exposure to hazardous substances that require medical attention must be reported in writing to the Laboratory Supervisor and the CHO. Reports should be filed within a week of the date of the accident. Written accident reports must be prepared following any of the following incidents: 1) Fires that require the use of a fire extinguisher or require sounding a fire alarm. 100
2) Injury requiring medical attention, no matter what the cause. 3) A spill of a hazardous substance that takes place outside a designated area. Any
accidental release of a particularly hazardous substance requires the filing of an accident report. This includes accidental release of untreated biohazardous materials down the drain.
Laboratory personnel should notify the Student Safety Coordinator or Laboratory Supervisor of any potentially hazardous situations or practices they are aware of in the laboratory. If any accidents are the result of an airborne material or gas, the IHP evaluates the situation and conducts air sampling if necessary to determine actual exposures. The results of all hazard evaluations and any air sampling data are available to all occupants of the affected areas. The Chemical Hygiene Officer or the IHP can be contacted directly for information. In addition, the results of any personal air sampling conducted are given to the individual as well as kept in the IHP's records. (Results are also added to the individual's medical records).
XII. Recordkeeping
XII. A. Authority for Chemical Use Specific chemicals require an authorization for purchase or use, including absolute ethanol, all controlled drugs/pharmaceuticals and a handful of extremely toxic chemicals that may be used in bioterrorism activity. A list of this last group may be obtained from the Biosafety Program. Those chemicals are controlled at BPEC through the use of an Authority for Chemical Use Form. The requirement of an Authority for Chemical Use Form is the decision of the Chemical Hygiene Officer. The training records are located in a file cabinet of the Chemical Hygiene Officer.
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Authority for Chemical Use Form plans to use the particularly hazardous (Researcher's Name) substance ______________________________________________________________ (common name for substance) in quantities of ___________________________________________________________ (container size and number of containers to be used in time period) for the period ____________________________________________________________ (calendar dates for use) ****************************************************************** This substance is a known _________________________________________________ (hazardous chemical classification) ****************************************************************** The signature of verifies that he/she has: (Researcher's Name - printed) SPECIAL PROVISIONS Read the relevant sections of the Chemical Hygiene and Safety Plan Facilities for
Particularly Hazardous Substance Use and Disposal (Section VII.) and the appropriate Standard Operating Procedure for this chemical.
TRAINING BEFORE USE Attended and completed the required training, safety seminars, physical exams, etc. as
specified by the BPEC Chemical Hygiene and Safety Plan, or SOP. EXPOSURE LIMITS Understood the permissible, safe exposure limits to the substance, both in terms of dose
and time. EQUIPMENT Learned how to properly use specific equipment for the transportation, handling, and use
of this substance; such as carrying containers, fume hood, biosafety cabinet, radiation shield, etc. Knows how to use and dispose of the material within its designated area.
DISPOSAL Understood the proper disposal procedures for the substance. EMERGENCY PROCEDURES
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Familiarized themselves with the emergency procedures to follow in case of accident, spill, or contact with the substance.
SPILLS/ACCIDENTS -CLEANUP Educated themselves on the procedures to follow in the event of a spill, including the
location of spill cleanup kits and associated personal safety equipment. HOUSEKEEPING Grasped the basic housekeeping tasks required to cleanup an area following the use of the
substance; such as decontamination of the biosafety cabinets, radiation check with the geiger counter, etc.
PERSONAL HYGIENE Learned the personal hygiene necessary to guarantee that, following use, the substance
does not contact his/her person, is not transported to another location or person, or is not carried out of the lab.
STORAGE AND LABELING Acquainted themselves with the proper storage facilities and labeling methods. (Signature of Researcher) (Date) (Signature of Chemical Hygiene Officer) (Date)
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XII. B. BPEC Safety Checklist and Test This document is designed to introduce new personnel to basic laboratory hygiene and safety procedures required in the BPEC laboratories. A copy of this document is given on the following pages:
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SAFETY CHECKLIST for training new researchers of the BIOTECHNOLOGY PROCESS ENGINEERING CENTER
GIVE A COPY OF THIS SAFETY CHECKLIST TO THE NEW WORKER. THE SAFETY TRAINER MUST TAKE THE NEW RESEARCHER ON A LABORATORY TOUR, POINT OUT AND DISCUSS THE FOLLOWING THINGS:
I. Emergency Procedure - POINT OUT THE LOCATION OF THE FOLLOWING:
• Lab exits, stairwell exits and fire alarms (at stairwells): MIT Policy in case of fire or emergency is: pull alarm and evacuate. Alternative is to call 100 and report nature of emergency - be sure to hang up the phone last.
• Fire extinguishers: You are not expected fight a fire unless necessary for your safety, or if the fire is very contained and obviously controllable. Pull pin on extinguisher to activate it.
• Fire blanket: Pull blanket from box and roll in it.
• Electrical box: In case of a flood in the lab, kill lab electricity.
II. Personal safety - POINT OUT THE LOCATION OF THE FOLLOWING:
• Lab coats & safety glasses: Always wear when conducting experiments with hazardous substances, cultures, and/or chemicals. Wearing a lab coat is mandatory in BL-2 areas.
