Cluster for Molecular Chemistry

GENERAL SAFETY RULES FOR WORKING WITH HAZARDOUS SUBSTANCES IN THE LABORATORY AREAS IN THE SL-2 (‘MEDIUM’) CATEGORY

Table of contents

1. Introduction
2. General safety
2.1. Important rules
2.2. Organization of Safety
2.3. Personal Protection Equipment (PPE)
2.4. Order, tidiness and hygiene
2.5. Orientation in the laboratories
2.6. Working alone, working outside regular working hours
2.7. End of the working day, night-time experiments
3. Hazardous substances
3.1. Preparing an experiment
3.2. Health risks
3.3. Threshold limits
3.4. Storage of chemicals
3.5. Waste
3.6. Compressed gases
3.7. Hazardous gases
3.7.1. Working with HF
3.7.2. Working with Cyanides
3.8. Cryogenic liquids
3.8.1. How to safely handle a cold trap
3.8. Chromatography columns
3.10. Distillation
3.11. Cleaning glass filters
3.12. Autoclaves
3.13. Biochemistry
3.13.1. Ethidium bromide
3.13.2. Acrylamide
3.13.3. Organic solvents
4. Faults, fire and accidents
4.1. General
4.2. Guidelines in case of a power failure
4.3. Guidelines in case of chemical spills
4.4. Guidelines for accidents
4.5. Guidelines in the event of fire
4.6 Guidelines for evacuation
5. Important information
5.1. List of abbreviations
5.2. Important internet links


GENERAL SAFETY RULES FOR WORKING WITH HAZARDOUS SUBSTANCES IN LABORATORIES IN THE CATEGORY SL-2 ('MEDIUM')

1. Introduction

As part of the Faculty of Science Policy on Hazardous Substances, approved by the Faculty Board on 8-11-2010, laboratories have been divided into a number of categories (Table 1). For more information see overzicht_gevaartekens_laboratoria_en_algemene_ruimten_jan_2013.pdf

Level

Risk category

Name

Description

Measure

0

None

SL-0

Offices and rooms where eating and/or drinking is permitted.

Lab coats prohibited.
Chemicals constituting a health risk prohibited.

1

Low

SL-1
(chemical laboratory with limited risk)

The rooms are used for work involving flammable, toxic, irritating or oxidising chemicals, or chemicals that could damage long-term health, in such small quantities that the health risks are limited.

A cotton lab coat, long trousers, sturdy shoes and safety glasses are compulsory while working with flammable, toxic, irritating or oxidising chemicals, or chemicals that could harm health in the long term.

2

Medium

SL-2
(standard chemical laboratory)

The rooms are used to work with flammable, corrosive, toxic, irritating, explosive or oxidising chemicals, or chemicals that may damage health in the long term.

A cotton lab coat, long trousers, sturdy shoes and safety glasses are compulsory, irrespective of the work being carried out.

2+

High

SL-2 + (chemical laboratory with increased risks)

The room is used to work with high-risk substances and/or processes.

In addition to a cotton lab coat, long trousers, sturdy shoes and safety glasses, adequate PPE (such as a face mask, gloves and gas mask) is also compulsory. Other clothing may be compulsory in specific situations.

B-1

Low

ML-1 (molecular biology laboratory)

Rooms used for work involving low-risk genetically modified organisms.

No entrance for unauthorized persons. To work in ML-1 labs training is required; contact the responsible officer (for labs 02.216 and 02.242: José Roelofs-Hendriks

2

Medium

L-2 (laser laboratory class 3B, 3R and 4)

Rooms used to work with high-risk laser beams More information on laser safety you can read on microscopyu.com.

To work at L-2 level, contact your Laser Safety Officer (for lab 03.221: Aigars Piruska)

 

The regulations stated below apply to laboratory areas in the 'medium' category (SL-2). These are areas were people work with chemicals that are flammable, corrosive, toxic, irritating, explosive, oxidising or otherwise dangerous to long-term health. The foremost safety rule is that lab coats and safety glasses are compulsory, irrespective of the work being carried out. Departments will alert new staff/interns starting work in laboratories in the 'high' category to the additional rules that apply there.
These safety regulations for working with hazardous substances are a more specific version of the general Faculty of Science safety regulations and recommendations issued by the Faculty of Science Internal and Housing Affairs department.1 We also refer you to this document for information on the Working Conditions Act, electricity, Ionising radiation, lasers, environmental protection, working with machines, hoisting and transport, working at heights, and working with vacuum applications. The faculty regulation for waste management for was drawn up with the Radboud University Department of Occupational Health & Safety and Environmental Service (also available in English)2.
Included in the policy on working with hazardous substances is the commitment to strictly monitor the compliance with the regulations in question. This means that the supervisors in the Bachelor's practical courses can refuse to admit students to the labs if they are not dressed according to the regulations, or send them away if they do not observe the guidelines relating to personal protection equipment or order, tidiness and hygiene; the same applies to Safety Stewards and work group supervisors in the labs of the research institutes in relation to Master's students, staff and guests. Environment, Health & Safety is a recurring subject in annual interviews between staff and managers. Repeated violation of safety regulations will result in sanctions.
Additional to the chemical labs, the molecular chemistry cluster makes use of some laboratory spaces with a higher safety level regarding the use of GMOs or lasers. Special safety officers are employed to administer the safety in those rooms. This manual does not include safety regulations on GMOs or lasers; contact your safety officer before starting to work in those labs.

