Safety guideVer. 1.2.1

2011

Faculty of Agriculture
Graduate School of Bioresource and Bioenvironmental
Science
School of Agriculture
Institute of Tropical Agriculture
Biotron Application Center
Bio-Architecture Center
Material Management Center

Foreword
In order to carry out research projects, teachers, researchers, and graduate and undergraduate students
in Faculty of Agriculture, Graduate School of Bioresource and Bioenvironmental Science, and Schoolof Agriculture should understand the risk of chemicals and devices, and the research activity
environment. It is necessary to secure one’s own safety, and social confidence of the university.
“Safety Guide Working Group”, which is designed under the safety and health department in Faculty ofAgriculture, created a safety guide called “Safety Guide in Faculty of Agriculture.” It showscompliance rules for safety when doing experiments and practice. It requires that not only teachers
but also students should have enough knowledge of various laws about safety and try to prevent
accidents, and comply with the rules because accidents which happen in the University became eligible
for criminal punishment, after transformation into an independent administrative institution in 2004.
This booklet was newly rewritten based on the “Safety guide in Faculty of Agriculture in 2008
(Japanese version).” First of all, we reorganized the contents of the chemical list to be more
understandable. The contents of the chemical list has been severely restricted by various laws such asFire Defense Law, Ordinance on Prevention of Organic Solvent Poisoning, Ordinance on Prevention ofHazards due to Specified Chemical Substances, and the Poisonous and Deleterious Substances Control
Law. Moreover, because of the new content, we tried to add security precautions in physics systemlaboratory, biological system laboratory, and chemistry system laboratory and outdoor activities. It
covers the basic part of research activities done at the agriculture research academy. We would like
you to share enough understanding concerning security with others by reading the related part of thisbooklet when doing a research project. Moreover, teachers and students should always keep thisbooklet alongside when performing research and try to do their best to follow security management
procedures. In order to revise the previous edition, we referred to the chemical coterie publication“Conducting safety experiment, seventh edition in 2006”, chemical coterie publication in Tokyo“Chemical experiment safety guide for students in 2003”, Graduate School of Engineering, Division ofAdvanced Science and Biotechnology, Department of Biotechnology, Osaka University “Basic
operation for experiment of applied biology -to experiment safely and correctly-“, Graduate School ofengineering, Department of applied chemistry, Kyushu University “Guidance of Safety and health,environmental management in 2007 ”, Graduate School of Science , Kyushu University ”Guidance ofsafety in Faculty of Science in February, 2006”, Laboratory for waste water treatment, Kyushu
University “Utilization Guidance in February, 2008.” Furthermore, although the English version of thesafety guide was already published online in 2009, it has finally become a printed booklet with thecooperation of the G30 Lecturers of the Faculty of Agriculture. Lastly, we would like to express ourappreciation to the teachers who contributed to this booklet.
February, 2011
Safety and health department, Faculty of AgricultureCONTENTS
1.	
  General preparation for safety working every day

1

1.1. Preparation for school life
1.2.	
  Preparation for laboratory works
1.3. Fire prevention
1.4.	
  What to do in an Earthquake
1.5.	
  Traffic safety measure
1.5.1. Attention concerning driving car
1.5.2. Avoidance of bicycle accidents
1.6. Theft and burglary
1.7. Overseas travel notes
1.8. Disaster and accident insurance
1.8.1. Disaster and accident insurance for student education and research
1.8.2. General liability insurance

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2.

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Laboratory emergency plan

2.1. Preparation for emergency and contact system
2.1.1. Occurrence of emergency situation
2.1.2. Emergency contact
2.2. Action plan when fire starts
2.3. First-aid treatment
2.3.1. Gas poisoning
2.3.2. Accidental ingestion of poisonous substance and absorption to skin2.3.3. Getting burned
2.3.4. Injuries
2.3.5. Electrical shock
2.3.6. Cardiopulmonary resuscitation
2.4. Evacuation Procedures

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3.

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Safety handling of chemicals

3.1. Precautions
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3.1.1. General rules
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3.1.2. Notes on the Industrial Safety and Health Law
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 3.2. Hazardous substances leading to fire and/or explosion (Fire Service Law object chemicals) 20
3.2.1. Pyrophoric chemicals (3rd group hazard materials under Fire Service Law)
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3.2.2. Water reactive chemicals (3rd group hazardous materials under Fire Service Law)
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3.2.3. Potentially explosive chemicals and water reactive chemicals (2nd group hazardous
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materials under Fire Service Law)
3.2.4. Oxidizers (Solid)(1st group hazardous materials under Fire Service Law)23

3.2.5. Oxidizers (liquid)(6th group hazardous materials under Fire Service Law)
3.2.6. Flammable materials (4th group hazardous materials under Fire Service Law)
3.2.6.1. Special flammable materials
3.2.6.2. High inflammable materials (class 1 petroleums, alcohols)
3.2.6.3. Medium inflammable materials (class 2 petroleums)
3.2.6.4. Low inflammable materials (class 3 petroleums, class 4 petroleums,
animal fat and vegetable oils)
3.2.7.
Potentially explosive substances (5th group hazardous materials under Fire
Service Law, Explosives Control Law)
3.2.8.
Regulation for maintaining hazardous materials
3.3. Poisonous substances which need strict management
3.3.1. Poisonous and Deleterious Substances Control Law
3.3.2. Specified chemical substances (Ordinance on Prevention of Hazards due
to Specified Chemical Substances, Industrial Safety and Health Law)
3.3.3. Organic solvents (Ordinance on Prevention of Organic Solvent Poisoning,
Industrial Safety and Health Law)
3.3.4. Pollutant Release and Transfer Register (PRTR) Law
3.3.5. Globally Harmonized System of Classification and Labeling of Chemicals (GHS)
3.4. Measurement of working environment
3.5. Online chemical control system

4.

Handling liquefied gas and high-pressure gas

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4.1. Gas property
4.1.1. Combustibility
4.1.2. Poisonous property
4.1.3. Oxygen’s susceptibility of substances to burn4.1.4. Oxygen starvation
4.2. Handling high-pressure gas
4.2.1. Container indication
4.2.2. Transportation
4.2.3. Storage
4.2.4. Confirmation of container
4.2.5. Notes on handling pressure regulator and valve
4.2.6. Other notes
4.3. Cryogen (cryogenic liquefied gas)

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5.

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Electric safety planning

5.1. Treatment for electric shock
5.1.1. Electric shock result from indoor distribution line
5.1.2. Electric shock result from high voltage equipment

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5.1.3. Electric shock result from electric/information devices
5.2. Electric disaster and its countermeasure
5.2.1. Overheat
5.2.2. Electric spark
5.3. Notes on electric devices.
5.3.1. OA equipment
5.3.2. Equipment in pause
5.3.3. Blackout, all-night service
5.4. Check for accident avoidance

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6.

Handling glassware

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6.1.
6.2.
6.3.
6.4.
6.5.
6.6.
6.7.
6.8.
6.9.

Safety precaution
Insert glass tube into a rubber plug (pipetter)
Remove glass tube from a rubble plug
Hold a large glass container or dangerous reagent bottle with one’s hands
Opening the lid of glass container
Opening ampoule
Clean up after damaged
Other notes on handling glassware
Notes on cleaning

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7.

Notes on physical system laboratory

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7.1. General notes
7.2. Tools
7.3. Safety handling on wood processing machine
7.4. Safety handling on agricultural machines
7.4.1. Work environment
7.4.2. Vehicle
7.4.3. Walking machine
7.4.4. Fixed mount type machine.

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8.

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Notes on biological system laboratory

8.1. Introduction
8. 2. Experimental equipment
8. 2. 1. Autoclave
8.2.1.1. Autoclave structure
8.2.1.2. Operation
8.2.1.3. Safety precautions
8.2.1.4. Other precautions
8.2.2. Centrifuge

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8.2.2.1. Precautions concerning maximum rotating speed
8.2.2.2. Precautions concerning centrifuge tube
8.2.2.3. Precautions when balancing
8.2.2.4. Precautions concerning sample leakage
8.2.2.5. Precautions during operation
8.2.2.6. Precautions concerning ultracentrifuge use
8.2.2.7. Precautions concerning swing-type centrifuge (clinical centrifuge)
8.2.2.8. Know-how for centrifuging
8.2.3. Thermostat Bath (Incubator)
8.2.3.1. Water bath incubator
8.2.3.2. Incubator, dryer and dry heat sterilizer
8.2.4. Reducing Pressure
8.2.4.1. Cautions concerning apparatuses
8.2.4.2. Notes on operation for reducing pressure
8.2.4.3. Rotary evaporator
8.2.5. Ultraviolet Lamp
8.2.6. Clean bench/Safety cabinet
8.3. Handling Reagent
8.3.1. Examples of reagents to handle with care
8.3.2. Cautions in purchasing
8.3.3. Precautions for use
8.3.4. Handling of carcinogens
8.3.5. Treatment of carcinogens
8.3.6. Extraction of nucleic acid with phenol/chloroform
8.4. Disposal of wastes
8.4.1. Main wastes and waste disposal method (Refer to 11.6.2)
8.5. Guidelines for Research Involving Recombinant DNA, Microorganisms,
Animals, Radioisotopes and Radiation generator
8.5.1. Recombinant DNA
8.5.2. Microorganisms for research
8.5.3. Animal experiments
8.5.4. Radioisotopes and Radiation generator

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9.

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Precaution against chemical system laboratory

9.1. Clothes
9.2. Notes on experiments
9.2.1. Glassware
9.2.2. Organic solvent and reagent
9.2.3. Device/Reaction
9.2.4. Others

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10.

Measures for safety in the field work82

10.1. General rules
10.2. Research in the sea and river
10.3. Shipboard, dive, snorkeling10.4. Research at the mountain.
10.4.1. Research at the mountain
10.4.2. Research in urban neighborhood and farming and mountain village
10.4.3. Research in the disaster10.4.4. Other notes
10.5. Overseas Research
10.5.1. Introduction
10.5.2. Plan and notification
10.5.3. Stay
10.5.4. After returning to Japan
10.5.5. Others

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11.

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Disposal of wastes

11.1. Classification of the laboratory wastes
11.2. Disposal of inorganic liquid wastes (special wastes)
11.2.1. Acidic and alkaline liquid wastes
11.2.2. Liquid waste with inorganic mercury (special liquid waste)
11.2.3. Organic mercury
11.2.4. Liquid waste with cyanogen and arsenic (special liquid waste)
11.2.5. Liquid waste with fluorine (special liquid waste)
11.2.6. Liquid waste with heavy metals (special liquid waste)
11.2.7. Liquid waste with heavy metals and organic substances (special liquid waste)
11.2.8. Photographic effluent
11.3. Disposal of the special liquid wastes
11.3.1. Method for fractionation and pool
11.3.2. Method of disposal processing
11.4. Disposal of organic liquid wastes
11.4.1. Notes about the separated storage of the organic liquid wastes
11.4.2. How to request the disposal of organic liquid wastes
11.4.3. Notes at the request
11.5. Chemicals being discarded
11.5.1. Chemicals being discarded (Class A)
11.5.2. Solid wastes with hazardous substances (Class B)
11.5.3. How to request the outsourcing disposal of “chemicals being discarded”
11.6. Separated refuse generated in the laboratory and daily life

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11.6.1.
11.6.2.
11.6.3.

12.

General wastes from business activities
Separated refuse in the laboratory
Daily separated refuse

VDT operation and computer safety management,
network security

12.1. VDT operation
12.1.1. Operation environment
12.1.2. Working hours
12.1.3. VDT equipment
12.2. Computer safety management and network security
12.2.1. Notes on handling the hardware of the computer
12.2.2. Precaution in using computer
12.2.3. Precaution in using e-mail12.2.4. Precaution in using Web browser
12.2.5. Precaution in using server
12.2.6. Precaution as an administrator

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1. General preparation for safety working every day
For activities in school, it is necessary to pay attention to avoiding the disaster and accident, andcreate secure surroundings. This chapter contains (a) general preparedness for daily school life, (b)preparedness for laboratory operation, (c) protection against fire, (d) countermeasures against
earthquake, (e) preparedness for traffic safety, (g) countermeasure against burglar (h) overseas traveling,(i) adequate insurance.

1.1. Preparation for school life
(1) Check the name tag of the section or laboratory, and clarify whereabouts at arriving and leaving.
(2) Check electricity, gas, water service and air-conditioning every day. If you finish your work or leave
laboratory for a long time, you must put the power switch off and close the main cock of the gas
and water service, and confirm the safety of locking the door.
(3) Confirm the installation location of the fire extinguisher, the fire-plug, the fire alarm, the emergency
telephone, and the fire escape equipment, and well informed of these operation methods.
(4) Smoking is limited in the specified places and deposit cigarette butts in the proper containers.
Moreover, never smoke in "Fire strict prohibition" place.
(5) Fix the laboratory equipments that are likely to fall during the earthquake.
(6) Obtain permission from your professor when you go to the laboratory 10:00 PM or later. (Sunday,
Saturday and national holiday are all days).
(7) Lock the doors when going in and out building and laboratory. Moreover, when you leave
laboratory for a long time, doors should be locked to prevent things within the room from being
stolen.
(8) Keep all the room clean and organized.

1.2. Preparation for laboratory works
This section contains a general preparation for doing research experiments in a correct and safe way.
(1) Well-organized, Checks, and Settlement in mind(a) Inessential things should be not put around the laboratory units, and things you need forexperiment are prepared in order.
(b) Keep the doorway of the laboratory for shelter, walkways, distribution board clean.
(c) Check the security in the laboratory regularly, and confirm no abnormality.
(d) Put up warning signs (eg. ”Danger Zone”) and alert people to take precautions against the1hazardous experiment device and its surrounding.
(e) Chemicals are kept in a specified place, and never left on laboratory table. Dangerous
chemicals must be managed safely and kept in a specific cabinet, and taken out only the amountnecessary for the experiment.
(f) Fix a chemical shelf and a large-scale equipment as not to fall down in case of earthquake.
(g) Use a table tap or an outlet that endures the electric power used in the laboratory. Don’tconnect several table taps to one power source. Moreover, set up a breaker if necessary, andreplace the corrupted or old electric wire with new one.
(h) Put on appropriate working wears and safety shoes for experiment. If necessary, wear the hard
hat, gloves, eye protector, earplugs, and gas masks for safety.
(i) After experiment, make a final safety check for gas, electricity, and water service. The waste
fluid and products are safely disposed. Appropriate working clothes and protective equipment
are used.
(2) Careful preparation for experiments
Experiments with incorrect design and incomplete devices likely cause the accidents. Before
doing experiments, check the normal operation equipment and make careful preparations.
Experiment should not be conducted alone especially at the risky experiment and nighttime. If
you make an experiment with an all-night unattended operation, please obtain the teacher’s
approval in the experiment beforehand. Moreover, take care of worker's health because worker’s
loose power of attention may cause an accident.
(3) Sincerely manage to do an experiment
While doing an experiment, it is necessary to stay calm and work on experiment seriously.
Moreover, you always pay attention to surrounding circumstances so as not to cause an accident.
(4) Confirm the measures you will take when an accident occurs before starting experiment.
In preparation for an emergency situation, it is necessary to know location of the main cocks,
switches, fire extinguishers, and escape routes. Furthermore, first aid measures, emergency
contact number and how to use the fire extinguisher also should be aware.

1.3. Fire prevention
The disaster with high possibility of encountering in experiment and daily life is a fire accident.
Here, fire prevention methods that should be noted in normal circumstances are summarized(1) Check the following matters regularly.
(a) Fire extinguishers (P. 3-4), fire-plugs (Handling method: P. 5), protective gear (gas masks, etc),
and emergency equipment (emergency ladder and survival kit, etc).
(b) Keep the following places clean: emergency staircase, fire door, fire-plug, corridor andbalcony.
(c) Table for the products causing fire or smoke should be made by noncombustible materials, andbe separated from surrounding combustible matters.
(d) Use undamaged rubber pipe and vinyl chloride tube for experiment.
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(The one with bends and cracks are improper.)(e) Separate electric code and gas pipe.
(f) Do not use non-standard fuse for power supply. Moreover, do not make many electric wiringloads on the same electrical outlet.
(2) Make a suitable arrangement of devices and chemical shelves in laboratory for emergency escapewhenever an accident occurs. Furthermore, keep the laboratory always clean and tidy.
(3) For chemical experiments, use only necessary amount of combustible solvent.
(4) Notes(a) Do not put the inflammability, flammable and combustible materials near the fire.
(b) Confirm the action taken when igniting. Make sure of installation location of gas valve, whichshould be turned off and power supply and fire extinguishers and escape routes, etc.
(c) Keep the escape routes clear when it breaks out.
(d) Put out fire after doing experiments.

Fire extinguishers
There are no effective extinguishing methods for all kinds of fires, so that an appropriate extinction
method is adopted according to the kind and the level of a fire.
Knowing fire extinguisher’s location, kinds and handling methods is always helpful for you.
Moreover, participation in the fire practice regularly and learning the manner of operation are necessary.
The types of fire extinguisher corresponding to the kind of a fire are shown here.

A Fire (Ordinary Fire)B Fire(Oil Fire)C Fire(Electric Fire)The fire extinguishers correspond to the above-mentioned fire.
There are three kinds of fire extinguishers in Department of Agriculture in the following.
Powder ABC fire extinguishers 10 (it is being put by each laboratory)
Powder ABC fire extinguishers 50 (it is being put by each building electric room)
Carbon dioxide fire extinguishers 5 (it is being put by a part of laboratory)3

ABC fire extinguisher
How to use a fire extinguisher- Stay about 2 m away from flame and use, please. Especially, in the case of oil fire, it is possible

to scatter oil and enlarge the range of a fire when approaching too much.
- Straightly target the bottom of fire and slowly approach to the fire to extinguish.
- Extinguish fire from the windward side when a wind existence.
- Radiate water without stopping to the last minute even if the flame disappears.
- Keep fire extinguisher upright when using.
- Grab the lever of the fire extinguisher firmly. When you cannot grasp the grip due to

weak-handed, once you put it on the ground or floor, and grasp firmly while strongly pushing
the lever.

Attention after using a fire extinguisher
1. Turn off gas pipe immediately after extinguish a blaze related to city gas or propane gas.
2. In case of fire related to electricity, switch off the power supply immediately and consult a qualified
person of the electric installation.
3. Confirm a fire whether there is an ember after extinguishing fire.
4. After extinguishing fire, ventilate the room promptly and evacuate to outdoor not to inhale neither
chemicals nor generated gases due to a fire.
5. Contact a doctor immediately when you feel unpleasant or abnormally in your body by inhalationof fire-extinguisher.
6. Do not eat foods attached to fire-extinguisher.
7. Request dealers to refill the fire-extinguisher promptly after using, even if a little volume used.
48. Consult manufacturers or dealers to dispose fire extinguisher. Do not throw away or radiate it
recklessly since it is very dangerous.
The

The indoor fire hydrant operation points
1.
2.
3.
4.
5.
6.

Confirm a fire.
Push the warning and start-up button.
Open the box and pull out the hose.
Extend the hose so as not to be twisted.
Open the valve and turn on water.
Hold the nozzle head and aim at the base of the fire.

warning

andstart-up bottom
The warning lamp
The warning speaker

1.4. What to do in an Earthquake
Earthquake is unpredictable. Moreover, there is no perfectmeasure against earthquake because it is hard to know thescale and it causes an unexpected secondary disaster. However,conducting an emergency drill against the unpredictabledisaster and establishment of the measures might help the damage lightened. In this section, themeasures taken when the earthquake happened in the laboratory are shown.
(1) Storeroom for books
Do not put a tall thing in surroundings of the desks usually used. Put the heavy load on the lower ofthe storage. Storerooms which are not frequently used are locked as much as possible. Moreover,
enforce the resistance to earthquake about the computer monitor.
(2) Chemicals
Chemical bottles are kept in the steel shelf that lock system is installed. The shelf of two or more isfirmly fixed between upper and lower part. Chemical storage room is attached to the wall and fixed
firmly. When the chemical shelf is set up on the board of an experiment, the bridge is built acrossthe chemical shelves, and each laboratory table is connected mutually or fixed with the frame
firmly.
The sliding door should be installed in the shelf. To prevent the chemical bottle from falling,wooden or pipe crosspiece of which width is several centimeters is installed under the front side of
the shelf in each shelf. Chemical storage rooms with lock systems installed strongly withstandearthquake.
(3) Cylinder
The chain for the gas cylinder fixation comes off foundation in case of the severe earthquake, and it
falls with the support stand of gas cylinder. When fixing with the chain, wrap not only one placenear the head, but also the middle part to fix the bottom with iron belt. The chain was tied to theclasp buried under a concrete wall. There is no space between the cylinder and the wall. Don’t fix
few gas cylinders together. Each cylinder is fixed individually. When the gas cylinder is connectedwith Gas Chromatograph, it fixes so that both should not slip down at the same time. The gas
cylinder which is not being used is always covered with the cap.
5(4) Glassware and sample
Glassware which is being used for distillation should be installed in the frame which fixed firmly
on the laboratory table or the wall. When keeping it in the shelf, it puts in the shelf with the sliding
door as well as the case of the chemicals and it packs it so that there is no space. Use a wide shelfboard so that there is no space between doors with the shelf board. The sliding door must be closed
after use, as well as the chemical shelf.
(5) Measuring instrument
Measuring instrument should put on the rubber mat or it’s foots are covered with rubber. Do not put
it on a slippery concrete stand. Surroundings of a fixed stand are raised a little. It is efficient to use
prevention tool for slip drop. It often moves to the heavier one at the earthquake.

1.5. Traffic safety measure
Regardless of school or outside school, it is necessary to note the traffic safety enough, to observe a
traffic rule, and to try to prevent the transportation disaster.
It is necessary to observe the following matters about the traffic of vehicles such as the car,motorcycles, and bicycles in school to secure safe traffic of the pedestrian and, keeping a tranquil
environment in school.
(1) Follow the traffic sign, and pay attention to an excessive speed.
(2) Do not park except the place designated as the parking. Especially, do not park near the fire-plugand other no parking places so as not to disturb urgent activities such as fire engines andambulances.
(3) Do not do obstructive activities such as rumbling engine noise or a horn. It obstructs the researchand educational activities. Practicing about the motorcycle and the car, idling away and idling for along time also disturb the neighbors.
(4) When causing a traffic accident in school, it first gives priority to the injury person's treatment, andit notifies the police afterwards. In addition, inform advising teachers, and the student in charge.
1.5.1.

Attention concerning driving car

Please join voluntary and liability insurance when you drive a car. There are two kinds of
insurance in Japan. One is compulsory automobile liability insurance, the other is voluntary insurance.
It doesn't suffice in the insurance of the compulsory automobile liability insurance at a lot of accident
amends. The parking lot for the car is also necessary.
1.5.2.

Avoidance of bicycle accidents

There are so many bicycle accidents reported. There is a case to have to pay a large sum of
indemnity money when becoming an assailant, and take care not to cause the accident not meeting withan accident, please.

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-Prohibition of drunken driving-Prohibition of driving with no lamplight at night.
-Prohibition of double driving.
-Do not use umbrella when you bike.
-Do not use a cellular phone when you bike.
-Prohibition of excessive speed.
- It takes a ride on a bicycle while breaking down.

1.6. Theft and burglary(1) Don’t walk alone as possible for the prevention of accident.
(2) Contact a police at once when becoming a victim of the theft or the burglar by any chance. In thecase that happened in school, please contact a related school officer such as the Komatsu gate post,
the advising teachers, and the officers in charge of general affairs for school. Moreover, it is usefulfor the arrest when assailant's characteristic is remembered at that time.
(3) Contact the Komatsu gate post when you find bike gangs to crowd in school.
Komatsu gate post ・・・call 642-2196 extension 8888
General affairs in Faculty of Agriculture・・・call 642-2796

1.7. Overseas travel notes
As provided in the admission rules of Department of Agriculture, students should give notice to thestudent office beforehand in spite of its purpose or reasons when students go abroad. When goingabroad as a group of Department of Agriculture, the representative should inform. When you hope tostudy abroad, you should submit appropriate document to give permission from the president
beforehand. Please refer to the details of Chapter 10.5 in this booklet.

1.8. Disaster and accident insurance
1.8.1 Disaster and accident insurance for student education and research
Kyushu University recommends for students to sign up for disaster and accident insurance forstudents education and research. This insurance covers the injury and the disaster happened in theclass, during an experiment, a practice, and outdoor activities and various university events including
the internship and the volunteer work. Moreover it covers an accident of movement between theuniversity facilities. Students have to join insurance positively because the number of accidents
increases every year. Students who are employed as a research assistant could not be covered byinsurance. (They will be covered by the workers accident compensation insurance.)7

(1) Insurance fee
Duration of insurance

Insurance fee（Science and technological）

One year¥1,200

Two years¥2,100Three years¥3,050

Four years¥3,900(2) Procedure to insurance
Junior year or more and graduate student must inquire of the student life division in school affaires
section, and receive a payment form at the office. Pay it in the nearest post office.
Inquiries
The students life division in school affaires section
Call: 642-2261
1.8.2 General liability insuranceThis insurance covered the liability for damage and accident happened by making others injured, andother’s fortune destroyed during school events.
(1) Insurance fee ¥340 a year
(2) Procedure to insurance
Bring insurance fee to the student life division in school affaires section.
You must join above-mentioned insurance (disaster and accident insurance for student education
and research) beforehand.

82. Laboratory emergency plan
2.1. Preparation for emergency and contact system
It is important to ensure personal safety and settle down.
2.1.1.

Occurrence of emergency situation(1) When an accident occurs, notify the accident with full voice to the person who is close to you.
(2) You should not take measures against an accident by yourself.
(3) The injured person should be moved to the safe place.
(4) If possible, try to minimize the accident.
- Switch off electric power.
- Close the main cock of the gas.
(5) In case of fire
- Mind an appropriate initial extinction.
- Push the button of fire alarm.
- If it is an uncontrollable fire, take shelter promptly and ensure emergency contact.
If the flame reaches the ceiling, it is uncontrollable.
(6) In case of gas poisoning-Do not enter the area carelessly.
-Close main cock of the gas-Open windows.
(7) In case of electric shock-Switch off power supply.
-Do not touch the person getting an electric shock, carelessly.
-Use dry stick, clothe or insulation gloves.
-If necessary, give artificial respiration and heart massage (Call ambulance?).
(8) In case of massive bleeding-Raise the hemorrhage part more than the heart.
-In order to stop bleeding, press an artery that is closer to the heart than the wound.
2.1.2. Emergency contact(1) Fire station (Higashi-ku) 119 or 641-1307(a) When(b) Where(c) What kind of accident. Fire or an accident resulting in injury or death.
(d) How’s situation-Situation of the occurrence-Possibility of expansion-Casualties presence(e) Caller’s name and phone number9(2) Emergency contact in the campus(a) Komatsu gate: Emergency call 8888 or 642-2196(b) Person in charge or laboratory: Professor, Associate Professor, Assistant Professor.

2.2. Action plan when fire starts(1) Call out loudly when there is a fire.
(2) Keep away surrounding combustible materials such as curtain from fire. Close the main cock of
the gas and switch off power supply. Close the main cock of the supply source of the fuel suchas gas cylinders.
(3) Calm down and try an initial extinction with a fire extinguisher or a water bucket. Be careful
not to water when chemicals catch a fire.
(4) Try an initial extinction with a fire extinguisher if wall burns. When a ceiling burns, extinction
with the fire extinguisher is difficult.
(5) Take shelter promptly and call an emergency phone when fire reaches to ceiling or your initialextinction becomes difficult.
(6) In case someone’s clothing catch fire, do not let him/her run or keep standing. It will make theflames higher. Take off his/her clothing immediately, or drop down to the floor and roll his/herbody (otherwise, other people must help to do so.) Then douse the fire directly or over theblanket rolled around the body to extinguish a fire.
Remove heat with cool water or ice packs until tissue around burn feels normal to touch.
Cover injured person to prevent shock.
(7) In case of a fire in the fume hoods, generally speaking, stop the ventilating because of theprevention of fire expansion to the upper side and the effects of extinction of fire. However,when a fire generates smoke and/or poisonous gas, ventilation should be kept. Judge theventilation based on the materials burned and the situation.
(8)When the ignition by the gush of the flammable gas from gas cylinder happens, surrounding
combustible materials should be removed without extinguishing fire. Next, pour water to thegas cylinder until it cools.
(9)When the flammable gas gushes without fire, remove the ignition source such as cutting themain cock of the power supply and the gas. Next, open the windows. If possible, close a nozzle
of gas.
(10) In case of fire accompanying poisonous fumes, use protective equipment such as hazard
masks for extinguishing the fire or extinguish it from the windward side.
10

(11) When you find a fire in other laboratory, first secure your safety and then run to the room witha fire extinguisher.

2.3. First-aid treatment
In proportion to the accident’s size, treat it as follows until carrying to the hospital.
Please refer to guidelines of this web site: http://www.healthcentral.com/
2.3.1. Gas poisoning
Do not enter a contamination area carelessly.
In case of gas leak, close the main cock of gas with caring fire and open the windows. Get theindividual out into fresh air and ventilate the area. Call an ambulance immediately. Inhale oxygen
into the individual as a first-aid treatment. Artificial breathing may need to be administered if
individual appears to stop breathing.
Occurrence of following poisonings is considered.
（１） Carbon monoxide poisoning（２） Halogen poisoning（３） Hydrocyanic acid poisoning（４） Sulfur dioxide poisoning（５） Hydrogen sulfide poisoning（６） Organic solvent poisoning
2.3.2. Accidental ingestion of poisonous substance and absorption to skin
The countermeasure is different according to swallowed material. Please refer to “Safety guide ofchemical experiment for students”, or “To make an experiment safety.” Those booklets are listed inreference. When consulting a doctor, you should bring MSDS (Material Safety Data Sheet) of theoriginating chemical.
(1) Ingestion of poisonous substancesFirst of all, vomit a poison by putting your fingers in throat when you have a clear sensorium.
However, the submucosal lesion may be promoted when forcibly vomiting in the case of ingestionof strong acid, alkali, gasoline, kerosene, insecticide for atomization, and bleach. Do not let vomit.
In such cases, try to protect mucous membrane by giving milk or albumen.
(2) Chemical Exposures to the Body
Quickly remove all contaminated clothing and footwear. Immediately flood the affected bodyarea in cold water for at least 15 minutes. Remove jewelry to facilitate removal of any residualmaterial. Wash chemical off with clean water only. Do not use neutralizing chemicals, cream,
lotions or salves.
Get medical attention promptly and call for assistance.

11

It should be noted that some chemicals (phenol, aniline) are rapidly adsorbed through skin. If alarge enough area of skin is contaminated an adverse health effect (systematic toxicologicalreaction) may occur immediately to several hours after initial exposure depending on the chemical.
If more than 9 square inches of the skin area exposed to a hazardous chemical, seek medical services
after washing the material off. If the incident involves hydrofluoric acid (HF), immediately seek
medical attention. Provide the physician with the chemical name.
(3) Chemical Splash in Eye
Irrigate the eyeball and inner surface of eyelid with plenty of cool water for at least 15 minutes.
Use eyewash or other water source. Forcibly hold eyelids open to ensure effective wash.
Check for and remove contact lenses. Get medical attention promptly.
2.3.3. Getting burned
Wash in volumes of clean water and cool for over 30 mins to 2 hours with ice-wrapped wet towel.
Do not cool the same part too much. In case of slight burn: Aerating a slightly burnt part, the victim
might cause pain, thus this part should be pasted with Vaseline, the sterile gauze, or the first-aid
adhesive tape. For the medium degree of burn: Wash in clean water, cover with the clean towel or sheet,and medical attention should be received. In case of airway burn occurred, ice should be held in one’s
mouth, examination of the medical institution should be received.
2.3.4. Injuries
If hemorrhage is a little, wash the wound parts with clean water and with 3% hydrogen peroxide.
If the wound parts are dirty with oil, oil should be wiped off with alcohol. The wound is disinfectedwith antiseptic solution at a slight injury, and covered with sterilized gauze, a first-aid adhesive tape, oran application bandage.
A vicious wound and a deep wound must receive doctor's treatment early.
The hemorrhage part is promptly maintained higher than the heart in case of the serious loss ofbloods, and the part of an artery that is nearer the heart than the wound should be pressed with thefinger or gauze. Stopping the flow of the blood with the bandage is not recommended, since the strong
binding with it might destroy organization of the wounded parts. A bruise is painted with iodine tincture
and covered with a cold compress. When you hit the head, you must see a doctor, even if outside
abnormality is not found.
In the case of fracture, a splint should be applied to the fracture and seek a doctor. The fixation of
the splint bandages at the top and bottom of the fracture site, and avoids the fracture part.
2.3.5. Electrical shock
When someone gets an electric shock, turn off the switch or liberate person from electric circuit bydestroying apparatus. If removal of the electric wire is necessary, use a completely dry stick, cloth, or
insulation gloves. When victim’s breathing stops, start artificial respiration. Next, if pulse does not start,
do the heart massage for a long time. For the heart massage, lay the victim down on his back, put yourboth hands on the victim’s heart, and push by the rhythm of once per second while putting weight.
122.3.6. Cardiopulmonary resuscitation
When an unconscious person doesn't react to calling, the following measures should be done.
Automatic external defibrillators (AED) are set up in the main entrance hall of 1st, 4th, and 6thbuildings of Faculty of Agriculture. In case of emergency, use this.
(1) Artificial respiration
1) Securing the airway
Place your hand on the victim’s forehead and tilt the victim’s head back as far as it will reasonably
go, essentially securing of the airway. Bring your face close to the chest of the patient with the
airway secured. Bring your cheek close to the patient’s mouth and nose, confirm the sound ofbreath, and feel the breath. Pay attention to vertical motion of the patient’s chest and abdomen.
This examination should be conducted within ten seconds.
2) Artificial respiration
Begin artificial respiration if the patient does not breathe. Secure the airway of the patient and blow
your breath twice into the patient’s mouth with closing the patient’s nose and paying attention toleak of air. If impossible, close the patient’s mouth and do artificial respiration by mouth to nose.
When you cannot blow smoothly, lift the neck of the patient, keep the airway, and blow in again. If
you hesitate to do artificial respiration by mouth to mouth or nose, you may use handkerchief for
the artificial respiration. It is convenient to carry the simple type of artificial respiration mask. If
the patient has hemorrhage around mouth and nose, you may apply only heart massage.
(2) Cardiopulmonary resuscitation (CPR)
Close your ear to the patient’s mouth, and then check the following symptoms.
- Does the patient breathe? (Check the movement of the chest or listen the sound of breath)- Does the patient cough?
- Does the patient have any movement in the body?
Examine the above within ten seconds.
When these symptoms are not seen or they are not clear, begin the cardiopulmonary resuscitation
soon. When either of symptoms is seen, the circulation is maintained, and it is judged that it is not
an arrest cardiac. When the circulation is not maintained, begin the heart massage at once.
If any signature of the circulation is not seen after it practices artificial respiration twice, the heart
should be massaged. Trace the edge of the lib of the patient using forefinger and middle finger andplace your hand at the center of a right and a left lib. The hand that has traced the rib is piled up
with the other hand. Extend the elbows, weigh straightly, and press the center part 15 times in apalm root. Press with 3.5-5 cm depth at the speed of 100 times per a minute.
(3) Automatic external defibrillator（AED）
AED is a machine of the heart electroshock that can judge the state of the heart for which theelectroshock is necessary. Most of the sudden death is due to a heart failure. It is called a cardiac
sudden death, and the sickness of ventricular fibrillation is a major cause of the death.

13

Ventricular fibrillation cause convulsion of heart and the heart would not play as the pump. In
such a case, survival is missed by about 10% per 1 minute and the almost person will die in ten
minutes. The only method of restoring this ventricular fibrillation to normal state is a defibrillation(electrical shock to the heart). AED judges whether the defibrillation is necessary, and directs alifesaving procedure by the voice. AED is made to easily do a lifesaving act including thedefibrillation.

Japan Red Cross Society : http://www.jrc.or.jp/study/safety/kinkyu/index.html
14

2.4. Evacuation Procedure(1) When escaping from the danger, close the main cock of gas. If possible, after processing dangerous
substances and confirming that there is no person, leave the room and close the door. But, do notlock it.
(2) Run away to the windward seeing the movement of smoke when there is no information on theannouncement for the selection of the escape route in the passage.
(3) Do not use an elevator because it will be stopped in the case of an emergency such as fire.
(4) The stairs become the passages of smoke, and there are a lot of dangers. Think about the escaperoute usually, and examine the construction of a building and the fire exit.
(5) When there is a lot of smoke, cover the mouth with the washcloth and take shelter by low posture.
Act calmly because it will take considerable time for smoke to fall to the floor level.
(6) When you could not utilize the escape stairway and the shelter ladder, please escape from thewindow in the terrace. However, please note that there might not be handrail in the terrace.
(7) Even though rooftop is a safe place, do not use it as an evacuation place except for emergency.
Confirm that there is no person inside before closing the fire protection door. You can open it bypushing or pulling it strongly.
(8) When issued evacuation advisory due to fire or earthquake, the evacuation route is Kaizuka
playground.

15

Method of operating&foreside'1)&downside'
4)

Remove the cabinet

Please descend from the foot.

2)

5)

Let down the main body of bag

Bend knees a little
And go down the slide on the hips

3)

6)

Pull out the entrance frame

Go above the ground without any impact

Perpendicular escape equipment

16

Method of operating&foreside'&downside'1)

1)

Remove the cabinet and throw a rope

Eliminate the rope and fix the catch of theforehead of bag to anchor point.

2)

2)

Throw the main body of bag and pull out

Hitch the catch to anchor point and pull out block

entrance frameline to extend the bag

3)

3)

Descend from the foot.

Fix the block line.

Descent escape equipment
17

3. Safety handling of chemicals
3.1. Precautions
3.1.1. General rules
A careless experiment accompanies risks. Whenever you do experiments, do not lose your guard. As
for the shock of accident, it does not abide in material, in the person concerned physically, and theinfluence is large also in mental respect. In addition, the influence is exerted to surrounding people.
For avoidance of damaging yourself and rolling in others, it is necessary to carry out an experimentwith full attention to the given instructions.
Researchers should bear the following points in mind upon the experiments.
(1) To prevent an accident and use laboratory efficiently, keep laboratory bench clean and arrange
stuffs orderly inside and outside.
(2) All the things kept in the laboratory must be secured safely to be prepared for unforeseen
circumstances like earthquakes. Flammable materials such as organic solvents which are more thanrestriction amount, and hazardous substances designated under the law must be classified. Keepthese materials in the dangerous material-keeping-cabinets specified under the responsible person
in an orderly manner and lock the cabinet.
(3) Laboratory custodian should handle a deleterious substance, a poison and an explosivenessmaterial. The custodian should confirm the operating suggestions carefully before use. Not only
deleterious substances, but also many materials strictly managed by other regulations (Deleterious
Substance Control Law, PRTR law, Ordinance on Prevention of Organic Solvent Poisoning,
Ordinance on Prevention of Hazards Due to Specified Chemical Substances) should be carefully
handled and have the use records using records.
(4) When handling a dangerous or harmful material, first investigate risk of ignition, explosion,
combustion (fire point, flash point, and range), and hazardous properties (tolerable quantity andfatal dose) enough. Second, examine assumed measures or treatment provided against emergency,
and then prepare necessarily available internal- and external-medicines and apparatus beforehand.
Distributors of the chemicals must be obligated to present MSDS to their customers. Therefore,
you can obtain the MSDS （Material Safety Data Sheet）beforehand. You can check it anytime.
MSDS is often converted into CD version in each reagent company. Moreover ”Vermont Safety
Information Resources, Inc” also provides the following things. （http://siri.org/msds/index.php）
Regarding Japanese domestic laws and regulations for chemicals, you should ask Japanese
members in your lab.
(a) Fire fighting measures against flash point(b) First aid measures, accidental release measures(c) Purifying method of volatile bad odor and acrimonious materials

18

(5) Use an explosion-proof type when you keep a low boiling point-inflammable material in the
refrigerator.
(6) Notes when other materials are used.
(a) While doing experiments, protective spectacles (safety glasses) should be used without fail.
Moreover, use the protective equipments such as gloves, disaster prevention masks, hazard
masks, wire nets, and screens appropriately.
(b) Confirm location of the fire extinguisher in preparation for emergency. Do not conduct anexperiment with materials which have a possibility of igniting in an ordinary laboratory.
(c) When chemical contacts to eyes and skins, start flushing the area immediately with runningwater over 15 min.
(d) In the experiments with generation of a smell malodorous substance or an irritating material,
observe the followings.
a. Perform the experiment in a draft, and use devices blocked outlet.
b. Lead the smell malodorous or the irritating material exhausted from the experimental
device to the outside through appropriate wet scrubbers.
(8) Working with two or more people whenever you conduct risky experiments.
(9) When you handle the chemicals, wear the white robe and fasten a button (Prevent it fromblowing down to the equipment). The white robe contacted to the chemicals should be washed.
Do not go outside laboratory with the white robe on.
(10) Conducting an experiment with wearing sneakers will help to evacuate easily in emergency
situation. Prompt actions cannot be taken with slippers.
(11) Carefulness when you conducting experiments with a gas burner there are possibility to
explosions occurrence and serious burn occur. Fix your hair carefully before doing anexperiment.
(12) The following website will be helpful to handle the chemicals and make an experiment safely.
Health and safety
The University of Melbourne: http://safety.chemistry.unimelb.edu.au/Chemsafety.php
MIT: http://web.mit.edu/environment/ehs/about_ehs.html
University of South Carolina: http://ehs.sc.edu/
Harvard University: http://www.uos.harvard.edu/ehs/
Yale University: http://www.yale.edu/oehs/

193.1.2. Notes on the Industrial Safety and Health Law
The universities in Japan receive the application of the Industrial Safety and Health Law. Whenan accident happens in the laboratory, penalty (penal servitude, penalty, and experiment stop andemergency measure instruction, etc.) is imposed on people who are in charge of the safety
management.
This law has several ordinances including Ordinance on Prevention of Organic Solvent Poisoning
and Ordinance on Prevention of Hazards Due to Specified Chemical Substances as rules that should beconsidered in conducting experiments of the chemistry system. Pertinent reagents are provided about
each rule. Moreover, when these are used, the working environment measurement is applied.
Industrial Safety and Health Law ⇔ Working Environment Measurement Law
Enforcement Order of the Industrial Safety and Health Act Cabinet Order
Ordinance on Prevention of Organic Solvent Poisoning
Refer to 1st, 2nd, 3rd, Table 3-10
Ordinance on Prevention of Hazards Due to Specified Chemical Substances
Refer to 1st, 2nd, 3rd, Table 3-9

3.2. Hazardous substances leading to fires and/or explosions (FireService Law object chemicals)
The substances with ignited danger are numerous. Therefore, their classification is based on several
Japanese domestic laws as shown in Table 3-1. In general, there are two types of combustible
substances, one ignited by heat or impact, and the other ignited by contact and mixture. Besides, thesesubstances are classified into 6 groups by their characteristics.

Table 3-1. Hazardous materials under several laws
Characteristics of chemicals

Hazardous
materials
under the fire
defense law

Under
the
high pressure
gas safety law

Under
the
industrial safety
and health law

Under thegunpowderlaw

Oxidation
natureFlammableSolidmaterials
areoxidized and the Liquid
fire explosion is Gas
caused.

1st group̶Oxidation
nature̶

6th group

̶

̶

̶

○

̶

It ignites in air, Solidand the fire Liquid
explosion
is Gascaused.

2nd group

Flammability

̶Flammability̶th

4 group̶

Flammability

̶

̶

○

Flammability
gas

̶

20

Pyrophoric,
Water reactive
Autoreactive

React to water Solidor airLiquidThe explosion
are caused byheat and theimpact.

Other

3rd group̶Flammability̶rd

3 group

̶

̶

Gas

̶

○

̶

Solid

th

5 group̶

Liquid5th group

̶

○

Gas

̶

○

̶

Inactive
gas

̶

○

̶

Explosivenature○Table 3-2. Hazardous materials categorized by the Fire Service Law
TypeAspectChemicals

1 Group

Oxidizers (solid)

Salts of following acids (chloric acid, perchloric acid, chlorite, bromic
acid, nitric acid, iodic acid, bichromic acid, permanganic acid), inorganic
peroxide

2nd Group

Combustible solid

phosphorous sulfide, red phosphorous, sulfur, iron powder, metal
powder, magnesium, solid alcohol

3rd Group

Pyrophoric liquid
or solid

metal potassium, metal sodium, alkylaluminum, alkyllithium, yellow
white phosphorus, alkali (earth) metal, metal hydride, metal phosphide,calcium carbide, alminum carbide

4th Group

Flammable liquidhighly flammable substances, petroleum
alcohols, animal oil and plant oil

5th Group

Explosiveor solid

liquidorganic peroxide, nitrate esters, nitro compound, nitroso compound, azo
compound, diazo compound, hydrazine dericative

6th Group

Oxidizers (liquid)

perchloric acid, hydrogen peroxide, nitric acid, bromine trifluoride,bromine pentafluoridest

substance (class 1-4),

3.2.1. Pyrophoric chemicals (3rd group hazardous materials under Fire Service
Law)
Pyrophoric substance is easily ignites or generates heat only by air touching. The representativeof these are metal organic compounds such as alkylaluminum, alkyllithium or nickel and palladium reduced.
3.2.2. Water reactive chemicals (3rd group hazardous materials under Fire
Service Law)
Water reactive chemicals react with water and release a gas that is either flammable or presents a
health hazard. The representatives of these are Alkali metals such as natrium and kalium, metaric
carbide, lithium, and many organic metallic compounds.

21

Table 3-3. Representative substances of pyrophoric substances/ Water reactive chemicals
Substances

CompoundReact number, gas, toxic, corrosive

Reaction

Alkali metal

Li【４】, H2 gas【１】spontaneously catch fire

Na, K

【１，２】, H2 gas, corrosive to skin.

Alkali earth metal

Ca, Ba【２】, H2 gas

Alkylaluminum(CH3)3Al,(C2H5)3Al,

on exposure to air.
【２】react with water to catch
fire, blow up at times.

【１，２】, CnH2n+2 gas【３】There is a possibility to(C3H7)3Al

blow up when mixing with

(C2H5)2AlCl,【４】, CnH2n+2 gas

oxidizing agent.

(i-C4H9)3Al,【４】There is a possibility to

C2H5AlCl3,

catch fire when reacting(C2H5)2AlH

with water to produce

Alkyllithium

CH3Li, C2H5Li【１，２】, CnH2n+2 gas

Zinc alkyl(C2H5)2Zn【１，２】, C2H6 gas

Metal hydrideLiH【４】, H2 gas

NaH,CaH2【３，４】, H2 gas, have toxic consequence.

Metal phosphide

Ca3P2【２】, PH3 gas, have toxic consequence

Metal carbide

CaC2【２】, C2H2 gas

Al4C3【２】, C2H4 gas

Phosphorus

P 4,yellow

white

combustible gas【５】Produce poisonous gas
during combustion.

【１，５】, toxic, corrosive

phosphorus
Chlorinatedsilicon

SiHCl3【１，３，４】, HCl gas, toxic

compounder

3.2.3. Potentially explosive chemicals and water reactive chemicals (2nd grouphazardous materials under Fire Service Law)
Of low degree ignition materials, metallic powder and phosphorus pentasulfide such as magnesium
and aluminum are well known. Generally, they are strong reducing reagents and ignite by heating,
impact and friction when mixing with an oxidizing substance.
Table 3-4. Low degree ignition materials
Substances

CompoundReact

number,toxic

Reaction

corrosive
Phosphorus

P 4S 3, P 4S 7

【３，５】

P 2S 5

【５】
Al,

Zn,

Mg

【１】There is a possibility to blow up when mixing
with oxidizing agent.

Metallic

Fe,【 ２ 】 There is a possibility to blow up when

powder（including ribbon base）

OthersPn, red phosphorus【１，２】, toxic corrosive【３】Catch fire by strong friction.

Sn【１．２，４，５】【４】spontaneously catch fire on exposure to air,

【１，２】

igniting dust powder.

oilcloth or cutting trash.
【５】React with water or produce poisonous gas
during combustion.