• Gloves: Wear when handling chemicals or cultures. Wearing gloves when working in BL-2 areas is mandatory.
• Goggles: Wear when handling acids, bases or potentially explosive materials.
• Safety shower: Use in case of chemical spill on the body.
• Eyewashes: Use to rinse out the eyes.
• Bandages
• No eating, drinking, smoking, or storing food is allowed in the lab. Always wear protective footwear in the lab.
III. Spills
• Point out location of spill kits, used to clean up chemical spills. Open up the spill kit and show contents to new researcher. Focus especially on the biological spill kit.
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• Mention that all lab workers are responsible for cleaning up their own spills, unless it is an unknown or toxic substance in which case EH&S Office (x2-EHSS) or Industrial Hygiene Program (x3-2596) should be called.
• Biohazardous spills should be treated with bleach before cleaning up. Clean-up rags go in biohazard bags. Show and open biological spill kit at end of benches 13/14.
IV. Safe Use of Laboratory Equipment and Facilities
Autoclave
• Indicate protocol posted next to the large autoclave and procedure for logging autoclaving of wastes, and disposal of biohazard wastes.
• Open autoclave door slowly (avoid rush of hot steam), following completion of the autoclave cycle.
• Stay aware that autoclaved liquids can superheat and boil over.
Compressed Gas Cylinders
• Tilt and roll on side to move the cylinder - move only with protective cap on, or use the BPEC cylinder cart.
• Stored cylinders should always be chained to a wall or clamped to a bench. Empty cylinders must be promptly returned.
Biosafety cabinets (BSC)
• Lights in hood are ultraviolet: turn them off when working in the hood.
• Always use caution when using a flame in the hood.
POINT OUT LOCATION OF THE FOLLOWING:
• Section of lab where radioactive work is being conducted.
• Cabinet where flammable solvents are stored.
• Location of liquid nitrogen.
• Biosafety cabinets used for BL-2 work.
• Biosafety cabinet used for adenovirus work.
• Biosafety cabinet used for microbial cultures.
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V. Waste Handling - POINT OUT LOCATION OF THE FOLLOWING:
• Biohazard bags: for disposable materials that have come in contact with cultures.
• Bleach buckets: for spent medium.
• Sharps gray container: for razor blades, Pasteur pipettes, contaminated broken glass.
• General trash container: for trash that has not been in contact with cultures.
• Sink: for bleach bucket bleach (or lysol), NOT for medium disposal.
VI. Aseptic Technique
Demonstrate proper techniques for working aseptically in biosafety cabinets and at bench.
VII. Chemical Hygiene and Safety Records - POINT OUT THE FOLLOWING IN 16-436, 16-476, and 16-463:
• Copies of BPEC Chemical Hygiene and Safety Plan.
• Names of Chemical Hygiene Officer and Student Safety Coordinators listed in Chemical Hygiene Plan.
• Standard Operating Procedures.
• Print outs of the chemical inventory (copies are located in 16-436 and 16-463).
****************************************************************************** AT THE CONCLUSION OF THE TRAINING SESSION, GIVE THE NEW RESEARCHER A
COPY OF THE SAFETY REVIEW THE CHEMICAL HYGIENE AND SAFETY CLEARANCE FORM. FILL OUT CHEMICAL HYGIENE AND SAFETY CLEARANCE FORM AND SUBMIT TO THE SAFETY OFFICER.
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SAFETY TEST
To Be Completed by All New Laboratory Personnel at the Biotechnology Process Engineering Center
1. What do you do if you see smoke or fire? Where are the locations of fire extinguishers? Can these extinguishers be used for paper, electrical, or chemical fires?
2. What would you do if your clothes caught on fire?
3. What would you do if you splashed chemicals on your body and clothes?
4. What would you do if you came in the lab to find a major flood?
5. The following are ways to maintain personal safety in the lab (check required safety measures):
________ wear gloves when handling hazardous chemicals or biohazardous samples ________ wear a lab coat when working in BL-2 areas ________ wear sandals or nothing on the feet on hot days ________ wear goggles when handling acids, bases or potentially explosive materials
________ wear safety glasses when entering into any lab areas
6. The following practices are NOT allowed in the laboratory:
________ eating lunch or drinking a beverage ________ reaching into the grey sharps bins
________ bringing plants or flowers ________ leaving doors of BL-2 areas (e.g. 16-436 and 16-463) open or ajar
________ leaving dirty labware in the sinks ________ storing food or beverages ________ bringing bicycles
________ going outside laboratory areas with latex gloves ________ leaving the 4th floor of Building 16 with a lab coat on ________ storage of glassware in sinks
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SAFETY TEST (continued)
7. a. Who is responsible for cleaning up a spill?
b. Where are the spill kits located, and what are in them?
c. If you spill a biohazardous material outside of the biosafety cabinet (on the floor or the bench), how do you clean it up?
d. If you spill something toxic and don’t know how to proceed, who should you call for help?
8. Who do you call in an emergency situation?
9. Which of the following waste chemicals should you put in properly labeled containers and call the MIT Environmental Management Program to have them pick the waste up and dispose of it? How would you otherwise deal with the waste chemical?
