Laboratory Standard Operating Procedure
(For the Use of Hazardous Materials or Equipment)
Name of Procedure: ______Phenol – Chloroform Extraction__________________
Prepared by: ________________________
Revision Date: ___________
LOCATION- This procedure may be performed in the following location(s):
Building Room #-
HAZARDS- The following materials and equipment associated with this procedure present exposure or
physical health hazards. The following hazards require safety precautions:
Phenol is readily absorbed through the skin and can cause severe burns to the eyes and skin. Phenol has a local
anesthetic effect, so that no pain may be felt on initial contact. Systemic effects include damage to the liver and
kidneys.
Chloroform is a skin and eye irritant, and it is a suspected human carcinogen and reproductive hazard. Adding
chloroform to phenol enhances the ability of phenol to be absorbed by the skin.
Ethyl ether is a mild skin and eye irritant. The primary hazard associated with ethyl ether is that it is extremely
flammable and it forms explosive peroxides after prolonged exposure to light and air. When kept for prolonged
periods after it has been opened, ethyl ether forms peroxides that may explode when the container cap is removed or
when it is concentrated during laboratory activities.
Wash hands thoroughly immediately after working with these chemicals.
Refer to a material safety data sheet (MSDS) for more detailed information on these and other laboratory chemicals.
ENGINEERING CONTROLS: Prior to performing this procedure, the following safety equipment must
be accessible and ready for use: (ex. Chemical fume hood, biological safety cabinet, laminar flow hood)
Perform all procedures involving greater than 10 ml of phenol or chloroform or any amount of ether in a chemical
fume hood
Grasp both the tube and the cap when vortexing to prevent the cap from opening, causing a splash or aerosol to
escape.
Use sealed safety cups when centrifuging phenol: chloroform, and wait at least 10 minutes before opening the
centrifuge to prevent exposure to aerosols.
PROTECTIVE EQUIPMENT- Prior to performing this procedure, the following personal protective
equipment (PPE) must be obtained and ready for use: (ex. Acid resistant gloves, safety eyewear, lab coat,
chemical splash apron)
Eye Protection: Wear safety glasses when performing phenol: chloroform extraction, even when working in a fume
hood.
Phenol can cause severe burns to the eyes. Wear chemical splash goggles when there is a splash hazard (for
example, when pouring phenol).
Gloves: Wear disposable nitrile gloves to protect against accidental hand contact. If accidental contact occurs,
remove and discard contaminated gloves immediately. (The breakthrough time for a 4 mil nitrile glove is
approximately 3 minutes for chloroform.)
Protective Clothing: Wear standard laboratory clothes. (Lab coat, long pants and closed toe shoes.)
WASTE DISPOSAL- This procedure will result in the following regulated waste, which must be
disposed of in compliance with the REHS Hazardous Waste Policy:
Collect all waste from phenol chloroform extraction (including contaminated pipette tips) and label as hazardous
waste and discard via REHS. (Note: most labs put the liquids into the waste stream but not the pipets or tips, which
should have minimal amounts.)
ACCIDENTAL SPILL- In the event of a spill of hazardous materials, follow these procedures:
Small Spill
ASSESS spill size, chemical type and available cleanup
equipment.
EVACUATE the immediate area.
WARN other personnel.
WEAR appropriate PPE.
CLEAN UP spill using appropriate spill kit if trained to
do so.
CONTAIN spilled material in a sturdy plastic bag or
sealed drum. Apply a hazardous waste label and contact
REHS for disposal.
Large Spill
ATTEND to injured persons, seek appropriate medical
attention.
EVACUATE area/building.
EXTINGUISH ignition and heat sources.
PREVENT spill from spreading.
CONTACT REHS for assistance. After hours call
Campus Police.
FILE a formal incident report
PRIOR APPROVAL- This procedure is considered hazardous enough to warrant prior approval from
the PI or department safety coordinator: YES/NO
NO