Standard Operating Procedure
Dimethyl Sulfoxide (DMSO)
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
X Hazardous Chemical
☐ Hazardous Class
Purpose
DMSO is used at 10-15% final concentration to freeze viable cells.
Physical & Chemical Properties/Definition of Chemical Group
CAS#: 67-68-5.
Class:
Molecular Formula: (CH3)2SO
Form (physical state): Liquid
Color: clear
Boiling point: 372.2°F (189°C)
Potential Hazards/Toxicity
Toxicological Data on Ingredients: Dimethyl sulfoxide: ORAL (LD50): Acute: 14500 mg/kg [Rat]. 7920
mg/kg [Mouse]. DERMAL (LD50): Acute: 40000 mg/kg [Rat].
Potential Acute Health Effects:
Slightly hazardous in case of inhalation (lung irritant). Slightly hazardous in case of skin contact (irritant,
permeator), of eye
contact (irritant), of ingestion, .
Potential Chronic Health Effects:
Dimethyl sulfoxide (DMSO).
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Slightly hazardous in case of skin contact (irritant, sensitizer, permeator), of ingestion. CARCINOGENIC
EFFECTS: Not available. MUTAGENIC EFFECTS: Mutagenic for mammalian somatic cells. Mutagenic
for bacteria and/or yeast.TERATOGENIC EFFECTS: Not available. DEVELOPMENTAL TOXICITY: Not
available. The substance may be toxic to blood, kidneys, liver, mucous membranes, skin, eyes. Repeated
or prolonged exposure to the substance can produce target organs damage.
Personal Protective Equipment (PPE)
Respirator Protection
No respirators used in this lab
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
Double latex (rubber) gloves.
Refer to glove selection chart from the links below:
http://www.ansellpro.com/download/Ansell_8thEditionChemicalResistanceGuide.pdf
OR
http://www.allsafetyproducts.com/glove-selection-chart-chemical-breakthrough-ratings.html
OR
http://www.showabestglove.com/site/default.aspx
OR
http://www.mapaglove.com/
Eye Protection
Wear safety glasses, goggles, or face shield if risk of splash exists.
Skin and Body Protection
When working withDimethyl Sulfoxide(DMSO) all persons shall wear a lab coat, nitrile gloves (for
limited exposure only- NOT FOR prolonged exposure), long pants and closed toe shoes, and safety
glasses if risk of splash exists.
Hygiene Measures
Wash thoroughly after handling. Remove contaminated clothing and wash before reuse. Do not get in
eyes, on skin, or on clothing. Do not ingest or inhale. Use only with adequate ventilation or respiratory
protection.
Engineering Controls
Engineering Controls: Provide exhaust ventilation or other engineering controls to keep the airborne
concentrations of vapors below their respective threshold limit value. Ensure that eyewash stations and
safety showers are proximal to the work-station location.
Dimethyl sulfoxide (DMSO).
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First Aid Procedures
If inhaled - remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult, give
oxygen. Get medical attention.
In case of skin contact - Wash with soap and water. Cover the irritated skin with an emollient. Get
medical attention if irritation develops. Cold water may be used.
In case of eye contact - Check for and remove any contact lenses. Immediately flush eyes with running
water for at least 15 minutes, keeping eyelids open. Cold water may be used. Get medical attention if
irritation occurs.
If swallowed - Do NOT induce vomiting unless directed to do so by medical personnel. Never give
anything by mouth to an unconscious person. Loosen tight clothing such as a collar, tie, belt or
waistband. Get medical attention if symptoms appear.
Special Handling and Storage Requirements
Precautions for safe handling
Avoid inhalation of vapour or mist. Keep away from sources of ignition - No smoking. Take measures to
prevent the build up of electrostatic charge.
Conditions for safe storage
Keep container tightly closed in a dry and well-ventilated place. Store under inert gas. Hygroscopic
Spill and Accident Procedure
Small Spill: Evacuate the spill area. 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.).
Absorb with an inert dry material and place double bag as dry waste. Finish cleaning with soap and water
and wipe up with paper towels. Double bag dry waste and label with online waste tag. Take to chemical
waste pick-up.
Large Spill: Evacuate the spill area. 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.).
Combustible material. Keep away from heat. Keep away from sources of ignition. Stop leak if without
risk. Absorb with an inert material and put the spilled material in an appropriate waste disposal. Finish
cleaning by spreading water on the contaminated surface and allow to evacuate through the sanitary
system. Double bag dry waste and label with online waste tag. Take to chemical waste pick-up. If spill is
too large to handle call EH&S Hotline at X59797 (310-825-9797, or cal 911 from a campus phone
Chemical Spill on Body or Clothes – If spill on skin is small, wash with large amount of water and
soap. If large volume is spilled remove clothing and rinse body thoroughly in emergency shower for at
least 15 minutes. Seek medical attention if necessary. 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 if
necessary. Notify supervisor and EH&S at x59797 immediately.
Dimethyl sulfoxide (DMSO).
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Medical Emergency Dial 911 from a campus phone or x52111 (310-825-2111) from
cell phone
IN CASE OF ACCIDENT :
Incident Report & Referral for Medical Treatment forms are located in the Lab Safety
Maunual. Fill out Incident Report and seek medical attention at :
1) Employees - Non-Life Threatening Emergency, go to Occupational Health Facility ( OHF is
located at Rm# 67-120 Center for Health Sciences. This is on the 6th floor, 7th corridor, Room
120 in CHS. [Enter through the School of Dentistry on Tiverton Drive and proceed to the “O” elevator to
the 6th floor.] OHF phone # is 310-825-6771, Open 7:30 a.m. - 4:30 p.m. Monday - Friday. OHF
is closed on holidays and weekends.
2) Students - Arthur Ashe Student Health and Wellness Center , 221 Westwood Plaza, Los
Angeles, CA 90095 (310) 825-4073
3) After-Hours Care and Serious Injury Care - If injury or illness occurs when OHF is closed,
or urgent immediate medical attention is required, treatment can be obtained at the Ronald
Reagan UCLA Medical Center Emergency Medicine Center, x52111 (310-825-2111).
OR if necessary : Call 911 from campus phone or 310-825-1491 from cell phone
IMPORTANT NOTE : Supervisor must be notified ASAP after an accident. EH&S must be
notified within 8 hours of serious accident at EH&S Hotline, 310-825-9797.
Needle stick/puncture exposure (as applicable to chemical handling procedure) – Wash the affected
area with antiseptic soap and warm water for 15 minutes. For mucous membrane exposure, flush the
affected area for 15 minutes using an eyewash station. Page the needle stick nurse by dialing 231 from a
campus phone, enter 93333 when prompted and then enter your extension. Hours: M – F, 8:00 a.m. to
4:00 p.m. At all other times report to Ronald Regan UCLA Medical Center (emergency room) at x52111.
Note: All needle stick/puncture exposures must be reported to EH&S at x59797 within 8 hours.
Decontamination/Waste Disposal Procedure
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
Dimethyl sulfoxide (DMSO).
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o
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
Consult waste pick-up schedule
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 - 1 ml
Conditions covered by this SOP : -196oC to 25oC
General : DMSO is used to freeze viable cells. DMSO is diluted in heat inactivated bovine serum. The final
concentration of DMSO in the prepared freezing medium is 10%. Concentrated DMSO must be handled
carefully using nitrile gloves, lab coat and eye protection if splash hazard exists (if used outside of a fume
hood or outside of a biosafety cabinet without a front window shield). All personnel using DMSO must and
have lab specific training on this SOP. All concentrated DMSO must be disposed of as chemical waste by
placing it in a container with an online waste tag and taking for disposal within 90 days.
Example : To Freeze down 5 vials of viable cells – With proper PPE in place DMSO is opened in the
Biosafety Cabinet and 0.5 ml of DMSO is placed in a test tube with 4.5 ml of heat inactivated fetal bovine
serum and the cells and mixed thoroughly . The vials are then placed in a Nalgene Cryofreezer
(cryofreezer must have isopropanol up to fill line) and the cryofreezer is placed in deep freezer ( -70oC to
-80oC) for at least 24 hours. Frozen vials can them be transferred to liquid nitrogen container. Use
appropriate PPE for using liquid nitrogen.
To Thaw vials of Frozen Cells - Remove vials from liquid nitrogen freezer wearing appropriate PPE for
Liquid Nitrogen.Transfer to 37oC water bath just until thawed, then with proper PPE for DMSO dilute cells
at least 1/10 with cell culture medium, or PBS and pellet cells. Remove supernatant and dispose of
diluted DMSO in - A) for human or transfected cells -15 to 20% bleach for at least 20 min and then into
regular liquid waste stream; B) for normal mouse cells - into regular liquid waste stream. Resuspend
cells and continue processing as needed.
NOTE - Any deviation from this SOP requires approval from PI. Any new protocol should be
shown to lab safety officer, approved, signed and dated by PI and user, and attached to this SOP
Any extended use of DMSO must be done with heavy latex gloves and approved by the PI.
Documentation of Training
Dimethyl sulfoxide (DMSO).
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
Prior to conducting any work with Dimethyl Sulfoxide (DMSO), designated personnel must provide training to
his/her laboratory personnel specific to the hazards involved in working with this substance, work area
decontamination, and emergency procedures.

The Principal Investigator must provide his/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 his/her laboratory personnel have attended appropriate laboratory
safety training or refresher training within the last one year.
Principal Investigator SOP Approval
Print name __________________________Signature___________________________
Approval Date:
I have read and understand the content of this SOP:
Name
Dimethyl sulfoxide (DMSO).
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Signature
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Dimethyl sulfoxide (DMSO).
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