22

3.2.4. Oxidizers (Solid)(1st group hazardous materials under Fire Service Law)
Oxidizing agents include chloric acid, perchloric acid, permanganic acid, nitric acid, inorganic
peroxide, and organic peroxide. Generally, they are easily decomposed by heating, impact and friction
and release oxygen to burn off inflammable materials. Oxidizing agents can blow up or catch fire whenmixing with organic substances or reducing materials, especially when mixing with strong acid.
Organic peroxide itself has possibility to blow up to react with heating, impact and friction.
Table 3-5. Oxidizers solid
Substance
Salt of Chloric acid

Salt of Perchloric acid
Salt of Chlorite
次 Chlorite
Salt of Bromic acid
Salt of Iodic acid
Salt of periodate
Salt of Permanganic acid
Salt of Bichromic acid
Salt of Nitric acid
Salt of Nitrous acid
Inorganic peroxide

Salt of Peroxosulfuric
acid
Peroxyborate
Oxidated products of Cr・
Pb・I
OthersCompound
NaClO3KClO3NH4ClO3, Ba(ClO3)2, Zn(ClO3)2, Pb(ClO3)2,AgClO3, HgClO3
NaClO4, KClO4, NH4ClO4NaClO2, KClO2, Cu(ClO2)2, Pb(ClO2)2Ca(ClO)2, bleach powder
KBrO3
NaBrO3, Mg(BrO3)2, Ba(BrO3)2
NaIO3, KIO3, Ca(IO3)2, Zn(IO3)2
NaIO4
NaMnO4, KMnO4, NH4MnO4Na2Cr2O7, K2Cr2O7, (NH4)2Cr2O7
NaNO3, KNO3, Ba(NO3)2
AgNO3
NH4NO3
NaNO2, KNO2
Li2O2, Na2O2, K2O2
MgO2, CaO2, SrO2, RbO2, CeO2
BaO2, PbO2
(NH4)2S2O8
NH4BO3

React number【1,2,3,5】【1,2,3,toxic】【1,2,3】

CrO3
PbO2,
I 2O 5
C3N3O3Cl3, trichloroisocyanuric acid
HIO4・2H2O, metaperiodic acid

【2,3, corrosive】【toxic】
【1,2】
【2】
【2】

【1,2,3】
【2,3】
【2】
【1,2】
【2】
【2】
【2】
【2,3】
【2】
【2】
【2, corrosive】
【1,2】
【2】
【2,4, corrosive】【2】【2,toxic】
【2】
【2,3】

Reaction【１】substance itself isunstable, it blow
up because ofheating, impact andfriction.
【２】It blow up whenmixingwith
substances which
are susceptible tooxidation.
【３】add strong acid toblow up【４】React with waterand heat up. Thereis a possibility toblow up.
【５】It is a deliquescentmaterial.
There
is
a
possibility to blow
up after penetrating
into paper or tree
and dry.

3.2.5. Oxidizers (liquid)(6th group hazardous materials under Fire Service Law)
As a strong acidic substance, fuming nitric acid, fuming sulfuric acid, concentrated nitric acid,concentrated sulfic acid, and chromic anhydride are known. These materials might generate heat, ignite
when mixing them with organic materials and reduced materials. There is a possibility to explode whenmixing it with strong acid.

23

Table 3-6. Oxidizers liquid
CompoundReact number

Reaction

HClO4【1b, 2a, 3, corrosive】【1】（a）metallic powder, （b）inflammable material such as

H 2O 2【1a, 1b, 4, corrosive】

alcohol, （c）catch fire or blow up when mixing it with amine

HNO3, fuming sulfuric【1c, 3, toxic, corrosive】

and hydrazine

acid
BrF3, BrF5

【2】react with water, （a）generate heat, （ｂ）produce oxygen【2b, 5, toxic, corrosive】【3】Spontaneously ignite when touching organic substancessuch as sawdust.
【4】hydrogen fluoride reacts with many substances to produce
fluoride.

3.2.6. Flammable materials (4th group hazardous materials under Fire Service
Law)
Flammable materials burn easily when there is an ignition source near by though it does not ignite
only by touching air. Note that a material having a low flashing point is dangerous and that even if amaterial has a high flashing point, when you heat it more than the flash point is dangerous, too.
Among the flammable materials, there are special flammable materials, high degree inflammablematerials, medium degree inflammable materials, low degree inflammable materials. The handling
method is described about each category.
3.2.6.1. Special flammable materials
The aspects of these materials are liquid at 20°C or liquid at from 20 to 40°C. The ignition
temperature of the materials is below 100°C. Their flashing points are below -20°C, and their boilingpoints are below 40°C. The typical materials are follows: diethyl ether, carbon disulfide, acetaldehyde,
pentane, iso-pentane, propylene oxide, divinyl ether, nickel carbonyl, and alkyl aluminum. Bear inmind that a fire is the most frequent accident in a chemistry laboratory. When these materials arehandled, the following attentions are necessary.
（１）Take care of discharge of gas and steam.
（２）Know the flash point, the fire point, and the explosion limit.
（３）Do not put needless solvent on the bench.
（４）Turn off the nearby naked light when you use the materials. Because their ignition temperature andflash point are extremely low, they ignite easily.
（５）Do ventilation well because the boiling point is low and the explosion limit is wide.
（６）It will be continued explosively when igniting once and fire is not extinguished easily.
（７）The space over the solvent container is often in the range of explosion. The container must be
sealed up. Take extra care of the ignition source when you use a little solvent.
（８）Do not keep voluminous solvent in the laboratory for a long time.
（９）Confirm location of a fire extinguisher. Especially, prepare it beforehand near yourself when thereis fear of the ignition.
（１０）The powder extinguisher, the carbon dioxide extinguisher or sand for fire prevention are good for
extinction. Although pouring water is not suitable for inflammable materials, it is good for theextinction of a peripheral fire.
24

3.2.6.2. High inflammable materials (class 1 petroleums, alcohols)
Substances which have high flammable characteristics at room temperature and their flash points
below 20°C are considered as high inflammable materials. The typical materials are petroleum ether,gasoline, petroleum benzine, ligroin, hexane, heptane, octane, pentene, and benzene, toluene, ando-xylene, alcohols (methylic-, pentyl-), dioxane, acetone, methyl ethyl ketone, ester of formic acids(methylic-, pentyl-), acetic esters (methylic-, pentyl-), and acetonitrile, pyridine, and chlorobenzene.
When these materials are handled, the following attentions are necessary.
（１）Know the flash point, the fire point, and the explosion limit.
（２）Do not put needless solvent on the bench.
（３）Because the inflammability of these materials is high, do not place the materials nearby fire,electric spark, and burners nearby, and do not heat the materials by direct flame.
（４）Be sure to do ventilation and not to stay in a low place because specific gravity of steam is heavy.
（５）The powder extinguisher, the carbon dioxide extinguisher or sand for fire prevention are good for
extinction.
3.2.6.3. Medium inflammable materials (class 2 petroleums)
Substances which have high flammable characteristics at intermediate temperature range and their
flash points between 20°C and 70°C are defined are medium inflammable materials. The typical are as
kerosene, light oil, turpentine, xylene, styrene, aryl alcohol, cyclohexanol, benzaldehyde, formic acid,and acetic acid. When heating it in the container with the opened mouth, it is necessary to note theaccumulation of steam of these materials within the container. The powder extinguisher or the carbon
dioxide extinguisher is good for extinction of flame caused by these types of materials.
3.2.6.4. Low inflammable materials (class 3 petroleums, class 4 petroleums,
animal fat and vegetable oils)
Substances generating inflammable gas while being heated at high temperature, and their flash
point over 70°C are considered as low inflammable materials. The typical materials are crude
petroleum, creosote oil, spindle oil, turbine oil, transformer oil, tetralin, ethylene glycol, diethylene
glycol, ethyl acetate, ethanolamine, nitrobenzene, aniline, o-toluidine, heavy lubricant (gear oil andmotor oil, etc.), and animal- and vegetable oils (canola oil, soybean oil, palm oil, etc.). These
substances are not easily ignited, because their flash points are high. However, it will be difficult to
extinguish easily once they catch fire. Be sure not to heat them over their flash points and take care
not to flash off steam that generated from the heated materials. The powder extinguishers or the carbon
dioxide extinguisher are good for extinction.
Table 3-7 shows properties of general solvent that is flammable or combustible.

25

Table 3-7. Properties of general solvents
Boiling

Flash

Explosion limit

spontaneous

Vapor

Permissible

point

point

Upper

Lower

ignite point

ratio

level

℃

℃

limit

limit

℃

（air）

consistency％％

Toxic potency

mg/m3

Pentene

30

–49

1.4

8.0

309

2.48

1800

(N)

Hexane

69

–23

1.2

6.9

260

2.97

180

(N) (R)

Heptane

98

–4

1.2

6.7

233

3.45

1600

(N) (R)

Benzene

80

–11

1.4

8.0

538

2.77

80

(S) (R)

Toluene

111

4

1.3

7.0

552

3.14

370

(ND)

Xylene

138

25

1.1

7.0

496

3.66

435

(N) (R)

Methylene

40

̶

12.0

19.0

662

2.93

1740

(N) (ND)

Chloroform

61

̶

–

̶

̶

4.12

50

(N) (ND)

Carbon tetrachloride

77

̶

–

̶

̶

̶

65

(N) (R)

Ethylenedichloride

82

21

6.2

15.9

449

3.35

200

(ND) (S)Methanol

65

18

6.0

36.5

470

1.11

260

(N)

Ethanol

78

16

3.3

19.6

399

1.59

1900

(N)

2̶propanol

82

21

2.5

5.2

456

2.07

980

(N)

Diethyl ether

34

–45

1.8

48.0

180

2.55

1200

(N)

Tetrahydrofuran

66

–14

2.3

11.8

321

2.50

590

(N）(ND)

Dioxane

101

12

2.0

22.0

180

3.03

180

(ND)

Acetone

57

–18

2.6

12.8

538

2.00

2400

(ND)

Ethyl acetate

77

4

2.7

11.5

482

3.04

1400

(ND)

Acetic acid

118

43

4.0

16.0

426

2.07

25

(ND) (S) (R)

140

54

3.0

10.0

380

3.52

20

(ND) (S) (R)anhydride

80

6

4.0

16.0

524

̶

70

(ND) (S) (R)Acetonitrile

46

–30

1.0

44.0

100

2.64

30

(ND)

Carbon bisulfide

95

95

2.6

42.0

215

–

–

(S)

DMSO

58

58

2.2

15.2

445

–

30

(ND) (S) (R)

DMF

105

105

–

–

6.18

–

(ND) (S)

Acetic

acid

– 105

1800

(R)

HMPA

（R） Respiratory obstruction（N） Narcotic action（S） Skin lesions（ND） Nervous disorder
3.2.7. Potentially explosive substances (5th group hazardous materials underFire Service Law, Explosives Control Law)
Potentially explosive substances are ignited or exploded by fire, impact, and friction. Nitric ester,
nitro, nitroso compound, diazo compound, diazonium salt, and compound, hyperoxidation combination
thing, and halogen acid conductor and acetylene heavy metal salt agitation are known.
In general, risk becomes high like the one to have a lot of uniting materials such as N̶O,N＝O,N
̶N,N＝N,O̶O,O̶halogen, N̶S,N̶halogen. Moreover, substances itself are not dangerous to
explode. There is a reagent that can produce peroxide while storing it and causes the explosion. You
26

have to be careful to handle dioxane, tetrahydrofuran, and diethyl ether that have been preserved for along time, or used. These substances should not be concentrated without discretion.
Table 3-8. Self-reacted materials
Substances

CompoundReact number

Reaction

Organic peroxide

C6H5CO•OO•COC6H5, benzoyl peroxide【1, 2, 3】【 1 】 It explodes by

CH3COC2H5•O2, methyl ethyl ketone peroxide【1, 4】

heating,

CH3NO3, C2H5NO3【6】

friction, and lightC3H5(NO3)3 nitroglycerin【1, explosive】【 2 】 It explodes byCell̶(NO3)2~3 nitrocellulose【1, 4, explosive】

contact with the strong(NO2)3C6H2OH picric acid, (NO2)3C6H2CH3【1, 3, 6, explosive】

acid

Nitric ester

Nitro compound

trinitrotoluene
Nitroso compoundimpact,【3】It explodes by the

C5H10N6O2【1, 2, 3】

N,N-dinitrosopentamethylenetetramine
Azo compound

C8H12N4

Diazo compoundC 6H 2N 4O 5

Hydrazine

NH2NH2•H2SO4 hydrazine sulfate

Hydroxylamine

H2NOH

and salt（H2NOH）2H2SO4 hydroxylamine sulfate

azoisobutyronitrile
diazodinitrophenolhydroxylamine,

mixtures such as theorganism, the halogen,【corrosive】

and sulfur.

【1, explosive】【 4 】 The spontaneous【corrosive】

decomposition

H2NOH•HCl,

NaN3 sodium azide

might

be caused to ignite andexplode. 【 5 】 The
cracked

Metallic azide

the【5, corrosive】

Othersgas

might

cause the explosion.
【6】The inflammability(NH2)2C=NH•HNO3 guanidine sulfate【1】

is large, and it is likely
toexplodeduringburning.

3.2.8. Regulation for maintaining hazardous materials
Chemicals described above are categorized as “hazardous materials” according to the Fire Service
Law. Based on their potential hazardousness, “Specified Quantities” of them are defined (Table 3.9.).
Each material can be stored less than 20% of the “Specified Quantity” in a compartment partitioned by
firedoors. In case any item listed on “hazardous material” is required to be kept more than 20%, it must
be stored in a specified storage under the supervision of safety department staff.
Table 3-9.

Specified quantities of hazardous materials

Group

Item

1

st

nd

Group

2 Group

Property

Specified quantity

Class 1 oxidizing solids

50 kgClass 1 oxidizing solids

300 kgClass 1 oxidizing solids

1,000 kg

Phosphorus sulphide

100 kg

Red phosphorus
SulphurClass 1 combustible solids
Iron powders

100 kg
500 kg27

Class 2 combustible solidsrd

3 Group

500 kgFlammable solids

1,000 kg

Potasium

10 kg

SodiumAlkyl aluminium
Alkyl lithiumClass 1 spontaneously combustible materials and

10 kg

water prohibitive materials
Yellow phosphorus

20 kgClass 2 spontaneously combustible materials and

50 kg

water prohibitive materials
Class 3 spontaneously combustible materials and

300 kgwater prohibitive materialsth

4 GroupSpecified

flammable

50 L

materials
Class 1 petroleums

Non-water soluble liquid

200 L

Water soluble liquid

400 L

Alcohols

400 L

Class 2 petroleums
Class 3 petroleums

Non-water soluble liquid

1,000 L

Water soluble liquid

2,000 LNon-water soluble liquid

2,000 L

Water soluble liquid

4,000 LClass 4 petroleums
Animalfats

6,000 Land

10,000 L

vegetable oilsth

5 Groupth

6 GroupClass 1 self-reactive materials

10 kg

Class 2 self-reactive materials

100 kgOxidizing liquid

300 kg3.3. Poisonous substances which need strict management
There are a lot of poisonous (toxic) substances of which are not defined the allowable
concentration. It is difficult to obtain information about toxicity of new compounds caused by reaction.
Thus, you should be careful when you handle reagents or new compounds. There are two cases of thesymptom when you touch toxic substances. One is that the symptom appears in a short time. The otheris that the symptom appears after repeating exposure to small amount of the substances. When directly
affected the eyes or skin, there are cases to receive stimulation with rapid pain such as acid or alkali toeyes or skin, or to have skin irritation later.
Among known chemicals, there are toxic gas (permissible density is less than 200 mg/m3), toxic
substance (oral fatal dose is less than 30 mg / kg body weight), and deleterious substance (oral fatal
dose is over 30 to 300 mg / kg body weight). Besides these acute toxicities, be careful to handle thechronic toxicity and the carcinogen. The following attention in handling is necessary. Note: The fatal
dose is not necessarily a standard of classification for toxic and deleterious substances.

28

（１）Investigate the allowable concentrations and recognize them beforehand.
（２）From the general knowledge of chemicals, deleterious inorganic substances are easily figuredout whether they are harmful or not, however, organic substances are very difficult to figure out.
Some of them have strong toxicity without anticipation. Use them after you examine themthoroughly.
（３）Wear the protection tools such as protective spectacles, the disaster prevention masks, hazard
masks, and gloves.
（４）Experiment has to be in the draft. Do not leave the toxic or deleterious substances in the draft
after experiment.
（５）Be careful not to leak the materials with the low allowable concentration.
（６）Use materials after obtaining approval from the person in charge of the laboratory. Moreover, letother people in the room know the use.
（７）Be careful especially when you handle a large amount of the substance.
（８）Even if you handle the deleterious materials wearing protective equipments, have first-aid
immediately in the case of accidental ingestion or adhesion to the eyes or skins. Generally, wash
at least 15 min or more with a large amount of stream when adhering to eyes and the skins.
Have a medical examination depending on the situation. In the case of inhalation of thesubstances, move to another place and keep the body warm with the blanket.
In the case of difficult breathing, artificial respiration or oxygen inhalation should be done. At
the same time, arrange the medical examination promptly by calling 119.
（９）When you use deleterious substances which are kept in the locked cabinet labeled with “poison”,
please receive permission from the teacher in the laboratory. Moreover, keep a record of the date,
the name of the user and the quantity consumed on the user’s list.
3.3.1. Poisonous and Deleterious Substances Control Law
For handling poisonous or deleterious substances, it is necessary to obey "Poisonous andDeleterious Substances Handling Rules in Kyushu University " besides "Poisonous and Deleterious
Substances Control Law" and other laws. (Rule Article 1) The obligation of the administration manager
in the department, the management representative, and the users provided in rules of this University are
written as follows.
（１） Obligation of the administration manager in the department (Rule Article No. 3)
The management head in the department must summarize the management of poisonous anddeleterious substances and take steps to prevent any accidents.
（２） Obligation of the management representative (Rule Article No. 6-11)
1) The cabinet for poisonous and deleterious substances should be hard, made of metallic, and be
separate from one for general chemicals.
２) It is necessary to take measures to prevent the cabinet from falling down and to prevent thecontainers for poisonous and deleterious substances from dropping from the shelf of thecabinet, due to disaster such as earthquake.
３) It is necessary to lock the cabinet surely. The management representative has the responsibility
to manage the key.
４） It is necessary to label the character “poisonous substance quasi drugs” on the cabinet and29bottle. In the case of poisonous substances, write “Poisonous” by white line on red-coloredbase. In the case of deleterious substances, write “Deleterious substances” by red line on
white-colored base.
５） The poisonous or the deleterious substances kept for a long term and without prediction of use
should be discarded promptly.
６） When the poisonous or the deleterious substances are discarded, you must obey to the standardof the abandonment provided by laws, ordinance, and the Kyushu University exhaust water
and waste management rule.
７） It is necessary to provide with the use list of the attached paper style, and to understand thevolume of inventories and the quantity consumed.
８） The user’s list should be crosschecked regularly with the quantity of poison and deleterious
substances kept in bottle.
９） In case of loss or theft of poisonous or deleterious substances, notify the administration
manager immediately.
（３） Users’ obligation (Rule Article No. 5, 9, 11)１） People who use the poisonous and deleterious substances must obey to the instruction by theadministration manager in the department or the management representative. Have enoughknowledge of the security and be careful to prevent an accident.
２） Every time you use the poisonous and deleterious substances, you should record in the user’s
list.
３） If
you
use
the
web-system“chemical
designforlaboratory
ver.3.0(http://chem.jimu.kyushu-u.ac.jp/default.aspx)” and records on the web-system, you do not haveto write the use on the paper. However, user should obey to instruction by the management
representative.
４） If you find a loss or theft of poisonous or deleterious substances, you should notify theadministration manager immediately.
Poisonous and deleterious substances user list

No.

SubstanceUnit

g ・ mL ・（）

Storage area
When

Total
Purchased

Used

Remaining
amount30

User namePurpose for Othersuse3.3.2. Specified chemical substances (Ordinance on Prevention of Hazards due
to Specified Chemical Substances, Industrial Safety and Health Law)
As for the trouble by the Specified Chemical Substance, the organ (target internal organs) in which
substances accumulated was dependent on the kind of substances. As for the symptom, cancer, hepatic
damage (icterus, hepatic cirrhosis), kidney problem (kidney inflammation, hematuria), lesions ofhematopoietic organs or blood problem (hemolysis, anemia) are known. The hazardous properties ofmain specified chemical substances and all specified chemical substances are listed as follows. The useof type 1 and type 2 specified chemical substances are subject to the Working Environment
Measurement Law. Parenthetic values show the management density. Refer to MSDS for otherhazardous properties.
Moreover, when the Specified Chemical Substance is used, the application exclusion regulations seenin Ordinance on Prevention of Organic Solvent Poisoning are not admitted. Thus, the limited exhaust
ventilation must be used during handling the Specified Chemical Substance.
Type 1 substances
Seven kinds of chemicals specified for the type 1 substance that is confirmed to be carcinogenic should
basically refrain from the use. Consider the use of the alternative.
Type 2 substances
The substances must be used in the limited part exhaust device.
Table 3-10. Specified Chemical Substance classification
Type 1 Substances
１.dichlorobenzidine and its salts

５ dianisidine and its salts

２ ∂-naphylamine and its salts

６ beryllium and its compounds

３ chlorinated biphenyl

７ benzotrichloride

４ ortho-tolidine and its salts
８ Substance contains 1-6 materials over 1% by weight. Or substance contains 7 over 0.5% by weight.
Type 2 Substances１

acryl amide

２０ methyl bromide２

acrylonitrile

２１ diachronic acid or its salts

３alkyl mercury compounds

２２ mercury or its inorganic compounds

４

asbestos

２３ trilene-diisocyanate

５

ethleneimine

２４ nickel carbonyl

６vinyl chloride

２５ nitroglycol

７chlorine

２６ P-dimethylaminoazobenzene

８

auramine

２７ p-nitrochlorobenzene

９o-phthalodinitrile

２８ hydrogen fluoride

１０ cadmium

２９ ß-propiolactone

１１ chrmic acid or its salts

３０ benzene

１２ chloromethyl methyl ester

３１ pentachlorophenold (PCP) or its sodium salts

１３ vanadium pentoxide

３２ magenta31

１４ coal tar

３３ manganese or its compounds

１５ arsenic trioxide

３４ methyl iodine

１６ potassiumcyanide

３５ hydrogen sulfide

１７ hydrogencyamide

３６ dimethyl sulfide

１８ sodiumcyanide37

１９ 3-3’-dichloro-4-4’-diaminodiphenyl methane

３8 Substance contains 1-37 and is designated by Department

formaldehydeof Labor.
Type 3 Substances
１ ammonia

６

phenol２ carbon monodioxide７phosgene

３ hydrogen chloride

８

sulfuric acid

４ nitric acid９

Substance contains 1-8 and is designated by Department

５ sulfur dioxideof Labor.

3.3.3. Organic solvents (Ordinance on Prevention of Organic Solvent Poisoning,
Industrial Safety and Health Law)
Although approximately 500 kinds of the organic solvents are used industrially, 54 kinds of thosereceive the application of “Ordinance on Prevention of Organic Solvent Poisoning” as listed in Table
3-9. These solvents are confirmed to harmful against human body and used comparatively widely.
“Ordinance on Prevention of Organic Solvent Poisoning” is providing the rule to prevent poisoning andthe troubles by organic solvents for workers.
(1) Notes in general characterization of organic solvent and handling
Organic solvents have generally high volatility. It is easy to stay in a high density when handling it in
an insufficient place to ventilate because the specific gravity of steam doesn't diffuse easily more
greatly than air.
If you might not notice a high-density drift in experimental room, there is a case that it caused
exposure when you are sitting down.
Thus, if you handle the organic solvent which is over acceptable concentration, you should make anexperiment in the limited part exhaust device (the fume hood). Furthermore, it is desirable to ventilate
an experimental room by taking enough fresh air from outside to reduce the concentration of organic
solvent.
Moreover, most organic solvents are inflammable and become explosive gas by mixing volatilized
solvent with air as described in the paragraph of “Fire Service Law.” Therefore, when handling organic
solvents, you should pay careful attention not only to the health but also to the fire risk.
(2) Acceptable consumption of organic solvents
Health problems by organic solvent depend on concentration in the air. It is rare for university toconsume large amounts of organic solvents unlike private factory.
About type 1 organic solvent:
It is excluded that the consumption of organic solvent in laboratory is below the “permissible
32consumption” calculated by the next expression.
“Permissible consumption” for type 1 organic solvent (g/h): (1÷15)× laboratories capacity (less than
150 m3)
About type 2 organic solvents:
It is excluded that the consumption of organic solvent in laboratory is below the “permissible
consumption” calculated by the next expression.
“Permissible consumption” for type 2 organic solvent (g/h): (2÷5)× laboratories capacity (less than 150
m3)
As far as the chemicals under “Ordinance on Prevention of Organic Solvent Poisoning” go, you do nothave to handle the chemicals in the limited part exhaust device when the chemicals are used within
permissible consumption. However, the chemicals with a strong smell or with a dangerous should be
handled in the fume hood naturally.
（３）At the risk of harming human body
The trouble with an organic solvent often chiefly appears in the central nervous system, the liver, andthe kidney. The hazardous property of the main solvents is shown in the following. The management
density is shown within the parentheses. The hazardous property of the class 1 solvent is higher thanthat of the class 2 solvent as understood from the management density.
Moreover, the Working Environment Measurement (refer to paragraph 3.5.) is applied to the use ofclass 1 and class 2 organic solvent. But it is not applied that of the class 3.
Class 1 organic solvents-Chloroform（3 ppm）→Strong narcotic. Liver and renal lesion.
-Carbon tetrachloride（5 ppm）→Cell poison (cutaneous absorption) to liver and kidney.
Class 2 organic solvents
・Methylene chloride （100 ppm）→Skin and mucosal irritation. Opiate potency.
・Aceton （750 ppm）→Skin and mucosal irritation, headach, and vomiting.
・Methanol （200 ppm）→Loss of sight due to optic nerve disease.
・1-butanol （25 ppm）→Irritaing odor. Stimulation of respiratory organ. Liver and renal lesion.
Percutaneous absorption.
・2-butanol （100 ppm）→Few odor. Dermatitides. Keratitiss・isopropanol （400 ppm）→Skin and mucosal irritation, Liver and renal lesion
Ethyl ether（400 ppm）→＞Dizziness (over 2000 ppm)＞lie unconscious(over 35,000 ppm)＞Dead (over 100,000 ppm)
・Ethyl acetate （400 ppm）→Skin and mucosal irritation Anesthetic action
・xylene（100 ppm）・toluene（50 ppm）→Anemia, trouble of hematopoietic system
・DMF（10 ppm）→Skin and anterior eye mucosal irritation. Liver problem
・THF（10 ppm）→Mucosal irritation in skin, eye, nose, throat. sick feeling,dizzy.
・ n-hexane（50 ppm）→multiple neuritis（sensory paralysis, difficulty in walking）

33

Table 3-11. Industrial Safety and Health Act appendix 6.2 and Ordinance on the Prevention of Organic
Solvent Poisoning 1st.
Class 1 organic solvents

Chloroform
Carbon tetrachloride
1,2-dichloroethane
1,2-dichloroethylene
1,1,2,2-tetrachloroethane
Trichloroethylene
Carbon disulfide

Class 2 organic solvents

AcetonIsobutyl alcohol
Isoproryl alcohol
Isopentyl alcohol
Ethyl eter
Ethylene glycol monoethyl ether
Ethylene glycol monoethyl ether acetate
Ethylene glycol monobutyl ether
Ethylene glycol monomethyl ether
O-dichlorobenzene
XyleneCresolChlorobenzene
Isobutyl acetate
Isopropyl acetate
Isoamyl acetate
Ethyl acetate
n-butyl acetaten-propyl acetaten-amyl acetate
methyl acetate
Cyclohexanol
Cyclohexane
1,4 dioxane
Dichloromethane
N,N-Dimethyl formamide
StyreneTetrachloroethylene
Tetrahydrofuran
1,1,1-trichloroethane
Toluenen-hexane
1-butanol
2-butanol
Metanol
Methyl isobutyl ketone
Methyl ethyl ketone
Methyl cyclohexanol
Methyl cyclohexane

34

Methyl-n-butyl ketone

Class 3 organic solvents

gasolinecoal tar naphtha
petroleum ether
petroleum naphta
petroleum benzin
oil of terebinth
mineral spirit

Mixtures consisting of the chemicals listed in above column

3.3.4. Pollutant Release and Transfer Register (PRTR) Law
The PRTR law has aimed at "The improvement of independent management of chemicals by theentrepreneur is promoted, and the obstacle in the environment maintenance is prevented beforehand".
In PRTR law, about the chemicals of one ton or more a year (The first designated specific chemical
substances are 0.5 tons or more) among the first designated chemical substances (354 items), each
campus should calculate the amount of emission to the atmosphere, the public water area, the soil andthe sewers. It reports to the Minister of Education, Culture, Sports, Science and Technology.
Our university record and calculate the transaction volume, the amount of emission, anddisplacement about 7 substances of the following 9 substances except for ethylene oxide gas used in
specific place and dioxin.
Table 3-12. Chemical substances applied to the PRTR law in Kyushu University
PRTR
CAS
Substances
Specific Toxic or Ratio
Steamnumber number
deleteriouspressurechemicalmmHg

Solubility
ｇ／L

1275-05-8Acetonitrile

Deleterious

0.79

88

High

63

1330-20-7

Xylene（o-, m-, p-）

Deleterious

0.87

8

0.1

95

67-66-3

Chloroform

Deleterious

1.48

197

8.0

145

75-09-2

Dichloromethane

1.33

435

13.0

227

108-88-3

Toluene

0.87

28

0.5

299

71-43-2

Benzene

0.88

95

1.8

310

50-00-0

Formaldehyde

4275-21-8Ethylene oxide

Deleterious○○

Deleterious

Formalin fluid

Deleterious

−

Gas

at

High

normaltemperature
179—

Dioxides

○

–

Notes

(1)“specific”：The first specific chemical substances(2)“steam pressure”, “solubility”：25°C property. Pay attention to the emission into air about thesubstances that have high steam pressure, and the displacement into sewerage
about the substances that have high-solubility.
（３）Ministry of the Environment website（http://www.env.go.jp/chemi/prtr/risk0.html）describes
the PRTR law and property.
35

3.3.5. Globally Harmonized System of Classification and Labeling of Chemicals(GHS)
In Japan, hazard classification of several chemicals is carried out based on the domestic laws
described above. In other countries, several laws or regulations exist. Through variations in definitionof hazards, a chemical may be considered flammable in one country, but not another. A lot of chemicals
are traded globally and imported into Japan. Different aspects of hazard for a chemical are notpreferable.
In 2003, United Nations Conference on the Environment and Development proposed adoption of
the Globally Harmonized System of Classification and Labeling of Chemicals (GHS) for all countries(http://www.unece.org/trans/danger/publi/ghs/ghs_rev00/00files_e.html). Through the GHS,
we are able to recognize physicochemical hazards of chemicals and hazards of chemicals to health andenvironment. Even if one chemical is not designated as substances regulated by Japanese laws or
regulations, we should pay attention to the following GHS pictograms labeled on the chemical bottles
or MSDS.
GHS pictograms and hazard classes(Physicochemical hazard)
GHS pictogram

Hazard statement

Explosives

Flammables

Self-reactives

Self reactives

Peroxides

Pyrophorics

Oxidizers

Gases

under

pressureSelf-heating
Emits flammable gas
O rg a n i c p e r o x i d e s(Hazard to health and environment)
GHS pictogram

Irritant

Carcinogen

Dermal sensitizer

Respiratory

HazardAcutestatement(harmful)

toxicity

Acutetoxicity(C a t e g o r y 1 - 3 )N a r c o t i c e ff e c t s

Corrosives

sensitizer

Environment

Reproductive

al toxicity

toxicity

Respiratory tract36

Acute oral toxicity
LD50 (mg/kg)

Category 1

Category 2

Category 3

Category 4

Category 5

< 5

< 50

< 300

< 2,000< 5,000

GHS—

pictogram
Signal word

DangerHazardFatalstatement

swallowed

Dangerif

FatalDangerif

Wa r n i n g

To x i cswallowed

ifswallowed

HarmfulWa r n i n g
ifswallowedMaybeharmfulifswallowed

3.4. Measurement of working environment
Experimental laboratory which handle the chemical substances under Ordinance on Prevention ofOrganic Solvent Poisoning (OSP) or Ordinance on Prevention of Hazards due to Specified Chemical
Substances (SCS) becomes the subject of the measurement of working environment as a rule.
Measurement of working environment is that measure concentration of the substances twice a year inexperimental laboratory which certified as indoor working area which handle harmful substances.
Measurement of working environment follows the working environment measurement standard that thehealth minister defied the method of gathering the samples. It is necessary to preserve the record of theworking environment measurement for three years.
Table 3-13.
Law

Substances applied to measurement of working environment
Chemical substances

Management density
ppm

Boiling
mg/m O25áP pointOáP
3

Stream pressure
OhPa 25áPSCS Type2

Formaldehyde

0.1

0.12

OSP Class2

Methanol

200

262

65

154

OSP Class2

Acetone

500

1,186

56

304

OSP Class1

Chloroform

3

14.7

62

267

SCS Type2

Acrylamide

—

0.1

193

1.2

OSP Class2

Isoproryl alcohl

200

491

83

58.7

OSP Class2

n-Hexane

40

141

69

200

OSP Class2

Ethyl eter

400

1,212

35

713

OSP Class2

Ethyl acetate

200

720

77

129

OSP Class2

xylene

50

217

140

10.7

OSP Class2

Dichloromethane

50

174

40

581

OSP Class2

Toluene

20

75.2

111

37.0

OSP Class2

Tetrahydrofuran

50

147.5

65

243

SCS Type2

Benzene

1

3.2

80

125

OSP Class2

N, N – Dimethyl formamide

10

30

153

4.5

37

3.5. Online chemical control system
Chemical control system is to manage chemicals by registering
personal computer, and it has been used in whole university
since 2006.
Control system is used only in the computer of university as
right-chart indicates. If you register the chemicals on the system,
you get unique chemical number. You can manage chemicals by
writing the number on the bottle or attaching bar-cord to the bottle.
The advantages of control systems are not to buy the extra chemicals by searching in stock online.
This leads to decrease the amount of stock. Moreover, regulation of chemicals and safety information(MSDS) can be obtained(1) Installation Guide
1. Personal computers which connected with LAN on campus are needed. Bar-code reader andbar-code printer are not necessary.
2. You need “attending Login ID” provided each laboratory. If you don’t know it, please ask the
Facilities Affairs Section, Faculty of Agriculture (Nogakukei Hozen-kakari).
3. You can download first step guide and operating manual from website
http://chem.jimu.kyushu-u.ac.jp/.
4. Refer to the first step guide and the operation manual, and resister user information and initial
setting of your attending (laboratory, minimum unit for chemical management).
5. Log on by user ID, and register the chemicals.
(2) The method of control
1. If you buy new chemicals, you should register it on the control system before use.
2. Every time you use poisonous and deleterious substances, you should register the amount of its
use on the control system. This called “usage control.”3. Other chemicals except for poisonous and deleterious substances are managed by “usage control”
or registering the date of purchase and the end of usage date. This called “number control.”“Usage control” is desirable.
4. You may manage the box that contains a lot of ampoules and small bottles or a variety of kits inunits or in kits.
5. The system doesn’t cover medical products, radioactive substances, and fuel such as heavy oil,and other materials in retention tank.
6. Synthetic materials and decomposing materials are also managed in this system.
(3) Other notes
1. If your research department is divided into two campuses, you handle it as another department.
2. When you move to other campus, you should ask the Facilities Affairs Section, Faculty ofAgriculture (Nogakukei Hozen-kakari) to change contents you registered.

38

4. Handling liquefied gas and high-pressure gas
High-pressure gas and liquefied gas are sometimes used in research works in University. We use
dry ice, liquid nitrogen or liquid helium to decrease the temperature of devices and samples. These
substances generate high-pressure gas if someone handle them wrongly and cause rapid evaporation.
It is possible for high-pressure gas to be poisonous or combustible such as chlorine gas or hydrogen gas.
Therefore the safety measurement against high-pressure gas needs multiple especially in University.
4.1 “Gas Property” indicates main gas types used in laboratory and physical and chemical propertiesof gas. It presents to pay attention to gas property such as heavy or light, combustible or thesusceptibility of substances to burn, poisonous, and deficiency of oxygen. Table 4-1 shows properties ofgases.
4.2 “Handling high pressure gas” indicate a manual how to handle a cylinder in laboratory. Whenyou handle high-pressure gas, you should have a certain level of gas savvy. For example, it isnecessary to check the container and the seal of cylinder for before using high pressure gas. Close avalve tightly after use.
4.3 “Handling low temperature liquefied gas” indicate the notes on liquefied gas used as cryogen.

4.1. Gas property
Various gases are used in laboratory. Information on chemical properties, pressure, lowtemperature and liquefied of gas are needed depending on the intended use. Gas exists in gaseous state,liquid state, and solid state (only dry ice is used practically). Table 4-2 shows the characters ofcompression gas in common use.
Table 4-1. The characters of gases in common use
Gas

FormulaMolar weight

Gravity

Flammability(g/mol)（air＝1）（vol %）○4~75

Toxicity

Odor

Hydrogen

H2

2.016

0.070

Helium

He

4.003

0.148

Methane

CH4

16.04

0.555

○5~15

Ammonia

NH3

17.03

0.60

○5~27

Neon

Ne

20.18

0.696

Acetylene

C2 H2

26.04

0.91

○2.2~85△

None

Hydrogen cyanide

HCN

27.03

0.969

○5.6~40○10 ppm

Yes (almonds)

Nitrogen

N2

28.01

0.967

Carbon monoxide

CO

28.01

0.966

○12.5~74○50 ppm

None

Ethylene

C2 H4

28.05

0.975

○3.1~32△

Yes (banana flavor)

Oxygen

O2

32.00

1.105

△

Fluorine

F2

38.00

1.31

△

Chlorine

Cl2

70.90

2.46

△

Others

None
None
None
○25 ppm

Yes (irritant)
None

None

39

None

Liquid is powder blue

○0.1 ppm

Yes

Wear protected gear

○0.5 ppm

Yes (irritant)

Yellow green

Argon

Ar

39.95

1.38

Carbon dioxide

CO2

44.01

1.52

Propane

C3 H8

44.10

1.55

Ozone

O3

48.00

ButaneC4H10

Krypton

None
△

None

○2.2~9.5

△

None

1.65

△

○0.1 ppm

Yes (irritant)

58.12

2.16

○1.8~8.4

△

None

Kr

83.80

2.82

None

Chlorofluorocarbon

CCl2F2120.93

13.8

None

Xenon

Xe

131.304.53

None

Sulfur hexafluoride

SF6

146.05

5.10

△

None

By 0.5％Liquid is dark blue

By 0.1％(1) Gravity・・・Show at 0℃ and 1 pressure because ratio of air by gas leakage is connected with safety.
Ventilation technique will be decided by this.
(2) Flammability・・・Flammability gas is indicated by ○circle sign, and flammability range is indicated by
mix proportion （vol %）in the air. It will burn if there is a gas in the range of this mix proportion. Triangularsign △ indicates the susceptibility of substances to burn.
(3) Toxicity・・・Gas with toxicity is indicated by ○circle sign. These numbers show the limit you can work inthis atmosphere for 8 hours a day and 40 hours a week. Small numbers have stronger toxicity. Narcotic is
indicated by triangular sign △.

4.1.1. Combustibility
Table 4-1 shows combustible gases. Combustible substances are marked by ○ on the table. Itshows flammability range (%) when leaking to air. This range is called “detonation” when burning
speed is over acoustic velocity. For example, in case of hydrogen, the range of burning is 4~7%,
detonation is 18~59%. There is danger of ignition in the empty-container. Once igniting, gas decreases
and blows up.
In order to avoid an accident of combustible gas, it is necessary to be aware of the range of burning,
avoid leaking gas, and ventilation. In case of gas leak, open the window immediately and dilute it withcarbon dioxide or nitrogen. For ventilation, please be aware of gas gravity. Ventilate a room in upperside in case of light gas and lower side when gas is heavy. Do not leave a fire or spark near the place
where you handle gas. Once igniting, extinguish by fire extinguisher and water, (notes on chemical
reaction), then close the main cork of the gas.
4.1.2. Poisonous property
Carbon monoxide and ammonia are known as poisonous gases in laboratory. Be aware ofventilation, and use a gas mask to avoid taking a breath directly. In case of people losing consciousness,take them out the room and lay them on their side, then call an ambulance. Please consult a doctor
immediately when poisonous gas was inhaled.
4.1.3. Oxygen’s susceptibility of substances to burn
Oxygen itself is not combustible and poisonous. But pure oxygen is very dangerous. Almost
everything ignites in pure oxygen. A little grease attached to the cap of oxygen cylinder causes a fire
when gas is leaked. As nitrogen having low boiling point evaporates first in liquid air and then enrichedoxygen evaporates, we should notice the concentrated oxygen. Pouring liquid nitrogen into open bottle,
40

oxygen seeps in and reaches a high concentration.
4.1.4. Oxygen starvation
Leaking gas other than oxygen into room causes oxygen starvation. It leads to breathing difficulty.
Permissible limit of oxygen for people is 18%. People become unconsciousness when available oxygenis fewer than 7%. It is necessary to ventilate a room when leaking a large amount of gas into room andin case of liquid gas leakage evaporation is required. Pay attention to lack of oxygen with an oximeter.
In case of losing consciousness, take victims out of the room immediately, and call an ambulance orconsult a doctor. Be aware of further accident. Actually, there was an accident in which two students
died in University because a large amount of liquid nitrogen was handled in a bad ventilation condition.

4.2. Handling high-pressure gas
It describes the case of using a cylinder filled with high-pressure gas to reduce the compression.
Gas is filled in steel bottle called cylinder with reduced pressure. Usually gas is bought from the agency.
Gas is very dangerous and causes major accidents when mistakenly handled.

4.2.1. Container indication
According to High Pressure Gas Safety Act, it is mandatory to put a punch mark on container. If it
is difficult to put it, use a seal. Possessors should indicate it very clearly and put their name, address
and phone number. Change marking color to distinguish different kinds of gas. (Refer to Table 4-2)
4.2.2. Transportation
Use a special transportation car to transport cylinders with cap. Handle it very carefully duringtransportation. Take off the adjuster of cylinder and put a cap to protect valve. Container of acetylene
gas and liquid gas should not be lay down.
(1) Use a special hand barrow for carrying cylinder.
(2) Take off adjuster and close valve tightly with protect cap on valve before moving.
(3) When you do not use a hand barrow, tilt cylinder slightly and transport it carefully.
(4) When taking stairs, the cylinder should be carried by two persons or more. Do not cover the capwith rope.
4.2.3. Storage
When fixing cylinders in laboratory, shield direct light and fix it with special trestle inwell-ventilated space. If you do not use a special trestle, you should fix it in the laboratory table or thepole with chain or band. Keep it away from heater to avoid danger of blowing up safety valve. If thesafety valve blows up, keep the cylinder in a safe place and leave it as it is until the pressure decreases.
In case of combustible gas, be aware of surrounding fire and ventilate a room immediately.
(1) Cylinders should be fixed on the container trestle.
41

(2) If you don’t use a trestle, fix it in the laboratory table or the pole with chain or belt.
(3) Keep liquid gas or acetylene cylinders stand vertically to use.
(4) Shield the direct light and avoid a moisture place.
The temperature should be under 40á.
(5) Keep cylinders away from electric cable or earth cable.
(6) Keep cylinders away from emergency stairs.
(7) Close a valve tightly and cover it with a cap when you don’t use.
(8) Store cylinders separately for combustible and poisonous, as well as for filled and empty.
4.2.4. Confirmation of container
Confirm the display of the gas container before opening the valve. As shown in Table 4-2,
container is separated into different colors depending on the type of gas. The name of filled gas and its
properties and address are also displayed. %!$means combustible gas, “ ”means poisonous gas.
The holder’s name and address is also shown. As shown in Fig. 4-1, there is a punch mark on the upperside of cylinder, and it shows the details of cylinder and filled gas.

Fig. 4-1. Punch mark on cylinder
Punch mark and code
1. The symbol of a specific container.
2. The symbol of passing an inspection or the name of inspector.
3. Container manufacture or its code
4. Name of filled gas i. e., “He”
5. Container code or serial number i. e., “GK19564”
6. Volume i. e., “V 47.3”
7. Weight i. e., kg“53.5”

8. The date of pressure proof test i. e., month-year (two digits of the year) “7-80”
9. Pressure proof test pressure i. e., kg/cm2 TP250
10. Maximum filled pressure kg/cm2 i. e., FP150-In the case of passing pressure proof test.
11. The code or the name of inspector and the date of inspection. i. e., 12-83 and 4-87
42

12. Confirmation of mass and variance of mass i. e., 4.90.53.5KG O90 ^ 4 t 53.5 kgP
13. Holder registry number i. e., M369
Table 4-2. Color-coding and display of properties depending on the type of gas.
High-pressure gas

Container

Character

Character color indicatesGas exit screw

color

color indicates gas type.

properties of gas.

Oxygen gas

black

white

Hydrogenous gas

red

white

Liquefied carbon dioxide

green

white

Liquefied ammonia gas

white

red

Liquefied chlorine gas

yellow

white

Acetylene gas

bark

white

ìﬂíwhite

Combustible gas

gray

red

ìﬂíwhite

Combustible, Poisonous

gray

red

ìﬂíredPoisonous gas

gray

white

Other gas

gray

white

Nitrogen gas

gray

white

right

Helium gas

gray

white

left

right
ìﬂíwhite

left
right

ìﬂíredêì\íblackleftì\íblackrightjoint without screw

ì\íblack

ìﬂí
Combustible gas ,ì\ípoisonous gas
4.2.5. Notes on handling pressure regulator and valve(1) Pressure regulator (Fig. 4-2.)(a) Use a specific regulator for gas. Do not use it for another purpose. Especially use “no-oil” only
for oxygen gas.
(b) Understand the handling method because each regulator has different methods depending on
their types.
(c) Keep the connection between valve and regulator clean and use appropriate screw. Use new
packing appropriate for the connection.
(d) When installing hexagon cap nut of regulator, use a specific screw wrench. Install it carefully not
to break the packing or screw.
(e) Pressure meter is used 1.5-3 times as much as normal.
(f) Do not lubricate the regulator with oil too much. It causes a fire.
(g) Close valve tightly and take out all gas from the regulator before taking off the regulator.

Fig. 4-2. Pressure regulator(2) Handling methods of valve (Fig. 4-3)(a) Valve has two screws, right and left. (Fig. 4-2)
43

(b) Do not touch a safety valve.
(c) When opening and closing valve, stand on the side of pressure meter. Quickly opening cause thebreakage of regulator. At this time, use a special handle or wrench.
(d) Open valve fully two and half turns when using it. (Acetylene is one and half turns)(e) When leaking gas from valve, move its container into safe place.
(f) Handle the valve carefully. Do not displace gas in the container to another container.
(g) Close the valve with leaving a little gas in the cylinder and take off regulator, and put a protect cap
to the valve after you’ve done with it.

Fig.

4-3.

The structure of valve and valve with wrench(3) Other notes(a) Close not only regulator but also valve of cylinder, and take off the connection between
laboratory units and regulators when suspending use of gas.
(b) In the case of heating up regulators and valve, use heating pad or hot water the temperature
under 40á.
(c) Return cylinders immediately after you’ve done with it.
4.2.6. Other notes(1) Combustibility/susceptibility of substances to burn(a) Do not use a fire within 5m from facilitates in which you use combustible gas or oxygen. Do
not put inflammable and combustible substances at a nearby site.
(b) Remove combustible substances such as oil from devices before using oxygen.
(c)The susceptibility of substances to burn (oxygen) should not be used in the room wherecombustible gas is used.
(d) Oxidized ethylene should be used after replacing its contents with N2 or CO2.
(e) To prevent from decomposition explosion, acetylene should be used under 1 kg/cm2 pressures.
(2) Poisonous gas(a) To prevent from inhaling poisonous gasesOH2S, CO, Cl2Pone should conduct an experiment in
the draft. In the case of inhaling cyanide gas (hydrogen cyanide: HCN, sodium cyanide: NaCN),
it will cause some danger of cyanide intoxication (headache, dizzy, respiratory paralysis).
Silane gasO
SiH4Pis combustible and has the capability of causing a danger of fire easily.
ObPMake poisonous gas harmless with alkali absorbent before being disposed off.
(c) Poisonous gas should be handled carefully because it effects on human health.
44(d) To use poisonous gas, wear appropriate clothes, glasses and gas masks.
(e) To use poisonous gas, it is better to use a small amount of it as much as possible in terms ofsafety.

