Standard Operating Procedures

Working with MPTP, MPTP-treated Animals and MPTP Waste Products

Last updated: March 12, 2012

I. Introduction

MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) is a neurotoxin used to induce parkinsonism symptoms in experimental models of Parkinson’s disease (PD). MPTP can produce irreversible neurological damage indistinguishable from PD in humans and animals.  The inappropriate handling of MPTP may result in exposure and irreversible neurological damage to research staff.

MPTP is lipophilic which freely and rapidly crosses the blood-brain barrier. It is metabolized to MPDP+ which is oxidized to the active metabolite MPP+. Excretion of unmetabolized MPTP occurs mainly during first day post injection while MPTP metabolites are excreted up to 3 days post injection. Excretion is mainly in the urine and some has been found in bile. MPTP in urine is expected to be ionized and not volatile and be absorbed by the bedding.

Exposure to staff may occur via needlesticks and inhalation of vapors from MPTP solutions, or inhalation of aerosols from MPTP contaminated bedding. Exposure potential is greatest during preparation, administration, and in the bedding during the first 3 days after injection. The high risk period will be from preparation of MPTP solutions to 3 days post administration after the first cage change.

The purpose of this SOP is to help ensure the safe work practices and appropriate medical surveillance for people working with MPTP

II. Prior to beginning work with MPTP (All of the following items are required for employees who plan to work with MPTP or enter MPTP work areas up to 3 days post administration)

• Medical Management .
  • Prior to initiating work with MPTP, the Principal Investigator (PI) and his/her staff must contact the Occupational Health Physician, Dr. Dana Headapohl, at 406-329-5746  to schedule an appointment for evaluation and to receive the post-exposure prophylaxis (PEP) of selegiline hydrochloride (Eldepryl).  The Occupational Health Physician will prescribe the selegiline.
  • Selegiline is obtained prior to beginning work with MPTP. It must be kept in the white capped container labeled “FOR MPTP USE ONLY” and readily accessible to those working with MPTP. It must be regularly checked to ensure it is in its proper place and the use date has not expired.
• Respiratory Protection
  1. N-95 espirators are required when working with MPTP- treated animals. You MUST be enrolled the University’s Respiratory Protection Program prior to wearing a respirator. Contact EH&RM to initiate this process
• Employee Training
  
  1. The PI must train his/her staff on this standard operating procedure (SOP) and hazards of MPTP prior to beginning work. This training must be documented.

III. Accidents, Injuries, and Potential Exposures

• In the event of a recognized percutaneous or mucous membrane exposure to MPTP:
  • Immediately ingest four selegiline HCl 5 mg capsules as directed by the Occupational Health Physician
  • Initiate first aid at the worksite:
    • Wash contaminated skin with soap and water for 15 minutes.
    • Irrigate contaminated eyes and mucous membranes should be irrigated for 15 minutes, using normal saline or water.
  • Quickly contact Curry Health Center at 243-2122 between 8:00 a.m. and 5:00 p.m. M-F. If Curry Health Center is closed or not available, go directly to Saint Patrick Emergency Room (ER). Be sure to disclose your name, your exact location, and that you were exposed to MPTP.
  • Notify the PI at as soon as possible.
  • Report the incident on the UM Online Accident Reporting System at: http://www.umt.edu/research/eh/workerscomp/default.aspx
• In the event of a possible exposure to MPTP
  • Immediately initiate first aid at the worksite as outline above.
  • Quickly contact Curry Health Center at 243-2122 between 8:00 a.m. and 5:00 p.m. M-F. If Curry Health Center is closed or not available, go directly to Saint Patrick Emergency Room (ER). Be sure to disclose your name, your exact location, and that you were exposed to MPTP.
  • Notify the PI at as soon as possible
  • Report the incident on the UM Online Accident Reporting System at: http://www.umt.edu/research/eh/workerscomp/default.aspx
  • In the event of a spill outside of biosafety cabinet, evacuate people from the room and close off the area. Contact EH&RM at 243-2881

IV. Personal Protective Equipment

• Personal protective equipment must be worn whenever working with MPTP or MPTP-treated animals and their waste within 3 days of last administration. These activities include (1) preparing MPTP solutions, (2) administering MPTP to animals, (3) entering MPTP-animal room within first 3 days of administration, and (4) first cage change (must be no earlier than 3 days after last administration.
  • When preparing MPTP solutions, you must:
    • Work in a certified biosafety cabinet.
    • Wear a disposable jumpsuit or disposable lab coat
    • Wear two pairs of disposable gloves.
    • Wear chemical safety goggles.
    • Wear an N-95 disposable mask (respirator).
  • Cage changing: Must be done no earlier than 3 days after last administration. If an emergency occurs and cages must be changed within 3 days of last administration, use the above PPE
  • Frequent glove changes are recommended. Change gloves immediately if their integrity has been compromised (e.g., soiled, punctured, torn)
  • Protective clothing must be disposed of into biohazard bags before exiting SB028
  • PPE must be removed in a specific order to prevent contamination.  When removing PPE,
    • Remove outer gloves.
    • Removeisposable jumpsuit.
    • Remove chemical goggles,
    • Remove N-95 respirator
    • Remove inner pair of gloves
    • Put on a new pair of gloves, spray goggles, with 1% bleach (wipe dry)
    • Remove the new gloves, wash hands, remove booties and exit SB028.

V. MPTP Access Restrictions and PPE

VI. MPTP Preparation

• Order PRE-WEIGHED MPTP from the manufacturer.   MPTP must be purchased in hydrochloride or tartrate salt form rather than as a free base.
• MPTP and MPTP-solutions must be stored in labeled, tightly capped containers. The container label must include the identity of the hazardous contents (i.e., MPTP) and the appropriate hazard warning (i.e., neurotoxin).  The primary container for MPTP must be placed in a sealed, leakproof, unbreakable secondary container, which must also be labeled as above.
• MPTP, as received from the manufacturer/vendor in its original undiluted form, must be stored in a clutter free certified biosafety cabinet in a labeled, sealed, leakproof secondary container. The BSC must be posted with the red MPTP sign in Appendix 1.
• During preparation and administration of MPTP:  ALL of Room SB028 is restricted to only those staff authorized to work with MPTP
• The room where the MPTP biosafety cabinet is located is restricted to staff authorized to work with MPTP and MPTP-treated animals for a minimum of 3 days after last MPTP administration. All staff working with MPTP and MPTP-treated animals must follow all of the steps in the SOP.  PPE must be removed prior to exiting the room
• If emergency maintenance must be done during the first 3 days after the injection, stop all work and decontaminate the area. Ensure that all concentrated and dilute concentrations of MPTP are closed and properly stored. Please escort the worker into the space and remain in the room during the entire maintenance. The sign should remain on the door and biosafety cabinet to inform other staff of the MPTP and MPTP treated animals. After the 3 days post injection, no extra PPE is required in this space and any requirements for escorts will be determined by lab staff
• MPTP solutions must be prepared in a certified biosafety cabinet. Appropriate personal protective equipment (PPE) must be worn, as noted in Section III.  All work surfaces must be covered with absorbent, plastic-backed, disposable absorbent pads
• The red MPTP sign in Appendix 2 must be posted on door to SB028 for a minimum of 3 days after last MPTP administration
• Disposable labware should be used when preparing MPTP solutions.
• When all work with MPTP is complete, carefully remove all absorbent pads and dispose into red bags.  Wipe all surfaces with a 1% bleach solution and discard of all wipes and garbage into red biohazard bags (up to 3 days following the last injection of MPTP).

VII. Administration of MPTP to Animals

• All MPTP animal work must be conducted in SB028.
• Animals must be chemically or physically restrained prior to starting the MPTP administration.
• Administration of MPTP to rodents must be conducted in a certified biosafety cabinet (BSC).  Both the personnel administering the MPTP and those in the immediate vicinity of the procedure must wear appropriate PPE.
• All work surfaces must be covered with absorbent, plastic-backed, disposable absorbent pads.
• Use only needle-locking syringes or disposable syringe-needle units (i.e., needle is integral to the syringe). Used disposable needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated before disposal
• After each injection, immediately place the syringe-needle unit in a sharps disposal container located in the biosafety cabinet.

VIII. Animal Housing

• House MPTP-treated animals in SB028 and keep the door locked at all times.
• For the first 3 days post-injection, the door to SB028 must be posted in such a manner that it is clear that the biosafety cabinet contains MPTP-treated animals. The BSC may only be used for the MPTP work (preparing, administering MPTP and housing animals)
• The animal cages must also be properly labeled
• For the first 3 days post-injection, keep the cages covered with microisolator lids. Ensure the lids are secured to the cage
• Cage changes should not be performed for the first 3 days post-injection
• 3 days after the final MPTP injection, the animals may be transferred to clean, standard caging and the MPTP door signs and cage tags may be removed) 
  
  •  Cage change/dumping should be performed wearing appropriate PPE (section III) and in a BSC.
  • Work atop absorbent pads
  • All bedding should be disposed of into red biohazard bags that are frozen in SB028 for disposal by EH&RM
  • Dirty cages and racks should be wiped down with 1% bleach and moved to the cage wash for immediate cleaning
  • Reusable water bottles can be sprayed with 1% bleach and processed same as other water bottles The use of special PPE is not required after the animals have been transferred to clean cages (beyond 3 days post-injection

•  VIII. MPTP Waste

  • Unused concentrated reagent (including empty vials) is managed as hazardous waste through EHRM.  See. http://www.umt.edu/research/eh/materials/hazmat.aspximes 
  • Spill cleanup materials, soiled PPE, and absorbent pads contaminated with concentrated reagent should be double bagged and managed as hazardous waste through EHRM
  • All waste generated up to and including the first bedding change must be double bagged and managed as hazardous waste through EHRM
  • Animal bedding
    • Bedding from the first change (3 days after last MPTP administration) must be double bagged and managed as hazardous waste through EHRM
    • Bedding from the first change (3 days after last MPTP administration) must be double bagged and managed as hazardous waste through EHRM
  • Bedding from subsequent cage changes can be discarded through the regular waste stream
    • Animal Carcasses Carcasses that have received MPTP within the last 3 days must be double bagged and managed as hazardous waste through EHRM
    • Carcasses that have not received MPTP in the past 3 days can be discarded through the regular waste stream