________ organic solvents ________ concentrated acids or bases ________ hazardous inorganic or organic solids ________ heavy metals ________ controlled substances
10. What methods of sterilization are utilized in the BPEC laboratories?
11. If you need milliliters of a particular solvent which is not in the BPEC lab, would you purchase a small bottle of it to meet your needs, or a large one because it is much cheaper?
12. What particularly hazardous substances are used in your lab? What precautions must be taken? Are there designated areas for working with these substances?
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SAFETY TEST (continued)
13. Where do you find out any known hazards associated with a chemical?
14. What information must you put on the label of any cultures, chemicals or solvents you prepare?
15. Which of the following practices are required of all lab workers? 1. ________ adhere to a specific SOP for working in the cell/tissue culture rooms 2. ________ adhere to a specific SOP for using a biosafety cabinet in the main lab 3. ________ adhere to a specific SOP for using the autoclaves 4. ________ adhere to a specific SOP for using a chemical fume hood
5. ________ adhere to a specific SOP for getting and filing a MSDS 6. ________ adhere to a specific SOP for placing and accepting deliveries (especially
for large boxes) 7. ________ promptly reporting equipment problems or failures to the person
responsible for maintaining that piece of equipment, AND to the Lab Supervisor
8. ________ keeping reasonable noise levels (especially radios) 9. ________ cleaning up your own messes
16. What are the specific locations in lab 16-436 where projects are classified “BL-2”?
17. Why is there red tape on the floor around biological safety cabinet #5 and around bench #9?
18. What is the major difference between lab 16-476 and lab 16-436, in terms of the cell types used (where are animal and microbial cells used)?
19. Why are specific sections of the freezers and of the refrigerators located in the cell bank room locked?
20. Where are copies of the BPEC Chemical Hygiene and Safety Plan located?
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SAFETY TEST (continued)
Indicate the proper disposal method for the waste listed in left column:
bleach or lysol bucket
biohazard bag
Sharps graycontainer
general trash
sink other
contaminated disposable gloves contaminated disposable pipettes contaminated Pasteur pipettes uncontaminated Pasteur pipettes spent medium; cell culture samples contaminated tips contaminated paper products uncontaminated paper Bleach bucket (or lysol) razor blades
uncontaminated broken flasks; empty bottles
21. I (New Employee) have read and understand the “Common Procedures and SOPs Form” provided on the next page (YES or NO).
This certifies that ______________________________________ has completed (New Employee)
SAFETY TRAINING as required for employees of the Biotechnology Process Engineering Center,
on __________________by ____________________________.
(Date) (SafetyTrainer)
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SAFETY TEST (end)
Common Procedures and Standard Operating Procedures Researcher's Responsibilities
• Clutter causes accidents and impairs evacuation. Be neat! Do not leave items such as bicycles, boxes, buckets and trash bags in the middle of the floor or blocking exits. In fact it is not allowed to store bicycles in labs 16-436 and 16-476, or in the instrument rooms.
• Maintain BL-2 lab door closed at all times. • Remove undesired left-over beverages and food from the shelf, outside lab 16-436, on
a regular basis. • Complete the Safety Test, and have your UROP student or guest complete one ASAP.
Keys to the lab and permission to work in lab cannot be given until the safety tests are completed and approved.
• To get rid of large boxes and empty crates, email [email protected] to request that they remove them from the corridor space.
• Clean work areas when work is completed using recommended procedures. • Use gloves inside the biosafety cabinet. • Decontaminate common work areas such as biosafety cabinets and incubators after
use. • Restock common shelves, drawers and carts with depleted materials. • Ensure that all gas cylinders are secured to stationary supports with clamps, straps
and/or chains, or supported free standing with a cylinder base unit, and that the cylinders are capped with screw-on caps when in storage.
• Rinse dirty glassware but do not leave any glassware in the sinks. • Place all biohazardous wastes in proper containers and empty containers as necessary. • Place biohazard bags in a polypropylene or metallic bucket (or tray), prior to
autoclaving, + autoclave biohazard label along with the bag + log the bag and the label.
• Maintain neat and clean refrigerator and freezer spaces. Assist with routine defrosting of freezers and consolidation of cell banks.
• Immediately chemically disinfect and discard contaminated cultures. Disinfect any contacted surfaces or equipment as well.
• Do not clutter the fume hood and wear proper gloves for manipulating corrosive chemicals in it.
• Report equipment malfunctions to the researcher assigned to the faulty apparatus. • Report any violations to the Laboratory Safety Officers or the Laboratory Supervisor.
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Standard Operating Procedure For Autoclaves
To autoclave bio-hazardous trash:
1. Make sure to double bag the trash with an orange or a clear bag.
2. Pour about a cup of water into the trash bag.
3. Do not tie the top of the trash bag.
4. Open the door to the autoclave. You must first pull the lever down before opening if
using the big autoclave.
5. Place an autoclavable (e.g. polypropylene or metal) tray inside the autoclave with the
bag of trash inside it.
6. Shut the door. Pull the inner lever up and turn the wheel clockwise until tight if using the
big autoclave.
7. Select the correct setting button. If you are autoclaving trash please select the “dry”
setting.
8. Set time for sterilization and drying.
a. For non-biohazard trash, sterilization is 45 minutes and drying is 15 minutes.
b. For biohazard trash, sterilization is 90 minutes and drying is 15 minutes.
9. Press the red ON button.
10. When autoclaving is done, first crack the door open to allow venting of steam
11. Use a pair of thick gloves to open and remove the bin from the autoclave.
12. Document your use of the autoclave in the logbook inside the autoclave room.
13. The autoclaved bags should be placed inside a normal gray trash bag. Tie the tag around
the top of the gray bag, and place outside the lab doors.
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Standard Operating Protocol Sheet for Long-Term Experiment
Room #_____________Date: _________ Name of Researcher in Charge:_______________________________ Home Phone Number of Researcher to Contact: ________________________________
Currently the experiment in this area is running 24 hours a day. In case of a problem (e.g. spill) this information may be useful to the person discovering it:
1. Cell or virus (specify the general type):
_____________________________
2. Hazardous chemical used:
____________________________________
3. Potential electrical hazard:
____________________________________
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Rules for work in tissue culture room B (Contact the Griffith group for use of room A)
General Management of the Room (shared between the Lauffenburger and Hamel groups) • The maximum occupancy in the room is 4. • Only approved researchers can use the room. • If anyone among the approved users sees a situation that goes against our protocols (misuse of equipment, poor
protocols, etc.), one should help correct the situation, and voice out one’s concern immediately. • Minimize traffic in the TC room when carrying critical steps, such as passages and the preparation of new cell stocks.
When implementing these steps, consider “reserving” the room during specific times.
• Starting February 2002, each group is responsible to purchase their own supplies.
Protective apparel • In the TC room, wear only the lab coats with tight cuffs. Those lab coats are 100% polyester. The rest of the time, when
in lab 16-436, the other lab coats (the ones with the loose sleeves) should be worn.
• When manipulating in the biosafety cabinet (BSC), arms should all be covered. Furthermore, 70% ethanol should be sprayed starting with hands and including the elbow. For every 3 times one “exit” the BSC, one should re-spray.
Preparation and Use of the BSC • Avoid blocking the front and the back grids in the BSC. Place only what you need inside the BSC, and avoid
accumulation of pipettes, tubes, etc.
• When moving your hands out of the BSC, while an on-going experiment in the BSC (e.g. for passages), hands should be moved slowly and perpendicularly to the front of the BSC.
• When working at the BSC, minimize the number of times you have to get your hands out of the BSC. Plan your experiment well, so that most of what you need for an experiment is within or nearby the BSC.
• When working in the BSC, work as much as possible in the middle of the work area.
Specialized Equipment • Purchase only T-flasks with membrane caps. • Incubators
1. Disinfect incubators once every 2 weeks. 2. The inside of the incubators should be sprayed with 15% H2O2, including the shelves, the fan, but not the CO2 or the
O2 ports. Furthermore the rubber liner should be removed and cleaned with diluted bleach. 3. Autoclave the trays of the incubators every 4 weeks. 4. Shelves should be autoclaved in separate bags (i.e. one shelf per bag). 5. In case of contamination of an incubator, cultures should be handled as if they were contaminated, and should not be
transferred to another incubator, without permission of its users. • Except for perfusion-based projects, all media should be purchased in bottles of 1 liter or less. • Water from the water bath should be replaced weekly, and the bath should be cleaned with Clorox weekly (pay attention to algae sometimes located on the edges of the bath). After the water bath has been cleaned and refilled, add Clear Bath Algicide VWR Brath® and Sigmaclean® water bath disinfectant.
• Spray medium bottles with 70% EtOH before use. Then cover the cap of the bottle with parafilm, before placing it in the water bath. Before a bottle is transferred into the BSC, spray with 70% EtOH on the entire bottle and on the parafilm, then trash the parafilm before placing the bottle inside the BSC.
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XII. C. Chemical Hygiene and Safety Clearance Forms A Chemical Hygiene and Safety Clearance Form must be submitted by all permanent and temporary laboratory personnel before beginning work in the laboratory. This form certifies that the researcher has read and understood the BPEC Chemical Hygiene and Safety Plan (for permanent personnel), completed the BPEC Safety Checklist, understands all additional BPEC training on the materials and equipment with which they will be working, and has successfully completed applicable Institute-wide training programs for the hazardous materials or equipment they will use. The Clearance Form must be signed by the researcher, the student supervisor of the researcher (for temporary personnel), and the Laboratory Supervisor. A copy of this Clearance Form is given on the following page.
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Chemical Hygiene and Safety Clearance Form for the MIT Biotechnology Process Engineering Center
(to be returned to J.-F. Hamel, Room 56-483) This form certifies that has successfully (Researcher’s Name - printed) completed the BPEC Chemical Hygiene and Safety Plan Initial Training requirements and is approved to begin research in the BPEC laboratories. In completing the initial training requirements, the researcher has: 1. Met with the Laboratory Supervisor, upon arrival at MIT. 2. Read and understood the BPEC Chemical Hygiene and Safety Plan (CHASP) and any other
materials provided by the Laboratory Supervisor. 3. Reviewed the BPEC Lab Safety Checklist with a senior-level researcher and completed the
safety test. 4. Completed Institute-wide training programs on biohazardous materials, radioactive materials
and lasers as relevant to their research. Proof of completion of these programs (tests administered or attendance records) has been submitted with the BPEC Chemical Hygiene Officer.