4.3. Cryogen (cryogenic liquefied gas)(a) When using elevator for transportation, you should not take on it with liquid gas containers andcylinders. If the valve of cylinders would be broken, gas will be leaking in the elevator and people
might choke to death. In the case that elevator stops by blackout when gas may not be leaked but
one may choke to death from the vaporizing gas.
If you find cylinders or containers in the elevator, you should not get on the elevator.
(b) Do not touch cryogenic liquefied gas directly. Please use it in a well-ventilated place and handle itnear floor surface.
(c) Do not attach oil to the devices of cryogenic liquefied gas.
(d) Wear a leather glove when using cryogenic liquefied gas. Do not use a cotton glove.
(e) It is harmless that you inhale a little amount of inert gases (carbon dioxide, nitrogen, argon gas) andcryogenic liquefied gas. However, if you inhale them in large amount, it may cause the absence ofoxygen. One may be faint by the decreased level of oxygen fewer than 10%. When handling large
amount of them, use a well-ventilated place.
(f) When using liquefied gas such as liquid nitrogen, open the window even in the winter. Do not use it
in the place without window.
(g) After taking a lecture “cryogen user training session” presented by low temperature centerand understanding the guideline that gave out in the lecture, you can handle liquefied gas by
yourself.

45

5. Electric safety planning
5.1. Treatment for electric shock
Electric shock means that electric current run through the human body. It has a high fatality rateand causes several problems. It is highly dangerous for electric current to run through nerve center ofhuman body such as brain and heart. According to past statistics, over 40-60 V of electricity causes
death.
5.1.1. Electric shock result from indoor distribution line
(1) Be aware of working on floor surface which allows electric current through easily.
(2) Do not touch a distribution line directly with wet hand.
5.1.2. Electric shock result from high voltage equipment(1) Connect electric discharge without touching.
(2) Do not use inappropriate insulation (for example, phenol or resin).
(3) Work with two or more persons near high voltage equipment. One person has to monitor.
(4) To prevent from exposing high voltage, shut the devices tightly with metal cover.
(5) Emit a warning sound and lighting when generated high voltage.
(6) Earth the edge of high voltage equipment when touching it or not.
5.1.3. Electric shock result from electric/information devices(1) Some devices contain high voltage circuits(2) Capacitor is charged even though the power supply is cut.
(3) Many series of connected batteries are dangerous.

5.2. Electric disaster and its countermeasure
5.2.1. Overheat
Short circuit, excess current, earth leakage may cause a fire. Pay attention to the following points.
(1) Overloading an electric circuit(2) Bad connection and partial disconnected by heavy load.
(3) Use an electric line endured rated current.
(4) Use an appropriate fuse or a breaker.
(5) Set up ground fault interrupter.
(6) Use a heat-resistant code for conversion heater.

46

5.2.2. Electric spark
Combustible gas or vapor of inflammable liquid causes blowup or a fire by electric spark.
Liquidization of insulator or fast fluidization of powder cause spark and lead to blowup accident.

5.3. Notes on electric devices
5.3.1. OA equipment(1) Follow the rated current of power code and power strip.
(2) In the case of putting power code on floor surface people walk, protect it with a code protector.
(3) Cut the power supply when you are out.
5.3.2. Equipment in pause
If you reuse the equipments that had not been used for a long time, you should check the conditions
before using because it is highly likely that such an equipment has lack of lubricating oil or insufficientinsulation.
5.3.3. Blackout, all-night service
When the power comes back on, turn off all the equipment because the electric equipments areautomatically working when returning from blackout.
Confirm the schedule of blackout by regular checking.
5.4. Check for accident avoidance
To avoid the accidents, check the following points regularly.
(1) Feel a shock when touching the devices or pipe: It is very dangerous. Stop using it immediately andcontact agricultural maintenance personnel.
(2) Breakage of covering codes: Be aware of old codes.
(3) Loosing connected screw: It causes short out or heat.
(4) Abnormal sound: Electric leak and discharge or defect of motor.
(5) Abnormal odor: Electric leak and discharge, degradation of insulator by heat.
(7) Run off ground lead: Confirm whether you forget to put ground lead when transported or not.
(8) Confirm whether the equipments and hard wiring are covered with water, gas or chemical liquid.
(9) Confirm whether power codes are covered with heavy load or not.

47

6. Handling glassware
6.1. Safety precaution
There are many cases of injury caused by carelessness of handling glassware. Check the following
points to avoid accidents resulting from handling glassware.
(1) Dispose of glassware cracked.
Cracked glassware are easily breakable. Broken glassware causes injury and it is likely to leakorganic solvent and poisonous substances. If you find cracked glassware in the laboratory, you
should dispose it immediately.
(2) Handle cracked glassware appropriately.
If the edge of glassware is cracked, it should be burned to smooth down the edge. If you do nottake care of it, someone might be injured.
(3) Purchase plastic containers for large amount of capacity.
Glass beaker and measuring cylinder more than 1 L are easily broken. So, it is recommended to use
plastic containers unless there are special circumstances.
(4) Well-organized
Doing experiments in a narrow place is not only decreasing working efficiency but also exposing
you to danger. Troubles may be caused by knocking the container and spilling its contents or
breaking the glassware. Keep the laboratory table clean and take enough working space when you
make an experiment.
(5) Tip-resistant
Do not leave materials at the edge of laboratory table. Those materials may cause accidents whensomebody passing through. If you find something may cause an accident, you should move it to asafe place.
(6) Magnetic stirrer
When putting magnetic stirrer into glassware, skew the container and slide it. As stirrer has astrong magnet, glassware would be broken when it move with a high speed.

6.2. Insert glass tube into a rubber plug (pipetter)(1) Do not try to insert thick glass tube into a narrow hole.
(2) Dab water on glass tube for sliding well before inserting into rubber plug. However, do not use
water or vaseline when inserting measuring pipette into pipetter.
(3) Grab the point in 2 cm from the edge of glass tube with thumb, forefinger, mid finger and carefully
insert it while twirling.

48

Breakage of glass tube may occur in just a couple of centimeters from the edge of rubber plug.
If you grab the points far from the edge of rubber plug and try to insert glass tube, glass tube will be
subjected to bending stress. If you grab the glass tube with five fingers, bending stress is moreincreased. Grab the point in 2 cm from the edge of glass tube with thumb, forefinger, mid finger. If
you don’t use fourth finger and little finger, you can avoid accidents of breaking glass tube. Please
try to create an appropriate hole again if you could not insert with three fingers because the hole is
too small.
For a bad example, Fig. 6-1 indicates. Glass tube will be broken if a force in the point of arrow
applied to it. According to the left-fig 6-1, when squeezing the glass tube into rubber plug, a force
acts in a vertical direction by the little finger. Breakage of glass tube occurs in the base of rubber
plug. According to the right figure, glass tube will break between mid finger and fourth finger.

Fig. 6-1. Bad examples (Insert glass tube into a rubber plug)

6.3. Remove glass tube from a rubble plug
Accidents occur when removing glass tube from a rubber plug.
To prevent from an accident, some cautions are needed as follows.
-Grab the glass tube with three fingers and rotate it softly. It is helpful to drop little amount of water
between glass tube and rubber plug, and knead the rubber plug.
-If you could not remove the glass tube from a rubber plug, cut a part of rubber plug.

6.4."Hold a large glass container or dangerous reagent bottle with
one’s hands(1) Do not hold strongly a beaker over 500 mL in one hand. There is a case of cutting the nerve endings
and it will take one year to cure it.
(2) Hold a gallon bottle with one’s both hands. If you hold only a part of handle, the handle may often
torn by its weight. Handle is not for carrying.

6.5. Opening the lid of glass container
There is a need to be more cautious for handling vial container, test tube, and thin glass container.
49Remember that glass will be broken by strong force. To chill a glass or warm the lid are effective for
opening it. Even though jelly bottle has enough thickness to avoid an accident, vial container and test
tube doesn’t have enough thickness. Those are easily breakable.

6.6. Opening ampoule
Think the reason why test reagents are inside the ampoule. There are test reagents unlikely oxygen or
water, having strong odor and poisonous. Consider the reason why those reagents are inside ampoule,and prepare for handling it.
(1) If you do not use all reagents you opened, prepare for the appropriate container to preserve ittightly.
(2) In the case of volatile materials, chill it with icy water and take off the liquid.
(3) File down(4) Open it carefully. Grasp an ampoule with one’s left hand and break off it with right hand. Takecare of your hands. You can wrap up it with a dry cloth before breaking off.
Consult an experienced teacher regarding opening of poisonous reagents such as poisonous
materials, special flammable materials and explosive properties or ampoule over 5 ml.

6.7. Clean up after damaged
When the glassware is damaged, use a broom or vacuum cleaner. Do not use your bare hands because
broken piece of glasses are very dangerous. When you break glassware filled with solution, you wipe
off the solution with a paper towel. If you wipe it with a cloth, you will be injured when wringing out
the rag.

6.8. Other notes on handling glassware(1) Glassware.
1. Containers should fit in with the size of the cover. The cover and container should be numbered.
2. Do not keep the cover dry when screwing it. Glassware would be damaged.
3. Put a strip of paper between the container and cover after drying.
4. Do not use glassware for preserve of alkaline solution because glassware will be melted by alkaline.
(2) Magic marker
Do not write anything in a part of white letters and fogged glass.
(3) Take off a label and magic marker.
Take off a label and magic marker before putting it into drying machine. Pulling it by force leads may
cause accidental breakage.

50

6.9. Notes on cleaning(1) Secure working space.
Secure enough working space when cleaning. Start working in the half of sink with nothing on it.
(2) Do not leave any devices in the sink.
Professional household workers wash dishes immediately after use because washing dishes after longtime requires a lot of time and effort. If you could not wash dishes immediately after use, you soak thedevices in water and put it on your laboratory table. If you leave it in the sink, you will bother others in
the following reasons.
1. Other people do not know whether the device handled dangerous reagents or not.
2. Other people can not use enough space to wash.
3. There are no devices when other people would want to use.
(3) When leaving a measuring cylinder in the sink, please lay it down.
Measuring cylinder is easy to be damaged by a high gravity point.
(4) Take off a tape and magic marker.
Take off a tape and magic marker before putting it into drying machine. Pulling it by force leads may
cause accidental breakage.
(5) Do not use a cleanser.
Do not use a cleanser for disposal of grime and magic marker. Even though you use a cream cleanser, it
scratches the surface of glass.
(6) Proper brushing
Two or three times brushing is not enough to wash. Look at the shape of brush and confirm whether thedevices are brushed enough or not. Use a sponge for open-mouthed containers like a beaker. It is better
to use special glassware for quantitative determination of reducing sugars by using such as
phenol-sulfuric acid method and for micro scale analysis such as fluorescent analysis. For example, put
the devices in a large stainless-steel container, and wash them cleanly with mild detergent (0.5%).
(7) Notes on alkaline detergent.
When using alkaline detergent, wear the eye protector and glove. Glassware can be melted by strong
alkaline. Using alkaline detergent for a long time and heating lead to melting the surface of glass.
(8) Wipe water off on the floor.
Floor become slippery when wet. Wipe water off immediately with mop.
51

7. Notes on physical system laboratory
Accidents occur when machines and tools are handled carelessly or by disorganized workers. Whenusing tools and machines, follow the rules and pay attention to the foreseen accidents.

7.1. General notes(1) Workers should be trained and understand how to use machines beforehand. .
(2) Confirm the range of work and eliminate a blockage before boot.
(3) Suspend the machines immediately when you feel abnormal.
(4) Switch off the machines before maintenance.
(5) Cover a rotating part of gear, belt, shaft, and rubber.
(6) Do not stop machines run by inertia with bare hands, tools, and bar.
(7) When the power went down, switch off the machines.
(8) After working hours, check the machines completely before switching off all machines.

7.2. Tools
Place emphasis on the general tools such as driver, spanner, puncher, and file.
(1) Treat tools with care.
(2) Use appropriate tools for work.
(3) Dispose of defective tools.
(4) Set tools to rights.
(5) Use an eye protector.

7.3. Safety handling on wood processing machine
Wood processing machines such as band saw and circular saw are used for sawing or grinding woods.
Understand that wood processing machines are very dangerous tools because the knife rotates at highspeed. Pay attention to the following points.
(1) Wear a long-sleeve tighten clothes. Do not wear clothes which have risk of getting sucked into suchas a white coat.
(2) Wear a pair of athletic shoes. Do not wear a pair of slippers or sandals.
(3) Do not use a pair of gloves.
(4) If necessary, use an eye protector and other protectors.
(5) Confirm the fixing of cutting blade before work.
(6) Turn on the switch and confirm the speed of rotating becomes constant, then start working.
(7) If it is difficult to insert woods into machine, please do not force them into.
(8) Dispose of wood chips after machine stopping completely.

52

7.4. Safety handling on agricultural machines
People who work on the farm should take careful note of safety and daily check of agricultural
machines to avoid an accident. When using machines or tools, check the safety device and protective
cover in advance and understand how to use them. If you find some problems on the machines andtools, please take necessary steps to settle this issue.
7.4.1. Work environment(1) Wear an eye protector and a dust mask when working under an environment where dust occurs.
When working indoors, enclose source of release with curtains or suck in with suction machines.
When working outside, stand in the wind.
(2) The noise may disturb the acknowledgement of alarm or contact between workers, and cause an
accident of agricultural work. Moreover a hearing loss and compromised physical function may becaused by the noise.
(3) Vibration for a long time may cause an accident and compromised physical function. Take a rest or
take turns properly. Prevent workers from working continuously.
(4) Evacuate to a safer place immediately at the bad weather.
7.4.2. Vehicle(1) In an emergency, confirm how to stop an engine or operating machine.
(2) Adjust your seat and wheel position and seat suspension for your physical constitution. In the caseof tilt steering wheel, column is needed to be fixed.
(3) Handle a machine with power steering carefully because its handle is very lightweight and you may
sway dangerously.
(4) Understand that crawler machine change the axis of rotation and radius of turn due to the evolution
system.
(5) There are many accidents caused by falls. Use seatbelts and safety frame and protective cab.
(6) When you start a machine, confirm back and forth and around. Keep people away from the machine.
Get in the driver’s side and confirm gear lever, PTO lever, each control lever be in neutral position,and brake before driving.
(7) When you drive a heavy working machine direct equipped at the back, go slow down and use abalance weight if necessary. A machine with a right and left brake should be connected with rightand left brake pedal.
(8) As there is a possibility of going out of control, let out the clutch and maintain gear neutrality on asteep down-grade.
(9) A machine with right and left brake should be connected with right and left brake pedal.
7.4.3. Walking machine(1) Each machine has a different operation procedure. Please understand how to operate it beforehand.
(2) Neutralize control lever or turn off it before starting engine.
(3) Do not get your feet under the rotary casually. Do not move with machine rotating.
53

(4) When going back, there is a risk of getting people caught in the operation machine. Pay attention to
the rear things or the floor condition.
(5) On the road or steep ground, it is better to use steering than to use clutch.
7.4.4. Fixed mount type machine(1) Engine type machine.
Engine type machine should be used in well- ventilated space because there is a risk of being
poisoned by gas when you use it indoors. Be aware of a fire when taking on fuel. Spilｌed fuel should
be cleaned away.
(2) Motorized model machine
The breakage or corrosion of consent, plug for power supply, code of power supply, earth cable,
and switch box should be repaired immediately. Keep liquids away from power supply.
(3) Fixing should be in a flat and strong space. Consult a person who has a specific knowledge about
moving and fixing. Cover a moving part or set up a guard fence.
(4) Use an adequate fuel tank. Fuel tank should be connected with machine to avoid leaking fuel.
(5) To avoid being poisoned by gas and imperfect combustion, fix an air hatch when using it in a
blocked space. Use it in well-ventilated space.
(6) Keep an enough space around the machine because heat generated may influence on around.
(7) Keep control board away from water and dust. Secure an enough power supply for machine. Fix anearth cable and short circuit breaker to avoid a short circuit.
(8) Power supply code should not be bundled because it may generate heat. Install wiring to keep awayfrom water, oil, and high temperature place, and sharp corner. Keep the aisle clear to avoid
disconnected wiring by stomp. Cover wiring with metal pipe in which there is a risk of the breakage bymouse.

54

8. Notes on biological system laboratory
8.1. Introduction
Ethidium bromide is a carcinogen and usually used at high concentration for equilibrium
density-gradient centrifugation (consider that it is more dangerous than RI). If it is spilled, be sure toreport it to the instructor and conduct proper decontamination. In particular, the Beckman
ultracentrifuge in the common equipment room in the basement can easily become contaminated at thetime of heat sealing the centrifuge tube. Check whether there is contamination using a built-in
ultraviolet (UV) lamp before and after sealing. Under short-wavelength ultraviolet irradiation in a darkroom, an orange glow indicates contamination. Of course, short-wavelength ultraviolet must not be
irradiated directly onto the skin or looked at with the naked eye.

8. 2. Experimental equipment
8. 2. 1. Autoclave
8.2.1.1. Autoclave structure
Among the many possible injuries that can occur in the laboratory, a burn caused by the use of an
autoclave is common as well as laceration from glass. Use the autoclave bearing in mind that its
operation has risks.
Figure 8-1 shows the structure of the most common autoclave in the field of biochemistry. The time tomaintain the obtained temperature after the temperature reaches the set value (the range indicated by
the arrow in Figs. 8-2(A) and (B)) is set on the timer and the standard time is 10 – 20 minutes at 121°C.
8.2.1.2. Operation
Packing(1) Check that the autoclave is filled with water up to
Drain
the bottom plate level.
Pressurevalve
sensor(2) Check that the drain valve is closed.
(3) Fasten the lid moderately.
If the lid is fasten insufficiently, the vapor will leak, Temperature
Bottom plateand if fastened too tightly, the packing will sensor
deteriorate fast, also causing vapor leakage. It is
Heaterbetter, as a rule, to turn the handle using theforefinger to the end, and then fasten it by turning
Drain
Boil-dry protection
the handle by 90 degrees with both hands.
outletsensor(4) Set the time and start the sterilization.
(5) Wait until the temperature decreases to at least
Fig. 8-1. Structure of autoclave
70°C.
(6) Check that the temperature is 70°C or less and thepressure is zero. Actually, there was a case inwhich the thermometer was broken and the spewing vapor caused a heavy burn on the upper bodyof the victim. Be sure to check both values.
55

(7) In addition, open the drain valve and check that no vapor spews, and then open the lid (someautoclaves have no drain valve or automatic open and close system).
8.2.1.3. Safety precautions
Do not leave the laboratory unattended until the timer cuts out. This is because no-water burning will
be caused if the vapor leaks caused by the aged deterioration of the packing. If vapor leaks, the sample
will continue to be heated up because the temperature cannot reach the set value and the timer cannot
start. Finally, all of the water will be exhausted, causing no-water burning (refer to Fig. 8-2(C)).
(B)

(C)

(D)

Temperature

！

Time

Time

Time

Temperature(°C)

(A)

120
100
80
Time

Fig. 8-2. Change in temperature in autoclave with time(A) A small amount of medium is sterilized, (B) a large amount of medium is sterilized, (C) when the vapor leaks,and (D) the deviation of the real temperature of difficultly cooled liquid (solid line) from the value indicatedon the thermometer (dashed line)(1) Do not leave the laboratory before the timer is shut off. This rule should be strictly applied
when everyone leaves the laboratory for a magazine meeting or seminar meeting. Do notplace too much confidence on no-water burning protection sensor because it is not assured toalways correctly function and whether the sensor correctly functions or not cannot bechecked.
(2) There is a deviation (difference) of the real temperature of the autoclaved liquid from the value
indicated on the thermometer (the thermometer part is cooled earlier than the liquid because theoutside of the autoclave is first cooled). The real temperature of agar medium, antiforming
agent, highly viscous liquids, such as sugar at high concentration and glycerin, and a large
amount of solution may be more than 100°C even though the thermometer indicates atemperature less than 80°C (Fig. 8-2 (D)). When taking out such a sample, sudden boiling may
occur, causing a burn. When it is autoclaved, wait until the value indicated on the thermometerdecreases to 60 – 70°C, take it out to avoid sudden boiling, and never shake up the solution justafter it is taken out.
(3) Do not put agar medium into a container exceeding half the container’s volume. For example,
when using a conical flask with a volume of 500 mL, the agar medium limit is 250 mL. If moreagar medium is used, the risk of sudden boiling increases, causing problems when mixing after
autoclaving (the concentration of agar around the bottom after autoclaving is high because agarsinks). If agar is mixed just after autoclaving, sudden boiling may occur. Leave it to cool at room
temperature for a few minutes.
(4) Do not autoclave a sample with a low boiling point, such as organic solvents. To do so may cause
fire or explosion; in addition, the bad smell produced will disturb others. The concentration of the56

sample will markedly change because of evaporation.
8.2.1.4.

Other precautions(1) If the apparatus is to be autoclaved, be sure to check that it can be safely autoclaved by referring to
the catalogue and other information sources. For example, the Pipetman of Gilson cannot be
autoclaved. Many plastic apparatuses cannot be autoclaved.
(2) A volatile strong acid (hydrochloric acid, nitric acid) can damage the autoclave. Sterilize it using afilter made of appropriate material.
(3) If a strong alkali is autoclaved, it will dissolve glass. If a glass container is repeatedly autoclaved
many times, the glass will progressively become thinner, causing breakage (pay attention because
glass components dissolve in alkalis).
(4) If a polyallomer centrifuge tube carrying organic solvent such as phenol and chloroform after DNA
or RNA extraction is autoclaved, it will be melt. Remove the solvent completely before
autoclaving.
(5) If the jar fermenter is autoclaved, do not open the lid until the temperature decreases to 60°C orless as a rule. It is difficult to cool a large amount of medium and it not only has a risk of sudden
boiling, as shown in Fig. 2 (C), but also may damage the pH sensor and dissolved oxygen (DO)concentration sensor owing to the change in its pressure.
(6) Be sure to loosen the lid of the container to be autoclaved. If sealed, the pressure outside thecontainer increases prior to the inside as temperature increases, and decreases as temperature
decreases. This pressure difference may cause the breakage of a glass container or the deformationof a plastic container.
(7) If a plastic container, such as a centrifuge tube, is autoclaved, the lid will be pressed closely against
the body when the container is cooled having a negative pressure after autoclaving, and will be
deformed when the container is further cooled having a lower inner pressure. Therefore, sterilize
the container with the lid removed, or wrap the container with aluminum foil while the lid is being
sufficiently loosened so that the lid cannot be pressed closely against the body.
(8) An uncracked medium bottle resistant to approximately one atmospheric pressure can be sealed
and autoclaved. However, it cannot be sterilized unless a drop of distilled water is put in the bottle(it takes at least three hours at 160°C to sterilize the sample under a dry condition).
(9) Overheating may transform the sample. If compounds with amino groups and reducing sugars,
such as yeast extract and glucose, are autoclaved at the same time, the Maillard reaction occurs;
therefore, the medium turns brownish and the sample strain may be prevented from growing. The
longer the heating time and the higher the pH (pH>6), the more marked this reaction. Moreover,this reaction occurs more markedly when a large amount of medium is autoclaved (Fig. 8-2 (B))
than when a small amount of medium is autoclaved (Fig. 8-2 (A)).
(10) If the sample is spilled (boiled over) in the autoclave, clean it up. Even if no sample is spilled,change the water once a week and clean up the autoclave every two to four weeks.
Examples of accidents caused by using the autoclave are as follows.
• A bottle, 90% filled with agar medium, was autoclaved with the lid closed. When it was takenout at 90°C or less, sudden boiling occurred and the flying lid caused a burn on the face of thevictim.
• When a jar fermenter was taken out at 90°C or more, sudden boiling occurred, causing a heavy
burn on the upper body of the victim (a keloid remains).
An antiforming agent was autoclaved in a conical flask. Although the antiforming agent filled a quarterof the flask, sudden boiling occurred when it was taken out at nearly 90°C, causing a burn that required57

a month for recovery.

(1) Do not leave the laboratory unattended until the timer shuts off.
(2) Do not open the lid until the temperature decreases to 70°C or less.
(3) A sample with a low heat conduction (high viscosity or a large amount) has a risk of sudden
boiling even at 70°C or less.
(4) No organic solvent, strong acid and strong alkali should be autoclaved.

8.2.2.

Centrifuge

First, please answer the following questions (the answers are on the next page).
Q1. Is it permitted to use a combination of differently shaped centrifuge tubes if they arebalanced with the scale?
Q2. When there is only one sample, can the sample be balanced with another centrifuge tube of
the same shape filled with water?
Q3. When there is only one sample, can the sample be balanced with another centrifuge tube of
the same shape filled with a solution, such as sucrose, that has the same specific gravity as
the sample?
Q4. Is it unnecessary to switch on the cooler when centrifuging at room temperature?
Q5. Why is the moisture wiped before balancing when the outside of the centrifuge is wet?
Q6. What might happen when a sample leaks from a tube broken during centrifugation?
Q7. Why should the centrifuge rotor be left upside-down after use?
Q8. Why is the cover of the chamber closed during cooling condition and open after use?

Did you answer all the questions correctly? The centrifuge is rotated at a high speed and its kinetic
energy is high. If used in a wrong way, a serious accident may result and the lifetime of the machine
may be markedly decreased. Know the correct method of use and understand the reasons why such amethod is used.
A1. No (There is no balance of moment.)
A2. No (There is no balance of moment.)
A3. No (If separated, the sample will have an imbalance of moment.)
A4. No (There is an increase in temperature due to the heat generated by the motor and thefriction between the rotor and air.)A5. Water on the outside of the centrifuge tube is transferred to the bottom of the rotor by
centrifugation, causing an imbalance of moment.
A6. (1) Machine damage (spindle, motor and rotor) due to imbalance.
(2) Biological contamination, chemical contamination, corrosion and fire (explosion) caused
by wet sample.
A7. To avoid imbalance owing to dust dropping on the rotor and precipitating water.
A8. Frozen ice in the chamber falls off by the wind pressure generated during centrifugation anddamages the rotor.

588.2.2.1. Precautions concerning maximum rotating speed
Use a centrifuge at 80% or less of the maximum allowable rotating speed as a rule. Because eachrotor of the centrifuge has a specified maximum allowable rotating speed, be sure to check the speed
before use. Never use the centrifuge at a speed exceeding the maximum allowable rotating speed
for any reason. The maximum allowable rotating speed indicates the allowable rotating speed of therotor, which has no contamination or scratches, used correctly with regular maintenance. Therefore,
normally use the centrifuge within 80% of the maximum allowable rotating speed of the rotor.
If the rotating speed is decreased by 20%, an almost equivalent centrifuging effect can be obtained byincreasing the running time by 50%.
8.2.2.2. Precautions concerning centrifuge tube
(1) Check the allowable centrifugal force and proof-solvent performance
Be sure to check to what extent the centrifuge tube to be used can withstand centrifugal force. If
solvent or a strong acid is centrifuged, also check whether the material of the centrifuge tube canwithstand them. Refer to Table 8-2 and the web sites below (it is advisable to bookmark them).
The allowable maximum centrifugal force is the value obtained when the rotor shape and centrifuge
tube shape are properly fitted. For example, a centrifuge tube with sharp edge is used with around-bottom rotor, the centrifuge tube may be broken even with centrifugal force within theallowable range.
http://www.hitachi-koki.co.jp/himac/support/m-tube.htm: endurances of centrifuge tubes made byseveral companies (in Japanese)
http://www.nalgenunc.co.jp/html/info.shtml: endurance, proof-solvent performance, cleaning
method, sterilization method (in Japanese)
http://www.assist-sar.co.jp/: endurances of several plastic products (in Japanese)(2) Do not use a deformed or cracked centrifuge tube
The centrifuge tube inevitably deteriorates as it is used. Do not use a deformed or cracked centrifuge
tube. Note that some centrifuge tubes may be deformed when a small amount of sample iscentrifuged even at less than the allowable centrifugal force, as is the case with Teflon centrifuge
tubes (80 - 90% of the centrifuge tube must be filled with solvent). Note that a disposable centrifuge
tube is not designed for repeated use, as its name suggests.
8.2.2.3. Precautions when balancing
For centrifugation, not weight balance but moment balance (weight × turning radius of center ofgravity) is required. This can be easily understood by comparing the case of a piece of clay attached tothe center of a string (a) with that of a piece of clay with the same weight on the end of a string (b)shown in Fig. 8-3. It is clear that the force given to the arm when swinging the string is higher in (b).
(a)(b)
Fig. 8-3. Gravity balance and momentbalance.

59

Table 8-2. Maximum allowable centrifugal force of disposable centrifuge tube
Manufacturer

VolumeMaterial

Maximum(mL)
Assist50

15, 13

Corning

50

centrifugal

force (g)Polypropylene

4,000

Polystyrene

4,000Polypropylene

4,100

Polystyrene

1,800

Improved polystyrene (Cat.No.430304)

1,800

Polypropylene

6,000(Cat.No. 430290, 430291, 430828, 430829, 430522)
15

Improved polystyrene

1,800(Cat.No.430053, 430055, 430788, 430789)
Polypropylene

6,000(Cat.No. 430052,430766, 430790, 430791, 430630)(1) There is no balance of moment between centrifuge tubes with different shapes (different centers ofgravity).
(2) There is a balance of weight but no balance of moment between a solution with a specific gravityof 1.2 and water with 1.2-fold volume (specific gravity: 1.0) (the distance from the center ofgravity to the rotating axis is different).
(3) For example, when a 50% suspension of fungus with a specific gravity of 1.2 (gravity as asuspension: 1.1) is balanced using saline with a specific gravity of 1.1, there is a balance of
moment before centrifugation. After centrifugation, however, the fungus with a specific gravity of
1.2 will be concentrated on the bottom of the tube (the center of gravity moves away from therotating axis), meaning that there is no balance of moment.
(4) If the centrifuge tube is balanced with the moisture on the outside of the tube or with water
precipitated in the rotor, there is no balance, either.
Correct balancing involves equally dividing a sample into centrifuge tubes of the same shape(material), the lid of which should also be of the same shape. The moment imbalance caused by the
specific gravity is practically allowable to some extent (for example, the case that Escherichia coli
culture solution is balanced with water). However, if a sample is centrifuged at the maximum allowablerotating speed, and ammonium sulfate, sucrose and glycerol solution at high concentration or a fungal(cell) suspension at high concentration are centrifuged, the same sample is equally divided forbalancing.
8.2.2.4. Precautions concerning sample leakage
In the case of an angle rotor, the liquid surface becomes vertical (parallel to the rotational axis)
during centrifugation (Fig. 8-4(A)). If the liquid surface is over the edge of the centrifuge tube, thesample will leak with poor sealing between the centrifuge tube and the lid. If the sample leaks,
imbalance will result, which will not only damage the machine and rotor but also cause biologicalcontamination (strain harmful to the human body and phage harmful to other experiments), chemical
contamination (for example, carcinogens, and poisonous and deleterious substances), corrosion (for60

example, ammonium sulfate, if left, heavily corroding aluminum alloy rotors), and at worst, fire andexplosion (in the case of organic solvent). Therefore, considering the case of the incomplete sealing of
the lid, do not put a sample into a centrifuge tube so that the liquid surface during centrifugation
is over the edge of the centrifuge tube. In particular, when a sample containing solvent with a low
flash point (for example, ethanol) is centrifuged, never completely fill the tube to a level where theliquid surface during centrifugation (rotation) is above the edge of the centrifuge tube. In
addition, never use a cracked or deformed centrifuge tube. If a dangerous reagent is spilled, besure to report it to the instructor and then, treat the spill. Even if the sample does not leak when theclosed centrifuge tube with the sample is pressed by the finger, there is no assurance that the lid has
been sealed completely. Assume that the liquid surface is 1 cm nearer the side of the rotational axis
from the edge of the centrifuge tube in Fig. 8-4(B). The pressure of a 10 m water column is 1
atmosphere under the condition of 1 g. In the case of centrifugation under the condition of 10000 ×g,
the pressure of a 1 cm water column corresponds to 10 atmospheres.

A

B
rmin
θ

90°-θ

rmax
Fig. 8-4. Liquid surface during rotation (A) and calculation of safe amount of liquid (B).

(1) Examine the angle θ of the rotor to be used.
(2) Tilt the centrifuge tube full of water by 90 - θ.
(3) The remaining amount of liquid is the maximum allowable.

8.2.2.5. Precautions during operation(1) Attach the rotor completely
Check the meshing direction of the tab on the bottom of the rotor with that of the rotating axis,
and then set the rotor completely (some centrifuges have no tabs). Take care not to bend or break thetab. After setting, check that the rotor is completely set by slightly turning it by hand. If you
remember how many turns the lid of the rotor requires for fastening it, you can find the incorrect
setting when you turn the lid less than usual.
(2) Check the attachment of packing
Set the packing between the rotor and lid. The screw of the rotor lid is cut in the direction inwhich the screw is fastened by acceleration. Conversely, it is loosened when the rotor isdecelerated (the rotor is decelerated, but the lid tries to rotate by inertia force). If packing is notcorrectly attached, the screw cannot be sufficiently fastened and the lid may become loose and fall
off during deceleration. If the lid flies off during rotation, it is generally very expensive (more thana few hundred thousand yen) to repair it. To our knowledge, there were three accidents presumed
to have resulted from such a default previously in our department.
(3) Monitor the centrifuge until the running state becomes steady
Do not leave the centrifuge unattended until the rotating speed increases up to the preset
value and becomes steady. Otherwise, appropriate action cannot be taken if there has been a
mistake in setting the centrifuge tube or in balancing, or if breakage of the centrifuge tube should61

occur. If something abnormal (abnormal sound) occurs, immediately push the stop button or set
the timer to zero. Take refuge and keep anyone away from the centrifuge until it completely stops.
(4) Never open the cover during running
Do not open the cover carelessly during running even if there is an abnormal sound. If thecentrifuge tube has been broken, the broken pieces will scatter at a high speed and can cause
serious injury, such as blindness.
(5) Never stop the rotor by hand
The hand may get caught in the rotating rotor, causing a serious accident, such as a fracture.
The rotational axis may be bent and the lifetime of the centrifuge may be decreased markedly.
“Because I am in a hurry” is not a valid excuse. Be patient and wait until the centrifuge stops
completely.
(6) Always keep the cooler switched on
The temperature of the centrifuge will be increased by the heat from the motor and from thefriction between the rotor and air. Therefore, keep the cooler switched on even during
centrifugation at room temperature.
(7) Cover of the chamber
While the cooler is switched on, keep the lid of the centrifuge closed to prevent condensationand freezing in the chamber. Inversely, when switching off the cooler, open the lid to dry the insideof the chamber. If a large amount of ice is generated, ice crystals may fall off with draft pressureduring centrifugation and scatter, damaging the rotor and chamber.
(8) Rotor after use
Remove the rotor and check that there is no contamination by possible sample leakage. Lay theremoved rotor downward to prevent losing balance the next time centrifugation is carried out,
owing to falling dust and condensed water collecting in the rotor. If the sample leaks, rinse therotor immediately. The rotor contamination causes not only imbalance but also corrosion of therotor and breakage. If a dangerous sample (recombined organism, carcinogen, poisonous anddeleterious substances, corrosive substance, and flammable substance) leaks, be sure toreport it to the instructor and then, treat the leakage properly.
8.2.2.6. Precautions concerning ultracentrifuge use(1) Use the ultracentrifuge in the presence of an instructor
The ultracentrifuge rotates at an ultrahigh speed of around 100,000 rpm at maximum. Takespecial care as the rotor may fly out and cause a serious accident if balancing is forgotten or the
sample leaks. If the rotor crashes through the chamber, the rotor will fly around the laboratory at ahigh speed and destroy the laboratory. If we consider the normal centrifuge as analogous to anormal car, the ultracentrifuge would be a racing car (F1 car). Even a small mistake may directly
lead to a serious accident. Use the ultracentrifuge in the presence of an instructor until you
sufficiently acquaint yourself with its use. Do not use it alone until you are recognized to be
sufficiently familiar with its use by the instructor.
(2) Precautions concerning plasmid purification
When purifying a plasmid DNA by cesium chloride – ethidium bromide equilibrium
density-gradient centrifugation, take special care of the temperature setting. If a density gradientis formed, the concentration of cesium chloride at the bottom of the centrifuge tube will increase.
Therefore, if the temperature is wrongly set at ‘4°C,’ the concentration of cesium chloride will
exceed the saturated concentration and its crystal will precipitate. Because the density of the crystal
is much higher than that of the solution, the moment balance will be lost and the rotor will flyout. Strictly observe the setting of temperature and time designated in the protocol.
62

Ethidium bromide is a carcinogen and usually used at high concentration for equilibrium
density-gradient centrifugation (consider that it is more dangerous than RI). If it is spilled, be sure
to report it to the instructor and conduct proper decontamination. In particular, the Beckman
ultracentrifuge in the common equipment room in the basement can become easily contaminated at
the time of heat sealing the centrifuge tube. Check whether there is contamination using a built-in
ultraviolet (UV) lamp before and after sealing. Under short-wavelength ultraviolet irradiation in adark room, an orange glow indicates contamination. Of course, short-wavelength ultraviolet mustnot be irradiated directly onto the skin or looked at with the naked eye.
(3) Heat sealing
When sealing the centrifuge tube by melting its rim through heating, any solution adhering to
the rim will cause incomplete sealing (increase in inner pressure by the centrifugal force → liquidleakage → imbalance → very serious accident). Carefully read the instruction manual and pay
attention to the following when sealing.
1) Pour a sample up to the line shown in Figure 8-5. If a larger amount is poured, the tube willtend to be sealed incompletely, and if smaller, the centrifuge tube will be deformed by thepressure due to the centrifugal force, also causing imbalance.
2) Completely wipe the sample solution adhering to the rim of the tube with Kimwipe. Because
the temperature at the part of the tube with the attached solution will not increase even whenheated by a heater (the tube will not melt), the tube will be incompletely sealed. When thesolution adhering to the rim dries up, the solute dissolved in the solution will remain in adried-up state, also causing incomplete sealing.

Completely wipe any
adhering solution
immediately after
pouring the sample.

Fig. 8-5.

Approximate
2:1

Appropriate sample
volume8.2.2.7. Precautions concerning swing-type centrifuge (clinical centrifuge)
When running a swing-type centrifuge, in which the test tubes can be centrifuged, mount the sameassemblies in all four arms. Even if centrifuging one sample (two samples for balancing), mount four
identical assemblies. In general, when using an assembly in which eight test tubes can be placed, themaximum rotating speed is 2,000 rpm (be sure to check the value yourself because it depends on thetype of centrifuge).
8.2.2.8. Know-how for centrifuging(1) Points for deriving the centrifugation effect from the theoretical formula
Assuming that the Stokes radius and specific gravity of the particle to be centrifuged are R and ρ,
respectively, the rotation speed is ω, the specific gravity and viscosity of the solvent are ρ0 and η,
the distance between the rotational axis and the liquid surface is rmin, and the distance between therotational axis and the bottom of the centrifuge tube is rmax, the time for the deposition of theobjective particle is given as
63.
Therefore, the following apply.
1) The centrifugation effect is proportional to the product of centrifugal force (rotating speed
squared) and time. Therefore, if the rotating speed increases by 1.2-fold, 1.44-fold the effect isobtained (here, strictly observe the maximum allowable rotating speed). If the rotating speed
cannot be increased, rotation time can be increased.
2) When particles in a solution of salt or sugar at high concentration are centrifuged, it takes a longtime to centrifuge because ρ−ρ0 is low. It is desirable to centrifuge such solutions after diluting
with water or buffer solution if possible. For example, when a sample with a specific gravity of
1.21 in a solution with a specific gravity of 1.20 is centrifuged, the difference in specific gravity
between the two is 0.01. However, if a solution is diluted to a specific gravity of 1.10 using anequivalent amount of water (specific gravity: 1.00), the difference in specific gravity will be
0.11 and the sample will be centrifuged for a time ten-fold shorter than the original. Moreover,dilution has an effect of decreasing the viscosity of a solution.
3) The viscosity of a solution decreases as its temperature increases. When increasing preset
temperature or diluting a solution, the viscosity of the solution will decrease, which enables
shortening of the centrifuging time.
4) Using an angle-type rotor with a low rmax/rmin, a sample is centrifuged more rapidly than whenusing a swing-type rotor with a high rmax/rmin.
(2) If there are problems with suspension of precipitate
In most cases, the objective is not centrifugation at a given rotating speed but precipitatecollection; therefore, it is desirable to decrease rotating speed. Moreover, when resuspending aprecipitate, loosening (softening) the precipitate to which the solution has not been added yet using
a vortex mixer will facilitate suspension.
(3) Precooling of rotor
In principle, mount the rotor, set the temperature and precool it for 0.5 – 1 hour before use. If
the rotor is required to cool rapidly, it is desirable to properly mount the rotor and lid and then toidle the rotor at 1,000 – 2,000 rpm for 5 – 10 min.
About normal centrifuge:(1) Balance the centrifuge with respect to moment not weight.
(2) Strictly observe the maximum rotating speed and usually use the centrifuge at 80% or less.
(3) Check that the centrifuge tube withstands the given centrifugal force and solvent.
(4) Sample leakage = risk (imbalance, biological contamination, chemical contamination, explosion andfire).
(5) Be sure to use a cooling device if equipped.
(6) Mount the same assemblies to all the arms of a swing-type rotor.
(7) Supervise the rotor until its rotation becomes stable.
(8) Do not open the cover of the chamber during rotation.
(9) Never stop the rotor by hand.
(10) Report sample leaks to the instructor.

64About ultracentrifuge:(1) Use an ultracentrifuge in the presence of an instructor until sufficiently acquainted with its use.
(2) Strictly observe temperature during equilibrium density-gradient centrifugation.
(3) Take special precautions not to contaminate the sample with ethidium bromide.

8.2.3. Thermostat Bath (Incubator)
The thermostat bath is generally used without special care but has a risk of causing large-scale
accidents such as fires. In our department, there have been small fires caused by boiling the water bath
dry and by heating of the oil bath, and foul odor emissions of melting plastic on the verge of fire in thedryer. An incorrect operation of the thermostat may cause inaccurate control of temperature, resulting
in the failure of the experiment even though no accident is caused.
8.2.3.1.Water bath incubator(1) Caution concerning overnight running and unattended running
Never run a water bath overnight without the boil-dry protection device. Even in thedaytime, unattended running is prohibited. At each use, check that the boil-dry protection device isfunctioning correctly before carrying out the experiment. In the case of the boil-dry protection
device, which stops the current flow when the float descends to a certain level due to a decrease inwater level, if water residue accumulates on the float, it will not move and will fail to function
properly when the water level descends. Do not neglect the maintenance of the bath to prevent
residue accumulation around the float.
(2) Water supplement
Water in the incubator (thermostat bath) evaporates. Be sure to maintain a sufficient amount ofwater in the bath. Because there is a large difference between room temperature and watertemperature and a low air humidity in winter, pay particular attention to increases in evaporation.
When running the bath all night, devise an automatic water supply system or prevent evaporationby covering the bath with aluminum foil. In particular, when using the bath at a high temperature,
examine the water level decrease per unit time and check whether a safe water level can be
maintained until your arrival at the laboratory the next day. It is dangerous to place plastic balls
or foam polystyrene on the water surface. They may cause fire if they come into contact with theheater.
(3) Check of water level
The last person leaving the laboratory must check the water level of a water bath left to run
overnight and add water if necessary even if it is not their own experiment.
(4) Do not overly rely on the safety device
In the user instruction of the water bath equipped with the boil-dry protection device, there is anote “check the operation of the safety device at each use.” The operation is not assured unlesschecked properly. If the safety device is activated, the power will be shut off and thetemperature will not be maintained, resulting in the failure of the experiment. The presence ofa safety device is not an excuse to neglect the addition of water even for a water bath equippedwith the safety device.
(5) Caution concerning shaking
When using the water bath with a shaking apparatus, adjust the water level and shaking speed sothat water is not scattered during shaking. In particular, when a sample requiring sterilization isshaken, devise a way to keep dirty water (more than 107 bacteria/mL is not unusual) away from the65cotton stopper and silicon stopper.
8.2.3.2.

Incubator, dryer and dry heat sterilizer(1) The incubator, dryer and dry heat sterilizer have no explosion-proof
construction.
Never put organic solvent or flammable gas into them.
(2) Do not put any flammable materials into the dry heat sterilizer
Do not put combustible material, such as plastic and paper, into the dry heat sterilizer. The only
exception is the cotton plug.
(3) Fix the temperature setting dial with tape
If the thermostat bath has a temperature setting dial, fix the dial with tape so that the preset
temperature cannot be shifted by contact with other materials. In our department, there has been acase in which a plastic instrument melted due to an inadvertent shift of the temperature dial
(fortunately, fire was prevented because of detection of a foul odor).
(4) Check that any instrument used in the dryer or dry heat sterilizer can withstandthe preset temperature
In general, plastic instruments (including the Pipetman) cannot withstand the temperature in
the dry heat sterilizer (160°C). If a chip, Eppendorf tube or plastic beaker is dry-heat-sterilized, it
will melt and flow into the high-temperature heater, causing a fire.
(5) Do not block air circulation
Do not place the sample or instruments on the outlet of the fan. Do not place large objects in themachines and do not fill the machines with too many objects. In general, the temperature sensor isinstalled in the upper part of the machines and the heater in the lower part. If air circulation(convection) is prevented, the temperature cannot be controlled. The heater continues working
until the temperature in the upper part reaches the preset value, which causes the temperature in thelower part to become too high. There has been an incident of fire due to a plastic instrument
melting in the high-temperature heater and flowing into the heater part.
(6) Do not remove the bottom plate
The board at the bottom of the machines separates the bottom plate (the temperature of which
will become high) and the sample to ensure air circulation. Do not remove the bottom plate evenwhen attempting to put in a tall sample, otherwise fire may result.
(7) When the sample is spilled
If the sample is spilled into the incubator, wipe it off immediately. Otherwise, it may corrode the
incubator and cause contamination.
Basic precautions(1) Display the user name and scheduled termination time for overnight running.
(2) Unattended (overnight) running of the machines without the safety device is prohibited.
(3) Check that the temperature has attained a steady state.
(4) Never use a socket multiplier.
(5) Pay attention to the electric capacity when using an extension cord.
(6) The control unit must be kept dry.
Water bath

(1) Is the amount of water sufficient?
(No water + nonfunctional safety device = fire)66

(2) At each use, check that the boil-dry protection device functions correctly.
(3) The last person leaving the laboratory must check the water level regardless of whose experiment
it is.
Incubator, dryer and dry heat sterilizer(1) Do not use organic solvent or flammable gas.
(2) Do not block the air circulation in the machines.
(3) Do not place combustible material in the dry heat sterilizer.

8.2.4.

