Standard Operating Procedure
Example: Research using BrdU
PI:
Lab Location:
Issue Date:
Revision Date:
___
Prepared by:
Approved by: ___________
_
Background:
Bromodeoxyuridine (BrdU) is used to label cells undergoing S-phase synthesis. It is a thymidine
analogue and gets incorporated in DNA. Because BrdU has cytotoxic, teratogenic, and
mutagenic properties, exposure presents potential health and safety hazards to laboratory staff,
animal handlers and other personnel who may be accidentally exposed to BrdU
Hazard Identification:
Toxic Properties
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Acute effects:
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Chronic effects:
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Systemic effects:
Wheeze and cough, shortness of breath, burning in the mouth, throat, or
chest.
Reproductive disorders and genetic alterations. BrdU is incorporated
into tissue DNA in place of thymidine in animals. BrdU-substituted
DNA replaces normal DNA and chromosomal proteins are altered
through: chromosome lengthening, chromatid breakage, and changed
sister chromatid exchange frequencies. Meiosis and mitosis are
affected.
BrdU integrates into DNA and affects the colon, stomach, bone
marrow, and spleen.
Experimental Procedures (EXAMPLE):
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Wear appropriate PPE: gloves, lab coat, eye protection and mask when working with BrdU
Preparation and mixing of BrdU will be performed in a fume hood.
Make a working solution of BrdU in PBS at 10 mg/ml.
Method of administration: At the animal facilities, we administer a single 100ul (1ml) dose
of BrdU solution to live mice using the I.P. (intraperitoneal) injection. The amount we
administer is1mg per mouse. BrdU is not expected to have any negative effects on the health
of the animals at these doses.
Dispose the empty, uncapped needle and syringe directly into a yellow chemotherapeutics
sharps container.
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COLLECT ALL RESIDUAL LIQUID FOR DISPOSAL AS HAZARDOUS CHEMICAL
WASTE. DISPOSE THE RESIDUAL MATERIAL IN YELLOW BAGS AS
CHEMOTHERAPEUTIC WASTE.
Handle all BrdU solutions over plastic backed absorbent sheets and dispose sheets in yellow
chemotherapeutic waste bags when work is completed.
Control Plan:
General Controls:
 Any work with BrdU needs to be done over disposable absorbent benchtop spill pads.
 Any laboratory equipment or surfaces that have come in contact with BrdU will be
decontaminated or disposed of. This can be done by wiping them down with soap and water
and paper towels (see Waste Management Procedures for instructions on towel disposal).
 Non-porous materials, such as glassware, will be decontaminated by soaking in bleach for 24
hours.
Engineering Controls:
 To prevent inhalation, solid/crystalline forms of BrdU will be handled and weighed out in a
chemical fume hood or similar exhausted enclosure. (If air currents making handling
difficult, a safety shield may be positioned in front of the balance to divert air around the
equipment.)
 Note: If you do not have a fume hood, you must wear an N-95 respirator and participate in
the University Of Minnesota’s Respiratory Protection Program.
 To reduce the chance of accidental injection, syringes and IV sets with Luer-Lok fittings
will be used for preparing and administering hazardous drugs.
TM
Required PPE:
 To prevent skin exposure, appropriate personal protective equipment (PPE) will be worn
when handling any amount of BrdU. Wear a lab coat, disposable gown, or coveralls. Ensure
that all exposed skin is covered. Shorts and sandals are not allowed in the lab!
 Use two pairs of powder-free, disposable nitrile gloves, with the outer one covering the gown
cuff whenever there is risk of exposure to hazardous chemicals.
 Wear ANSI Z-87 approved safety glasses.
 Wear goggles and a face shield when splashes to the eyes, nose, or mouth may occur.
Control Procedures for animal care staff:
 RAR Area Supervisor will be informed of our intent to use BrdU two weeks prior to use
 Cages of animals injected with BrdU will be clearly labeled as such
 All bedding changed during chemical administration and for 72 hours beyond final
administration must be collected in yellow bags for disposal
 Follow PPE and handling requirements specified in the Standard Animal Husbandry
Chemical Hazard SOP (attached)
Waste Management Procedures:
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Excess BrdU solids: Dispose as hazardous chemical waste, following procedures detailed in
the University’s Hazardous Waste Guidebook.
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Disposable lab supplies (e.g., gowns, gloves, benchtop pads, etc.,): Collect in yellow bags
for incineration.
Needles and sharps: Must be disposed of in separate sharps container that is labeled “BrdU
Sharps”, sealed and placed in a yellow bag for incineration.
Contaminated bedding: Dispose of in yellow bags for incineration.
Spill cleanup materials: Place in leak proof container and box for disposal as hazardous
waste following procedures detailed in the University’s Hazardous Waste Guidebook.
For more information:
http://www.dehs.umn.edu/PDFs/Chemo%20Waste%20Disposal%20fact%20sheet.pdf
Spill and Accident Procedures:
Spill Cleanup
 Solid: If source container drops and breaks (1-100 grams max), avoid raising dust – do not
sweep up dry material. Evacuate spill area, restrict other from entering the area and from a
safe spot, call 911 and request AHERPS.
If a small amount (<250 mg) spills outside of spill containment, ensure personal protective
equipment(PPE) is appropriate (an N-95 respirator if outside of the hood, double-nitrile
gloves, goggles, and a lab coat), cover material with paper towels, spray lightly with bleach
water solution (10% bleach) and carefully scoop material and paper towels into container for
appropriate disposal (see Waste Management Procedures) For amounts in between, use
prudent judgment to decide whether to attempt cleanup, or call for spill response.
 Solution: If BrdU solution should spill, prevent liquid from entering sewer. Apply
absorbent pads for larger amounts of liquid and, using appropriate PPE, carefully scoop
wetted absorbent material into container for appropriate disposal (see Waste Management
Procedures).
Accidental Exposures
 Wash exposed skin with soap & water, flush eyes for 15 min.
 Call 911 or seek medical attention.
 For urgent care, employees may go to HealthPartners Occupational and
Environmental Medicine (M/F day time or Urgent Care after hours), or UMMCFairview Hospital (24 hrs). You may seek medical attention at the closest available
medical facility or your own healthcare provider.
 Follow-up must be done by HealthPartners Occupational and Environmental
Medicine.
 Report the incident to your supervisor as soon as possible, fill out the appropriate
documentation.
 Employee First Report of Injury
 Supervisor Incident Investigation Report
 Note: It is important to fill out all of the appropriate documents to be eligible to collect
workers compensation should any complications from the hazardous exposure arise in the
future.
Report all exposures to supervisor as soon as possible!
References:
BrdU MSDS from Sigma Aldrich
http://www.sigmaaldrich.com/MSDS/MSDS/DisplayMSDSPage.do?country=US&language=en&product
Number=B5002&brand=SIGMA&PageToGoToURL=http%3A%2F%2Fwww.sigmaaldrich.com%2Fcata
log%2Fproduct%2FSIGMA%2FB5002%3Flang%3Den
UCLA: EH&S Staff. “5-Bromo-2-Deoxyuridine FACT SHEET”
http://biosafety.ucla.edu/docs/factsheet%20bromodeoxyuridine.pdf
Animal Husbandry Chemical Hazard Notification
PI Name:
PI Phone:
IACUC Protocol Number:
Emergency Contact Information
Name:
Work Phone:
After-hours Phone Number:
Name of Hazard:
Hazard Type: (e.g., carcinogen, reproductive toxin, cytotoxic, neurotoxin):
Start date and end date of Hazard:
PPE for entering room and for handling animals:
 disposable gown, lab coat and/or scrubs
 disposable shoe covers or waterproof boots that must be rinsed prior to leaving the room.
 safety glasses with side shields or face shield
 gloves
 disposable head cover
 N95 or PAPR
 Other:
Instructions for handling bedding:
 Don PPE listed above.
 All bedding changed during chemical administration and for 72 hours after the last administration
must be collected in yellow chemical waste bags for disposal.
Euthanasia (Adhere to the euthanasia method approved in your protocol)
For optimal safety, CO2 euthanasia of chemically-treated animals should be done in the
following manner:
1. Use an empty, clean box bottom with no bedding in it. Place yellow bag inside as a liner.
2. Follow all steps for humane CO2 euthanasia as posted on the instruction sheet. This sheet is
located near the euthanasia station.
3. After confirming the animal is dead, seal the yellow bag, and place it in the nearest cooler.
Carcass disposal
All carcasses collected during chemical administration and for 72 hours after the last administration
must be disposed of in yellow chemical waste bags.
NOTES:
The above procedures must remain in place while a hazardous chemical is being administered to
animals AND for 72 hours after the final administration.
Research staff must alert RAR at least two weeks prior to the start of administration of any
chemical hazard to animals.
Additional instructions: