1 / BSL1 Bleaching procedure
Effective January 1, 2009
Revision: #1

TITLE: Biosafety Level 1 (BSL1) Bleaching procedure

SCOPE: All Staff and Students within the Undergraduate Chemical Biology program

RESPONSIBILITY: Lab Coordinators and Technical Staff

PURPOSE: To Outline the Proper Procedures for Bleaching BSL1 contaminated plates, equipment and supplies.

  1. PURPOSE
  1. To outline the proper procedures for safely decontaminating reagents, supplies and equipment within the Undergraduate Chemical Biology Lab.
  2. Reduce the risk of exposure of staff and students to biohazardous agents within the lab.
  1. RESPONSIBILITY
  1. Lab coordinator:
  1. Appropriate personal protective equipment (PPE) is available.
  2. MSDS are accessible and available to staff and students.
  3. Safety practices have been communicated to relevant personnel.
  1. Staff and students:
  1. Read, understand and follow the procedure described.
  2. Wear appropriate PPE and review MSDS prior to implementing procedure.
  1. PROCEDURES

The following procedure is applicable for E. coli BSL1 cultures.

  1. 10% Bleach:
  1. Assemble all required reagents and supplies to complete the task.
  2. Obtain commercial bleach (concentration 6%).
  3. Dilute commercial bleach at a 1:10 ratio in distilled water.
  1. 1 part bleach 6% to 10 parts distilled water.
  2. Solution will last at this concentration for only a few hours. It will be beneficial to make the solution just prior to its use.
  1. Store solution in a small vessel 4L cubitainer and affix it with a tap.
  2. Dispense diluted bleach into 250ml labeled squeeze bottles and set out on bench top for staff and or students to use.
  1. 70% Ethanol:
  1. Assemble all required reagents and supplies to complete the task.
  2. Solution can be made with 95% or absolute ethanol.
  3. Dilute the ethanol as per the following dilution.
  1. For 95%: 736ml of ethanol/1L of distilled water.
  2. For absolute: 700ml of ethanol/1L of distilled water.
  1. Store diluted solution in a sealed vessel until use.
  2. Dispense 70% solution into 250ml labeled squeeze bottles and set out on bench top for staff and or students to use.
  1. Decontaminating:

Disenfecting Agent: E. coli BSL1

Volume: 1 part culture to 0.5 part bleach.

Time: 5minutes

  1. All work surfaces are to be decontaminated after each use with 10% bleach solution followed by a 70% alcohol rinse.
  2. All supplies, solutions and active cultures are to be soaked in a 10% bleach solution for at least 5 minutes to ensure culture has been ‘killed’.
  1. Add one (1) part culture to one half (0.5) part bleach.
  2. Soak in the provided waste container or a large wash bucket.
  3. Decant off liquid waste into aqueous bio waste bottle.
  1. After soaking supplies, solutions and active cultures are disposed of via the following procedures.
  1. All solids and disposable supplies (culture dishes, pipettor tips etc.) are to be bulked into appropriate bio waste bag and put into bio waste stream.
  2. Follow Environmental and Occupational Health Services (EOHSS) biomedical waste storage and disposal procedure.
  3. Reusable supplies (poly spreaders) can be rinsed with distilled water, dried and stored for subsequent use.
  4. All liquid waste is considered to be disinfected and can be bulked into large waste drums in the basement and put into chemical waste stream.
  1. All equipment is to be wiped down after each use with disinfectant wipes and allowed to dry.
  2. After disinfecting PPE is to be removed and discarded appropriately.
  1. Gloves are to be placed into bio waste bags.
  2. Lab coat is to be placed in a bag and washed via campus laundry procedure.
  3. Goggles/glasses can be wiped down with disinfectant wipes.
  1. Hands are to be washed with disinfectant soap prior to leaving the lab room.

Standard bleaching procedure for Undergraduate Labs /