________________________ (Signature of Researcher) (Date) ________________________ (Signature of Assistant Chemical Hygiene Officer (Date) or of Chemical Hygiene Officer)
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XII. D. Exit Clearance Forms On termination of a research project or completion of a theses, graduate students, post-doctoral fellows, visiting scientists, visiting students/scholars need to clear all of their supplies from the lab, office and computer room and complete and Exit Clearance Form. The Exit Clearance Form must be signed by two coworkers, the Laboratory Supervisor and the researcher’s advisor to obtain exit clearance. The researcher’s advisor should not sign off on the researcher’s thesis or related final document or forms until the Exit Clearance Form is completed and submitted to the CHO. A copy of this form is given on the following page.
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Exit Clearance Form for the MIT Biotechnology Process Engineering Center
This form certifies that has successfully (Researcher’s Name - printed) completed the BPEC Chemical Hygiene and Safety Plan requirements for checking out of the laboratory, office and computer room space assigned to the researcher. Completing these requirements means the researcher has: 1. Accounted for all of their chemicals, by disposing of them, or reassigning them to another
researcher, or a combination of both. In doing so, they followed all MIT chemical disposal regulations.
2. Discarded or reassigned any personal equipment. Put commonplace equipment belonging to
them such as pipettors, glassware, small reactors, timers, etc. into the lab common supplies. 3. Removed personal files from the computers and any personal diskettes from the computer
room. Cleared data from excess diskettes and added the diskettes to the common diskette box in the computer room.
4. Cleaned out their desk and related office space; file cabinets, bookshelves, storage closets,
refrigerators, etc. To obtain exit clearance, this form must be signed by two coworkers, the Laboratory Supervisor, and the researcher’s advisor. The researcher’s advisor should not sign off on the researcher’s thesis or related final document or forms until this form is completed and submitted to the CHO. ________________________________________ ______________________________ (Signature of Researcher) (Date) ________________________ (Signature of Researcher) (Date) ________________________ (Signature of Laboratory Supervisor or of Assistant Chemical (Date) Hygiene Officer) ________________________ (Signature of Researcher’s Advisor) (Date)
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XII. E. Large-Scale Batch Fermentation Run Logs Large-Scale Batch Fermentation Run Logs are required for all microbial or animal cell culture fermentations over 10 liters in volume. Presently, no such work is done in BPEC.
XII. F. Medical Surveillance and Accidents All records pertaining to employee medical surveillance and accidents must be kept for 30 years. Any records of surveillance performed by the MIT Medical Department are kept in the Medical Department files if they are confidential. All accident reports will be filed in the BPEC CHASP files under the name of the individual involved, or the location if no individuals were involved.
XII. G. MSDS Material Safety Data Sheets, (MSDS) are bulletins prepared by manufacturers to summarize the health and safety information about their products. MSDS come in many formats and present the information in different ways Federal law requires that suppliers send an MSDS to MIT the first time a compound is ordered by a student or employee. The EH&S Office maintains a file of MSDS’s for MIT. Any member of the Institute can refer to this file and if desired obtain copies of Material Safety Data Sheets, through the Web links offered by MIT’s Safety Program.
XII. H. Radioisotope Work Radiation badges are assigned to users of radioisotopes if the amount and energetics of the radioisotopes employed dictate their use. This determination is made by the Radiation Protection Program (RPP). The results of the badge monitoring are kept on file at the RPP. Similar to medical files, these files must be maintained for 30 years.
XII. I. Standard Operating Procedures An example BPEC Standard Operating Procedure format is given on the following page, and is followed by a SOP actually in place related to long-term experiments.
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Standard Operating Procedure (Chemical Hazard Classification or Equipment Name) ****************************************************************************** Biotechnology Process Engineering Center Identification Code: Revised: (CHO Signature) ****************************************************************************** I. Description of Hazardous Chemical Classification II. Required Procedures Before Use III. Special Ordering Procedures IV. Protective Equipment V. Precautions VI. Handling VII. Exposure Limits (mrem) VIII. Disposal IX. Spills/Accidents X. Storage and Labeling XI. Transportation XII. Housekeeping After Use XIII. Personal Hygiene After Use XIV. Types of Chemicals (or specific names of hazardous chemicals) that Fall Under this
SOP XV. List of BPEC Chemicals that Fall Under this SOP
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XII. J. Training Attendance records for the Chemical Hygiene and Safety Initial Training Meetings and Updates are filed with the Chemical Hygiene Officer. Training documents on biohazardous material, lasers and radioactive material are provided by the Institute-wide Training programs. Attendance to Chemical Engineering Department hygiene and safety seminars, for those BPEC students that are in this department, is kept by the Chemical Engineering Department.
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XIII. References The following references either appear in this document, or are additional useful sources of hygiene and safety information. Biosafety in Microbiological and Biomedical Laboratories, Second Edition, U.S. Department of
Health and Human Services, Washington, D.C., 1988. The BPEC Safety Manual, 2000.
Catalog of Teratogenic Agents, Seventh Edition, T.H. Shepard, John Hopkins University Press,
Baltimore, 1992. Chemical Hazards of the Workplace, Fourth Edition, N.H. Proctor, J.P. Hughes, and M.L.
