CHEMICAL HYGIENE PLAN
Set A
Biology Chemical Handling and Introductory Courses
Chemical Storage, Chemical Handling, and Preparation of Chemical Solutions for
Biology Laboratories
Chemical Storage:
All chemicals must be added to the Chemical Inventory upon receipt and deleted from the
inventory when the containers are empty and/or properly disposed of. All chemical
containers must be clearly and legibly labeled with the chemical NAMES and/or
chemical descriptions of the contents, along with appropriate warning labels.
Most dry chemicals will be stored in MSC107, arranged alphabetically. The exceptions
to this are:
Acids/acidic solutions: Acidic solutions will be stored in screw-top sealed
containers in the ACID cabinet in MSC 107.
Bases/ alkaline solutions: Alkaline solutions will be stored in screw-top sealed
containers the BASES cabinet in MSC 107.
Corrosives/corrosive solutions: Corrosives and corrosive solutions will be stored
in screw-top sealed containers in either the ACIDS or BASES cabinet in MSC
107.
Irritants: Dry chemical irritants may be stored with the general chemicals on the
shelves in MSC 107. Irritant solutions and volatile irritants should be stored in
the fume hood in MSC 107.
Metabolic poisons: Known metabolic poisons should be double-sealed in plastic
bags and stored in the fume hood in MSC 107.
Neuroactive and vasoactive chemicals: Neuroactive and vasoactive chemicals
should be stored in the locked cabinet or in the locker refrigerator/freezer in MSC
102D (vivarium surgical room).
Chemical Preparation and Disposal:
All chemical preparation for introductory biology classes should be conducted in the
Biology Chemical Prep room MSC 107. The single current exception to this will be
microwaving agarose gel to liquefy it.
Safety glasses and gloves should be worn during all chemical preparation
Aqueous salt solutions not involving heavy metals: Simple salt solutions (e.g. Ringer’s,
physiological saline, buffer solutions, sucros solutions, etc.) posing no health or
environmental threats may be freely mixed with not further precautions. Excess solution
may be rinsed down the drain.
Solutions containing metal salts: All solutions containing metal salts must be disposed of
as waste in appropriate containers in a designated Satellite Accumulation Area.
Acidic and alkaline solutions: These solutions should be prepared with due caution. Any
spills should be neutralized and cleaned up immediately. Simple acid and base solutions
may be disposed down the drain AFTER CHEMICAL NEUTRALIZATION TO pH 6-8.
Organic solvents: All solutions containing organic solvents must be disposed of into
appropriately labeled hazardous waste containers in a designated Satellite Accumulation
Area.
Volatile, corrosive, irritant, poisonous, neuroactive, vasocative, and/or teratogenic
chemical solutions: These solutions should be prepared only by trained faculty of staff
personnel, NOT BY STUDENTS. Each solution and all rinseate from storage containers
must be properly disposed of in a labeled container in a designated Satellite
Accumulation Area.
Container Disposal:
All empty original containers should be triple rinsed and have the label defaced or
removed prior to disposal in solid trash containers. Weigh boats used for measuring dry
chemicals should be similarly treated. Any rinseate containing hazardous substances
must be collected, labeled, and processed as hazardous waste.
All glassware used in the preparation of chemical solutions must be triple rinsed, then
cycled through the dishwasher (if practical). Any rinseate containing hazardous
substances must be collected, labeled, and processed as hazardous waste.
Broken, chipped, hazardous, or unwanted glassware and glass containers should be
disposed of in a designated glass waste box. Each such container should be triple rinsed
(if this can be safely done) prior to disposal. Any rinseate containing hazardous
substances must be collected, labeled, and processed as hazardous waste.
BIO103 Human Biology
Course Description – This is an introductory non-majors class and laboratory in general
biology, offered 1-2 times per year. Enrollment for each section is typically 20-24
students.
Potential Waste Sources: Many laboratory exercises involve models, microscopy slides,
or computer simulations and generate little to no waste. Specific waste-generating
exercises involve a variety of household chemicals, chemical test reagents, and gel
electrophoresis of DNA (using minute quantities of ethidium bromide).
E-Waste: College Computer Services will dispose of outdated/replaced/
nonfunctional computers, monitors, printers, and computer peripherals according
to the standard college e-waste disposal protocols.
Classroom Waste: Paper trash will be disposed with general trash. Used fluorescent
bulbs will be treated as universal waste, collected by custodial and/or Physical
Plant staff, then packaged and stored at the central campus universal waste site for
disposal by a licensed carrier.
Broken Glass: Unless otherwise specified, broken glass will be stored in the
designated broken glass containers for disposal by a licensed carrier.
Sharps: Unless otherwise specified, all used syringes, hypodermic needles, pins, and
scalpel blades will be stored in the designated Sharps containers for disposal by a
licensed carrier.
Specified Laboratory Waste:
Anatomical Models: All anatomical models are constructed of plaster, metal,
and/or stable, non-toxic plastics. Irreparably-damaged models will be
disposed in the general trash. In the past 14 years no models have been
disposed of.
Microscopy Slides: Microscopy slides include commercially prepared and
mounted slides, as well as wet mounts of plant tissue and human cheek cell
scrapings. All broken slides will be disposed as glass waste in standard broken
glass containers. Methylene blue in normal saline is used as a stain for wet
mounts of cheek cell scrapings. Waste methylene blue and rinseate (< 20ml
per year) will be collected to a container of waste stains for commercial
disposal.
Earthworms: 20-40 live earthworms are purchased from a local bait shop.
Survivors of the lab are killed by freezing, then disposed of in the dry trash.
Household Chemicals and Foodstuffs: The following nontoxic household
chemicals are used in small quantities (<2 lb. total waste) in dry form or as
pureed aqueous solutions for two laboratory exercises. All waste from these
chemicals are disposed of as dry trash and/or by flushing down the drain.
flour, cornstarch, laundry detergent, baking soda, milk, orange juice, vinegar,
honey, peanut butter, tuna, refried beans, raw hamburger, lettuce, vegetable
oil, after-shave lotion, Alka-Seltzer, household ammonia, lemon juice, vinegar
Chemical and Biomolecular Standards: Simple aqueous solutions of glucose,
starch, albumen, and 1% hydrochloric acid are made up fresh for a single
laboratory exercise. Excess solutions are flushed down the drain at the end of
the lab (~1.5 L total).
Biomolecular Test Reagents: Bromothymol blue, iodine solution, Benedict’s
solution, biuret solution, and Sudan IV solution (<2ml each) are used as
chemical test agents, in solutions of the biomolecular standards and foodstuffs
listed above. There is no practical way to reclaim these solutions, so they are
flushed down the drain, or disposed of in the paper trash (Sudan IV test on
filter paper).
Electrophoresis of DNA: A single lab exercise involves preparation of agarose
gels, impregnated with <0.1% ethidium bromide (200ml total voume), a
loading dye (12uL total volume), TAE buffer solution (200ml total volume),
lambda phage DNA (Carolina Bological - 120uL), and three restriction
endonuclease samples in individual micro Eppendorf vials (Carolina
Biological – 36uL total). Agarose gels contain a non-toxic level of less than
0.1% ethidium bromide, so they will be placed in laboratory trash.
Electrophoresis buffer contains <10ug/ml ethidium bromide and will be
released to the drain. Stock solutions of 10ug/ml ethidium bromide are used
until empty, then reconstituted in the same containers.
Spills: Small-scale (<1L) spills will be contained and flushed down the drain. Although
there is little potential for large-scale (>1L) spills, such spills will be contained using
the spill kit in the adjacent Biology Prep Room MSC125, then packaged, labeled,
transferred to the Hazardous Waste Accumulation Room MSC132 for disposal by
licensed hazardous waste carriers. Mercury spills will be contained using the spill kit
in the fume hood in this room (MSC 128).
BIO110 Principles of Biology I
Course Description – This is an introductory class and laboratory in general biology,
offered as two sections once year. Enrollment for each sections combined is typically
20-24 students.
Potential Waste Sources: Several laboratory exercises involve microscopy slides, and/or
computer simulations and generate little to no waste. Specific waste-generating
exercises involve a variety of household chemicals, chemical test reagents, gel
electrophoresis of DNA (using minute quantities of ethidium bromide), catechol and
phenylthiourea solutions, a tyrosinase enzyme solution, thin-layer chromatography
involving ethyl ether, petroleum ether, and N-propanol, and pond water collected
from Foster Lake on the Wesleyan campus,
E-Waste: College Computer Services will dispose of outdated/replaced/
nonfunctional computers, monitors, printers, and computer peripherals according
to the standard college e-waste disposal protocols.
Classroom Waste: Paper trash will be disposed with general trash. Used fluorescent
bulbs will be treated as universal waste, collected by custodial and/or Physical
Plant staff, then packaged and stored at the central campus universal waste site for
disposal by a licensed carrier.
Broken Glass: Unless otherwise specified, broken glass will be stored in the
designated broken glass containers for disposal by a licensed carrier.
Sharps: Unless otherwise specified, all used syringes, hypodermic needles, pins, and
scalpel blades will be stored in the designated Sharps containers for disposal by a
licensed carrier.
Specified Laboratory Waste:
Microscopy Slides: Microscopy slides include commercially prepared and
mounted slides, as well as wet mounts of plant tissue and human cheek cell
scrapings. All broken slides will be disposed as glass waste in standard broken
glass containers. Methylene blue in normal saline is used as a stain for wet
mounts of cheek cell scrapings. Waste methylene blue and rinseate (< 20ml
per year) will be collected to a container of waste stains for commercial
disposal.
Earthworms: 40-60 live earthworms are purchased from a local bait shop.
Survivors of the lab are killed by freezing, then disposed of in the dry trash.
Household Chemicals and Foodstuffs: The following nontoxic household
chemicals are used in small quantities (<4 lb. total waste) in dry form or as
pureed aqueous solutions for two laboratory exercises. All waste from these
chemicals are disposed of as dry trash and/or by flushing down the drain.
flour, cornstarch, laundry detergent, baking soda, milk, orange juice, vinegar,
honey, peanut butter, tuna, refried beans, raw hamburger, lettuce, vegetable
oil, after-shave lotion, Alka-Seltzer, household ammonia, lemon juice, vinegar
Chemical and Biomolecular Standards: Simple aqueous solutions of glucose,
starch, albumen, and 1% hydrochloric acid are made up fresh for a single
laboratory exercise. Excess solutions are flushed down the drain at the end of
the lab (~3 L total).
Biomolecular Test Reagents: Bromothymol blue, iodine solution, Benedict’s
solution, biuret solution, and Sudan IV solution (<4ml each) are used as
chemical test agents, in solutions of the biomolecular standards and foodstuffs
listed above. There is no practical way to reclaim these solutions, so they are
flushed down the drain, or disposed of in the paper trash (Sudan IV test on
filter paper).
Electrophoresis of DNA: A single lab exercise involves preparation of agarose
gels, impregnated with <0.1% ethidium bromide (200ml total voume), a
loading dye (24uL total volume), TAE buffer solution (400ml total volume),
lambda phage DNA (Carolina Bological - 120uL), and three restriction
endonuclease samples in individual micro Eppendorf vials (Carolina
Biological – 72uL total). Agarose gels contain a non-toxic level of less than
0.1% ethidium bromide, so they will be placed in laboratory trash.
Electrophoresis buffer contains <10ug/ml ethidium bromide and will be
released to the drain. Stock solutions of 10ug/ml ethidium bromide are used
until empty, then reconstituted in the same containers.
Enzyme Activity Lab: A single exercise involves colorimetric tests of the
enzymatic activity of tyrosinase on solutions of catechol. All waste catechol
(< 300ml at <1% concentration) and tyrosinase (<20ml) will be collected and
stored in a designated “Catechol Waste” container in the satellite
Accumulation Area in MSC 128, or eventual disposal by a licensed carrier
Thin-Layer Chromatography Lab: A single lab exercise involves extracting plan
photosynthetic pigments in acetone (250ml) and petroleum ether (10ml),
running thin-layer chromatography with a solvent composed of 90%
petroleum ether and 10% n-propanol (600ml), then re-extracting the pigments
in ethyl ether (300ml). Waste acetone, chromatography solvent, and ethyl
ether are separately saved for disposal in designated containers in the Satellite
Accumulation Area of MSC 128. Waste chromatography strips (silica on
plastic) are disposed of in the general dry trash.
Lake Ecology: In this laboratory exercise biological samples are collected from
Foster Lake on slides and as bulk water samples. Water samples are
ultimately flushed down the drain. Slides are cleaned and reused whenever
possible, so minimal glass waste is produced.
Spills: Small-scale (<1L) spills will be contained and flushed down the drain. Although
there is little potential for large-scale (>1L) spills, such spills will be contained using
the spill kit in the adjacent Biology Prep Room MSC125, then packaged, labeled,
transferred to the Hazardous Waste Accumulation Room MSC132 for disposal by
licensed hazardous waste carriers. Mercury spills will be contained using the spill kit
in the fume hood in this room (MSC 128).
BIO112 Principles of Biology II
Course Description – This is an introductory class and laboratory in general biology,
offered as two sections once year. Enrollment for each sections combined is typically
10-15 students.
Potential Waste Sources: Laboratory exercises generating little to no waste involve
models, microscopy slides, preserved specimens for examination only, and/or
computer simulations waste. Specific waste-generating exercises involve dissection
of preserved animals and a single physiology lab involving pithed frogs.
E-Waste: College Computer Services will dispose of outdated/replaced/
nonfunctional classroom/laboratory computers, monitors, printers, and computer
peripherals according to the standard college e-waste disposal protocols.
Classroom Waste: Paper trash will be disposed with general trash. Used fluorescent
bulbs will be treated as universal waste, collected by custodial and/or Physical
Plant staff, packaged and stored at the central campus universal waste site for
disposal by a licensed carrier.
Broken Glass: Unless otherwise specified, broken glass will be stored in the
designated broken glass containers for disposal by a licensed carrier.
Sharps: Unless otherwise specified, all used syringes, hypodermic needles, pins, and
scalpel blades will be stored in the designated Sharps containers for disposal by a
licensed carrier.
Specified Laboratory Waste:
Anatomical Models: All anatomical models are constructed of plaster, metal,
and/or stable, non-toxic plastics. Irreparably-damaged models will be
disposed in the general trash. In the past 14 years no models have been
disposed of.
Bones and Skeletons: All animal bone and skeletal samples are permanent, stable
preparations with no hazardous properties. In the event that disposal of a
sample is needed, it would be packaged, labeled, and transferred to the
Hazardous Waste Storage Room MSC132 for disposal by a licensed carrier.
In the past 14 years no bones have been disposed of.
Invertebrate and Vertebrate Study Collections: Specimens in this collection are
cataloged and stored in either formalin, ethanol, or Carosafe, or are
permanently imbedded in plastic. These specimens will not be removed from
their storage containers. Specimens designated for disposal will be packaged,
labeled, transferred to MSC132 for storage, and disposed of by licensed
hazardous waste carriers.
Microscopy Slides: Microscopy slides include commercially prepared and
mounted slides, as well as wet mounts of live invertebrates. All broken slides
will be disposed as glass waste in standard broken glass containers. No stains
or other potentially hazardous materials are used in the wet mounts.
Dissections: Several laboratory exercises involve dissection of approximately 30
earthworms preserved in ethanol and 20 rats injected with latex and preserved
in Carosafe, a non-toxic preservative. All solid dissection products, including
the finished carcasses will be double-bagged, labeled, and stored frozen in
room MSC132 for disposal by a licensed carrier. Used gloves will be
disposed in the general trash. Excess Carosafe (<0.5L/year) and ethanl
(<.1L/year) in rinseate from specimens, gloves, dissection instruments, and
dissection trays will be flushed down the drain.
Live Vertebrates: A single exercise involves the use of two live frogs Rana
utricularia. Frogs will be maintained in the primary aquatics room of the
Vivarium MSC 102-G. Aquarium water, as well as rinseate from containers
and rocks will be flushed down the drain. Euthanized frogs will be doublebagged, labeled, and stored frozen in MSC132 for disposal by a licensed
carrier. The only solution used with the frogs will be standard Frog Ringers
(<100mL), containing sodium chloride, potassium chloride, calcium chloride,
magnesium chloride, and sodium bicarbonate salts, as well as glucose.
Unused Ringers solution will be flushed down the drain.
Spills: Small-scale (<1L) spills will be contained and flushed down the drain. Although
there is little potential for large-scale (>1L) spills, such spills will be contained using
the spill kit in the adjacent Biology Prep Room MSC125, then packaged, labeled,
transferred to the Hazardous Waste Accumulation Room MSC132 for disposal by
licensed hazardous waste carriers. Mercury spills will be contained using the spill kit
in the fume hood in this room (MSC 128).
BIO203S Research Methods in the Natural Sciences
Course Description – This is an intermediate-level class offered in one section per year.
Enrollment is typically 10-20 students.
Potential Waste Sources: This course involves standard training exercises, “practicums”
based on exercises from the existing upper-level courses, and student independent
projects. Only the student independent projects generate novel waste not covered
under the protocols for upper-level courses in the department.
E-Waste: College Computer Services will dispose of outdated/replaced/
nonfunctional classroom/laboratory computers, monitors, printers,
electrophysiological stimulating and recording equipment, and computer
peripherals according to the standard college e-waste disposal protocols.
Classroom Waste: Paper trash will be disposed with general trash. Used fluorescent
bulbs will be treated as universal waste, collected by custodial and/or Physical
Plant staff, packaged and stored at the central campus universal waste site for
disposal by a licensed carrier.
Broken Glass: Unless otherwise specified, broken glass will be stored in the
designated broken glass containers for disposal by a licensed carrier.
Sharps: Unless otherwise specified, all used syringes, hypodermic needles, pins, and
scalpel blades will be stored in the designated Sharps containers for disposal by a
licensed carrier.
Specified Laboratory Waste:
Live Fish: An introductory exercise involves simple observation of live fish
purchased from local pet shops – either Bettas (Betta splendensI) I or glass
knifefish (Eigenmannia virescens). All fish are either maintained by the
department for future use, or given to students as pets. Only commercial
dechlorinating solutions such as Stress-Coat will be used. Aquarium water, as
well as rinseate from containers and rocks will be flushed down the drain.
Dead fish and organic litter will be disposed in the general trash.
Live Amphibians and Reptiles: Live amphibians and reptiles may be used for
student projects. All amphibians and reptiles will be maintained in the
primary laboratory space MSC 101/103 and/or aquatics room of the Vivarium
MSC 102. Organic litter will be disposed in the general trash. Only
commercial dechlorinating solutions such as Stress-Coat will be used.
Aquarium water, as well as rinseate from containers and rocks will be flushed
down the drain. Dead amphibians and reptiles will be bagged, labeled, stored
frozen in MSC 128, and disposed of by a licensed carrier.
Standard Saline Solutions: Standard saline solutions are aqueous solutions
containing some or all of the following chemicals: sodium chloride, sodium
phosphate, sodium bicarbonate, sodium isothionate, potassium chloride,
potassium bicarbonate, potassium isothionate, calcium chloride, magnesium
chloride, glucose, sucrose, dilute sodium hydroxide, dilute hydrochloric acid.
None of these chemicals, as used, are listed hazardous wastes. Waste of all
such solutions will be flushed down the drain.
Teratogenic, Bioactive, and/or Neuroactive Chemicals: Student projects may
involve minimal (<10mmoles) amounts of such potentially hazardous
chemicals in dilute (<10mM) aqueous or saline solutions. An examples from
past course offerings would be the application of dilute thyroxine to promote
amphibian metamorphosis. As much as possible all hazardous chemicalcontaining wastes, along with any rinseate from the storage containers, will be
separately collected to labeled containers within the satellite storage site in
MSC107. These materials will be ultimately transferred to the Hazardous
Waste Accumulation Room MSC132 for disposal by licensed hazardous
waste carriers.
Spills: Because of the extremely dilute nature of all chemical solutions, there is little
potential for hazardous spills in this course. Any spill involving potentially
teratogenic, bioactive, and/or neuroactive chemicals will be contained using the spill
kit in the adjacent Biology Chemical Prep Room MSC107, then packaged, labeled,
transferred to the Hazardous Waste Accumulation Room MSC132 for disposal by
licensed hazardous waste carriers.