1These safety regulations can be found on http://www.radboudnet.nl/fnwi/fnwi/arbo/ (in Dutch).
2The faculty waste management regulation was devised with the RU Department of Occupational Health & Safety and Environmental Service and can be found on https://www.radboudnet.nl/veiligengezond/duurzaamheid/afvalscheiding/ (in Dutch).

2. General safety

2.1. Important rules

There are several important obligatory rules.

Safety glasses should be worn at all times in SL-2 laboratories, irrespective of the work being carried out. Safety glasses can be obtained from Peter van Dijk. If you have prescription glasses you can ask Peter van Dijk to order a pair of prescription safety glasses at the cluster's expense (not for undergraduate students).
Lab coats should be worn at all times in SL-2 laboratories, irrespective of the work being carried out. If you work in the department for more than two weeks (Bachelor internship or longer) a lab coat is provided by the department. It is strictly forbidden to work alone in the lab. Always make sure that there is a PhD student, post-doc or staff member within seeing or hearing distance from the lab. Undergraduate students are not allowed to work in the lab outside working hours ( in the evenings and weekends) without direct supervision. Students have only access to the wings between 08:00 and 18:00.
It is not allowed to work with genetically modified organisms (GMOs) in the SL-1, SL-2, and L-2 laboratories of the Cluster of Molecular Chemistry. GMO work should be carried out in the Molecular Biology labs assigned for this purpose (e.g. ML-1). Working in such labs can only be done with permission from the responsible co-workers. It is not allowed to take the products of work with GMOs out of these labs (and into other labs) unless one is absolutely sure that the cells are dead. Discuss this carefully with your mentor, and with the responsible people in the GMO-authorized laboratory.

Make sure you know how to handle in case of an emergency (see sections 4.2-4.7).

Before starting an experiment, always make sure you know the safety risks involved and what measures should be taken regarding safety and environment (waste management, see section 3.5).

Work tidy and clean up the workspace(s) when you are finished. Always close your fume cupboard completely when you go away. Report any defects to equipment to the responsible person immediately.

Fill in the appropriate overnight experiment card when you conduct overnight reactions (see section 2.7).

Ask for help from the responsible co-worker or an experienced PhD student, post-doc or staff member when using unfamiliar equipment or techniques, in particular:

Hazardous gases (see section 3.7)
Schlenk lines
Distillation setups
Cold traps (see section 3.8)
Vacuum setups (desiccators, freeze-dryers, etc.)

2.2. Organisation of safety3

The following staff fulfil an important role in the faculty/departmental safety policy:
Mentor: Every new person starting in a laboratory (student, intern, PhD student, post-doc researcher, support & management staff, academic staff, hereinafter to be referred to as laboratory worker or staff and students) will be assigned a mentor (usually his/her academic supervisor) to be his/her first point of contact in relation to safety questions.
Lab Steward: Every laboratory area has a Lab Steward (list per department), who is responsible for the day-to-day running of the lab. The Lab Steward (or someone appointed by him/her) checks the laboratory area every evening to make sure that: all equipment is switched off, all solvent containers have been returned to the appropriate ventilated cabinets, and all taps have been turned off. If the Lab Steward is not the last person to leave the laboratory at the end of a working day, he/she must delegate this checking duty to another person. The Lab Steward is also responsible for the experiments conducted at night (see relevant section).
Safety Steward: In addition to a Lab Steward, every laboratory area also has a Safety Steward who keeps an eye on the safety in the lab. The Safety Steward is in touch with the departmental environment, health & safety officer, and in combination with him/her and the other Safety Stewards, they form a departmental Safety Committee which meets every month to discuss the current state of affairs. The Safety Steward can always be contacted for questions related to safety.
Environment, Health & Safety Officer: Every department with laboratories in the ‘medium’ category or higher has an environment, health & safety officer (PAM-mer), who acts as the contact person for the Department of Occupational Health & Safety and Environmental Service (university level) and the environment, health & safety coordinator (faculty level).
Environment, Health & Safety Coordinator: This person is the first point of contact for cross-department and department-level health and safety matters.
Responsibilities: As stated in the faculty safety regulations, managers are responsible for guidelines and monitoring. The director of business operations is responsible for the policy and its enforcement throughout the faculty.
Drills: Every laboratory worker must take part in a fire extinguishing drill and safety instruction session at least once a year (organised together with the Department of Occupational Health & Safety and Environmental Service).

3Every member of staff may adopt one or more of these roles

2.3. Personal Protection Equipment (PPE)

Personal protection equipment (lab coat and safety glasses) is compulsory at all times in labs in the ‘medium’ category or higher. This also applies to staff and visitors present in the lab areas but not actually handling hazardous substances; other experiments taking place in the lab can also constitute a hazard. Pegs for lab coats and cupboards for safety glasses are located at the entrance to the lab. Coats and safety glasses are also available for visitors. They must be put on when entering the laboratory and taken off and hung up when leaving. People may only enter/leave the labs via these entrances; all other exits are reserved for emergencies.

A complete overview of Personal Protection Equipment (PPE) is available in the general Faculty of Science safety regulations and recommendations4. The most relevant PPE for working with hazardous substances in lab areas in the ‘medium’ category are shown below.

4For the links, see paragraph 5.2 at the end of this document.

2.4. Order, tidiness and hygiene

As will be explained in more detail in 3.1. Preparing an experiment and 3.2. Health risks, these safety regulations revolve around reducing risk when working with hazardous substances by using the Personal Protection Equipment described in 2.3 . If during preparations for an experiment it becomes evident that one or more of the substances being used is highly toxic, the person conducting the experiment should consider whether replacing the substance with a less toxic alternative would affect the results. Alternatively, consider using smaller amounts of the substance. Whatever the circumstances, all experiments involving chemicals must be carried out in a fume cupboard, all solutions, reagents and products must be clearly and correctly labelled, and proper PPE should be worn at all times.

Safety regulations to prevent ingesting hazardous substances: Safety regulations to prevent absorption through the skin: Safety regulations to prevent entry via the eyes (and eye damage): Safety regulations to prevent inhalation: Safety regulations to prevent accidents:

The main message of the detailed safety regulations shown above is that good personal hygiene and meticulous, careful, orderly and tidy working practices will help to prevent accidents. This is not only the responsibility of the person carrying out the work, but also of the people working around him/her. Many accidents are not caused by the victim, but by someone else working in the same area.

A large proportion of accidents in a laboratory are caused by falling, stumbling, or bumping into things. Order and tidiness in the lab are vital. Never work on the edge of the bench or fume cupboard. Place heat sources (burners, heating mantles and hot plates) where they are least dangerous. All laboratory workers must conduct experiments in a way that will keep the fire risk to a minimum. When working with flammable substances, welding or other activities with a high fire risk, extinguishers should always be within easy reach. Make sure that benches and fume cupboards are clean. Clear up spillage immediately (including water). Label all spray bottles. Wash and tidy up equipment after use. Make sure that the lab is clean and tidy at the end of every working day. Plan weekly and six-monthly cleaning sessions together with other workers in the laboratory.

Every laboratory worker must ensure that: 2.5. Orientation in the laboratories

Before embarking on experiments, new laboratory workers must feel confident in the laboratory and be made aware of a few important aspects.

Every laboratory worker must be (and remain) familiar with the location of and where applicable, how to use: 2.6. Working alone, working outside regular working hours

Working alone means that a person is working in a situation whereby he/she cannot continually be seen or heard by others. Situations like this can occur during regular working hours; laboratory workers should try to anticipate periods in which most of the group is attending a conference, for example. The guidelines for working alone are as follows:

Figure 1: Overtime clock.

 

Laboratory, practical or workshop activities are not allowed outside regular working hours unless explicit permission has been given by the manager concerned; working alone is still prohibited, even if permission for working outside regular working hours has been granted. The group leader is authorised to make decisions in these matters and is therefore ultimately responsible.

2.7. End of the working day, night-time experiments

  1. Close the fume cupboard at the end of the day and switch off the lights.
    IMPORTANT: fume cupboards must be properly closed as they only work at 30% of their capacity at night!
  2. Switch off all equipment at the end of the day.
  3. Take special care that all water cooling is turned off if it’s not needed for an experiment. In case you do need water cooling, please take care that all your water hoses are properly secured (only use transparent/plastic ones!) and that the water flow is not too high. Be aware that water pressure might increase significantly during the evening and at night.

The Lab Steward (see 2.2) is responsible for experiments that continue into the night; regulations may vary per department. For each overnight experiment a card that shows information about the specific risks of the experiment must be placed at the fumehood window. This card can be red, green or yellow, depending on the risk level of the experiment. The general rules regarding the cards are as follows:
Green card: for reactions that do not require heating/reflux, pressure, vacuum or hydrogen
Red card: for reactions that require heating/reflux, pressure, vacuum, or hydrogen; also an overnight running vacuum pump requires a red card.
Yellow card: for continuously operating setups (drying liquids under nitrogen or argon.

Also the filling in of the cards should happen according to strict rules:

If the researcher concerned has not left one of these cards or the appropriate card at the fume cupboard containing the experiment, if the card is filled in incomplete, or the reaction set-up causes a safety hazard, the Lab Steward is authorized to stop the experiment during his evening rounds. The cards are collected and placed in the rack near the entrance to the laboratory area so that the Internal Emergency Team and fire service, as well as other people that make use of the lab, can quickly assess the risks and locations of experiments in the area in the event of an emergency.

IMPORTANT: laboratory workers carrying out night-time experiments are personally responsible for taking the card back to the fume cupboard with the experiment the next morning. The Safety Steward can withdraw the card (or cards) if the laboratory worker misuses them.

3. Hazardous substances

3.1. Preparing an experiment

Before every experiment, examine the specific risks of the chemicals you are using by looking at the label, consulting Chemwatch5 and reading the supplier’s information. Do not simply take note of the risks if everything goes according to plan, but look at what can happen if something goes wrong. Think about the possible repercussions, or what would happen if the agitator, reaction flask, cooling or heating were to break or fail. Precautions must be taken for these risks.
The safety regulations shown in the previous section (2. General safety) are aimed at limiting all the risks involved in working with hazardous substances. The following paragraph (3.2. Health risks) shows exactly why everything must be done to minimise the risk of a hazardous substance being ingested, absorbed via the skin, penetrating the eyes or being inhaled. If the risk inventory indicates that one or more of the substances being used in an experiment is highly toxic, the laboratory worker should consider whether less-toxic chemicals could be used without affecting the results. Another useful measure is to keep the amount of the substance used to a minimum. Whatever else, all experiments involving chemicals must be carried out in a fume cupboard, all solutions, reagents and products must be correctly labelled, and good personal protection equipment must be worn.

5For links, see paragraph 5.2 at the end of this document.

3.2 Health risks

The GHS-CLP (Globally Harmonised System of Classification, Labelling and Packaging of Chemicals) system5 introduced throughout Europe in 2009 distinguishes between the following hazard risks and categories of hazardous substances and mixtures, using the pictograms shown below:

Any particular substance may belong to several risk classes and categories.

Figure 2: GHS categories for hazardous substances and the relevant pictograms.

A GHS label comprises the following elements:

Home-made mixtures and solutions of chemicals should be labelled according CLP (Classification, Labelling and Packaging) regulation. The bottle or container for general use should contain:

For Personal use also: If for solutions no GHS symbols are applicable, a blank GHS sticker should be used as a proof of checking its properties. If GHS classification of newly synthesized compounds is unknown, no stickers should be used. Stickers with GHS symbols as well as blank stickers are available from Peter van Dijk.

Posters showing GHS symbols and the H and P statements are available from the Department of Occupational Health & Safety and Environmental Service. The measures per hazard category shown below correspond with those in section 2.3. Personal protection equipment and 2.4. Order, tidiness and hygiene.

The hazard categories are:
Flammable:
Solids, aerosols, gases and liquids (such as organic solvents with a high vapour pressure, e.g. diethyl ether, ethanol) are flammable.
Measure: Avoid open flames in a lab where these substances are used, and beware of heat sources, sparks, electric discharge etc. that may cause ignition.

Corrosive:
Corrosive (aggressive) substances (such as concentrated acids, concentrated alkali, strong oxidisers) are dangerous if they come into direct contact with the skin, eyes and mucous membranes.
Measure: Avoid contact by using PPE.

Toxic:
Toxicity: It is best to assume that all chemicals are potentially dangerous to health, even though for some of them (e.g. water) the concentration would have to be very high to pose a serious risk. In chemistry, the term toxic substances is used if even small doses of a substance could damage the body. Toxicity is therefore relative and depends on the dose. This in turn depends on:

The effect of the dose also depends on personal factors such as sensitivity and general condition.
Measure: Avoid contact by using PPE; do not eat or drink in the labs and wash your hands when leaving the lab.

Irritating, Sensitising, Harmful:
Irritating substances are mildly toxic and single exposure causes minor irritation to the skin, eyes or airways without lasting damage. In the case of repeated exposure without time to recover, the cumulative effect can cause a chronic inflammation. Examples: high or low pH, dissolving skin oil due to organic solvents, specific reactions to epoxide resins, for example, with protein in the skin or mucous membranes. Sensitising substances are allergens that cause an allergic reaction by activating the immune system. This can lead to inflammation and allergic conditions. Alongside reactions to biological-based substances, an allergic reaction can also be caused by latex, for example. Take care when using gloves (see Appendix 1).
Measure: Avoid contact by using PPE; do not eat or drink in the labs.

Long-term health risk:
This includes all the CRM substances: carcinogenic (causes cancer), reprotoxic (dangerous to unborn foetus) and mutagenic (causes permanent changes to genetic material).
Measure: Avoid contact by using PPE; do not eat or drink in the labs. If substances in this category are ordered in Labservant, a warning is generated by Chemwatch stating that this is a CRM substance; in extreme cases, certain orders will initially be blocked. The Internal & Housing Affairs department and the head of the department concerned will first look for an alternative to this substance; if this is not available, a permit for working with the substance will have to be applied for via the Department of Occupational Health & Safety and Environmental Service.
IMPORTANT: Pregnant women must avoid all contact with reprotoxic substances. Tell your manager in plenty of time if you are pregnant. Strictly speaking, all laboratory workers (m/f) planning a family should stay away from reprotoxic substances.
Compounds that damage the airways when inhaled (such as asbestos and silica) form another important category and can lead to silicosis (Potter’s rot). Asbestos should only be processed by specialist companies; chromatography columns should be filled in a fume cupboard with good exhaustion (see 3.6).
Measure: See the PPE for breathing protection (dust and gas masks) in the faculty safety regulations6.

Hazard to aquatic environment:
Toxic to water organisms.
Measure: Avoid releasing these substances into the environment by for example discharging them through the drains (see also Appendix 3).

Explosive:
Substances and mixtures that may undergo a powerful exothermal discharge without contact with oxygen (air), with a rise in pressure resulting from the accumulation of gases.
Measure: Avoid heat, shocks and high concentrations. Use the fume cupboard when working with substances of this kind.

Oxidising:
Helps to ignite flammable substances.
Measure: Avoid contact with flammable substances.

Compressed gases: see section 3.6

6For links, see paragraph 5.2 at the end of this document.

3.3. Threshold limits

Threshold limits have been set for hazardous substances:
The limit of the exposure concentration is linked to a specific duration. The combination of threshold limit and duration should be chosen so that a laboratory worker would not suffer damage to his/her health even after having worked with the defined threshold limit of the substance concerned for forty years. For this reason, the threshold value is defined as a Time Weighted Average over a period of eight hours, the length of an average working day; this is the so-called 8-hour TWA. The time-weighted average for acutely toxic substances is defined as the 15-min TWA. Specific cases require an instantaneous exposure threshold limit, the so-called ceiling value (reference time practically zero). The laboratory worker is at liberty to determine the reference time for the threshold limit, as long as the basic principle is realised: exposure at the threshold limit during an employee’s entire working life must not cause damage to his/her health; this also applies to his/her children (see table 2 for the example of acetone).

Threshold limits can be found in Material Safety Data Sheets. However, the extent of exposure in the lab is hard to estimate. Therefore, always work in your fume cupboard to prevent inhalation.

Table 2. Threshold limits for occupational exposure to acetone.


Legal threshold limit (Netherlands)

Acetone

Threshold limit 8-hr TWA

1210 mg/m3

Threshold limit 15-min TWA

2420 mg/m3

 

EU (directive 2000/39/EC)

Acetone

Indicative threshold limit 8 hr

500 ppm

1210 mg/m3


3.4. Storage of chemicals

Chemicals are separated according to their danger categories. In case of multiple categories for a single compound, the priority is:

  1. Flammable
  2. Oxidizing
  3. Toxic
  4. Corrosive
  5. Dangerous for the aquatic environment
Dangerous combinations can be found in Figure 3. The fifth category is normally combined with the other categories, so no extra precautions are needed.
Don’t store Flammable and Oxidizing compounds together at any time.



Figure 3. Dangerous and "safe" combinations of the different GHS categories.

Solvents: All solvents have to be stored in the safety cupboards ("plofkasten"), in different drip trays according to their safety label. This also applies to the solvent stock. Each drip tray should contain chemicals of only one danger category; its volume should be at least 110% of that of the largest bottle.
Liquid commercial chemicals should also be stored in these safety cupboards. Each tray is assigned to a single danger category and its volume should be at least 110% of that of the largest bottle. Acids and bases: have to be stored in separate cupboards under the fumehoods (this also applies to acids such as SOCl2). However, if they are flammable (such as KOH in EtOH) bases and acids should be stored in the safety cupboard. This does not apply to HCl solutions as they are corrosive; its ethylacetate and dioxane solutions go in the waste fume cupboard. Nitric acid (oxidizing) should be stored completely separate from all other liquids.
Solid commercial chemicals: Personal chemicals (only solids, registered in your name) can be stored under your own fume hood, as long as they are categorized as mentioned above in different drip trays.
Shared solid chemicals, such as salts, bases and silica, can be stored per category in the apothecary cupboard.
Toxic chemicals should not be stored in the lab but in the logistic centre. If they are used in the lab they should be returned to the logistic centre as soon as possible.
Oxidizing chemicals, however, have to be in a separate drip tray in the safety cupboard.
Freezer/Fridge: Commercial and "home-made" chemicals should not be in the same freezer/fridge. All the different categories have to be in different drip trays.

Liquids of contents 250 ml or less and solids of contents 1 kg or less hasn’t to full fill these rules. However place them always in a drip tray.

During ordering of a chemical a location has to be given were it will be stored. This location should be in agreement with the above rules. After use of a chemical it should be returned to its location that is given on the label that can be found on the container.

"Home-made" chemicals: Home-made mixtures and solutions of chemicals should be labelled according CLP (Classification, Labelling and Packaging) regulation. The bottle or container for general use should contain:

For Personal use also: If for solutions no GHS symbols are applicable, a blank GHS sticker should be used as a proof of checking its properties. If GHS classification of newly synthesized compounds is unknown, no stickers should be used. Stickers with GHS symbols as well as blank stickers are available from Peter van Dijk.

3.5. Waste

For waste originating from our laboratories the following main categories are distinguished: liquid waste, solid waste, glass waste, sharp objects. For a full overview, please refer to the faculty regulations mentioned in the introduction (also available in English)7 and see the AMD website7 on "Afvalstoffenregeling FNWI"/"Waste Management Regulations" and "Overzicht van de indeling voor gevaarlijke stoffen RU"7.
On the whole, paper/cardboard that has not been in contact with chemicals, and the majority of special waste products such a ink cartridges, batteries and adhesives are collected per department in special containers in the technical area near to the lifts.

Labelling: All waste containers must be properly labelled; appropriate stickers are available in every lab area. See faculty waste disposal regulations) for an overview. IMPORTANT: the old labels must be removed or rendered illegible on all empty containers ready for disposal before being taken to the Logistic Transfer area; this also applies to glass bottles and jars in the glass waste.

Liquid waste should be divided into different categories according to the flow chart in Appendix 2 and disposed in properly labelled 5-litre or 10-litre jerry-cans. The main categories relevant for our labs are: inorganic acids (10-litre jerry-can), inorganic lyes (10-litre jerrycan), non-halogenated organic solvents, halogenated organic solvents, ethidium bromide solutions, toxic liquids, watery solutions originating from laboratories.

Solid heavy metals, to prevent ignition, are collected in liquid waste containers according to the liquid waste flow chart (see Appendix 2).

Solid waste, including silica from the chromatography columns, soiled paper and glass contaminated with chemicals, but with the exception of heavy metals, needles and special waste products (see above) such as ink cartridges, batteries and adhesives, are collected in a brown container with a beige lid. The appropriate labels should be attached to the container (label 5.12 and ADR label toxic).

Glass when contaminated with chemicals, can be collected separately in a similar brown container with beige lid or in a solid waste container (See above). The appropriate labels should be attached to the container (label 5.12 and ADR label toxic). Clean glass is collected in a glass waste container in the laboratory. Rinse glass first if it has been contaminated with chemicals, or let solvents evaporate in the fume cupboard. IMPORTANT: remove the original labels or render them illegible.

Needles and other sharp metal objects must be collected in a special waste container, a so-called "SharpSafe" (yellow with a white lid and a narrow opening – make sure they are properly attached). Preferably, this container is only used for needles without the covers, as putting the cover back on the needle is an additional risk and the covers take up extra space. To detach a needle from a syringe in a safe manner, the incisions in the SharpSafe lid can be used. A SharpSafe should NOT be used for glass; neither for syringes, tubing or any other non-sharp thing you might attach to a needle.

Waste is removed from the transfer rooms in the laboratory wings by the Logistic Centre, but only if their instructions as given here are strictly followed up:

Waste, such as paper, plastic, batteries, ink cartridges, glass, can be placed in containers in room HG03.004 or HG3.081. Polystyrene boxes can be placed on the floor in those rooms.

7For the links, see paragraph 5.2 at the end of this document.

3.6. Compressed gases

Pipes for hydrogen (10 bar), argon, nitrogen, carbon dioxide and helium have been fitted on the lab benches and/or in fume cupboards in line with expected use in the lab areas. If another gas (or hydrogen> 10 bar) is needed regularly for long periods, a large cylinder can be placed in one of the cylinder cupboards outside the lab, and connected using the interchangeable gas feed. Large gas cylinders of this type are available from the Logistics Centre or must be ordered.
IMPORTANT: the gas cylinder must be secured in an upright position. If a cylinder falls (for whatever reason) and the reduction valve is damaged, the cylinder can be launched like a torpedo. Be aware that if large volumes of nitrogen or helium (or any other relatively ‘safe’ gas) suddenly escape from a damaged cylinder or container in a closed, badly ventilated room, this can lead to an oxygen shortage and ultimately to suffocation.
If at all possible, use small cylinders (the so-called) ‘lecture bottles’ in the fume cupboard. Keep these small cylinders in the fume cupboard until they can be removed via the Logistic Transfer area.
Before using either large or small cylinders, check that the inspection date has not elapsed. Listen carefully to your mentor’s instructions about using the reduction valve.
The Matheson Gas Data Book (Library: Geert Grooteplein 15 Application number: MB 56 b 10) is useful reading if you want to know more about the properties of various gases.

3.7. Hazardous gases

Hazardous gases we define as gases or vapors that can cause serious injuries or even death upon inhalation and/or direct exposure to skin. A list of highly toxic gases can be found in paragraph 5.2. When you are planning to perform an experiment involving the use of a hazardous gas, comply with the following rules:

3.7.1. Working with HF

Hydrogen fluoride (HF, g) is very hazardous to handle because it is highly corrosive and can cause deep tissue damage and systematic toxicity after exposure. Therefore an indicator card should be present on the fume hood where the experiment is performed and at all the lab entrances. Also be sure to warn everybody present on the lab at the start of the experiment.

If there is a need to work with HF (g) always contact Dennis Löwik (office: 03.016) for supervision and accompaniment during all of the experiments performed.

3.7.2. Working with Cyanides

Cyanide is a very toxic substance that is present in salts (eg NaCN), in acid (HCN, which in gas phase smells like almonds) and as reagents (eg cyanogen bromide, CNBr). Hydrogen cyanide is lethal at a concentration of 270 ppm and a concentration of 180 ppm is life threatening within several minutes.

When working with any form of cyanide (salt, acid or reagents) be sure to:

If an accident with cyanides occurs be sure to:

3.8. Cryogenic liquids

Vessels of liquid nitrogen can be ordered from the Logistics Centre and will be delivered to the department via special transport. IMPORTANT: Be aware that liquid nitrogen can escape from a vessel very quickly (if a Dewar container is damaged and loses its cooling capacity). In a closed room, this can cause a lack of oxygen and ultimately suffocation. The laboratories where cryogenic liquids are used must therefore be adequately ventilated. Smaller rooms can be fitted with an oxygen sensor to warn if the oxygen level drops.

3.8.1. How to safely handle a cold trap

Another potential danger when using liquid nitrogen is the fact that oxygen from the air can condense into it. This should be taken into account when handling a cold trap. A cold trap is commonly used to condense solvents, thereby preventing harmful gasses from entering the attached pump (Figure 4). It is important to know how to install and more importantly, remove the cold traps, as formation of liquid oxygen (bp -183°C) and explosion thereof can cause serious injury.

Installation:

Make sure to NOT attach the vacuum pump to the top adapter as this will cause the vacuum pump to suck up condensed solvents when it reaches the level of the internal tube.
Make sure the installation is airtight (close the system).
Switch on the vacuum pump.
Immediately place/fill the dewar flask with liquid nitrogen.

Removal:

Remove the dewar flask with liquid nitrogen.
Allow air into the system and switch off the pump.
The cold trap is now safe to be disconnected with appropriate gloves to protect you from the cold.

DANGER: when air is allowed into the cold trap whilst still being cooled by liquid nitrogen, oxygen will condense inside the trap to form extremely dangerous mixtures of organic material and liquid oxygen which can potentially explode.

Figure 4: Cold trap.


3.9. Chromatography columns

Chromatography using silica in glass columns should be carried out in a fume cupboard. Silica is made up of minute particles which can penetrate the lungs causing silicosis (occupational lung disease) (see also hazard category Irritating/Sensitising/Damaging in paragraph 3.2); columns should be filled and emptied in the fume cupboard. Inspect the glass for cracks before it comes under pressure, and protect yourself from glass splinters by packing the column in a net. Use a pressure valve if the column is to be pressurised.

3.10. Distillation

Listen carefully to your mentor‘s instructions every time you distil a new solvent; check that the correct drying agent has been used, and whether the solvent needs to be pre-dried. Make sure that the cooling is switched on, never turn off the nitrogen taps, and let the solvent cool to room temperature before refilling. If in doubt, ask your mentor. All distillation set-ups must be constructed so that the heating mantle can be taken out from under the set-up in an emergency by removing the lab jack.

3.11. Cleaning glass filters

Cleaning glass filters with a mixture of sulfuric acid and hydrogen peroxide can cause an explosion if the filter is rinsed with an organic solvent such as acetone and then not properly dried. This makes it a very risky procedure, which should only be used if it is the only possible way to clean the glass filter. Even then, it should only be applied by experienced staff.

3.11.1 Procedure for cleaning glass filters with sulfuric acid and hydrogen peroxide
The last few years a number of incidents occurred during cleaning glass filters with a mixture of sulfuric acid and hydrogen peroxide. To avoid such incidents use the following procedure.

Use this procedure only if the glass filter can not be cleaned by any other method and if excess waste is removed. Make sure that all the glassware used in this procedure is free of acetone or any organic solvent. Only a very small amount of acetone is enough to cause a big explosion.

Procedure:


3.12. Autoclaves

Materials that need to be sterilised before use should be sterilised in a special pressure cooker or, if large amounts need sterilising, in an autoclave. All materials originating from safe microbiological procedures must also be sterilised after use in line with the strict regulations governing safe microbiological procedures. Depending on the nature of the materials, they can be sterilised using dry sterilisation (only in the autoclave) or they can be post-sterilised, whereby the temperature and the times are adjusted to correspond with the amount and nature of the materials.
Place all objects to be sterilised in drip-trays or pans. Check glass bottles carefully for tiny cracks before starting. In the case of liquids that are being sterilised in closed bottles check that the screw top has not been screwed on tight, so that the pressure caused by heating can escape. Beware of delayed boiling after the post-sterilisation process. Liquids can easily boil over, particularly if the bottles are full and the autoclave or pressure cooker is opened while the temperature is still high.

3.13. Biochemistry

Biochemistry and cell biology involve fewer safety risks than organic chemistry, but for some biochemical assays, hazardous chemicals are required and thus it is important that you know how to work with them. Compounds that alter biochemical structures are often toxic or carcinogenic. Moreover, chemicals might be needed for purification or other experimental purposes. Always be aware that biochemical experiments bring possible risks even though performed at SL-1 safety level, and make sure you know how to safely handle the compounds you need.

3.13.1. Ethidium bromide

Ethidium bromide is a DNA stain which binds to DNA through interchelation and can therefore act as powerful mutagen. Always work on the designated bench and make use of gloves when handling ethidium bromide. Only touch contaminated materials with your gloves and discard your gloves immediately after use. Consider wearing only one glove in case you have to handle both contaminated and non-contaminated materials. Ethidium bromide-containing waste needs to be discarded into the solid waste container.

3.13.2. Acrylamide

Acrylamide is widely used as a cross linking agent for electrophoresis separation procedures (e.g. SDS-PAGE). Acrylamide is easily absorbed by the skin and acts as a neurotoxin and (possibly) as carcinogen. When handling acrylamide powder, always work in a fume hood. Chances of ingesting or absorbing acrylamide are limited when working with ready-made solutions of the compound (common in most labs). Nonetheless, one should always wear gloves when working with acrylamide and discard your gloves immediately after use. It is recommended to use a dedicated part of the lab for working with acrylamide in order to minimize the risk of cross-contamination. Once FULLY polymerized, acrylamide gels are significantly less toxic. They can be discarded in the solid waste containers.

3.13.3. Organic solvents

When using organic solvents for e.g. staining/destaining of protein gels or trizol/phenol purification, handle them in the fume hood as much as possible even when working with low quantities. Also always make use of the proper liquid waste container (halogen rich, i.e. compounds containing F, Cl, I or Br, or halogen poor). Contaminated waste materials should go into the solid waste container. If you are not sure how to handle a certain chemical, consult someone who does.

4. Faults, fire and accidents

4.1. General 4.2. Guidelines in case of a power failure

During a power failure there is not enough ventilation inside the labs and surrounding areas to provide a safe working environment. But also after a power failure safety cannot be immediately guaranteed. Everything might look normal inside the labs while certain systems are not running. For example, point ventilation and air-conditioning can be still non-operational, which causes dangerous situations even when the fume hoods are running again.

So in case of a power failure, even if everything seems to be back operational immediately, handle as follows:

4.3. Guidelines in case of chemical spills

For chemical spills, the ‘Hazardous Laboratory Chemicals Disposal Guide’ can be consulted here: Hazardous Laboratory Chemicals Disposal Guide. A supply of the Bentonite absorber material frequently recommended in this Guide is available on each wing, please make sure of its location before you start chemical work.

There are several classes of chemical spills:

4.4. Guidelines for accidents

The guidelines on how to deal with an accident are shown below:

4.5. Guidelines in the event of fire

There are various ways of raising the alarm in the event of fire,:

  • By telephone: ring the alarm number 55555.
  • Manually: push the button on the manual fire alarm (red box with glass front, see Figure 5).
  • Automatically: the smoke alarms are activated.

 

Figure 5. Manual fire alarm

The alarm centre (manned 24 hours per day, 7 days per week for the entire university and the hospital) will respond to the alarm. An automatic emergency call will be put through to the Internal Emergency Team during working hours (8.30-16.30 hrs.), and outside office hours, the Nijmegen fire service will be alerted. If necessary, the evacuation alarm will sound on the floor of the wing where the alarm was raised and any other wings/floors affected. The emergency escape route doors will automatically unlock; they can always be opened by activating the nearest green box. The partitioning fire doors along the central corridors will close automatically if necessary.

The guidelines for what to do if you discover a fire are as follows:

First phase:

Second phase:

Third phase:

This phase is not directly relevant to laboratory workers

4.6. Guidelines for evacuation

Evacuation can be initiated by an alarm or by members of the Rapid Intervention Team/Internal Emergency Team. The evacuation alarm is a continuous signal (a "slow whoop"), which is tested every first Monday of the month at 12:00h.

In the event that the premises must be evacuated, the following supplementary guidelines apply:

IMPORTANT: it is vital that all staff and students forced to evacuate a lab during a fire register their names at the assembly point so that the emergency services know who has been brought to safety.

Figure 6. Floor plan of the 3rd floor of the Huygens building, showing the fire-proof partitions around the fire compartments in red, as well as fire alarm buttons, hose reels and the flight routes to the emergency stairways.

5. Important information

5.1. List of abbreviations

Table 3. List of abbreviations

Abbreviation

Explanation

AMD

Arbo- en MilieuDienst - Department of Occupational Health & Safety and Environmental Service

ARBO

ArbeidsOmstandigheden - Working Conditions

BHV

Bedrijfs HulpVerlening - Internal Emergency Team or Company Emergency Response Organization (CERO)

CERO

Internal Emergency Team or Company Emergency Response Organization (BHV)

CLP

Classification, Labelling, and Packaging

CRM

Carcinogenic (causes cancer), Reprotoxic (dangerous to unborn foetus) and Mutagenic (causes permanent changes to genetic material

EHBO

Eerste Hulp Bij Ongelukken - First Aid

FNWI

Faculteit Natuurwetenschappen en Informatica - Faculty of Science

GHS

Globally Harmonised System of Classification and Labelling of Chemicals

HP

Hazard and Precautionary statements

HR

Personeel- en Ontwikkelings Afdeling (P&O) - Human Resources Department

IHZ

Interne en Huisvestingszaken - Internal and Housing Affairs

OC

OnderdeelCommissie - Representative Council (Faculty)

OR

OndernemingsRaad - Workers Council (University)

PAM-mer

Preventiemedewerker Arbo en Milieu (formerly: ARBO en Milieu-contactpersoon) - Environment, Health & Safety Officer

PBM

Persoonlijke Berschermingsmiddelen - Personal protection equipment

PPE

Personal protection equipment

RU

Radboud Universiteit Nijmegen - Radboud University Nijmegen

SEH

SpoedEisende Hulp - Hospital Emergency Aid Department (A&E unit)

TWA

TijdGewogen Gemiddelde (TGG) - Time Weighted Average

VMT

Veilige Microbiologische Technieken - Safe Microbiologal Procedures

. Important internet links

Information concerning Arbo at the intranet from FNWI
http://www.radboudnet.nl/fnwi/fnwi/arbo/.

Information concerning Health, Safety and Sustainability at the intranet from AMD
http://www.radboudnet.nl/amd/english/.

Laser safety
http://www.microscopyu.com/articles /fluorescence/lasersafety.html.

Waste management
The faculty waste management regulation was devised with the RU Department of Occupational Health & Safety and Environmental Service and can be found on
https://www.radboudnet.nl/amd/english/sustainability/environment/

Information on substances (Chemwatch)
http://jr.chemwatch.net/

Accidents
Accident report form for reporting accidents and incidents to the Department of Occupational Health & Safety and Environmental Service
https://www.radboudnet.nl/amd/english/safety/occupational/
Don't forget to inform the PAM’mers.

Overview of highly toxic gases
http://en.wikipedia.org/wiki/List_of_highly_toxic_gases

Hazardous Laboratory Chemicals Disposal Guide (advice in case of chemical spills)
Hazardous Laboratory Chemicals Disposal Guide.pdf

Durability at the University
http://www.ru.nl/duurzaamheid/duurzaamheid/duurzaamheidsagenda
http://www.ru.nl/sustainability/

Appendix 1 Breakthrough times for various glove materials.

For latex gloves supplied by VWR: VWR Chemical Resistance Gloves Chart.pdf
For nitrile gloves supplied by Kimberley: Resistance Guide for Kimberly Clark Nitrile Gloves.pdf

 

Appendix 2. Flow diagram for liquids in laboratories; decision chart to determine whether a substance may be discharged into the drain