Fischman, 1996. Dangerous Properties of Industrial Materials, Seventh Edition, N. Irving Sax, Van Nostrand
Reinhold Company, New York, NY, 1989. Destruction of Hazardous Chemicals in the Laboratory, by G. Lunn and E.B. Sansone, 1990.
Developing a Chemical Hygiene Plan, J.A. Young, W.K. Kingsley and G.H. Wahl, Jr.,
American Chemical Society, Washington D.C., 1990. Laboratory Health and Safety Handbook; A Guide for the Preparation of a Chemical Hygiene
Plan, R.S. Stricoff, John Wiley & Sons, Inc., New York, NY, 1990. The Merck Index, Eleventh Edition, M. Windholz, Editor, published by Merck & Co. Inc., 1989.
The MIT Chemical Engineering Department’s Chemical Hygiene Plan, 2001.
The MIT Chemistry Department’s Chemical Hygiene Plan and Safety Manual, 2001.
The MIT template for Chemical Hygiene Plans, 1990.
National Safety Council’s R&D Newsletter, Jan-Feb., 1987.
Prudent Practices in the Laboratory, prepared by the National Academy of Sciences’
Committee on Prudent Practices for Handling, Storage, and Disposal of Chemicals, Hazardous Substances in the Laboratory, National Academy Press, Washington, D.C., 1995.
The Sigma-Aldrich Library of Chemical Safety Data, Second Edition, R.E. Lenga, Ed., 2
volumes, 1988. What Every Chemist Should Know About Teratogens, R.E. Beyler and V.K. Meyers, J. Chem.
Ed., 59, 759-763, 1982.
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XIV. Appendices
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Air Monitoring Results Within 15 days of receiving monitoring results, the Chemical Hygiene Officer/Industrial Hygiene Program shall notify the employee in writing either individually or in writing. All air monitoring results are kept on file by the Chemical Hygiene Officer and also at the Industrial Hygiene Program. They are available for review upon request. Copies of air monitoring results for laboratory areas covered by this Chemical Hygiene and Safety Plan shall be stored here and the copy of this plan kept in EH&S/IHP.
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Laboratory Exhaust Hood Annual Surveillance Data and Certificate
The Industrial Hygiene Program surveys fume hoods annually. The survey consists of measuring the face velocity of the hood and using a smoke stick to visually check its containment effectiveness. If the hood passes both the face velocity and smoke containment tests, the hood certification label is updated. If the hood does not pass the survey and the problem is so severe that the fume hood is unsafe for use, then it is labeled with a "DO NOT USE" sign. If the problem if less severe so that the fume hood can still be used even though it does not pass the fume hood survey, it is not updated and a list of problem hoods is sent to the chemical hygiene officer along with a description of the problem.
If the problem is one that physical plant can correct (e.g. a slipping fan belt, cracked duct work, etc.) than IHP submits a work order to physical plant to have it repaired. Physical plant notifies IHP when the repairs have been made and the fume hood is resurveyed. If it is a problem that relates more to users (e.g. a cluttered hood) then IHP notifies the Chemical Hygiene Officer. It is the responsibility of the Chemical Hygiene Officers to get the problem corrected and to contact the IHP to resurvey the fume hood. It is the responsibility of the Chemical Hygiene Officer to notify IHP if a fume hood does not have a current certification date (not more than one year old).
The average face velocity criteria used for most hoods at MIT is 100 feet per minute. The hood face is divided into nine equal areas and face velocity is measured in the center of these equal areas. Each measurement must be within 20 percent of the accepted average face velocity criteria. The nine readings are averaged and the face velocity at the fully opened sash height is indicated on the survey label. If the face velocity average is less than 100 feet per minute than the sash height that does produce a 100 feet per minute average is found and the hood is labeled with a line that indicates the maximum safe operating sash height. The sash will not be lowered below a reasonable working height (usually 20 inches). Instead, an increase in airflow is recommended.
Once the face velocity measurements are completed, the containment tests are conducted on the hood with smoke sticks. The hood face is traversed with a smoke stick to observe the air flow patterns. No back flows that result in release of smoke from the hood are permitted. If they exist the hood is not updated and goes on the list of uncertified hoods with a description of what IHP believes is causing the containment problem.
The type of hood and the physical condition is noted on the hood worksheet. If parts of the fume hood are missing such as air foils or side panels this is noted. Removal of air foils usually produces a hood with unacceptable containment.
Biological safety cabinets are also surveyed on an annual basis if they are vented to the outside. This survey also consists of a face velocity measurement and a smoke test and an updating of the IHP biosafety cabinet label. It is not the NIH biosafety cabinet Certification that requires an aerosol challenge of the HEPA filters. The IHP recommends that biosafety cabinets be certified on an annual basis and whenever moved. It is the responsibility of the Chemical Hygiene Officer to see that they be done as required.
If there is any question about a fume hoods operation the IHP should be called immediately. When a new fume hood is installed, it is the responsibility of the Chemical Hygiene Officer to see that no hazardous substances are used in the hood until it is surveyed and labeled by IHP. If any changes of any kind are made to the fume hood system, IHP should be notified so a hood survey can be conducted.
