Standard Operating Procedure
Colcemid
This is an SOP template and is not complete until: 1) lab specific information is entered into the
box below 2) lab specific protocol/procedure is added to the protocol/procedure section and
3) SOP has been signed and dated by the PI and relevant lab personnel.
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Laboratory Safety Manual and Chemical Hygiene Plan.
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Type of SOP:
☐ Process
☒Hazardous Chemical
☐ Hazardous Class
Purpose
Colcemid, N-deacetyl-N-Methylcolchicine, is related to colchicine, but animal studies found it to be much
less toxic. It is a microtubule depolymerizing agent that arrests mitotic cultured cells in metaphase. Its
mechanism is similar to that of the anti-cancer drug vinblastine. It should be treated carefully since it is
mutagenic, tumorigenic, and teratogenic. Colcemid is recommended for use in chromosome analysis
during karyotyping and in cell synchronization.
Physical & Chemical Properties/Definition of Chemical Group
CAS#: 477-30-5
Class: Toxic
Potential Hazards/Toxicity
Colcemid
UCLA- EH&S
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Colcemid is a hazardous substance that can exert toxic effects from ingestion. Acute side effects may
include loss of appetite, nausea and vomiting, allergic reaction, and local irritation. Gout and renal failure
can occur. While colcemid is not considered an eye, skin or respiratory irritant, exposure by these
modalities may still potentially cause irritation and respiratory distress.
There is some concern that long-term exposure may cause mutations, but the data is limited.
Toxicity
Intravenous (rat) LD50 1.7 mg/kg
Parenteral (rat) LD50 1.7 mg/kg
Oral (mouse) LD50 25.53 mg/kg
Intraperitoneal (mouse) LD50 35 mg/kg
Intravenous (mouse) LD50 56 mg/kg
Intramuscular (mouse) LD50 87 mg/kg
Reprotoxin
US California proposition 65 – Reproductive toxicity
Personal Protective Equipment (PPE)
Respirator Protection
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
Gloves must be worn. Use proper glove removal technique to avoid any skin contact.
NOTE: Consult with your preferred glove manufacturer to ensure that the gloves you plan on using are
compatible with Lithium chloride.
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
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ANSI approved properly fitting safety glasses or chemical splash goggles. Face shield is also
recommended.
Skin and Body Protection
Flame resistant lab coats must be worn and be appropriately sized for the individual and buttoned to their
full length. Laboratory coat sleeves must be of sufficient length to prevent skin exposure while wearing
gloves. As outlined in UCLA Policy 905 personnel should also wear full length pants, or equivalent, and
close-toed shoes. Full length pants and close-toed shoes must be worn at all times by all individuals that
are occupying the laboratory area. The area of skin between the shoe and ankle should not be exposed.
Hygiene Measures
Wash thoroughly and immediately after handling. Remove any contaminated clothing and wash before
reuse.
Engineering Controls
Handle using a chemical fume hood with good ventilation and electrically grounded lines and equipment.
First Aid Procedures
If inhaled
Move into the fresh air immediately and give oxygen. 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 and inhalation. Keep away from sources
of ignition. Avoid heat and shock or friction when handling.
Conditions for safe storage: Air sensitive. Keep container tightly closed in a cool, dry, and well-ventilated.
Keep away from incompatible materials and conditions. Store in original container. Store away from heat
sources and in a flame proof area. Keep cool and protect from sunlight.
Spill and Accident Procedure
Chemical Spill Dial 911 and x59797
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UCLA- EH&S
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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.).
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.
Chemical Spill on Body or Clothes – Remove clothing and rinse body thoroughly in 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.
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
Use proper personal protective equipment and properly dispose chemical.
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
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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
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
Amount Covered by this SOP:
0 - 5 mg powdered
0 - 10 ml at up to 10 µg/ml solution
Temperature Range Covered
2°C - 38°C
Colcemid is most commonly used in cells for chromosome analysis. Colcemid is available in both
powdered and dissolved forms. Because of its toxicity all powders and solutions should be prepared and
used in a fume hood.
Preparation of 10 µg/ml colcemid stock solution
In a fume hood, weigh out 100 µg colcemid from a 5 mg stock bottle and dissolve with 10 ml PBS, HBSS,
or other media. Prepared solution should be labeled appropriately and stored protected from light at 2°C8°C.
Performing chromosomal analysis
1. Add colcemid (final concentration 0.01-0.02 ug/ml – a 500-1000x dilution of the 10 ug/ml stock) to
recently fed cell culture
2. Incubate culture for 30 minutes to 1 hour
3. During incubation prepare slides: Wash slides in fix (3:1 methanol : acetic acid) and then soak
them in ice cold water until ready to use. It is important for the slides to be both cold and wet.
4. Wash cells with PBS and trypsinize cells as usual.
5. Add media plus serum and resuspend the cells until a single cell suspension is achieved.
6. Spin down cells and remove supernatant.
7. Add 1 ml 0.56% KCl dropwise
8. Resuspend the pellet again aiming for a single cell suspension
9. Add 4 ml of 0.56% KCl
10. Incubate at room temperature for 6 minutes. This number is important as you are now swelling
the cells. Therefore this time should be adjusted for specific experiments
11. Spin down cells and remove supernatant
12. Add 1 ml fix (3:1 methanol:acetic acid) and resuspend pellet.
13. Add 4 ml fix and incubate for 5 minutes at room temperature.
14. Repeat steps 11-13 three more times. Finish with 1 ml total volume
15. Remove a slide from the water, blot edges to remove excess liquid
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UCLA- EH&S
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16. Drop the cell suspension (dropwise) from at least one foot above the surface of the slide (2-3
drops/slide). If the cells are dropped from too close a distance, they will not burst. So, make sure
you drop them from high enough.
17. Allow the slides to dry
18. Stain slides with Giemsa (1:20 dilution) (Giemsa Modified Stain - Sigma GS-500) for at least 15
minutes and wash slides with water
19. Observe chromosomes
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 colcemid, 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 or Lab Supervisor SOP Approval
Print name_________________________Signature___________________________
Approval Date:
I have read and understand the content of this SOP:
Name
Colcemid
UCLA- EH&S
Signature
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Date
Date: 9/16/2015
CW/Reviewed By: