Standard Operating Procedure
Glutaraldehyde
Department:
Date SOP was written:
Date SOP was approved by PI/lab supervisor:
Principal Investigator:
Internal Lab Safety Coordinator/Lab Manager:
Lab Phone:
Office Phone:
Emergency Contact:
(Name and Phone Number)
Location(s) covered by this SOP:
(Building/Room Number)
Type of SOP:
☐ Process
☒Hazardous Chemical
☐ Hazardous Class
Purpose
Glutaraldehyde is corrosive. It can cause irreversible eye damage. Can cause skin burns. Maybe fatal
if swallowed. Prolonged or frequently repeated skin contact may cause allergic reaction in some
individuals. Plastic container, if present, may cause static ignition hazard. Aspiration may cause lung
damage. Causes asthmatic signs and symptoms in hyper-reactive individuals.
Physical & Chemical Properties/Definition of Chemical Group
CAS#: 111-30-8
Class: Corrosive
Molecular Formula: OHCC3H6CHO
Form (physical state): liquid
Color: colorless
Boiling point: 100.5°C
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Potential Hazards/Toxicity
LD50 Oral:
116-206 mg/kg [Rat]
Primary routes of entry: Inhalation, ingestion, and skin and eye contact.
Eyes: Liquid will cause severe and persistent conjunctivitis, seen as excess redness and marked swelling
of the conjunctiva and profuse discharge. Severe corneal injury may develop, which could permanently
impair vision if prompt first-aid and medical treatment is not obtained. Vapor will cause stinging
sensations in the eye with excess tear production, blinking and possibly a slightly excess of redness of
the conjunctiva.
Skin: Brief contact will cause itching with mild to moderate local redness and possibly swelling.
Prolonged contact may result in pain, severe redness and swelling with ulceration, tissue destruction and
possibly bleeding into the inflamed area. Contact with solutions of Gluteraldehyde may cause harmless
yellow or brownish coloration of skin.
Ingestion: Moderately toxic. May cause moderate to marked irritation and possibly chemical burns of the
mouth, throat, esophagus and stomach. There will be discomfort o pain the chest and abdomen, nausea,
vomiting, diarrhea, dizziness, faintness, drowsiness, thirst, weakness, circulator shock, collapse, and
coma. Aspiration into the lungs may occur during ingestion or vomiting, resulting in lung injury.
Inhalation: Vapor is irritating to the respiratory tract, causing stinging sensations in the nose and throat,
discharge from nose, possibly bleeding from the nose, coughing, chest discomfort and tightness, difficulty
breathing and headache. Heating the solution may result in more severe irritant effects.
Chronic Exposure: Repeated skin contact may cause a cumulative dermatitis. May cause skin
sensitization in a small portion of individuals and present as an allergic contact dermatitis. This usually
results from contact with liquid, but occasionally there may be a reaction to vapor.
Personal Protective Equipment (PPE)
Respirator Protection
Use a half-face mask with organic vapor (US) respirator cartridges.
Respirators should be used only under any of the following circumstances:
 As a last line of defense (i.e., after engineering and administrative controls have been
exhausted).
 When Permissible Exposure Limit (PEL) has exceeded or when there is a possibility that PEL will
be exceeded.
 Regulations require the use of a respirator.
 An employer requires the use of a respirator.
 There is potential for harmful exposure due to an atmospheric contaminant (in the absence of
PEL)
 As PPE in the event of a chemical spill clean-up process
Lab personnel intending to use/wear a respirator mask must be trained and fit-tested by EH&S. This is a
regulatory requirement. (https://www.ehs.ucla.edu/ep/ih/resp)
Hand Protection
Use nitrile or butyl gloves when using Glutaraldehyde.
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Eye Protection
ANSI approved properly fitting safety glasses or chemical splash goggles. Face shield is also
recommended.
Skin and Body Protection
Lab coat, long pants and closed-toe shoes must be worn. Refer to UCLA policy 905.
Hygiene Measures
Wash thoroughly and immediately after handling. Remove any contaminated clothing and wash before
reuse.
Engineering Controls
Always use a certified chemical fume hood when handling Glutaraldehyde.
First Aid Procedures
If inhaled
Move into the fresh air immediately and give oxygen if breathing is difficult. If not breathing give artificial
respiration. Seek medical attention immediately.
In case of skin contact
Immediately flush skin with plenty of water for at least 15 minutes while removing contaminated clothing
and shoes. Wash any contaminated clothing before reuse. Thoroughly clean shoes before reuse. Seek
medical attention immediately.
In case of eye contact
Check for and remove any contact lenses. Rinse thoroughly with plenty of water for at least 15 minutes
and consult a physician. Seek immediate medical attention and continue eye rinse during transport to
hospital.
If swallowed
Do NOT induce vomiting unless directed by medical personnel. Never give anything by mouth to an
unconscious person. Seek medical attention immediately.
Special Handling and Storage Requirements
Precautions for safe handling
Avoid contact with skin and eyes. Avoid inhalation of vapour or mist.
Conditions for safe storage
Keep container tightly closed in a dry and well-ventilated place. Containers which are opened must be carefully resealed and kept upright to prevent leakage. Store
under inert gas. Air sensitive.
Spill and Accident Procedure
Chemical Spill Dial 911 and x59797
Spill – Assess the extent of danger. Help contaminated or injured persons. Evacuate the spill area.
Avoid breathing vapors. If possible, confine the spill to a small area using a spill kit or absorbent material.
Keep others from entering contaminated area (e.g., use caution tape, barriers, etc.).
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Small (<1 L) – If you have training, you may assist in the clean-up effort. Use appropriate personal
protective equipment and clean-up material for chemical spilled. Double bag spill waste in clear plastic
bags, label and take to the next chemical waste pick-up.
Large (>1 L) – Dial 911 (or 310-825-1491 from cell phone) and EH&S at x59797 for assistance.
N/A as we do not use Glutaraldehyde in quantities greater than 100mL.
Chemical Spill on Body or Clothes – Remove lab coat and clothing and rinse body thoroughly with
water (or in an emergency shower) for at least 15 minutes. Seek medical attention. Notify supervisor and
EH&S at x59797 immediately.
Chemical Splash Into Eyes – Immediately rinse eyeball and inner surface of eyelid with water from the
emergency eyewash station for 15 minutes by forcibly holding the eye open. Seek medical attention.
Notify supervisor and EH&S at x59797 immediately.
Medical Emergency Dial 911 or x52111
Life Threatening Emergency, After Hours, Weekends And Holidays – Dial 911 (or 310-825-1491 from
cell phone) or contact the Ronald Reagan UCLA Medical Center (emergency room) directly at x52111
(located at 757 Westwood Plaza, enter from Gayley Avenue). Note: All serious injuries must be reported
to EH&S at x59797 within 8 hours.
Non-Life Threatening Emergency – Go to the Occupational Health Facility (OHF), x56771, CHS room
67-120 (This is on the 6th floor, 7th corridor, room 120. Enter through the School of Dentistry on Tiverton
Drive and proceed to the “O” elevator to the 6th floor.)Hours: M - F, 7:30 a.m. to 4:30 p.m. At all other
times report to Ronald Regan UCLA Medical Center (emergency room) at x52111. Note: All serious
injuries must be reported to EH&S at x59797 within 8 hours.
Decontamination/Waste Disposal Procedure
Use proper personal protective equipment and properly dispose chemical. Clean area with soap and
water.
General hazardous waste disposal guidelines:
Label Waste
 Affix an on-line hazardous waste tag on all waste containers using the WASTe Online Tag
Program https://ehs.ucop.edu/waste as soon as the first drop of waste is added to the container
Store Waste
 Store hazardous waste in closed containers, in secondary containment and in a designated
location
 Double-bag dry waste using transparent bags
https://www.ehs.ucla.edu/hazwaste/management/containers
 Waste must be under the control of the person generating & disposing of it
Dispose of Waste
 Dispose of regularly generated chemical waste within 90 days
 Call EH&S at x61887 for questions
 Empty Containers
o Dispose as hazardous waste if it once held extremely hazardous waste (irrespective of
the container size) https://www.ehs.ucla.edu/hazwaste/types/extremely-hazardous
o Consult waste pick-up schedule
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o https://www.ehs.ucla.edu/hazwaste/management/pick-ups
Prepare for transport to pick-up location
 Check on-line waste tag
 Write date of pick-up on the waste tag
 Use secondary containment
Safety Data Sheet (SDS) Location
Online SDS can be accessed at http://msds.ehs.ucla.edu.
