THOMAS JEFFERSON UNIVERSITY
ANIMAL RESOURCES
STANDARD OPERATING PROCEDURES
Section
Page
I.D.10
1 of 6
Title:
Date Issued / Revised
Safe Practices Using the Neurotoxin MPTP in the
11/12/1997
Animal Facility
Purpose:
8/18/08
5/10/11
5/28/13_____
To establish safe practices using the neurotoxin MPTP in the animal facility.
Introduction: MPTP (1-methyl-4-phenyl-1,2,3,6 - tetrahydropyridine) can induce a permanent
Parkinsonian syndrome in humans. Handling of MPTP and MPTP - treated
animals has become an important issue for all personnel working with this
neurotoxin (1).
Responsibility:
All staff engaged in the use or handling of MPTP, or working within a laboratory using MPTP,
are responsible for understanding all hazards associated with its use, and for using appropriate
personal protective equipment (PPE).
The Principle Investigator (PI) is responsible for ensuring that his/her staff has been trained in the
use, storage and handling of MPTP. Appropriate training documentation must be maintained.
Use of MPTP is restricted to the specific staff members who have had training in proper use,
handling and storage. Documentation of this education must be kept by the PI in the laboratory
personnel files as well as with the Biological Safety Officer.
Administrative Approval
Prior to the use of MPTP in animals, an IBC-14 form must be completed and approved by the
Laboratory Safety Committee.
Training:
Prior to conducting any work with MPTP, the PI must provide training to his/her laboratory
personnel regarding the hazards involved in working with this substance, proper
transport/handling, decontamination, and emergency procedures.
The PI must provide his/her laboratory personnel with a copy of this SOP, the MPTP MSDS, and
the selegiline hydrochloride drug information sheet.
Annual training regarding the emergency treatment of exposures will be done in University
Health Services (UHS) at the time of the annual health screening. This will include the location
of the medication, the first aid measures to follow and the appropriate steps to follow in reporting
an exposure to UHS or to the TJUH Emergency Department. Annual in-services for all personnel
will cover the current policies and procedures to follow for exposures.
THOMAS JEFFERSON UNIVERSITY
ANIMAL RESOURCES
STANDARD OPERATING PROCEDURES
Section
Page
I.D.10
2 of 6
Title:
Date Issued / Revised
Safe Practices Using the Neurotoxin MPTP in the
Animal Facility
11/12/1997
8/18/08
5/10/11
5/28/13
Medical Evaluation
All laboratory personnel involved with the handling/storage/administration of MPTP must be
medically evaluated prior to working with MPTP and annually thereafter in UHS. As a
component of the evaluation process, fit-testing for N-95 respirators must be performed.
Administration to Animals
MPTP is used to create an animal model of Parkinsonism. Solutions are prepared by adding
sterile saline to a sealed vial of MPTP powder and then diluting this to a stock solution of 20
mg/cc. Alternately, MPTP may be received as a liquid stock solution. This stock solution is then
diluted to working concentration determined by the particular study (i.e., animal species to be
used). Solutions are administered to animals as i.m. injections (monkeys), i.v. injections
(monkeys) or subcutaneous injections (mice).
Injection of animals with MPTP is conducted with appropriate restraint of each species to
minimize the possibility of accidents. MPTP is diluted inside of a chemical fume hood or in a
Type II B1 or Type II B2 Biosafety Cabinet. Syringes are also loaded in these hoods. If a hood
of the correct type is not available in the animal facility, then the syringes must be loaded in the
lab and transported to the animal facility. When loaded syringes are transported to the animal
facility, they are placed in a tray and covered securely for transport (such as in a closed
Tupperware container) to the animal rooms. Safety syringes are used for injections. If safety
syringes are not used, documentation of the reason for the exception is required.
After injections are made, the syringe is placed directly into a sharps container that is labeled for
incineration. If any MPTP remains in the syringe, an equal volume of 1.0% bleach solution in
water is drawn up into the syringe before it is placed in the sharps container. Needles should not
be recapped or removed from the syringe. Any contaminated gauze should be disposed of in the
red waste bag system and marked for incineration.
The investigator is responsible for identifying each animal which receives MPTP. This will be
done by marking the cage card with the name of the compound (MPTP). The cage card labeled
MPTP will be removed by the investigator 72 hrs after final administration of MPTP.
Bleach Instructions
Common household bleach contains 5.25% sodium hypochlorite. A 1.0% bleach solution in
water with 5.25% sodium hypochlorite consists of 1 part bleach to 4 parts water (1 in 5 dilution).
Thus 20 ml of bleach can be mixed with 80 ml of water OR 10 ml of bleach can be mixed with 40
ml of water. A new 1.0% bleach solution must be made on a weekly basis.
THOMAS JEFFERSON UNIVERSITY
ANIMAL RESOURCES
STANDARD OPERATING PROCEDURES
Section
Page
I.D.10
3 of 6
Title:
Date Issued / Revised
Safe Practices Using the Neurotoxin MPTP in the
Animal Facility
11/12/1997
8/18/08
5/10/11
5/28/13
Intramuscular Injections
Monkeys - Cage backs are pulled forward and monkeys are securely squeezed against front wall
of cage for injection into the thigh muscle.
Intravenous Injections
Monkeys – For intravenous injections, light gas anesthesia (i.e.1-2% isoflurane) is used prior to
injection. Injections are made into the saphenous vein.
Subcutaneous Injections
Rodents-mice are held securely in one hand and against a tabletop while injected subcutaneously.
Prior to injection, a disposable protective barrier must be placed on the counter. After the
injection is completed, the barrier must be placed in a biohazard red bag and marked for
incineration. As an alternative, the mice can be held over the cage during the injection process.
Any feces or urine produced by the mouse will then fall into the cage and can be disposed of
during the cage changing process.
Personal Protective Equipment and Practices
Preparation of MPTP must be performed in a chemical fume hood or a Type II B1 or Type II B2
Biosafety Cabinet by personnel wearing double nitrile gloves, N-95 respirator, eyeshield and
disposable gown. All personnel injecting animals or working in rooms containing primates that
have received MPTP within the last 72 hours must wear nitrile gloves, N-95 respirator, eyeshield
and disposable foot covers and gown. Double gloving is recommended. Wearing of such
personal protective equipment in rooms housing rodents exposed to MPTP is not required except
when injecting MPTP, handling the cages or animals, because of the minimal exposure risk.
Chronic exposure to MPTP could cause Parkinsonism. Acute exposure to high doses of MPTP
by ingestion, inhalation, or injection could cause a rapid onset Parkinsonism. There are no
specific signs other than the development of Parkinsonism that would signal a break in technique.
THOMAS JEFFERSON UNIVERSITY
ANIMAL RESOURCES
STANDARD OPERATING PROCEDURES
Section
Page
I.D.10
4 of 6
Title:
Date Issued / Revised
Safe Practices Using the Neurotoxin MPTP in the
Animal Facility
11/12/1997
8/18/08
5/10/11
5/28/13
There is the possibility that minute quantities of unmetabolized MPTP could be excreted in urine,
feces or the respiratory process. Mice should be housed in cages fitted with wire bottoms to keep
them away from contaminated bedding material. Unmetabolized MPTP will be ionized and not
volatile and well-absorbed by the bedding. Unmetabolized MPTP has been shown to be less than
2% of the injected dose in large animals. Unmetabolized MPTP is not considered to be vapor
borne. Precautions should be taken to minimize contact with animal fur and excreta and avoid
inhalation of bedding dust. Gloves should be changed frequently using the appropriate technique
for de-gloving.
For spilled material, it will be blotted up immediately by personnel wearing protective gear as
described above, transferred to a hood, and treated with 1.0% bleach prior to disposal in a red
waste bag. A supply of selegiline will be kept in the laboratory and all personnel will have
access to this. Selegiline will also be available in the Jefferson Alumni Hall animal facility.
Supervisors and the veterinarians will have access to the selegiline. Immediate first aid measures
will be initiated in the department (15 minute lavage of any exposed mucosal surfaces, 15
minutes washing with soap and water of any wound). A supervisor will be contacted who will
then call USH (5-6835) or TJUH Emergency Department (5-6840) to advise that an exposure has
occurred. The supervisor will also contact one of the attending veterinarians for further
instructions in regards to Selegiline administration for the animal care staff. The veterinarian will
contact Dr. Ellen O’Connor, MD for permission to dispense Selegilline. The veterinarian or Dr.
Ellen O’Connor will then report back to the supervisor with instructions on the use of Selegiline.
The conversations will be documented and placed in employee’s file. Workers with exposure to
MPTP by needlestick, cutaneous contact, or inhalation may be requested to immediately take a
dose of selegiline kept in the department prior to contacting UHS or the TJUH Emergency
Department. The worker will be sent over immediately for evaluation of continued treatment
with selegiline. Since Selegiline is an MAO inhibitor, UHS will discuss drug/food interactions as
part of the evaluation. Each lab working with MPTP has an appointed chemical hygiene officer
who is fully versed in all laboratory procedures.
Animal Housing and Decontamination Procedures
Rodents - Mice and other rodents are housed in microisolator cages fitted with wire bottoms to
keep them away from contaminated bedding material. Rodents are transferred to fresh cages 3
days after the last MPTP injection. The cage and bedding sprayed with 1.0% bleach and allowed
to sit for 10 minutes prior to dumping the bedding in a red waste bag for incineration. The cage is
rinsed and washed in the cage or tunnel washer.
THOMAS JEFFERSON UNIVERSITY
ANIMAL RESOURCES
STANDARD OPERATING PROCEDURES
Section
Page
I.D.10
5 of 6
Title:
Date Issued / Revised
Safe Practices Using the Neurotoxin MPTP in the
Animal Facility
11/12/1997
8/18/08
5/10/11
5/28/13
Monkeys - Monkeys are housed in stainless steel primate squeeze cages. Because of the small
amount of MPTP given to monkeys, there is virtually no chance of any unmetabolized MPTP
being excreted. However, pans under monkey cages should be sprayed with 1.0% bleach solution
after MPTP is administered and after a 10 minute waiting period, waste material flushed into the
floor drain making sure to minimize aerosols.
Identification of Hazard
Appropriate signs with instructions for special entrance requirements will be posted on the door
to the animal room. A chemical hazard sign will be posted identifying MPTP as the agent in use.
Individual animal cage cards will be marked indicating animals receiving MPTP or those which
may be hazard. The facility color coded dot system may be used. A copy of this SOP will be
available in each MPTP room.
Waste Disposal
Liquid waste material is destroyed by adding bleach to a final concentration of 1.0% and then
labeled as MPTP waste and disposed of as hazard chemicals through Environmental Health and
Safety. Decontamination of cages will be accordance with the standard operating procedures
described in this document. All waste disposal such as dumping contaminated bedding should be
done in a ventilated waste disposal cabinet. Syringes and sharps are to be discarded in a sharps
container labeled for incineration. Animal cadavers are to be placed in biohazard red bags and
marked for incineration. If an animal dies or is to be necropsied prior to 72 hours after MPTP
administration, the individual handling the animal must wear full personal protective equipment
as previously described. Any instruments used in the process must be treated with 1.0% bleach
prior to further cleaning procedures.
THOMAS JEFFERSON UNIVERSITY
ANIMAL RESOURCES
STANDARD OPERATING PROCEDURES
Section
Page
I.D.10
6 of 6
Title:
Date Issued / Revised
Safe Practices Using the Neurotoxin MPTP in the
Animal Facility
11/12/1997
8/18/08
5/10/11
Contact Information:
Dr. Ellen M. O’Connor, MD
Cell: (215) 219-2808
Home: (610) 649-4090
Department of Environmental Health and Safety
(215) 955-7233
Biological Safety Officer
(215) 503-7422
Emergency Spill
811
University Health Services
(215) 955-6835
Department of Emergency Medicine
(215) 955-6840
References
1. Przedborski, S., Jackson-Lewis, V., Naini, A.B., Jakowec, Petzinger, G., and Akram, M.
The Parkinsonian Toxin 1-Methyl-4-Phenyl-1,2,3,6-tetrahydropyridine (MPTP): a
Technical Review of its Utility and Safety, Journal of Neurochemistry, Vol 76, Pp. 12651274, 2001
2. Journal of Neurochemistry, Vol 76, Pp. 1265-1274, 2001
3. Yang S.C., Markey S.P., Bankiewicz K.S., London W.T., and Lunn G.;
4. Recommended Safe Practices for using the Neurotoxin MPTP in Animal Experiments,
Lab Animal Science, Vol. 38, No. 5, October 1988
5. IBC-14 - Hazardous Chemicals and Toxins Form, Thomas Jefferson University
6. Safe Work Practices for the use of MPTP in the Laboratory and Vivarium, UMDNJ,
Environmental and Occupational Health and Safety Services, April 16, 2004.
7. Procedures for Working with MPTP or MPTP-Treated Animals, Division of Occupation
Health and Safety (DOHS), National Institute of Health
8. MD Consult: Selegiline
9. MPTP MSDS
Signatures:
Reviewed by:
Name
Title
Date
Name
Title
Date
Approved by: