Serkan SAYINER, DVM PhD. Assist. Prof.
Near East University, Faculty of Veterinary Medicine, Department of Biochemistry
Laboratory Safety and
Cleaning
Issue Date: 26.09.2019 | Revision Date/No: -/-
Laboratory Safety
Laboratory Safety
• It should never be forgotten that the laboratory is a dangerous environment with serious work.
• Laboratory safety is crucial in order to protect staff and the working material. It involves the use of certain laboratory rules, methods, infrastructure and equipment during the study.
• Laboratories should not be operated in a manner that could disrupt or cause hazard/danger. Staffs in these places must know the potential danger and what to do in an emergency. All rules, whether verbal or written, should be followed carefully, and the parts that are not understood should be asked to the laboratory supervisors.
Laboratory Safety
• We need to know that the chemicals used in the
laboratory can be toxic, carcinogenic, caustic, irritating or flammable. Many chemicals are harmful to health and the environment if used improperly or disposed of
improperly.
• Strong acids and bases, volatile, flammable, mutagenic and corrosive compounds, radioactive isotopes,
electrical and cutting objects are hazardous and must be used properly.
Laboratory Safety
• Why is it essential?
• Safety operation protects,
• Laboratory staff,
• General service officers,
• Visitors and
• Work and studies.
Laboratory Safety
• Risk analysis must be performed to ensure safety.
• Risk can be defined as the likelihood or probability of a given hazard of a given level causing a particular level of loss of damage.
• Hazards should be identified and risks assessed.
• All relevant data should be used.
• Control procedures should be established to minimize these risks.
• Assessments should be recorded.
• Control measurements should be made and evaluated.
• According to the section to be studied, special training on how to do this process should be taken.
Control Measures
• Less risky substances should be used.
• The safest form of the substance should be used (e.g. instead of powder use solution).
• Ensure good ventilation.
• Operations can be carried out in partially enclosed environments (such as a fume hood).
• Operations can be performed in fully
enclosed environments (such as laboratory glove box).
Control Measures
• Exposure time should be shortened, distance increased and volume reduced.
• Protective equipment must be worn.
• Glove
• Goggles, Protective mask
• Laboratory Coat
• Closed shoes
• Long hair should be tied.
Laboratory Gloves
• There are various types of laboratory gloves;
Latex, Nitrile, Neoprene.
• An appropriate type of gloves should be used according to the task.
• Chemical protective gloves should be selected according to the materials to be worked.
• Gloves must be removed before using
instruments, telephoning and leaving the laboratory.
Laboratory Hygiene
• Drinks, food and cigarettes should not be consumed in the laboratory.
• Cosmetic products should not be used.
• The face, lips and eyes should not be touched.
• Pens and ballpoint pens should not be taken into the mouth.
• Wash hands before leaving the laboratory.
Especially before consuming food.
Common Laboratory Hazards
• Fire
• Fracture of glass materials
• Needles
• Spillage
• Pressurized equipment and gas cylinders
• Extreme heat and cold
• Chemical Hazards
• Biological Hazards
• Radiation
Chemicals
• Information about the health hazards of all chemical substances must be learned by the laboratory staff.
• The laboratories must have a written chemical hygiene plan and the plan should include the procedures and practices to reduce the exposure of the personnel to hazardous chemicals.
• The rules or chemical written records should be revised whenever they change. In all areas where hazardous
materials such as chemical storage rooms and cold rooms are stored and handled, proper ventilation and discharge
channels must be provided.
Chemicals
• The laboratory should have a periodically controlled inventory of chemicals.
• New and old should be distinguished,
• Expiration dates should be followed,
• The critical stock level should be determined,
• Storage conditions and the area should be determined,
• Risk and protection methods should be determined,
• Records must be kept (Name, Catalog no, Lot no, CAS no, Volume, Hazard class, storage condition, storage location).
• MSDS Form and,
• Label information must be available.
Chemicals
• MSDS (Material Safety Data Sheet)
• MSDS is a document that contains information on the potential hazards (health, fire, reactivity and environmental) and how to work safely with the chemical product. It also contains information on the use, storage, handling and emergency procedures all related to the hazards of the material.
• MSDSs are prepared by the supplier or manufacturer of the material. It is intended to tell what the hazards of the product are, how to use the
product safely, what to expect if the recommendations are not followed, what to do if accidents occur, how to recognize symptoms of
overexposure, and what to do if such incidents occur.
Chemicals
• What information is on the MSDS?
• Product Information (product identifier
(name), manufacturer and suppliers names, addresses, and emergency phone numbers), Hazardous Ingredients, Physical Data, Fire or Explosion Hazard Data, Reactivity Data
(information on the chemical instability of a product and the substances it may react
with), Toxicological Properties (health effects), Preventive Measures, First Aid Measures, Preparation Information (who is responsible for preparation and date of preparation of MSDS).
Chemicals
• NFPA Labeling (National Fire Protection Association).
• The NFPA diamond provides a quick visual representation of the health hazard,
flammability, reactivity, and special hazards that a chemical may pose during a fire.
• The NFPA diamond consists of four color- coded fields: blue, red, yellow, and white.
• The blue, red, and yellow fields—which represent health hazard, flammability, and reactivity,
respectively—use a numbering scale ranging from 0 to 4. A value of 0 means that the material
poses essentially no hazard, whereas a rating of 4 indicates extreme danger.
• The white field is used to convey special hazards.
2 2
3 W
Chemicals
• HMIS/HMIG Labeling (Hazardous Materials Identification
System/Guide)
• This system allows staffs to identify at a glance the type and degree of hazards
associated with each product they use.
2 1 3 E
Classification of Chemical Hazards
• The Globally Harmonized System of Classification and Labelling of Chemicals (GHS) is an internationally
agreed-upon standard managed by the United Nations and Universal warning hazards pictograms are core
elements of GHS.
• Pictograms are warning signs indicating hazards.
• Knowing the pictograms is critical to alerting laboratory personnel and distinguishing specific hazards from emergencies such as fire or
explosion.
GHS Hazards Pictograms
GHS Hazard Pictograms
• GHS01: Explosive
• Explosives, unstable explosives, self-reactive substances and mixtures, organic peroxides.
• GHS02: Flammable
• Flammable gases, aerosols, liquids, solids, Self-reactive substances and mixtures; Pyrophoric liquids and solids;
Combustible solids and liquids; Self-heating substances and mixtures; Substances and mixtures, which in contact with water, emit flammable gases; Organic peroxides.
GHS Hazard Pictograms
• GHS03: Oxidizing
• Oxidizing gases, liguids and solids.
• GHS04: Compressed gas
• Compressed gases, liquefied gases, refrigerated liquefied gases, dissolved gases.
GHS Hazard Pictograms
• GHS05: Corrosive
• Corrosive to metals, skin corrosion, serious eye damage,
flammable gases, self-reactive substances or mixtures, organic peroxides.
• GHS06: Toxic
• Acute toxicity (oral, dermal, inhalation).
GHS Hazard Pictograms
• GHS07: Harmful
• Acut toxicity (oral, dermal, inhalation), skin and eye irritation, skin sensitization, specific target organ toxicity following single exposure (respiratory tract irritation, narcotic effects), harmful to ozone layer.
• GHS08: Health Hazard
• Respiratory sensitization, Germ cell mutagenicity,
Carcinogenicity, Reproductive toxicity, Specific target organ
toxicity following single exposure, Specific target organ toxicity following repeated exposure, Aspiration hazard.
GHS Hazard Pictograms
• GHS09: Environmental Hazard
• Acute hazards to the aquatic environment,
• Chronic hazards to the aquatic environment,
• Environmental toxicity.
Chemical Labels
• The identification of a chemical substance is essential.
• International regulations have made it necessary for each chemical to have a label describing the
substances/chemicals, risks and preventive measures.
• The label must include the content of the chemical, the manufacturer's name, hazard markings as well as the R and S codes.
Chemical Labels
• The labels must be read carefully and handled in accordance with the information given.
Chemicals
• Precautions should be taken against fires.
• Flammable materials should be used to a minimum.
• Must be stored in special cabinets.
• Experiments should be carried out using heating devices with temperature control.
• For example, a water bath should be used instead of a hot-plate or bunzen burner.
Chemicals
• Safety goggles should be worn when handling acidic and alkali solutions.
• Acids, caustic materials and strong oxidizing agents should be mixed in the sink.
• Acids should be diluted by adding slowly to water. Never add water to the concentrated acid.
• All reagent bottles should be labeled.
• The reagent's name, concentration, preparer's name and surname, preparation date and expiry date must be on the label.
Classification of Chemical Substances
• Corrosive Chemicals: Acids and Bases. They are
corrosive, toxic, flammable, caustic, reactive with water and unstable chemicals. Acids and bases are stored
separately. Absorbents or neutralizers should be used in the event of leaks or spills.
• Toxic Substances: They enter via oral, dermal or inhalation route. Acute and chronic effects occur depending on the level of exposure.
Classification of Chemical Substances
• Carcinogens: All kinds of substances that cause cancer cells to develop or grow. Examples are arsenic
compounds, benzidine, lead arsenate, chloroform and ethylene oxide. Most are aromatic amines used in
laboratory (such as benzidine).
• Reactive chemicals: They ara flammable and
inflammable chemicals. Examples include acetone,
benzene, diethyl ether, alcohols, heptane and toluene.
The NFPA label is important.
Classification of Chemical Substances
• Flammable Chemicals: Are stored in appropriate safety containers and cabinets. They must be kept away from oxidants.
• Oxidizers: Cause easily oxygen release. Chlorate, permanganate, inorganic peroxide and nitrate are examples. They accelerate the burning of organic
substances. They are stored in a cool and dry place.
They must be kept away from flammable, inflammable and reducing agents.
Classification of Chemical Substances
• Light Sensitive Chemicals: They are substances that react when exposed to light. It must be protected from light and stored in cool and dry places in coloured bottles.
• Peroxide Forming Chemicals: These are chemicals which can
inflamme against impact and heat. It has the ability to form shock- sensitive explosive peroxide crystals. Ex. Cyclohexane and ethyl
ether. Stored in airtight containers, dark, cool and dry places.
• Pyrophoric chemicals: They are liquids, solids, and gases that will ignite spontaneously in air at or below 54,4 °C. E.g. Lithium
amides, potassium. They are stored in a cool and dry place.
Biological Hazards (Biohazards)
• Blood and other sample types (such as serum, plasma, blood products, vaginal secretions, semen, CSF, synovial fluid) can be considered as potential infectious sources.
• Infectious agents are transmitted to personnel by accidental needle pricking, splashing of infectious materials or contamination of tabletop and floor,
centrifugal accidents, and contact of cut or injury sites with contaminated laboratory materials.
Biological Safety (Biosafety)
• It can be defined as the most suitable combination of
laboratory infrastructure, design, equipment, techniques and applications in order to ensure that human and
environmental studies are carried out with materials and infectious microorganisms containing potentially
pathogenic hazards for humans and environment.
• Its purpose is to protect laboratory staff, other people and the environment from potentially dangerous
microbiological agents.
Biological Safety (Biosafety)
• The most important element of biological safety is strict adherence to standard and/or microbiological practices and techniques.
• Each laboratory should identify and implement
biosecurity practices appropriate to the hazards and potential risks to which it may be exposed.
• In addition, risk assessment should be made and the most appropriate biosecurity elements should be
gathered together.
Biological Safety (Biosafety)
• The World Health Organization (WHO) divided microorganisms into four risk groups.
• Group 1: Any biological agent that is unlikely to cause disease in healthy workers or animals. E.g. Bacillus subtilis does not cause infection in humans.
• Group 2: İnsanlarda enfeksiyona neden olan, toplum sağlığı açısından riskin sınırlı olduğu mikroorganizmalar. Any pathogen that can cause human disease but, under normal
circumstances, is unlikely to be a serious hazard to the community. The disease is treatable or preventable. E.g. Streptococcus, Herpesvirus.
• Group 3: Any pathogen that usually causes serious human disease but does not ordinarily spread by casual contact from one individual to another. Cause serious disease in
humans. Treatments and vaccines for these diseases may exist. E.g. Yersinia pestis, HIV
• Group 4: Microrganisms cause deadly disease in humans, and they can easily travel from one person to another. No treatments or vaccines exist for these diseases. E.g. Ebola
virus, Lassa virüs
Biological Safety (Biosafety)
• Laboratory infrastructures are designed at four
different levels in terms of application and protection criteria in parallel with risk groups.
• Biosafety Level 1: If Group 1 microorganisms to be studied. Typical work area is open-type bench.
• Biosafety Level 2: If Group 2 microorganisms to be
studied. Biosafety cabinets or laminar flow cabinets are used.
Biological Safety (Biosafety)
• Biosafety Level 3: If Group 3 microorganisms to be studied. Class 3 Biosafety cabinets are used. It is also called isolation laboratory.
• Biosafety Level 4: If Group 4 microorganisms to be studied. Full isolation suits is used. Laboratories has maximum level of isolation.
Lab Glassware
• The correct techniques must be applied when using glassware.
• Glass material should never be used under pressure or vacuum unless it is designed for mentioned circumstances.
• Cracked or broken glass materials should be discarded. This can be a risk for staff.
• The broken glass should be disposed of in suitable waste bins (such as needle waste bins). Do not dispose of in general trash.
Spills
• Clean immediately after spilling liquid.
• This was determined as part of the risk assessment.
• Hazardous substances must be disposed as toxic waste.
Gas Tanks
• It should never be used without training.
• The lowest number should be used.
• If possible, it should be stored externally.
• Measure against falling or overturning (can be chained).
• It should only be moved with a gas cylinder trolley.
• Appropriate regulators and control equipment for the relevant gas should be used.
• Precautions should be taken against leaks.
Cryogenics
• Liquid gases are extremely cold and may cause burns.
• Liquid gases evaporate and can cause asphyxiation in many people.
• If cryogens need to be transported in the elevator, it should be noted that there are special procedures to follow.
• Special training is required to use cryogenics.
Electrical Equipment
• Before use, a visual inspection of the electrical equipment should be carried out and any significant abrasion or
defects should be investigated.
• All portable electrical equipment must have a current PAT test ”label.
• NEVER use malfunctioned equipment.
General Laboratory Practices
• Keep the work area tidy.
• Wastes should be cleaned, necessary materials should be washed, finished work should be put in place.
• Make sure everything is safe before leaving work.
• There should be a general cleaning procedure.
• A tidy laboratory prevents accidents for everyone.
Laboratory Equipments
• Laboratory equipment should not be used without training.
• Personnel must successfully complete the training of the
equipment. Then authorization is made.
First Aid
• Laboratory staff should receive first aid training.
• In chemical splashes, wash with plenty of water for 10 minutes.
• If there is bleeding, it should be controlled by direct pressure application. Caution should be paid to foreign bodies such as glass.
• All accidents should be reported to the laboratory manager.
Wastes
• How to dispose of laboratory waste as part of the risk assessment should be
determined.
• Solvents and oils should be separated into suitable bottles or drums.
• There should be procedures for the
collection and disposal of chemical and biological waste.
• It should not be disposed of with normal waste unless permitted.
Laboratory Safety
• Laboratory safety instructions and rules covering all processes must be established.
• The rules are posted in the workspaces.
• Trainings should be conducted at regular intervals to keep staffs' knowledge up to date.
• Staff should not hesitate to ask questions for unclear points.
• It should be noted that safety is a teamwork.
Emergency Response Guide
• It is always necessary to get ready for accidents that may occur in laboratories.
• Each laboratory must prepare an emergency plan and all personnel should be familiar with it in order to plan the actions to be taken in order to minimize the loss of property and life in case of accidents, disasters and fires (fire, chemical spill, injury, gas smell / electric leakage, earthquake and flood).
Emergency Response Guide
• Emergency plan should include officers and phone numbers.
• The emergency plan should include tasks being done by laboratory manager and staff in case of fire, chemical spill, treatment, gas odour/electric leakage, flood and earthquake.
Laboratory Cleaning
Cleaning Labware
• The cleanliness of the labware used is critical for the reliability of the analysis results.
• Any contamination in the labware (beaker, pipette, etc.) significantly affects the experiment or study done.
• It is investigated the effects of metal ions, organic
molecules and other chemicals involved in biochemical processes in many experiments. Therefore, the use of dirty labware leads to failure of the experiments.
Plastic Labware Cleaning
• Polyethylene plastic materials are generally used in laboratories.
• Polyethylene materials to be used for the first time are first cleaned with 8 M urea solution. It is then rinsed with
distilled water, washed with 1 N KOH solution, passed
through distilled water again and then washed with 0.001 M EDTA to remove metal ion contamination. Finally, rinsed with distilled water.
• Following this, it is sufficient to wash with 0.5% detergent and rinse with distilled water after each use.
Lab Glassware Cleaning
• Glassware is cleaned as soon as possible after use.
Residues adhere to the glass surface, as solvents will evaporate and residues are attached to glassware. It gets harder to clean.
• If it cannot be cleaned immediately, it must be left
running through tap water. In the cleaning of glassware, it is usually sufficient to wash with 0.5% detergent
water.
Lab Glassware Cleaning
• If water drops remain on the glass surface after cleaning, this mean the process is not sufficient. However, special washing may be required for a more precise work.
• For the removal of organic residues in glassware, bichromate cleaning solution has been used for many years. However, it is not recommended today because of the toxic effect of
chromium, its carcinogenicity and the intensive use of H2SO4.
• It is required to wash with concentrated nitric acid to
remove the ions. Glass pipettes should be soaked in 0.5%
detergent and then rinsed in running water.
Cleaning of Quartz and Glass Cuvettes
• Cleaning of quartz or optically polished glassware with KOH or strong bases prepared in
ethanol causes corrosion.
Instead, it is enough to clean with 0.5% detergent and rinse with distilled water.
Drying of Plastic and Glass Labware
• Glassware is dried in special ovens. We can also dry many plastic materials in these ovens. However,
cellulose nitrate centrifuge tubes (because they are explosive) should not be placed in the drying oven.
• If we are not sure of the type of plastic material, the drying oven should not be used. The glassware can also be dried with acetone. However, acetone-bound
pollution should not be used if it affects the experiment.
Sterilization
• Sterilization refers to any process that eliminates, removes, kills, or deactivates all forms of life.
• After sterilization, there is a possibility of 1 live in 1 million microorganisms in the environment.
• The acceptable limit for sterilization is considered to be at least 10-6 and the media is free from all
microorganisms, including endospores.
Sterilization
• Disinfection is a process that is designed to kill actively growing and vegetative microbial microorganisms to a certain level, and it does not, unless the disinfectant is classified as a sterilant, apply to bacterial endospores.
Disinfection is generally applied to inanimate surfaces or objects such as work areas, equipment.
• Decontamination is the process of decreasing
antimicrobial presence in an area or on a surface.
Sterilization is actually a type of decontamination along with disinfection and antisepsis. The aim of
decontamination is that the contaminated material is got ready for the next treatment.
Sterilization
• Physical and chemical methods are used in sterilization.
• Sterilization by physical methods is carried out by incineration, humid heat, tyndallization, dry hot air and filtration. Ex. Autoclaving.
• In chemical sterilization, sterilization is performed by applying various chemicals. Ex. Ethylene oxide, formaldehyde.
• The time required for sterilization depends on the type and the amount of material.
• For example; solids require longer time than liquids. Time, in sterilization of liquids, increases as the amount of volume increases. Sterilization
method is determined according to the type of materia.
References
• Altıntaş A, Fidancı UR, Sel T, Yılmaz G, Pekcan M (2015) Veteriner Laboratuvar Teknikleri ve Prensipleri. 4.baskı, Anadolu Üniversitesi Yayınları, Eskişehir.
• Beran JA (2013). Laboratory Manual for Principles of General Chemistry. 10th ed. Wiley
• Karagül H, Altıntaş A, Fidancı UR, Sel T (1999). Temel Biyokimya Uygulamaları. Medisan Yayınevi, Ankara.
• Kimyaca.com. Erişim: https://kimyaca.com/malzeme-guvenlik-bilgi-formu/ Erişim Tarihi: 26.10.2018
• Özerol E. Laboratuvar Güvenliği. Erişim:
http://www.tibbigenetik.org.tr/portalyonetim12//images/48/Dosyalar/tmp//201837161633.pdf Erişim Tarihi: 27.10.2018
• The International Genetically Engineered Machine. Erişim: http://2016.igem.org/Safety/Risk_Groups Erişim Tarihi: 28.10.2018
Revision Status
• Issue Date: 26.09.2019
• Revision Date/No: -/-