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Standard Operating Procedure For RADIOACTIVE MATERIALS (currently, no such SOP is in use at BPEC)
Biotechnology Process Engineering Center (Date) Identification Code: ___________ __________________________ Revised: ___________ (CHO Signature) ****************************************************************************** I. Description of Hazardous Chemical Classification
Radioactive materials are any materials that spontaneously emit alpha, beta or gamma ray by disintegration of its nuclei of atoms.
II. Required Procedures Before Use A. Obtain training and authorization from the radiation protection office. 1. Know amounts, types, and forms (liquid, solid) of isotopes to be used. 2. Know type of procedure and equipment to be used. 3. Get appointment, attend session, etc. B. Obtain specific training in laboratory from radiation safety officer or experienced worker. 1. Learn locations of disposal container, Geiger counters, shielding, storage
facilities. 2. Learn rules of courtesy, sign-up and clean-up forms, sign-out of equipment 3. Get training on specific equipment such as gamma counter. C. Begin actual work: 1. Check Geiger counter[exposure meter] for batteries, and proper setting. 2. Check work area for previous contamination with Geiger counter. 3. Minimize clutter near and on work area: avoid spill hazards, contamination of
apparatus carelessly placed in the way. 4. Replace blotting paper with multiple layers if necessary. III. Special Ordering Procedures A. Write the radiation authorization number [10-G-2] and the amount and type of isotope to
be procured on the requisition form. B. Obtain a radioactive purchase order number from the MIT Purchasing Office. These PO
numbers are processed late in the day, so the PO number cannot be obtained just by walking the requisition form over to the office.
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IV. Protective Equipment A. Clothing. 1. Lab coat at all times. 2. Double-gloves, vinyl or latex, to protect against skin exposure. 3. Eye protection; splash-resistant glasses or goggles. B. Shielding. 1. Shield larger quantities of radioactive material in appropriate container (lead pig,
etc.). 2. Use shielding barriers (lead sheets, or plexiglass face shield) to protect from
exposure from materials that are not well contained. C. Geiger counters. Periodically monitor effectiveness of shielding, check for contaminations. D. Exposure badges. If participating in the badge program, always use badges (ring badge for hand
exposure, clip badge for body exposure.) (See below for exposure criteria). V. Precautions A. Avoid direct contact or ingestion with any radioactive materials. B. Avoid and minimize spill danger by: 1. Minimizing clutter. 2. Aliquoting radioactive materials to several smaller amounts. C. Minimize exposure by: 1. Minimizing exposure time. 2. Maximizing distance from body (or body parts) to source. 3. Maximizing shielding. D. Other general precautions for the BL2 lab apply here. E. Whenever possible, perform a trial run with low activity materials to ensure the adequacy
of procedures and equipment. VI. Handling A. Use designated pipettes only. B. Eject pipette tips into a disposable container (plastic bottle, large T flask, etc.) to
consolidate solid waste. C. Work with small amounts of isotope at a time if possible to minimize exposure and spill
potential.
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VII. Exposure Limits (mrem) Weekly Quarterly Annual A. Whole body: 100 250 5000 B. Skin (beta) 575 7500 30000 C. Extremities: 1450 18750 75000
Each worker who may receive in excess of 25% of these amounts should be provided with radiation badges by the RPP.
Note: Beta particles with less than 70 keV will not penetrate the skin. Since the maximum energy from tritium is 18.6 keV, exposure is not a problem (although ingestion is still dangerous). Other common isotopes are of more concern. 14C and 35S put out beta particles at up to 167 keV. 32P up to 1700 keV. 125I emits gamma rays at only 35 keV, but gamma rays penetrate light barriers such as skin. Fortunately, 125I at 1 microcurie can be shielded with a few mm of lead or with lead-lined plexiglass.
VIII. Disposal
All materials (liquid, solid, sharps) should be disposed in the appropriate waste containers provided by the RPP. There is a solids waste can in 16-436 that is used for spent containers, gloves, contaminated paper, pipette tips, and any other disposable potentially contaminated solids. There is one liquid waste container, in 16-436. All of these containers have sign-out cards attached. These should be filled out after each use, to indicate the name of the worker, the date, the isotope, and the amount disposed. When containers are 2/3 full, the worker should contact the RPP for disposal.
A. The total amount of radioactivity in a waste container must be controlled such that
exposures do not exceed 2 mrem/hr at 1 foot from the container. B. Avoid reactive mixtures in liquid waste containers. Adjust pH to 4 to 10 prior to
disposal. C. Biohazardous, carcinogenic, or toxic material shall not be disposed without prior
authorization from the RPP. IX. Spills/Accidents
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A. Routine spills: When the radioactive lab bench in 16-436 is used it should always be fitted with multiple layers of absorbent, plastic backed blotter paper. In the event of a spill in this area, the contaminated area of blotter should be cut out and disposed in the solid waste container, taking care to avoid cross contamination of the worker or other equipment. If non-disposable surfaces (floors, equipment) are contaminated, they should be carefully cleaned with count-off and disposable tissues. The area should be checked for residual contamination using the Geiger counters after the cleanup.
