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CALIFORNIA STATE POLYTECHNIC UNIVERSITY, POMONA Environmental Health & Safety STANDARD OPERATING PROCEDURE Safe Work Practices: Proper Pipetting SOP GENERAL INFORMATION: In working with materials classified as a Biohazard, certain risks may be present which depend on the agent being used. A particular concern when working with Biohazards is the generation of aerosols, the act of dispersing a material into the air. This can be caused deliberately or inadvertently, the latter being of critical concern in order to limit accidental exposure while working. Infectious agents that possess a route of transmission through inhalation are the most dangerous as others are less likely to create a hazard in aerosol form however, safe work practices to reduce the generation of aerosols is a good and prudent industry practice. While there are many opportunities to generate aerosols using high-speed equipment, this guidance is focused specifically on pipetting and transfer of liquids in small volumes. All work involving biohazards, infectious or disease causing agents should ALWAYS be done inside a biosafety cabinet with a proper rating and current certification. Please refer to the Biosafety Cabinet Program and respective SOP’s for proper use and materials handling inside a biosafety cabinet. NEVER PIPETTE BY MOUTH (MOUTH PIPETTE). Under no circumstances is this ever an approved or accepted practice regardless of the material being used. Use “Mark to Mark” pipettes that do not require forcible expulsion of a final drop from the pipette. If “Mark to Mark” pipettes are not available, consider pipetting “by difference” to achieve the volume needed without creating an aerosol. Never blow the last drop from a pipette as this can create significant aerosol and inhalation risks. Always use filtered pipettes and pipette tips to protect equipment from contamination Never mix liquids or solutions by suction and expulsion from the pipette from contamination with aerosols. If possible, try to work immediately above a plastic-backed absorbent pad to minimize the formation of aerosols. When transferring liquids into a container, always dispense liquids against the sidewall and low to the liquid surface to prevent splashes and formation of aerosols. Never drop liquids from a height to a surface or to another liquid. 9/23/2013 3:18:02 PM 5 If using a vacuum source for aspiration of liquids, protect the vacuum source from any possible contamination by using a collection and overflow flask, protected by a HEPA or hydrophobic filter to prevent the formation of aerosols at the vacuum source and causing contamination. Avoid using glass Pasteur pipettes which creates puncture risks. Biohazards should always utilize plastic aspiration pipettes. Collect used pipettes within the biosafety cabinet inside a horizontal collection tray that allows the pipette to be completely immersed in a disinfectant. To decontaminate a pipette, draw disinfectant into the pipette in a vertical orientation and dispense gently in the horizontal collection tray. A minimum of 10 minutes in a disinfectant is required to be considered inactivated. 9/23/2013 3:18:02 PM 5