Laboratory Safety and Hazardous Waste Training

Lab safety identifies hazards and controls risks by:

using risk assessments, eliminating/substituting unnecessarily hazardous work with safer methods, engineering ways to reduce exposure, implementing administrative controls, training personnel to ensure awareness, using personal protective equipment

minimum PPE for all labs

gloves (nitrile), eye protection (glasses or goggles), lab coat (flame resistant for chemistry), closed toed shoes, long pants

experiments using biological materials are reviewed by the…

Institutional Biosafety Committee (IBC)

the Institutional Biosafety Committee (IBC) ensures safety and compliance with federal, state, and institutional standards including…

NIH (NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules), CDC/NIH (Biosafety in Biomedicatl and Microbiological Laboratories), OSHA (Bloodborne Pathogen Standard)

biohazard

an agent or material of biological origin that has the capacity to produce deleterious effects in humans, animals, or the environment

biohazard examples

bacteria/viruses/fungi/parasite/prions/human or animal cells/human or animal tissues/human or animal fluids/biological toxins/allergens/venoms/recombinant or synthetic nucleic acids

biosafety levels (BSLs)

indicator of the biological hazards associated with a laboratory

BSL values are determined based on…

biological agents, procedures, facilities, and PPE

routes of entry for an exposure

inhalation, inoculation, mucous membrane, ingestion, skin contamination

to minimize exposure to biohazards…

wear appropriate PPE, don’t eat/drink/apply cosmetics in labs, remove gloves before opening doors/using a phone or computer/touching exposed skin, don’t reuse gloves

safety data sheets (SDS) provide information on a chemical’s…

physical properties, fire and explosive hazards, reactivity, health hazard information, safe handling and use precautions, spill control guidance, waste disposal procedures

chemical classes: flammable liquids

organic chemicals with a flash point below 140°F (60°C)

flash point

temp at which a flammable liquid gives off enough vapor to flame

when working with flammables…

use higher flash point alternatives or purchase pre-diluted chemical stocks, conduct all work in a chemical fume hood and remove possible ignition sources, wear a flame-resistant lab coat, store chemical containers and waste in a flammables cabinet when not in use

chemical classes: corrosives

chemicals that have a pH that is below or above neutral; strong acids and bases can cause severe burns, compromise container integrity, and react violently when mixed

chemical classes: oxidizers

chemicals that are highly reactive with other chemicals or materials; strong oxidizers are capable of igniting organic materials on contact

chemical classes: peroxide formers

chemicals that form explosive peroxide compounds over time. Their bottles should be dated upon receipt and initial opening, tested annually, and have their results documented on the label

eye protection: safety glasses

impact protection, minimal splash protection

eye protection: splash goggles

impact and splash protection

contact lenses

permitted, but should be removed immediately in the event of exposure

eye protection must have an…

ANSI-Z87.1 stamp

LD50

the amount of a substance required to kill 50% of a population

LD50

indicates the toxicity of a given substance

Lower LD50

higher toxicity

Higher LD50

lower toxicity

LD50 < 5

super toxic

LD50 = 5-50

extremely toxic

LD50 = 50-500

very toxic

LD50 = 500-5,000

moderately toxic

LD50 = 5,000 - 15,000

slightly toxic

LD50 > 15,000

practically nontoxic

LD50 units

mg/kg

Dose =

chemical concentration x duration of exposure

glove permeation

the ability of chemicals to pass through a pair of gloves

types of gloves to avoid

latex and PVC

all chemical containers must be labeled with…

chemical name(s), date prepared, your name, “TOXIC” if toxic

toxic and hazardous materials should be stored…

below eye level

storage: each chemical class should be…

uniquely segregated—stored in secondary compartments with containers of the same chemical classification to prevent mixing in the event of a spill

container compatibility: corrosives in metal containers?

incompatible

container compatibility: strong acids or bases stored in conical tubes?

incompatible (conical tubes corrode, especially around the cap)

container compatibility: solvents in plastic containers?

incompatible

Store corrosives and organic solvents in…

glass containers

Rule of thumb: store chemicals in…

containers of the same material in which they were shipped

indicators of compromised containers

discoloration of liquid or cap, crusty residue buildup around the cap or on the container’s exterior, cracks or leaks, chemical odors or vapors, rusting metal shelves/brackets in the corrosives cabinet (acid containers are compromised)

dispose of all compromised materials as…

hazardous waste

do not store volatile chemicals in…

open flasks or beakers

if a protocol requires cold solvents, store in refrigerators/freezers/cold rooms for…

short periods of time, in quantities < 100 mL

corrosives stored in the fridge or freezer?

never under any circumstances

chemical fume hoods protect the user from…

vapors, gases, aerosols, dust

chemical fume hoods draw air in at a velocity between…

60-150 ft/min (80-120 fpm is optimal)

to maximize air flow in a chemical fume hood,

keep air slots at the rear of the hood clear

to ensure the best containment in a chemical fume hood,

work six inches inside the hood

to protect against spills and splashes in a chemical fume hood,

work behind the sash, keeping it below the maximum height of 18 inches and closing when not in use

secure gas cylinders with…

a chain or strap around the upper third of the container

store compressed gas cylinders in…

well-ventilated areas

when compressed gas cylinders are not in use,

close valve and secure shipping cap

compressed gas cylinders’ valves or regulators:

do not attempt to adapt or modify

_____ secure a compressed gas cylinder around the regulator

do not

keep a compressed gas cylinder’s regulator, pressure relief valves, and hose lines…

clear

make sure the labels on a compressed gas cylinder are…

visible and legible

when handling liquid nitrogen, wear…

heavy-duty, “loose” fitting gloves with long cuffs; a lab coat, closed-toed shoes, safety glasses, and a face shield

dispense and use liquid nitrogen in…

well-ventilated spaces only

following each use and tank change of liquid nitrogen…

firmly close dispensing valve

cryogenic storage: only store vials in…

gas phase (vials stored in liquid phase can explode if liquid nitrogen has permeated the vial and expands rapidly upon warming)

a material becomes a waste when…

it can no longer be used for its intended purpose, it has been abandoned, or it is deemed a waste by the owner

a material becomes hazardous when the material becomes…

corrosive, ignitable, reactive, toxic

3 steps of managing hazardous waste

label it, close it, store it in secondary containment

Containers for hazardous waste must have…

screw-top lids

if reusing containers for hazardous waste collection…

deface old labels and triple rinse

when picking a container for hazardous waste collection…

choose the smallest reasonable container (1 gallon maximum)

all hazardous waste containers must have ___ affixed as soon as waste is added

a completed EHS Hazardous Waste Label

step 1 of EHS Hazardous Waste Labelling

fill out lab information (PI and contact person)

step 2 of EHS Hazardous Waste Labelling

list all chemical constituents in the container, using full chemical names (no abbreviations or chemical formulas)

step 3 of EHS Hazardous Waste Labelling

estimate the percentage of each chemical in mixture and include pH if the container contains corrosives

step 4 of EHS Hazardous Waste Labelling

indicate the hazards by checking the appropriate box ro boxes

step 5 of EHS Hazardous Waste Labelling

contact EHS to schedule a waste pickup

color of hazardous waste labels for Dartmouth College laboratories are…

fluorescent green

to schedule a waste pickup,

email EHS, indicating the type, volume, and location of the waste

Hazardous waste don’ts:

evaporate, drain-dispose, throw into regular trash

solutions suitable for drain disposal

dilute sugar and salt solutions, buffer solutions, solutions with pH = 7-9

solutions unsuitable for drain disposal

organic solvents, toxic dyes or stains, solutions with heavy metals

avoid creating unknowns by…

thoroughly labelling all containers

do not ___ unknown chemicals

open or smell

upon discovery of unknown chemical containers…

contact EHS to request a waste pickup

Drug Enforcement Administration (DEA)

regulates pharmaceuticals and controlled substances

Paints, solvents, thinners, and other maintenance products are…

potentially hazardous

photographic chemicals…

must be collected, and cannot be drain disposed

lab glassware must be ___ before disposal

triple rinsed

Incompatible hazardous wastes must be…

collected separately and stored in a segregated manner

radioactive material (RAM)

substance comprised of unstable atoms that emit ionizing radiation during decay

ALARA

“as low as reasonably achievable.” principle that maintains low radiation levels

RAM stocks are housed in the laboratory, secured under…

lock and key

monochromatic radiation

emits light of a single wavelength

coherent/synchronous laser radiation

emits waves in phase

unidirectional laser radiation

emits waves in the same direction

caution laser radiation sign indicates…

laser use for which your natural blink response provides sufficient protection

danger laser radiation sign indicates…

high-powered laser use for which your natural blink response is not sufficient

laser pointers’ maximum output power should be

< 5 mW

the majority of needlestick accidents are due to…

recapping

before disposal, syringes must be…

empty