B. Emergency Spills: The RPP should be notified in the event of accidental release of
radioactive materials to the laboratory surfaces, drains, or ventilation system; in the event of ingestion of radioactive materials, or in the event of exposure to levels of radioactivity which exceed the quarterly limits described here. In the event of ingestion of radioactive materials, the RPP will perform bioassay tests.
NOTIFY RPP AT 3-2180 OR 3-2360; CALL 100 AFTER HOURS. REPORT RADIATION CONTAMINATION, ROOM, INJURIES, YOUR NAME
AND EXTENSION. Special procedure for containing spill: 1. Notify RPP or emergency (100). 2. Rope off or guard spill area against re-entry. 3. If the spill was of a powdered, volatile, or gaseous material, evacuate all
personnel, turning off critical apparatus if necessary. Close and post laboratory doors to prevent reentry.
4. Assemble potentially contaminated persons in one location in or near the laboratory, monitor for contamination, and detain them to prevent spread.
5. Wait for RPP personnel to arrive. X. Storage and Labeling A. All radioactive materials should be stored in designated areas (the work bench or in
designated refrigerators). B. The materials should be adequately shielded to minimize exposure to persons/chemicals in
the laboratory. C. All storage containers/facilities should provide adequate protection against fire,
explosion, or flooding; against accidental breakage; and against unauthorized removal. D. All materials should be labeled with a special radioactive substance label (yellow/orange
with radiation symbol and "CAUTION RADIOACTIVE MATERIAL"). The label should contain the name of worker, type and amount of isotope, form of isotope, date of receipt, and date of expiration. Expired materials should be disposed of promptly, and not accumulated in the storage area.
XI. Transportation A. Radioactive substances must be transported with appropriate shielding, to minimize
exposure to others and in a properly labeled, shatter-proof container.
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1. Shielding must reduce exposure to less than 50 mrem/hour at the container surface, or 2 mrem/hour at 3 feet from the container.
2. There should be no detectable contamination on the surface of the container as measured by a wipe test.
3. Only authorized persons may transport the material. B. Pedestrian transport of radioactive materials outside of the MIT complex is not allowed
without the permission and approval of the RPP. C. For further regulations regarding transport, refer to the MIT Required Procedures for
Radiation Protection. XII. Housekeeping After Use
The laboratory area must be cleaned and screened after use of radioactive materials. Equipment and waste containers must be stowed neatly. Blotter paper should be replaced if torn or soiled. Waste container sign-outs must be completed. The gamma counter log should be filled out indicating background counts before and after use, and the number of samples measured. Finally, the Geiger counters should be used to ensure lack of contamination of work area or tools, and the counter should be turned off to conserve batteries.
XIII. Personal Hygiene After Use
Gloves should be disposed in the radioactive waste container. Hands should be washed thoroughly. Exposed skin, hair and clothing should be surveyed for contamination. The RPP should be notified immediately if residual contamination is detected. Radiation badges should be stored in a safe place with only background levels of radioactivity and should be returned to the RPP at regular intervals.
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Section 8-10 Institute Safety Manual POLICY ON THE IDENTIFICATION AND DISPOSITION OF CHEMICAL, BIOLOGICAL, AND RADIOACTIVE SUBSTANCES (IN LABORATORIES OR OTHER WORK AREAS) Purpose: To assure that persons working with chemical, biological or radioactive substances properly label all containers of such substances. Requirements: Contents of all containers or apparatus containing or contaminated with such substances be identified by chemical name. All chemicals must be identified with the chemical name, not symbols or abbreviations. Enforcement: Supervisors, advisors, or other persons responsible for organizing and directing work are required to enforce compliance with the provisions of this policy by all persons whom they supervise. The supervisor of any person who is to vacate the laboratory (or other work area) should first arrange for the proper disposition of all chemical, biological, and radioactive substances. The supervisor shall require that all substances be identified, containerized and labeled before releasing or reassigning the laboratory or work area to the next occupant. Disposal: Unwanted substances not requiring inactivation in the laboratory shall be disposed of through existing Institute disposal procedures outlined in the MIT Accident Prevention Guide published by the Safety Program and in Part III subpart O (Waste Disposal) of this plan. Packaging, labeling, and disposal of radioactive materials are handled by procedures established by the Radiation Protection Program (3-2180). Publicity and Monitoring: Departmental Safety Committees will post, publish, circulate or otherwise announce annually the requirements of this policy to all affected personnel within the department. Departmental Safety Coordinators in their normal day-to-day activities will check for compliance with this policy and report to their supervisor violations in their areas of responsibility. Material Safety Data Sheets (MSDS): Material Safety Data Sheets, (MSDS) are bulletins prepared by manufacturers to summarize the health and safety information about their products. The Safety Program Web site provides resources to obtain MSDS.
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Material Safety Data Sheets come in many formats and present the information in different ways. Regardless of the format, the information that is required by OSHA includes: Product Identity Hazardous Ingredients Physical/Chemical Properties Special Precautions Protective Equipment Health Hazards Reactivity Hazards Fire and Explosion Hazards Spill Cleanup A User's Guide to Material Safety Data Sheets is available from the Safety Program. Consult with the Safety Program and the Industrial Hygiene Program to apply this general information to your work situation.
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