Protocol/Procedure
Quantities Covered by this SOP:
0 – 100 milliliters (mL)
Conditions covered by this SOP:
Storage: 4°C
Perfusion: 18° - 25°C (room temperature)
Use:
When perfusing mice for EM use, glutaraldehyde is necessary to preserve the plasma membrane of cells.
Specific Example:
Below is the procedure used to perfuse a mouse with 2% paraformaldehyde and 0.1% - 2.5%
glutaraldehyde and the recipe to make 2% paraformaldehyde and 2% glutaraldehyde.
Mouse perfusion using 2% Paraformaldehyde* and either 0.1% or 2.5% Glutaraldehyde
(Always perfuse under a ventilated chemical fume hood.)
1. Perfuse mouse with 4% paraformaldehyde* and either 0.1% or 2.5% glutaraldehyde (~2x body
weight in mL, for example 50 mL for a 25 gram mouse). We store the 4% paraformaldehyde in the
refrigerator at 4°C, but use the fix for perfusion at room temperature.
2. After perfusion, keep the animal with brain intact at 4°C (in refrigerator) for 1 hour.
3. Dissect out the brain and place in same fixative for 1 hour on shaker at room temperature (gentle
speed).
4. Rinse very thoroughly in 0.12M phosphate buffer (Millonig’s). Rinse the brains three times in room
temperature Millonig’s buffer for 30 minutes in each rinse on a shaker at moderate speed.
5. After the rinses are done, leave the brains in 30% sucrose (dissolved in Millonig’s) overnight on a
shaker at moderate speed. Leave the brains at room temperature for a few hours and then place into
the refrigerator overnight. The brains will float when they are first put into the sucrose, but will sink
when they are ready to be embedded.
6. Block the brains and embed in OCT on dry ice.
4% Paraformaldehyde* Fixative in 0.12 M Millonig's Phosphate Buffer
(The following steps should always be done under a ventilated chemical fume hood.)
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1. Place 600mL beaker on top of balance and zero it out.
2. Measure out 40.0 grams of paraformaldehyde* in beaker.
3. Add 400mL of ddH2O and heat the solution to 60º (no more). It is best to heat the solution to about
55˚, then turn off the heat. Let the solution continue to stir while it’s heating.
4. Add 10 N NaOH dropwise to clear solution (the solution will clear up a lot, but should still be a bit
“cloudy” as that is the nature of the fix. It will be clear after filtration.
5. Stir the solution until it has equilibrated down to room temp.
6. While heating Paraformaldehyde, mix the following solutions and add to cleared paraformaldehyde
solution (don’t mix the two solutions until the Paraformaldehyde has equilibrated down to room
temperature).
500 ml ddH2O
16.88 g Sodium Phosphate Monobasic
3.86 g NaOH
7. Add enough ddH2O to bring volume to 1 liter.
8. pH to 7.3 (7.2 - 7.4) and filter.
2% Paraformaldehyde* + 2.5% Glutaraldehyde in 0.12M Millonig’s Phosphate Buffer
(The following steps must always be done under a ventilated chemical fume hood.)
1. Pour 500mL of 4% paraformaldehyde* fixative in 0.12M Millonig’s phosphate buffer into a 1500mL
beaker.
2. Pour 500 mL of 0.12M Millonig’s phosphate buffer into the same 1500mL beaker.
3. Add 100mL of 25% Glutaraldehyde into fix solution.
4. Stir solution for 15 minutes.
5. pH to 7.3 (7.2 – 7.4) and filter.
*Paraformaldehyde is a regulated carcinogen.
gluteraldehyde fix and perfusing with it.
Read the SOP for paraformaldehyde before making
NOTE
Any deviation from this SOP requires approval from PI.
Documentation of Training (signature of all users is required)

Prior to conducting any work with Glutaraldehyde , designated personnel must provide training to her
laboratory personnel specific to the hazards involved in working with this substance, work area
decontamination, and emergency procedures.

The Principal Investigator must provide her laboratory personnel with a copy of this SOP and a copy
of the SDS provided by the manufacturer.

The Principal Investigator must ensure that her laboratory personnel have attended appropriate
laboratory safety training or refresher training within the last one year.
Principal Investigator SOP Approval
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Print name
__________________________Signature___________________________
Approval Date:
I have read and understand the content of this SOP:
Name
Glutaraldehyde
UCLA- EH&S
Signature
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Date
Date: 9/16/2015
CW/SH: