Chemicals of Concern
Contact Chemical Safety
Call 303-492-6025
Email chemsafety@colorado.edu
Ethidium bromide (EtBr) is a commonly used DNA stain in molecular biology, but it poses serious health and environmental hazards. As a powerful mutagen, EtBr intercalates into DNA and can cause genetic mutations, increasing the risk of cancer and other health effects with prolonged exposure. It can be absorbed through the skin, inhaled as a powder or vapor, or accidentally ingested. Because of its toxicity and resistance to degradation, EtBr must be disposed of as hazardous chemical waste, which adds to the cost and complexity of its use. These risks have prompted many laboratories to seek safer alternatives for nucleic acid staining.
SYBR Safe:
SYBR Safe DNA Gel Stain, developed by Invitrogen, is designed specifically as a safer alternative to ethidium bromide. It offers comparable sensitivity for detecting DNA in agarose gels but with significantly reduced mutagenicity. Studies have shown that SYBR Safe does not penetrate cell membranes easily, reducing its potential to interact with DNA inside living cells. It can be excited using either blue light or UV light, though blue light is preferred to avoid DNA damage. One of its key advantages is that gels stained with SYBR Safe can often be disposed of as non-hazardous waste, depending on local regulations. This makes it a practical and environmentally responsible choice for routine lab work, especially in educational or teaching labs where safety is a top concern.
GelRed:
GelRed, produced by Biotium, is another popular alternative that offers both high sensitivity and enhanced safety. Unlike ethidium bromide, GelRed is structurally modified to be membrane-impermeable, meaning it cannot enter living cells and bind to DNA, thereby greatly reducing its mutagenic potential. Independent lab tests have confirmed that GelRed is non-toxic and non-mutagenic under standard lab conditions. It binds nucleic acids more strongly than ethidium bromide, providing brighter and more stable fluorescence, and can be visualized using the same UV transilluminators or blue-light systems. Additionally, GelRed is stable at room temperature and compatible with a wide range of electrophoresis protocols. Its safety profile and high performance make it a trusted choice in both research and clinical settings.
Hydrofluoric acid (HF) is an extremely dangerous chemical used in various industrial and laboratory processes, including glass etching, metal cleaning, and semiconductor manufacturing. Unlike many acids, HF penetrates the skin deeply and can cause severe tissue damage and systemic toxicity without immediate pain, leading to delayed symptoms. Once absorbed, HF binds to calcium and magnesium in the body, potentially resulting in hypocalcemia, cardiac arrhythmias, and even death. Burns from HF can be deceptively minor on the surface while causing extensive damage beneath, making immediate and proper treatment essential.
Before working with hydrofluoric acid (HF), all personnel must contact Environmental Health & Safety (EH&S) for approval and guidance. EH&S will ensure that proper training, safety protocols, and emergency response measures are in place, including the availability of Calgonate® Gel and appropriate personal protective equipment. Due to the extreme hazards associated with HF, no work should begin until EH&S has reviewed and approved the procedures.
Calcium Gluconate Requirement for HF Labs:
Calcium gluconate is an essential first aid treatment for hydrofluoric acid (HF) exposure and is required in all laboratories where HF is used or stored. When HF contacts the skin, it can penetrate deeply and bind to calcium in the body, potentially causing severe tissue damage, nerve injury, and life-threatening systemic toxicity. Calcium gluconate works by neutralizing fluoride ions, limiting further injury and helping prevent serious complications such as hypocalcemia and cardiac arrest. It must be readily available in the form of a topical gel (such as Calgonate® Gel) and should be applied immediately after any suspected HF skin exposure. All lab personnel must be trained in its use, as rapid response is critical to minimizing harm.
Dichloromethane (DCM), also known as methylene chloride, is a volatile and toxic solvent commonly used in laboratories for cleaning, degreasing, and extracting compounds. While highly effective, DCM poses significant health risks. It is a potent central nervous system depressant and can cause dizziness, headaches, and nausea upon inhalation. Chronic exposure may lead to liver and kidney damage, and it is a possible carcinogen, linked to an increased risk of cancer with prolonged exposure. DCM is also highly flammable and should be handled with care, in well-ventilated areas, and with proper personal protective equipment (PPE).
If dichloromethane (DCM) is being used in your laboratory, it is essential to contact Environmental Health & Safety (EH&S) for guidance on safe handling, storage, and disposal practices.
EPA Requirements for Dichloromethane (DCM) Use:
The Environmental Protection Agency (EPA) mandates strict guidelines for the use of dichloromethane (DCM) due to its toxicity, potential carcinogenic effects, and environmental risks. Laboratories working with DCM must comply with a range of requirements aimed at minimizing exposure to workers and preventing environmental contamination.
- Exposure Assessments:
- Conduct regular monitoring of air quality to assess DCM concentrations in the laboratory.
- Ensure monitoring meets EPA guidelines for permissible exposure limits (PELs).
- Identify potential exposure pathways (inhalation, skin contact) and take appropriate corrective actions as needed.
- Exposure Control Plan (ECP):
- Develop a plan that outlines controls for DCM exposure, including proper ventilation (e.g., fume hoods), personal protective equipment (PPE), and engineering controls.
- Ensure the ECP addresses safe storage, handling, and disposal of DCM.
- Implement training for all personnel on DCM risks and safe handling practices.
- Reassessments Based on Initial Monitoring:
- Reevaluate exposure levels and the effectiveness of the Exposure Control Plan after initial monitoring events.
- Modify controls and practices based on reassessment outcomes to ensure continued compliance with EPA standards.
- Conduct periodic reviews and updates to the plan as new data or regulations become available.
Peroxide-forming chemicals are highly reactive substances that can form potentially hazardous peroxides when exposed to air, light, or heat over time. These peroxides can be extremely dangerous, as they may detonate without warning, especially if the chemical becomes concentrated. Common peroxide-forming chemicals include ethers, dioxanes, and some aldehydes and ketones. Due to the risk of peroxide formation, laboratories must adhere to strict protocols to ensure the safe handling, storage, and disposal of these substances.
Requirements for the Use of Peroxide Forming Chemicals in Laboratories:
- Dating the Chemical:
- Upon receipt and opening of peroxide-forming chemicals, laboratories must clearly date the containers with both the date of receipt and the date the container is first opened. This allows for proper tracking of the chemical’s shelf life.
- Testing for Peroxides:
- Peroxide-forming chemicals should be tested annually for the presence of peroxides, especially if they have been stored for an extended period or are not used frequently. This testing should be conducted using appropriate chemical tests or kits designed to detect peroxide formation.
- Disposal of Peroxide Forming Chemicals:
- Peroxide-forming chemicals should be disposed of promptly when they exceed their recommended shelf life or if peroxide formation is detected. Laboratories must follow established chemical waste disposal procedures, ensuring that peroxides and peroxide-forming chemicals are disposed of in a safe, controlled manner.
Piranha acid, a highly reactive mixture of concentrated sulfuric acid and hydrogen peroxide, is commonly used to clean organic residues from glassware in laboratories. Despite its effectiveness, it is extremely dangerous and must be handled with extreme caution. The mixture reacts violently with organic materials and can cause explosive reactions if prepared or used improperly. It generates intense heat and corrosive fumes, posing severe risks of chemical burns, respiratory damage, and even detonation.
Due to the extreme hazards associated with piranha solution, all personnel must contact Environmental Health & Safety (EH&S) before preparing or using it. Piranha solution is highly reactive and can cause violent, exothermic reactions, toxic fumes, and severe chemical burns. EH&S will review your procedures, ensure proper training is in place, and verify that all necessary safety controls—such as appropriate PPE, fume hood use, and waste disposal plans—are established. No work involving piranha solution should begin without prior EH&S approval.
Nano-Strip:
Nano-Strip is a commercially available cleaning solution designed as a safer and more stable alternative to piranha acid for removing organic residues from substrates such as silicon wafers and glassware. It typically contains stabilized sulfuric acid and hydrogen peroxide, offering strong oxidative cleaning power without the unpredictable and violent reactions associated with freshly mixed piranha solution. Nano-Strip is formulated for consistency and ease of handling, reducing the risk of spontaneous decomposition or explosive reactions. While still corrosive and requiring careful use, Nano-Strip provides a more controlled and manageable option for high-purity cleaning applications in research and semiconductor labs.
Pure-Strip:
Pure-Strip is a commercially formulated cleaning solution used as a safer, more controlled alternative to traditional piranha acid for removing organic contaminants from substrates such as silicon wafers, glassware, and laboratory equipment. Designed for high-performance surface cleaning, Pure-Strip offers strong oxidative power while minimizing the risks associated with manually mixing hazardous acids. Its stabilized composition enhances user safety by reducing the chances of violent reactions and fume generation. Although still corrosive and requiring appropriate PPE and ventilation, Pure-Strip provides a more consistent and manageable solution for labs seeking to maintain rigorous cleanliness standards without the extreme hazards of piranha solution.
AlnoChromix:
AlnoChromix is a specialized cleaning solution designed for the removal of organic residues and contaminants from laboratory glassware, metal surfaces, and semiconductor substrates. It is a safer and more stable alternative to traditional piranha acid, offering powerful oxidative cleaning properties without the violent reactions associated with piranha solution. AlnoChromix effectively cleans surfaces while reducing the risk of chemical burns, fume hazards, and instability, making it ideal for use in laboratories where strict safety protocols are essential. While still requiring proper handling and PPE, AlnoChromix provides a reliable and less hazardous option for high-purity surface preparation in scientific and industrial applications.
RCA Clean:
RCA Clean is a widely used cleaning solution in semiconductor and materials science labs, designed to remove organic and inorganic contaminants from silicon wafers, glassware, and other substrates. It typically consists of a mixture of deionized water, hydrogen peroxide, and ammonium hydroxide, offering an effective and safer alternative to highly reactive cleaning solutions like piranha acid. RCA Clean is particularly known for its ability to thoroughly clean surfaces without causing damage or highly exothermic reactions, making it a preferred choice for preparing substrates in microelectronics and surface science. While it still requires proper handling and PPE, RCA Clean is an essential solution in labs where precision and safety are paramount in cleaning procedures.
Contact EH&S for more information about Piranha acid alternatives.
Toxic gases can present acute physiological effects when inhaled. The International Fire Code (IFC) and the National Fire Protection Association (NFPA) define toxic gases separately based on the lethal concentration of the gas that kills 50% of the animal test population, known as the LC50. Hazard classification of the gas and the requirements for use depend on the LC50. The chart below highlights the gas classifications based on LC50 for the IFC and NFPA.
Due to the code and regulatory requirements, as well as the inherent risks associated with these type gases, use of these materials requires careful consideration to ensure they are used safely. EH&S and Facilities Management must be consulted prior to the use of toxic gases on campus.
Storage and Use Requirements:
- Highly toxic gases must be stored in an exhausted gas cabinet/enclosure, regardless of volume or concentration.
- Toxic gases in portable cylinders require local exhaust ventilation or collection systems.
- Exceeding MAQ triggers additional safety requirements and potential building occupancy changes.
Ventilation Requirements:
- Toxic gases may not be used or stored in rooms with recirculated air.
- If a catastrophic release exceeds 25% of IDLH, gases must be stored in an exhausted enclosure.
- Ventilation monitors are required for any exhausted enclosure with toxic gases.
- Certain situations may require interlocking ventilation with automatic gas shutdown.
Toxic Gas Monitoring:
- Monitoring systems required when gas release could exceed 10% of the IDLH.
- Gas monitoring must be continuous and near where gases are used or stored.
- Systems must detect at or below OSHA PEL or ACGIH TLV and be regularly calibrated.
Alarm Requirements:
- All monitoring systems must have visible and audible alarms for gas leaks.
- Alarms should be located in the lab and adjacent corridors.
- Alarm thresholds should be set to OSHA PEL or ACGIH TLV (whichever is lower).
- Monitoring system should be tied to the fire alarm control panel for building evacuation coordination.
Gas System Management:
- All regulators, valves, and lines must be chemically compatible with the gases used.
- Gas systems must be leak-tested before use.
- Regulators must match the size and type of gas cylinders.
- Purge vents must connect to an exhaust system discharging to a safe location.