Working With MPTP, MPTP-Treated Animals and MPTP Waste Products


Revised December 15, 2015


I.     Purpose

The purpose of this standard operating procedure is to outline the proper procedures for handling the neurotoxin, 1-methy-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP).  MPTP produces neurochemical deficiencies similar to the ones present in Parkinson’s disease.  This procedure describes safety precautions required for preparing MPTP, injecting MPTP into mice, working in animal rooms where MPTP is present, changing dirty MPTP cages, and disposing of MPTP-contaminated waste.

II.    Prior to beginning work with MPTP

A.    Keep a prescription of the drug selegiline hydrochloride (Eldepryl) readily available to those working with MPTP in the event of an accidental exposure (Principal Investigator must acquire a prescription from the OH&S physician).

1.    Keep the selegiline in a secure location which is known and readily accessible to those working with MPTP.

2.    Check the selegiline regularly to ensure it is in its proper place and the use date has not expired.

B.   Obtain medical clearance for wearing a respirator from the occupational health physician before a fit-test for a N-95 respirator is done by Environmental Health and Risk Management (x2881). This includes all personnel planning to work with MPTP, including Laboratory Animal Resources (LAR) Staff. 

III.   Accidents, Injuries, and Potential Exposures

A.    In the event of a percutaneous or mucous membrane exposure to MPTP:

1.    Immediately ingest four selegiline HCl 5 mg capsules

2.    Initiate first aid at the worksite:

a. Wash contaminated skin with soap and water for 15 minutes.

b. Irrigate contaminated eyes and mucous membranes for 15 minutes, using normal saline or water.

3.    Notify the Principal Investigator, a LAR staff member, and the Director of LAR as soon as possible.

4.    Call Environmental Health and Risk Management (x2881).

B.   In the event of a spill, evacuate people from the immediate area.

1.    Contact Environmental Health and Risk Management (x2881).

2.    Contact Campus Security (x6131) 

IV. Personal Protective Equipment and Respiratory Protection

A.    During the first 7 days of MPTP usage all personnel must wear personal protective equipment when working with MPTP or MPTP-treated animals and their waste, or when entering SB 028.

    1. Preparing MPTP solutions

a.  Working in a certified chemical fume hood

      • Wear two pair disposable lab gloves
      • disposable jumpsuit
      • safety glasses and
      • N95 mask (respirator)

 b. Working in a BSL-2 cabinet (if the MPTP bottle contents will be rehydrated by inserting a syringe through the rubber stopper a BSL2 may be used)

      • wear two pairs of disposable lab gloves
      • disposable jumpsuit
      • safety glasses N-95 mask (respirator)

2.    Administering MPTP and working with treated animals or with caging in SB 028.

a. Perform all work in the BSL-2 cabinet in SB 028

      • Wear two pairs of disposable lab gloves
      • disposable jumpsuit
      • safety glasses
      • N-95 mask (respirator).

3.    Performing routine tasks only and NOT working with MPTP, animals or caging material in SB 028.

a. disposable lab gloves

b. lab coat

c. N-95 mask (respirator)

B.   After the 7-day treatment with MPTP

1.    Personnel handling MPTP-treated animals, caging and bedding in BSL-2 cabinet in SB 028

a.    two pairs of disposable lab gloves

b.   disposable jumpsuit

c.    safety glasses

d.   N-95 mask (respirator)

2,    All personnel entering SB 028 for routine tasks must wear at the minimum:

a.   one pair of disposable lab gloves

b.   lab coat

C.   Change gloves frequently.

1.    Change gloves immediately if their integrity has been compromised.

2.    Wash hands with soap and water after removing gloves.

D.   Remove protective clothing immediately prior to exiting a room in which MPTP is in use.

1.    Remove garments first, then remove gloves, safety glasses, and mask (respirator), respectively.

2.    Place N95 mask (respirator), gloves and garments in a red biohazard bag located in the room.  Label bags (date and MPTP), tie bags closed.

3.    Wash safety glasses with a mild detergent and water and thoroughly wash hands.

4.    Take MPTP-labeled and dated biohazard bag to the hazardous waste disposal freezer in HSB 015A.  Environmental Health and Risk Management will be notified by LAR to pick up the waste. 

V.   Training and Notification

A.   The PI must train his/her staff on proper procedures for safely working with MPTP, MPTP-treated animals and MPTP caging materials prior to beginning work.

B.   72 hours prior to administering MPTP to animals, the PI must inform the LAR Facility Manager (Kelly Carrick, x4892) that an MPTP animal experiment is pending and the PI will provide:.

1.  The dates of the experiment

2     The number of animal cages to be used

3.    The names of personnel to be involved

4.    Assurance that all personnel have been properly trained

5.    Assurance that all personnel have read and understand the SOP and AUP

VI. MPTP Preparation

A.   Purchase MPTP in hydrochloride or tartrate salt form rather than as a free base.

B.   Store MPTP and MPTP-solutions in labeled, tightly capped containers.

1.    Label the container with the identity of the hazardous contents.

2.    Place the primary container for MPTP in a sealed, leakproof, unbreakable secondary container, which must also be labeled as described above.

C.   Prepare MPTP solutions.

1.    If weighing out MPTP, weigh and prepare solutions in a certified chemical fume hood (in investigator’s laboratory)

a. Wear appropriate personal protective equipment (see section IV).

b. Cover all work surfaces with absorbent, plastic-backed, disposable bench paper.

c, Use disposable labware when preparing MPTP solutions.

d. Collect and dispose of MPTP solutions as chemical waste.

2.    If diluting MPTP directly in bottle without removing rubber stopper, prepare in the BSL-2 hood in SB 028

a. Wear appropriate personal protective equipment (see section IV).

b. Cover all work surfaces with absorbent, plastic-backed, disposable bench paper.

c. Use a syringe with attached needle to add saline to MPTP through the rubber stopper. 

d. Dispose of needle and syringe into MPTP-labeled sharps container.

e. Dispose of MPTP bottle (and any liquid remaining in bottle) into red biohazard bag with other MPTP waste.  Take MPTP-labeled and dated biohazard bags, when full, to hazardous waste disposal freezer in HSB 015A. Environmental Health and Risk Management will be notified by LAR to pick up the waste. 

D.   When all work with MPTP is complete,

1.    Place sharps containers, MPTP bottles and any disposable lab ware into red biohazard bag.

2.    Carefully remove all bench paper and dispose of into a red biohazard bag.

3.    Wipe all surfaces with 70% ethanol

4.    Remove all personal protective equipment and place into the biohazard bag.

5.    Wash hands thoroughly with soap and water.

6.    Take MPTP-labeled and dated biohazard bags to the hazardous waste disposal freezer in HSB 015A.  Environmental Health and Risk Management will be notified by LAR to pick up the waste.

VII. Administration of MPTP

A.    Conduct all MPTP experimentation involving animals in SB 028.

B.    Cover all work surfaces with absorbent, plastic-backed, disposable bench paper.

C.   Administer MPTP to mice in the BSL-2 cabinet located in SB 028.

1.    Wear personal protective equipment (see Section IV.A.2).  This includes all personnel administering the MPTP and those present in the room.

D.   Restrain rodents either chemically or physically prior to starting the injection procedure.

E.    Use disposable needle-locking syringes or disposable syringe-needle units (i.e., needle is integral to the syringe).

1.    Do not bend, break, recap, remove from disposable syringes, or otherwise manipulate the syringe-needle unit before disposal.

2.    Place the used syringe-needle unit into a MPTP-labeled sharps disposal container.

3.    Enclose the sharps disposal container in a MPTP-labeled and dated red biohazard bag at the end of each day.  Dispose of the bag and sharps container appropriately when the experiment is concluded. 

VIII. Animal Housing

A.    House MPTP-treated animals in SB 028 and keep the door locked at all times.

B.    Post the MPTP-animal room (SB 028) with signage

1.    MPTP-treated animals present

2.    Entrance restricted to authorized personnel

3     PPE must be worn by all personnel entering or working in SB 028

a.    disposable gloves

b.   N -95 mask (respirator)

c.    safety glasses

d.   disposable jumpsuit (a lab coat may be worn by LAR personnel entering the room for routine tasks) 

4.    MPTP hazard sign

5.    SOP for MPTP

6.    AUP number.

C.   Place animals into clean caging immediately prior to MPTP injection.

D.    Label the mouse cages to indicate that hazardous material is present (MPTP).

E.   All personnel entering and working in SB 028 must wear personal protective equipment in the animal room at all times during MPTP injection and for 7 days following the final injection (see Section IV.A.).

1.    This includes LAR personnel performing daily tasks.

F.    7 days post-injection and after bedding is changed:

    1. Remove MPTP sign from door.
    2. To enter SB 028, personnel will put on a pair of disposable lab gloves and a lab coat – additional protective equipment is no longer required to enter the room.

 3. However, personnel must wear complete personal protective equipment at all times when handling the animals, bedding and caging in SB 028 (see Section IV.A.).

 4. Dispose of all personal protective equipment into MPTP-labeled and dated red biohazard bags in the hazardous waste disposal freezer in HSB 015A.  Environmental Health and Risk Management will be notified by LAR to pick up the waste.

G.  SB 028 final cage change and cleaning (7 days after injection) and room cleaning

1.    Wear personal protective equipment during cage change and cleaning (see Section F.3 above).

2.    In BSL-2 cabinet, change and manipulate cages and mice in SB 028.

3.    Place mice into clean cages for transport to HSB 016. 

a. HSB 016: euthanize and surgically manipulate mice (wear gloves and lab coat)

b. HSB 016: dispose of euthanized mice into dated and labeled bag and place inside red biohazard bag in HSB 015A biohazard freezer

c. HSB 016: take all mouse boxes used in HSB 016 to the cage washing room in HSB

4.    In BSL-2 cabinet, spray MB-10 on dirty bedding in cages to keep the dust down

5.    In BSL-2 cabinet, dump bedding and food into red biohazard bag

6.    In BSL-2 cabinet, spray the outside of the cages, bottles, hoppers, etc with MB-10.

7.    Fill sink in SB 028 with 1% bleach water and soak all emptied caging materials in the sink for 20 minutes.

8.    Rinse all caging materials with tap water.

9.    Dispose of all personal protective equipment, waste and disposable materials used in experiment into MPTP-labeled and dated red biohazard bags and take to the hazardous waste disposal freezer in HSB 015A.  Environmental Health and Risk Management will be notified by LAR to pick up the waste.

10.   Take all pre-cleaned caging materials from SB 028 to the SB dirty cage wash room for routine processing through cage washer.

11.   Make certain that the room and BSL-2 cabinet are completely clean at end of experiment and remove all MPTP-related signage from door of SB 028