Reducing Pressure

8.2.4.1. Cautions concerning apparatuses(1) Do not draw up benzene or halogen-containing solvent directly with thewater-jet pump.
Do not draw up benzene or halogen-containing solvent (chloroform, dichloromethane andchloroethylene) with the water-jet pump. Otherwise, halogen-containing solvent will bedischarged into the sewage and cause environmental contamination. Neither should they be drawn
up with the circulation water-jet pump (when replacing water in the circulation water-jet pump,water with the halogen-containing solvent will be disposed of into the sewage).
(2) Do not draw up acidic gas directly with the vacuum pump.
For example, hydrochloric acid, acetic acid and trifluoroacetic acid will corrode the inside of thepump and considerably reduce its lifetime. Use a special Teflon-coated pump or a pump with a trap
containing sodium hydroxide or other chemicals.
(3) Do not draw up organic solvent or solution directly with the hydraulic vacuum
pump.
Organic solvent and solution are less detrimental than acidic gas, but also reduce the lifetime ofthe pump. In addition, a sufficient vacuum level cannot be attained because of the vapor pressureof the drawn solvent or water. Draw them up through a cold trap cooled with liquid nitrogen or dry
ice - ethanol (methanol). If drawing up a solution, the silica gel column can be used alternatively.
In this case, replace the silica gel when its color changes (the gel becomes tinged with red).
(4) Break the vacuum condition before stopping the pump
Before stopping the pump, break the vacuum condition by using the three-way cock or detaching
the hose. Some vacuum pumps have no backflow valve. If such a pump is stopped before breaking
the vacuum, oil will flow back and ruin the sample. (Cleanup is very difficult.) The water-jet pumpnormally has no backflow prevention valve. If it is operated incorrectly, water will flow back andruin the valuable sample. In particular, for drying at a low pressure with a desiccator, backflowing
water will react strongly with the drying agent (phosphorus pentoxide, calcium chloride,
concentrated sulfuric acid, etc; silica gel is no exception), causing an accident. In fact, the use of
the water-jet pump for drying at a low pressure is incorrect (the sample cannot be dried completely
because of the vapor from the water flow).
8.2.4.2.

Notes on operation for reducing pressure(1) A container exposed to a low pressure must be resistant to a low pressure
Use only thick containers designed for use under a low pressure, such as a pressure-reduction
bottle, an aspiration bottle and a desiccator. Before each use, check that the bottle is not cracked or67

chipped. Be aware that a conical flask may break when the pressure is reduced. The broken piecesof the flask and contents scatter violently, which is very dangerous.
(2) Be ready for quick response to ‘sudden boiling’
When reducing the pressure of the solvent, attach a three-way stop cock to the pump to enable
immediate cessation of pressure reduction in the event of sudden boiling. Sudden boiling tends
to occur particularly just after starting pressure reduction, when starting to heat the sample, and justbefore the sample solvent is completely exhausted. Therefore always monitor the pump and be
prepared to take immediate action, such as keeping your hand on the stop cock in the event of
sudden boiling. Never leave the pump unattended just after starting pressure reduction.
8.2.4.3. Rotary evaporator(1) Start the exhaust slowly and carefully
Start the exhaust, keeping your hand on the stop cock to immediately break the vacuum
condition in the event of bubbling or sudden boiling. Never leave the pump unattended just after
starting pressure reduction.
(2) Do not regularly use the vacuum pump for pressure reduction
Generally use the water-jet pump, not the vacuum pump. If using the vacuum pump, draw up
solutions through the trap cooled with liquid nitrogen or dry ice – ethanol (methanol), as noted
above.
(3) Do not heat until the pressure has become sufficiently low and stable
Start heating after a sufficiently steady low pressure is obtained and no bubbling or sudden
boiling occurs.
(4) Stopping the flow of cooling water may cause a fire
Check that sufficient cooling water flows 5 minutes after starting pressure reduction and at thetime of leaving the evaporator unattended. The solvent may be not collected but exhausted, which
may cause an explosion and fire.
(1) Use a container designed for use under a low pressure.
(2) Stopping the flow of cooling water in the rotary evaporator causes explosion and fire.
(3) Drying agent + backflow water = danger (stop the pump after all operations)(4) The use of benzene or halogen-containing solvent with the water-jet pump is prohibited.
(5) Attach the proper trap to the vacuum pump.

8.2.5. Ultraviolet Lamp
Ultraviolet radiation, in particular, short-wavelength ultraviolet radiation, is harmful to the humanbody. When staining and observing DNA, short-wavelength ultraviolet radiation will damage the DNA,for example, by causing thymine dimer formation and DNA breakage.
(1) When turning on the lamp, even for a short time, always wear protective glasses.
(2) For operations taking more than a few seconds, such as collecting DNA fragments from agarosegel, wear long-sleeved clothes (lab coat), gloves and a full-face mask.
(3) The ultraviolet lamp attached to the clean bench emits harmful short-wavelength (normally 245 –254 nm) ultraviolet radiation. Although ultraviolet radiation can be blocked by glass or acrylic
plates, direct exposure of the skin or direct viewing with the naked eye must be avoided.
Furthermore, it differs from the long-wavelength lamp (generally 312 – 365 nm) used in the68transilluminator. Care must be taken not to mistake them when exchanging the lamp.
(4) When using the transilluminator, the wavelength of which can be varied, set the switch to the longwavelength for DNA excision (pay attention to H and L on the selector switch, which refer not tothe output power but to the wavelength).
8.2.6. Clean bench/Safety cabinet
Clean bench is a device which makes a space separated from the surrounding air. If you perform
experiments with the bench, you can avoid contamination of several organisms or substances fromoutside. Before experiments, the clean bench kills saprophytic bacteria with equipped ultraviolet lamps
in the device. During experiments, the device keeps positive pressure inside by incorporating clean air
filtered with a HEPA filter, leading to avoidance of bacteria’s invasion into the device. You should
handle reagents or perform experiments for cell culture in the clean bench or safety cabinet in order toavoid contamination. (The safety cabinet keeps a negative pressure inside to avoid leakage of biologicalmaterials such as virus etc. to the outside).
（１）Management of Bunsen burner
When you have done experiments or leave the clean bench even for a short time, you must extinguishput out the fire of a bunsen burner. You should also keep the bunsen burner off the wall surface of theclean bench. Keep it away from the place where the wall surface becomes hot.
（２）Notice for sterilization with ethanol
When you fire a spreader for sterilization, you should realize the risk of fire and solvent at the sametime. Ethanol should be kept in the metal box. The lid should be kept handy during operation. If bychance, ethanol ignites, you should put out the fire by covering the lid on the box.
（３）Even if you wash your hands, your hands are still the most dirty thing in theclean bench. Consider the sequential order of the operation and arrange thecomponents so as not to pass your hands over the non-contaminated components.
・ Non-contaminated items should be put in the inmost part of the operation space andpossibly-contaminated items should be put nearest your side.
・ As a general rule, you should put items, which are treated on the right hand, in the right side, andones which are untreated by the left hand, on the left side.
・An operation, such as opening the lid with your both hands, should be finished before starting
experiments.
（４）Keep the inside of clean bench clean
In the case of spilling medium, you should immediately wipe it up. The spilled medium
might lead to growth of bacteria and contamination of the experimental system.
（５）Wear clean lab coat during experiments and roll the sleeves up to your elbow.
Before experiments, you should sterilize your hands with 0.05-0.1% benzalkonium chloride solution
or 70% ethanol. Of course, you should sterilize apparatus used in the bench with the solution or
ethanol.

69

（６）Do not leave unnecessary items in the bench.
Do not leave apparatus such as an unused laboratory dish, a medium, a pipette or the mixer in theclean bench. The following problems occur:
・Work space becomes small.
・Parts under shadow and not exposed to the UV lights increase and those parts might not besterilized.
・Plastic parts deteriorate in particular, and the apparatus will be damaged by UV lights.

8.3. Handling Reagent
When using reagent, always wear protective glasses. If you get dangerous reagent (solution) in
your eyes, immediately (within a few seconds) wash the eyes with generous amounts of water for morethan 15 minutes, and consult a physician. In particular, if you get alkali or organic solvent in your eyes,
wash the eyes vigorously without delay. Every second you delay the higher the risk of blindness. It is
more important to sufficiently wash the eyes than to quickly consult a physician.
Before using reagent in your experiment, understand the handling instructions for all reagents.
Refer to a textbook, such as “Carrying Out Experiments Safely” (KAGAKU-DOJIN) to acquire generalbasic knowledge about “what substances are dangerous in what ways,” and “what should be done in theevent of an accident.” In addition, consult the Merck Index and make sure to acquire sufficient
background knowledge for each reagent to be used with respect to, for example, the following points:(1) How harmful is the reagent to the human body?
(2) Are there any risks of fire or explosion?
(3) What procedure should be followed when the reagent is spilled?
(4) Proper method of treating waste liquid(5) Proper storage method (refrigerating, freezing, protecting from light, nitrogen substitution, etc.)
8.3.1. Examples of reagents to handle with care
Some reagents, which are normally used without special care, may be unexpectedly dangerous.
Some familiar examples of reagents that are rarely used but should be handled with special care are
given below.
(1) Special flammable substances
Ether is an example. According to the Fire Defense Law, its danger level is ranked the highest,
and therefore, all open flames in the room must be extinguished before it can be handled. Here,“handling” includes “holding” the solvent bottle. Ether is flammable because of its extremely low
ignition temperature. Once ether catches fire, the fire expands explosively, is difficult to extinguish,
and therefore, utmost caution is required. The centrifugation of these solvents is strictly
prohibited (a special explosion-proof centrifuge, which is unavailable in our department, isrequired).
(2) Highly flammable substances
Ethanol, methanol, acetonitrile, hexane and acetone are examples. Similar to special flammable
substances, extinguish all open flames, such as that in a stove, before “holding” the bottle, in casethe bottle is dropped and broken. It is dangerous to dispose of these solvents in a sink even though
they are soluble. Because the amount of solvent vapor increases with liquid surface area, thesesolvents may catch fire from the water heater or a spark from the motor in the centrifuge and70explode.
(3) Poisonous and deleterious substances
Potassium cyanide, sodium cyanide, sodium azide, mercury compounds and arsenic compounds
are examples. Phenol, chloroform, acrylamide, sulfuric acid, hydrochloric acid, sodium hydroxide,
potassium hydroxide and acetonitrile are also examples. Store them in their designated lockablestorage cabinet and manage their mass by preparing a log. Upon recognizing a lost or incomplete
form in the log, immediately report it to the instructor.
When the ordered poisonous and deleterious substances are delivered, be sure to record them inthe log. When borrowing or lending such substances between laboratories, be sure to obtainapproval from the instructor.
(4) Other familiar dangerous substances• Hydrogen peroxide solution
If metal is mixed into this solution, it may react explosively (recall the science experiment togenerate oxygen from hydrogen peroxide solution using potassium permanganate as a catalyst,
performed in elementary and junior high schools). There has been a case of explosion due to thecontamination of a small amount of rust. Never use a metallic container or injector.
• Ammonium persulfate
This is used as a polymerization initiator of polyacrylamide gel. Because it occasionally may
react explosively with metal, do not use a metallic medical spoon when weighing theammonium persulfate. Use a plastic or bamboo spoon. When using the ammonium persulfate
for the preparation of polyacrylamide gel, the following procedure is recommended (10%ammonium persulfate solution for electrophoresis, although slightly more expensive, iscommercially available):
1) Purchase ammonium persulfate in units of 1 g.
2) Add 3–5 mL of pure water directly into the reagent bottle and dissolve the ammonium
persulfate.
3) Transfer the solution into a disposable tube with a volume of 15 mL and dilute to 10 mL.
4) Dispense an amount suitable for one-time use in the range of 100 – 150 µL into several
Eppendorf tubes.
5) Store it in the freezer below –20°C.
In most protocols, ammonium persulfate should be prepared at the time of use. However, there
is no problem with the use of ammonium persulfate dispensed from a stock kept in the freezer.
The error in the preparation by this method does not cause any problem when using ammonium
persulfate as a polymerization initiator of acrylamide gel.
• Acrylamide
This is neurotoxic. If it comes in contact with the skin, no problem arises if the skin is washedimmediately, but it may cause paralysis if the skin is left untreated.
• PhenolThis is a corrosive substance. If it comes in contact with the skin, it causes burns.
Precautions during the extraction of nucleic acid are given in 8.3.6.
• Chloroform
This is deleterious and corrosive and is designated as a specified toxic substance. If it comesin contact with the skin, although less harmful than phenol, it may still cause burns. Chloroform
is considered to cause liver cancer if inhaled over a long period. Be sure to use it within thedraft chamber. Precautions during the extraction of nucleic acid are given in 8.3.6.
• Ethidium bromide, nitrosoguanidine, ethyl (methyl) methane sulfonate
These are highly carcinogenic agents. Consider them more dangerous than isotopes. Usethem under the instructor’s guidance until you acquire full handling expertise. Ensure full71

•knowledge on the treatment for spills and the disposal method.
Phosphorus pentoxide, quicklime (calcium oxide), concentrated sulfuric acid,silica gel
These are also used as drying agents and reagents of these four substances react violently withwater. Even silica gel is dangerous because it bursts upon contact with water. Exercise sufficient
caution when disposing of the contents of a desiccator.

8.3.2. Cautions in purchasing(1) Acquire proper knowledge about the reagent to be purchased
Be sure to acquire knowledge about the reagent to be used, using information sources, such as the
Merck Index. It is necessary not only to obtain the proper knowledge concerning toxicity andexplosiveness but also not to fail in the experiment. For example, useful information, such as the
preparation method of the solution of a difficult-to-dissolve reagent (shifting its pH slightly or
dissolving it initially in another solvent) and the stability of the reagent, can be obtained.
(2) Purchase the reagent only in the smallest necessary amount
It is not uncommon for a reagent disposal fee to be tens or hundreds of times more expensive thanthe purchasing cost. Recognize that the remains of excessive purchases are not only wasteful but
require additional disposal fees.
Many reagents deteriorate in storage. Purchase reagents requiring special storage methods(hygroscopic reagent, reagent in ampoule and reagents to be protected from light, stored under nitrogen,refrigerated or frozen) in only the smallest necessary amounts. For example, 1-g bottles and 10-gbottles are commercially available; if 2 g of the reagent is required, it is often less expensive topurchase two 1-g bottles than one 10-g bottle.
8.3.3. Precautions for use(1) Label check
Check the label three times so as not to use an incorrect reagent. First check: when retrieving thereagent bottle from the reagent cabinet (always check the label). Second check: when measuring
the reagent (if placing numerous reagent bottles together near the balance, always check the label
before measuring the reagent). Third check: when returning the bottle to the reagent cabinet(always check the label when replacing the bottle back on the shelf). The following are examples
of commonly confused reagents.
1) Polyvalent anions, such as phosphoric acid, citric acid, EDTA and ATP
For example, tetravalent EDTA has five types of salt, namely, free acid and mono-, di-, tri- andtetra sodium salt. Phosphoric acid has several salts, such as sodium salt, potassium salt andammonium salt, and each salt has three types.
2) Number of bonded water molecules
For example, Na2HPO4 has three types of salt, namely, anhydride, heptahydrate anddodecahydrate. For mol concentration, the amount to be measured can be calculated from eachmolecular weight. However, note that the weight concentration is different depending on thetype of salt used (the number of bonded water molecules in the reagent used must be accurately
recorded in the experiment note).
(2) If reagent is spilled
If the reagent is spilled during measurement, be sure to clean it up. Otherwise, the spilled
reagent will come into contact with the back of the powder paper and contaminate the next reagent
being measured. Only the person who spilled the reagent knows its identity. If left untreated, it72cannot be properly cleaned up.
(3) Medical spoon
Before using the medical spoon to measure reagent, ensure that the spoon is washed well, rinsed
with pure water and dried completely. If a wet medical spoon is used, the reagent may deteriorate
due to moisture, and will not be measured accurately. It is not uncommon for experiments to failbecause of a very slight amount of impurity contamination. Note that a metallic medical spoon
should not be used for some reagents, such as ammonium persulfate.
(4) Be sure to return the reagent
Be sure to immediately return the reagent used with the label facing forward. If the label ispeeling off or worn away, take proper steps, such as reattachment or rewriting of the name. It ishighly expensive to dispose of reagents containing unknown substances.
(5) Opening container of refrigerated or frozen reagent
When using a reagent stored in a refrigerator or a freezer, be sure to bring it to room
temperature before opening the lid. If the lid of a cold bottle is opened, vapor in the air willcondense, making the reagent wet. When wet, the reagent may deteriorate and will no longer bemeasured accurately. The deterioration of a cold reagent, caused by exposure to room temperature
for a short time, is negligible compared with that caused by storing under a wet condition at a lowtemperature for a long time. However, (6) and (7) are exceptions.
(6) Ammonium water, hydrogen peroxide, corrosive carbohydrate solution
The pressure in a bottle of ammonium water and hydrogen peroxide may be high. If thetemperature in the bottle is high, cool it with ice water before opening the lid. When a medium
containing sugar corrodes in a sealed bottle and the pressure within a bottle filled with carbon
dioxide is high, be aware that the contents may scatter explosively if the lid is carelessly loosened.
Always wear protective glasses without fail.
(7) Opening an ampoule
Open an ampoule after cooling it with ice water. If the total amount of reagent is not used in theexperiment, prepare a container of suitable volume and material and which can be sealed, before
opening the ampoule. Because there is a reason for sealing the reagent in an ampoule (for example,
extreme sensitivity to oxygen or moisture, or a foul odor), it must again be sealed for storage after
opening. Note that Parafilm cannot be used to seal an ampoule, because of its breathability.
8.3.4. Handling of carcinogens
Ethidium bromide, nitrosoguanidine, ethyl (methyl) methane sulfonate are highly carcinogenic andshould even a slight amount of these substances be spilled and left untreated, all those present, not justthe experimenter, will be exposed to long-term risk of disease. Consider these substances moredangerous than isotopes because spilled substances are difficult to detect (ethidium bromide can bedetected by ultraviolet irradiation). When using these substances, as with the case of isotopes (or morethan with isotopes), exercise particular caution, as described below. Be sure to use them under the
instructor’s guidance because procedures differ depending on the reagent and the experimental aim.
(1) Always wear protective glasses and gloves. Never touch these substances with bare hands.
Consider used gloves to be contaminated, and do not carelessly touch other materials when
wearing gloves.
(2) Request the presence of the instructor when measuring these substances. Take sufficient care
not to spill any amount of the substance.
(3) Store the reagent bottle in a transparent plastic bag with the top closed.
(4) Dispose of the old bag used to store the reagent by the method described in (8).
(5) Place polyethylene-coated filter paper of sufficient size on the laboratory table to be used.
73

(6) If the substance is spilled outside the area of the filter paper, report to the instructor and
immediately neutralize it properly and adequately.
(7) Be sure to perform neutralization for all the instruments used (containers such as test tubes, chips,
medical spoons and powder paper).
(8) Dispose of used gloves, filter paper on the laboratory table and reagent bag, keeping them awayfrom people (not only those in the laboratory but also those involved in waste disposal). For
example, hold the bag to be disposed of while wearing gloves, then take the gloves off inside out
such that they envelop the bag and then bind them at the top. Moreover, put the entirety intoanother bag, bind the top of the bag and dispose. Be sure not to include any sharp objects, such as atip, which may tear the bag. Dispose of the tips after separate neutralization.
8.3.5. Treatment of carcinogens
The following are general precautions and not to be considered complete. Study and devise proper
suitable treatments by yourself.
(1) Ethidium bromide
This is often treated with sodium hypochlorite solution, however, its carcinogenicity cannot becompletely neutralized, and it is also said that its toxicity may in fact be increased. The oxidation
treatment with potassium permanganate described in “Molecular Cloning (2nd Ed., E8)” is also
incomplete. Therefore, hire a professional disposer to dispose of dilute solution, such as gel stain
solution, and concentrated solution used for equilibrium density-gradient centrifugation, treatingthem both as halogen-containing waste liquid. The dilute waste should be discarded as “H-b waste”
described in Chapter 11.4.
A spot contaminated with ethidium bromide shows orange luminescence when ultraviolet lightis irradiated in the dark. Although the detection sensitivity is higher when using short-wavelength
ultraviolet light, such light is itself carcinogenic. If ethidium bromide comes into contact with theskin (or there is a possibility of it having come into contact with the skin), check for orange
luminescence using the long-wavelength ultraviolet lamp and wash the skin with a brush and soap
until the orange luminescence is no longer seen.
(2) Nitrosoguanidine
Nitrosoguanidine is decomposed upon heating in an autoclave as an acid or alkali solution.
Take special caution never to spill this solution in a space where heat treatment is prohibited.
(3) Ethylmethane sulfonate (EMS)
Neutralize this solution with sodium hypochlorite or sodium thiosulfate solution. According to“Microorganism Genetics Experiment Method,” 9.8 mL of 6% sodium hypochlorite solution isused to neutralize 0.2 mL of 3% EMS. If EMS is spilled, wipe it thoroughly with sodium
hypochlorite solution wearing gloves.
8.3.6. Extraction of nucleic acid with phenol/chloroform(1) Wear protective glasses and gloves when treating with phenol/chloroform
When extracting nucleic acid with phenol/chloroform, use a tube made by a manufacturer with areliable sealing level. Depending on the manufacturer, some tubes that are insufficiently sealed andcause sample leakage may be included in the provided supply, which will require special attention
when mixing tubes using a vortex mixer. Be sure to wear protective glasses and gloves.
(2) When phenol or chloroform comes into contact with the eye or the skin74

If it enters the eye, immediately (within a few seconds) flush the eye with water for more than
15 minutes, and then, see a doctor. If it comes in contact with the skin, never wipe it withethanol. Skin oil will be removed by the solvent, which will cause an even worse burn.
Immediately wash it away with water, and then with soap.
(3) Collect the waste liquid
Be sure to collect waste liquid and hire the proper waste disposal firm for its disposal. Do notdispose of it in the tube. Tubes are disposed of as nonflammable substances, which are usuallyused as landfill. Therefore, tubes with waste liquid will contaminate the environment and pose a
danger to the worker.

8.4. Disposal of wastes
Materials including bacteria or touched by similar things are at risk of pathogenic infection.
Sterilize all of them before disposing of them appropriately. Artificial recombination of genes should betreated in the same way. Autoclave treatment (121°C, 20 minutes) is effective.
8.4.1. Main wastes and waste disposal method (Refer to 11.6.2.)-Containers for culture, aluminum foil, glass fragments.
Treat materials used for culture by autoclaving and dispose of as noncombustible. When liquid or
solid media are left in the container, separate and wash the container before disposing of it.
Example for waste: Containers used for cell culture and pipettes (plastic) are put in a plastic bag
for autoclaving and given an autoclave treatment before disposing of them as noncombustible
materials.
-Liquid wastes such as medium
Sterilize liquid wastes if possible in an autoclave container. If necessary, dilute them or neutralize
acids and alkali. They should be treated with fungicide including chlorine. Treated by autoclave, it isnecessary to neutralize them with sodium thiosulfate beforehand. Solutions which include chlorine giveoff poisonous chlorine acid gas. Handle them with care.
-Materials designated as an industrial waste, such as injection needles and syringes, are subject tospecial control. Dispose of them as quasi hospital wastes following the regulations of Kyushu
University.

8.5. Guidelines for Research Involving Recombinant DNA,Microorganisms, Animals, Radioisotopes and Radiation generator
8.5.1. Recombinant DNA(1)

AccordingtothesafetycontrolregulationsofKyushuUniversity
http://www.kyushu-u.ac.jp/university/office/kikaku-bu/kenkyusenryakuka/dna/index.htm), it isnecessary for all researchers planning Recombinant DNA experiments to submit their safety plan
document to the Recombinant DNA experiments committee in Kyushu University and to undergo
screening by the committee and receive the approval of the President of Kyushu University.
75Based on the combination of host and donor of nucleic acids, there are two categories. They are
institutional approved experiment and ministerial ascertained experiment, and require affirmationby the Minister of Education, Culture, Sports, Science and Technology.
(2) According to the safety control regulations of Kyushu University, researchers who perform
recombinant DNA experiments are required to be provided education and training and go through a
medical checkup before the experiment. They should use appropriate equipment depending on theexperimental level and considerations of safety in order to prevent accidents.
(3) By departmental regulations, it is required that researchers prevent proliferation of recombinantorganisms by transfer to research workers and other persons or proliferate to outside within
facilities (Physical containment). Researchers must carry out their experiments in special
facilities classified as follows; P1, P2 and P3 for microbiological experiments; LS-C, LS-1 andLS-2 for large-scale culture; P1A, P2A, P3A and specific rearing compartments for animal
experiments; P1P, P2P, P3P and specific isolation chambers for plant experiments. For alllaboratories and facilities except P1, advance approval must be obtained for the chief researcher
and facilities must bear a P2 (or other) level testing sign.
(4) All researchers must have regular medical checkups complying with a regulation prescribed by the
University.
(5) Principal investigator must submit a report to the President at the end of an experiment.
(6) For transfer of recombinant organisms, researcher must report to the President. In the event of any
assignment of recombinant organisms, advance approval of the safety director is required.
(7) The department for Recombinant DNA in the Faculty of Agriculture is the Strategic Planning
Section (Senryaku-kikaku kakari).
8.5.2. Microorganisms for research(1) When handling microorganisms for research one is required to give consideration to ensuring safety
and to acquiring the knowledge and skills necessary to handle pathogenic microorganisms.
(2) When handling microorganisms for research, the researcher is required to give notice to the head ofa department (level 2) or to receive the President's approval for use of those organisms through the
Dean (level 3, 4). These levels are governed by the management rule of the University(http://www.kyushu-u.ac.jp/university/office/kikakubu/kenkyusenryakuka/dna/index.htm).
For microorganisms categorized as level 1 which have no risk of causing severe disease in
humans and animals, the researcher is not required to notify, however, organisms having a risk ofopportunistic infection or disease should be treated based on level 2.
(3) According to the safety control regulations of Kyushu University, researchers who perform
microbiological experiments are required to receive education and training and to go through a
medical checkup before the experiment. They should use appropriate laboratories depending on theexperimental level (P2 laboratories for level 2 microorganisms; P3 laboratories for level 3
microorganisms).
(4) Microorganisms used for research must go through sterilization procedures depending on the level.
76

(5) All researchers must have regular medical checkups complying with the regulation prescribed by
the University, and they must have an extraordinary medical checkup in case of an accident.
(6) For conducting research using microorganisms categorized as level 3 or 4, a researcher must receive
the President's approval for use through the Dean.
(7) The department for Recombinant DNA in the Faculty of Agriculture is the Strategic Planning
Section (Senryaku-kikaku kakari).
8.5.3. Animal experiments(1)

Accordingtoregulationsforanimalexperiments
at
KyushuUniversity(http://www.kyushu-u.ac.jp/university/office/kikaku-bu/kenkyusenryakuka/animal/index.htm), allresearchers performing animal experiments are required to attend lecture(s) concerning animal
experiments, to be certified, receive a registration card, and then to register themselves as animal
experimental professionals. They should refer to the “Manual of animal experiment, Kyushu
University” when performing experiments using animals.
(2) All researchers are required to submit an application for animal experiments and to receive thePresident's approval for performing the research through the Dean.
(3) The rearing of experimental animals must be pursuant to the conditions that are approved. The
researcher must understand the physiology, ecology and habits of the animals, and they must
carefully and humanely rear animals and make efforts to contribute the body scientific knowledge.
The researchers must also house the experimental animals responsibly and must prevent damage tohuman life, personal injury, assets and contamination of the environment that might be caused byexperimental animals.
(4) Animal experiments using recombinant animals, infection experiments with pathogens andexperiments using dangerous chemicals must follow the guidelines for recombinant DNA
experiments, handling of microorganisms for research and this handbook, respectively. And theseexperiments must be carried out in an authorized animal room or animal experiment laboratory thatis available and provides physical containment for prevention of pollution and escape.
8.5.4. Radioisotopes and Radiation generator(1) It is impossible for us to perceive radial rays and X rays without using devices which will detect therays. We know, however, that a large amount of exposure can pose a serious health risk and genedisturbance. Therefore, use of this equipment must adhere to guidelines and rules to prevent the
exposure to the user and third persons.
(2) Only a researcher who is provided education and training and goes through a medical
checkup before the experiment and is registered in handling the facility (radiation controlledarea) can use radioisotopes (RI), ionization radiation and the radiation generator. In case ofmulti-year usage, the research is required to receive education and training and to goundergo a medical checkup.
77

(3) Every handling facility, RI caddy and radiation generator for ionization radiation or X-ray carry theradioactive sign (Red trefoil on yellow triangle).
Any persons who have not been registered as a user are not permit to enter to the controlled area or
use equipment indicated by the radioactive sign. It is prohibited to use the sign without permission.
(Use of unsealed radioactive material)(1) In general, a person who belongs to Faculty of Agriculture, Graduate School of Bioresource andBioenvironmental Sciences and School of Agriculture must use unsealed radioactive material at theRadioisotope Center (Laboratory of Hakozaki campus or Semi-high level laboratory). The researchmust be performed according to the rules concerning the prevention of radiation hazards due toradioisotopes and others of each facility.
(2) Because nuclear species and amount used are settled with respect to each facility, the user should
register a suitable laboratory according to each research plan.
(3) All users are required to apply for registration and receive the acceptance of the Dean and to certify
registration by the Director of the Isotope Center. The user planning to use these materials andfacilities during the next year should complete an application for renewal within the annual period.
(4) When a registered user of RI, he/she must obtain permission from the Director of the Isotope Center
before use. Because the maximum amount used per day is set with each nuclear species at facilities,
total usage in the facility must be kept at less than the max amount.
(5) When users are using RI, the user must wear a dosemeter and carry out the experiment under the
regulations of the facility to keep exposure at a minimum. User should perform the experimentwith an understanding of the principles for preventing exposure, which are distance, time andshielding.
(6) At the time of your first experiment, it is recommended that the user perform the experiment
without RI (perform a cold run) before undertaking the real experiment and confirm operatingprocedures, abilities of exposure and contamination. After the simulation, the user should devise
countermeasures for problems for using during the real experiment.
(Use of sealed radioactive material)(1) A person who belongs to the Faculty of Agriculture, Graduate school of Bioresource andBioenvironmental Sciences and School of Agriculture is permitted to use the equipment containing
sealed radioactive material (sealed source) only with permission from the education minister. The
research must be done according to the faculty rule concerning the prevention of radiation hazards
due to radioisotopes.
(2) All users are required to apply to register for acceptance from the Director of the Department and tocertify a registration by the Dean of Agriculture. The user who plans to use these materials andequipment in the next year should complete the application for renewal within the annual period.
(3) For use, permission of the Dean of Agriculture is required. User should submit the experimental
plan document and receive permission before use.
(4) Inexperienced users must not independently use the facilities but must carry out their experiment78

with an experienced person. Admission of visitors who are not included in the experiment isrestricted.
(5) In use, a user must wear a dosemeter and carry out the experiment so as to keep exposure under aminimum. The registry must be supplied.
(6) Because the installation location of the equipment containing sealed source is registered, allowing
the equipment and radiation source to move freely is prohibited. In case of redeployment or
temporal ablation of a radiation source is caused by repair, the user must get the permission of the
Dean through General Affairs Section (Shomu kakari) before moving the equipment.
(7) If you find or presume you have a risk of ionizing radiation injury, you must report this to the officeof Faculty of Agriculture or guard's room at Komatsu gate (after office hours). You should refer tothe network indicated near the equipment.
(Use of X-ray generator)(1) Persons who belong to Faculty of Agriculture, Graduate school of Bioresource andBioenvironmental Sciences and School of Agriculture are permitted to use the X-ray generator in adesignated area. Research must be performed according to the faculty rule concerning theprevention from radiation hazards due to radioisotopes.
(2) All users desiring to use this equipment are required to apply for registration with acceptance fromthe Director of the Department and to certify a registration by the Dean of Agriculture. The user
planning to use it during the next year should complete the application for renewal within theannual period.
(3) For use, permission of the Dean of Agriculture is required. A user should submit the experimental
plan document and receive permission before use.
(4) Chief administrator concerning the prevention from Radiation Hazards due to X-ray and others is
appointed for each service space of the X-ray generator. Authorization by the chief X-ray
inspection engineer is set for the equipment and is required for internal operation.
(5) Researcher must use the X-ray generator under the guidance of the authorized chief. User must
indicate the machine they will work with prior to use in front of the area. If a lamp for indication isequipped, user must turn on the lamp during use.
(6) When user operates in the controlled area, user must wear dosemeter and carry out the experimentin order to keep exposure at a minimum. The registry must be supplied.
(7) During running of the X-ray generator, admission of visitors who are not included in the experimentis restricted.
(8) The registry must be supplied when running the X-ray generator.
(9) If you find or presume you have a risk of ionizing radiation injury, you must report to the office ofFaculty of Agriculture or guard's room at Komatsu gate (after office hours). You should refer to the
network indicated near the equipment.

799. Notes on chemical system laboratory
It is possible to cause unforeseen accidents in chemical experiments, so that please pay attention to
all operations you do. You should study basic knowledge about chemical completely to do safety
experiment. Each basic operation of chemical experiment has meaning, so you examine it before doing
experiment. We will explain necessary knowledge on chemical experiment and indicate instructions to
avoid an accident. To avoid an accident, you should acquire the following knowledge before doing
experiment.

9.1. Clothes(1) If you do not wear glasses usually, please wear an eye protector because you may lose sight if
chemicals are put into your eyes. Do not perform experiments with the naked eyes in thelaboratory because chemicals might splash into your eyes from other tables. Moreover, avoid
wearing contact lenses because it is possible that contact lenses melt and stick to your face.
(2) When using dangerous reagents, you should wear appropriate protecting gloves to reagents.
(3) Wear comfortable shoes. Do not wear a pair of slippers, shoes with high heels, and boots.
(4) Wear a white coat. Do not wear fancy clothes.
(5) Do not wear accessories.

9.2. Notes on experiments
Organic chemistry experiments require sensitive handling because you use a large amount ofinflammable organic solvents such as ether, ethanol, and hexane.
9.2.1. Glassware(1) Check the cracks of glassware you use. Even though there is a small crack, it causes breakage when
you expose it to a vacuum.
(2) Do not expose thin and flat-base glassware to a vacuum.
(3) An accident by glassware is caused when you insert glass tube and temperature gauge into cork
stopper and rubber plug. Please refer to chapter six for the operation.
9.2.2. Organic solvent and reagent(1) Most of organic solvents are inflammable. These may catch fire even for lighter or little sparks andthere is a risk to cause a tragedy. Pay attention to hot organic solvents in the flask because there is arisk for boiling and catching fire immediately. Organic solvents may catch fire even quite far from afire on laboratory board because the ratio of steam solvents is heavier than that of air. Especially,
carbon disulfide is dangerous.
(2) There is a case that volatile solvents such as dichloromethane and ether (boiling point is under80

40°C) may blow out the plug by the evaporation even in sunlight.
(3) Dry the paper absorbed inflammable solvents and desiccating agents such as sodium sulfate andcalcium chloride in the draft before disposing of it in the specified area.
(4) Try not to inhale the vapor of solvents. If you feel dizzy when using solvents, you should stop theexperiment and take a rest in the fresh air. Chemicals should not be attached to the skin. Organic
solvents are easily absorbed from the skin. Pay attention to the sodium cyanide because it isabsorbed from a wound site. Do not take any chemicals in the laboratory into your mouth.
(5) Handle toxicity gas and fuming reagents in the draft. Devices are treated appropriately after using.
(6) Concentrated sulfuric acid and concentrated sodium hydroxide should be washed by a large amountof water because these damage not only clothes but also laboratory board. When you spill it in large
amounts, you make it cold and diluted, and neutralized. In the case of acid, it is treated by NaHCO3.
9.2.3. Device/Reaction(1) The devices should be assembled with instruction and firmly fastened after placing center of gravity
down.
(2) When heated, make a hole to prevent the pressure inside from increasing.
It is better to use a pressure buffer such as air ball. Moreover, when putting calcium chloride
drying tube on the reflux tube, be careful not to be aggregated by damp.
(3) When liquid boiled, boiling stone should be added. If you forget to add it, you should add it after
cooling the temperature under the boiling point. If you add a boiling stone at high temperature,
there is a risk that liquid may come to the boil and explode intensely. Moreover please do not add
an activated charcoal to the heated liquid. If it comes to the boiling point, liquid is ejected to theceiling.
9.2.4. Others(1) In the case of an accident, you rely on a reliable person who has judicial type of mind.
You should do experiments with others at all times.
(2) Keep your laboratory table clean. You should care about second accidents.
(3) You should predict an accident caused by chemicals you use before doing experiments.

8110. Measures for safety in the field work
There are various research and practical training in the field work of agricultural department.
Measures which are different for safety depend on their aim and content. Sometimes, field work
distracts attention away because you are in a different environment from a daily life. There is a risk thatcauses tragedy by the lack of experience. Even though you have enough experience in the field work,
you should not count on yourself.

10.1. General rules(1) Guide of practical training
In the case of field work, you should follow the instruction of teachers. Practical training has a broad
range of contents. The guidelines you should follow are different between practical training in the farm,forest and oceanfront training. If you have any questions, please do not hesitate to ask teachers
something to know before training.
(2) Clothes
You should wear a long sleeve shirt and trousers to protect against sun burn and insects even in summer
and wear a hat to protect against heat. It is effective to cover your neck by wearing a towel to protect
against solar heat. If you go to mountains in the hunting season, you should wear brightly-coloredclothes to prevent a misfire incident. In winter, you should dress warmly. In the extremely cold season,
it is better to use a jacket or coat than to wear several layers of clothing. Moreover it is necessary towear long socks to protect your ankle from dangerous animals.
You should not wear high heeled shoes or pumps. It is better to wear trekking shoes. Athletic shoes ortennis shoes should not be worn in the rocky place or mountain stream.
There are bees or woolly worms in the mountain and needles which hurt you. In the beach, there is arisk of fall and hurt your hand by oyster shell. Therefore, you should wear gloves.
(3) Baggage list
During the field work, you should keep your hands empty. If you have belongings, you use a backpack.
Do not carry a handbag or haversack. Even though you have research equipments, you should use only
one hand and keep your foot. When it is raining, you should wear a rain suit or have a folding umbrella.
In summer, it is better to carry water to avoid heat stroke. Moreover it is better to carry medical goods
such as band-aids and antiseptic substance.
(4) Act in the field work
You should follow the instruction of teacher during the field work. If you go from teacher’s sight, you
should act with others. The most important thing for fieldwork safety is acting slowly. Please be awareof the importance of safety on each training program by confirming with your supervisor.
82

(5) Lodgment
Observe lights-out time to prepare for working of next day and do not cause trouble withneighborhoods
There is a risk that causes lack of sleep, hangover and heat exhausted. It is better to avoid sleeping in atent. If you stay outside, you should research well beforehand.
(6) Dangerous animals
Please do not touch wild-animals in the field work without reason. There are lots of precious animals
that are in danger of disappearing. Also there are dangerous wild-animals such as hornet, adder, tiger
keelback and akajei. Please follow the instruction of teachers to protect such animals. You should
keep off carcass because there is a risk of catching infection diseases such as bird flu.

Fig. 10-1. Venomous creature(7) Graduation thesis research/Graduate school research
When you do a field work research in your graduation thesis, you have to pay attention to the safety
and act in a level-headed manner. Please do not overestimate yourself. Consult with a teacher andconsider adequacy and safety of the research plan. Moreover you should make an application for
special permission about collection of wild animals if necessary.
Observe rules in the laboratory-level and guidelines of each academic society. In the case of using an
adjunct facility, you make an application for special permission.
8310.2. Research in the sea and river(1) Clothes
It is necessary to wear a wet suit (Fig. 10-2 left) or life jacket. It will keep certain force of
buoyancy in deepwater. High boots make you go to deep water level for researches. Using a dry suit
(Fig. 10-2 right) makes you stand on your head in the water because of allowing air into the foot area.
So, you should evacuate the air before you go into the water. Shoes with felting bottom such as
Japanese style socks (Fig. 10-3 AYU-TABI) are effective and can avoid danger from the bottom of river.
Wearing gloves is also effective. It is necessary to wear a helmet in the cliffy area.

Fig. 10-2

Wet suit and dry suit

Fig. 10-3.

Japanese style socks (AYU-TABI)(2) Pay attention to safety ways in the river
Even in the shallow river, you can be swept away. Even in the slow river, it is possible that thedepth of water is increased. It is necessary to make preparations for safety.
You have to confirm the weather condition before starting research and doing researches even if
the weather is clear. When there is a dam in the upper of river, there is a risk of high water. Pay
attention to watering alarm.

84

(3) Pay attention to safety ways in the sea
In the sea, pay attention to the condition of waves. In high waves, it is not only dangerous but also
affects the working efficiency. You may be swept off your feet by waves on the beach or ledge. In caseof an earthquake, there is danger of tsunami. Bring a radio in the place where you can not listen tocommunity wireless system by autonomy. In case of an earthquake, you have to return to the beach andconfirm tsunami information and evacuate if necessary.
(4) Sea-tide
Pay attention to tides in the sea. Make a research plan in consideration of the tides condition.
Incoming tide makes waves lead from off-island to shore, and falling tide makes waves lead from shore
to off-island. At the dry beach, there is a risk that the tide comes in. If you research off-island, you haveto act with time to spare.
(5) Watch where you walkSome of the rocks in the mountain stream can be slippery by the moss. Shingle bottom in the
middle of river can be not only slippery but also may make you tumble over because shingle bottom iseasily crumbled by your weight. You may get in the sand in sand-clogged place just after work ofconstruction. Your feet may sink in the sand in silt-clogged estuary. If you get in the mud, you should
fall on your knees. Rocky shore can be slippery by marine plants. There are many dangerous andharmful things in the bottom of river and sea such as glass chip, oyster shell, and acorn shell.
(5) Others
There are many dangerous wild animals in Kyushu such as hornet, adder, and tiger keel back of theriver, and akajei, plotsus lineatus of the coast and estuary. Be careful of lightning. There was a case thatsome people were dead in the river and sea during research. You should not do research by yourself,
and you should call for help immediately in case of an accident.

10.3. Shipboard, dive, snorkeling(1) Shipboard
Wear a life jacket that has a certain amount of buoyant on shipboard and wear safety boots andcotton work gloves if necessary.
While the vehicle is moving, hold the handhold tightly. Do not get seated on the side of the ship. It
is necessary to be cautious when going by an oncoming ship due to the ship swings.
Unfixed observation equipments, brought from outboard, may be moving. Moreover, hat and towel
may get blown. While you work, be cautious not to get your hands, feet and clothes caught in wire,
cable and rope. Do not throw your body overboard when you use a rope and cable.
While you work on the shipboard, you should follow the instruction of captain and confirm theinformation of weather condition and ocean weaves. When going by an oncoming ship, the big wave
may come, so you should be cautious about handling. If you get sea sick, you should take turns
85working. Using the ship that each university owns, you should get permission of the university office.
Please contact the office beforehand.
(2) Dive and snorkeling
Diving operation needs a lot of experience. It is necessary not only to take a license of scuba
diving and diver but also to have a lot of experience of dive. Buddy system is necessary. Snorkelingdoes not need a license, so you should have self-management for safety because there are not enoughrules about snorkeling. It is better to have a license of scuba diving or diver. Diving is very special
operation, so read through “Kyushu University Diving Regulation” carefully.

10.4. Research at the mountain
10.4.1. Research at the mountain(1) Clothes
Wear a hat or helmet and long sleeve shirts and trousers to protect your head from fall of rock inthe mountain. Moreover wear mountain climbing boots or trekking shoes and carry rain apparel.
Heavy-duty backpack is recommended. It is necessary to carry leather gloves or cotton work gloves.
Cell phone, map, handy GPS, and field note are needed. According to circumstances, water, food, flash
light are needed. In case of camping or climbing a snow mountain, choosing appropriate clothes withextra caution is required.
As an emergency contact tool, cell phone is effective. But there are many places you can not use acell phone because of outside cell phone communication area in the mountain. You have to research thearea if you can use a cell phone beforehand. In case of an emergency contact by cell phone, it isnecessary to inform the disaster area clearly. Make sure the cell phone has not run out of battery.
(2) During research, before and after researchContact an administrator of the mountain beforehand and get permission to enter the mountain. If
you are late to arrive at lodgment, please contact an administrator of lodgment. Entering the mountain,
please carry an emergency contact number such as administrate office number and hospital number.

（川辺書林「森を育てる技術」より）

Fig. 10-4.

Clothes for working in the mountain86Collecting information beforehand and communicating with an administrator of mountain areneeded. Only persons who are well-seasoned can use a chainsaw for cutting trees or grass cutter. You
should learn how to use cutting machines before using them. You should enter a mountain with others.
If you have no other choice but to go into a mountain by yourself, you have to contact the management
office or research education institute often. Going by car to a research area, keep away from roadshoulders on the forest road because there are undeveloped crash barrier or collapse prone roadshoulders. Excessive speed may cause an accident. It is easy to slip on the dirt road and animals may
dash to the road. Do not drive at night if you drive on the forest road for the first time. Be cautious towalk on the forest road because there is a cliff or steep terrain on the side of the road. It is easy to getlost in the forest so please do not walk on the road by yourself. If you get in the forest at night foranimal research, please get permission of forest management office. Confirm the dangerous point in thedaytime beforehand and avoid walking alone. There are lots of dangerous animals in the forest, pleaseavoid getting in the bushed area. There are many cases to be dead from the bite of a hornet. Wasp
venom allergy must be checked beforehand and you should know how to deal with the problem in caseof the bite from a hornet.
Your skin may get poison ivy from anacardiaceous such as Rhus ambigua. This disease may be
preventable to cover your skin carefully. Get a permission of a forest management officer before
working in high place where you climb up a tree or steel tower. Wear a safety belt, helmet and harness
if necessary. When going up or down a slope, you grasp a rock or tree. Do not keep your weight on thedead wood or float stone. When going up or down a slope, do not stand on the slope vertically.
Especially when going down a steep slope, you go down slowly with facing a slope. Going down aslope is more dangerous than going up.
Please be cautious not to throw a stone on the upper side of a steep slope or a cliff. If there areworkers, houses and roads at the bottom of the cliff or steep slope, give your full attention to it.
If there are falling rocks, you should shout “Rakuseki” (Falling rock) .

Fig. 10-5.
Vespa ducalis
Weak aggression and poison comparatively.
Strong threatening conduct(Left : on the leaf, right: killed by insectcide)87

Fig. 10-6. Rhus ambigua
Left: mixed community with other leaf, right: juvenile leaf)
Pictured by 山と渓谷社「樹に咲く花」

10.4.2. Research in urban neighborhood and farming and mountain village
There are many accidents in urban neighborhood and faming and mountain village. Depending on
the condition of land or the purpose of research, clothes or devices you should wear for safety can bechanged. For example, you are recommended to wear comfortable shoes when walking for a long time
for the research. In the construction area or river area, it is better to choose clothes that may become
dirty. At the hearing research in farming and mountain village, wearing high heels or skirt is not
appropriate even though meeting place is an office because the research could be conducted in a field,
mountain forest, or plastic greenhouse. Sometimes, people working at the food processing institute or
cattle farmer do not like perfume and long nails. So, you should pay attention to your appearance.
Recently, as bird flu is especially prevalent in the world, researchers from outside are considered as apossible cause of flu. Please follow the instruction wearing a mask, hat and white robes and sterilizing
hand and shoes. You should not give neighborhoods troubles. As you meet with other people in the
research circumstance, it is assumed that your research devices may be stolen or you may hurt other’s
devices. Pay attention to your safety and other’s safety and prepare for accidents before doing research.
10.4.3. Research in the disaster
It is necessary to have portable GPS, helmet, and cotton work gloves in the research of volcano orslope collapse area. Moreover you should have a gas mask, gas detector, and goggle and wirelessapplication in the area of crater or eructation. Volcano gas is heavier than air and remains in hollow.
Please avoid going to the hollow when there is no wind.
In the research of national park or quasi-national park, please get permission from the environment
ministry or park management office beforehand. When doing research in the national forest, file proper
records with forest management office.
In off-limits area, you should get permission from each autonomous community and collect88

information about the weather and volcano beforehand. Deploy people in the safety area and contact
them when you enter and leave there. When you handle the electricity at the disaster site in the districtof residence, you should wear rubber gloves. Wearing trekking shoes is better. When you carry heavythings, you should wear safety shoes. Especially, collapsed buildings and wasted materials containedged tools, so it is better to wear safety shoes with iron in the shoe sole. During disaster-relief service,
you should not do research in the area. In case of an earthquake or sediment disaster, there may be asecond disaster. Collect enough information before going to the research area. In case of sedimentdisaster, if you are not specialist of it, you should be away from a dangerous slope. During the summer,
you should be careful of heat stroke.
10.4.4. Other notes
Be careful of lightning strike in a mountain or large grass field during the summer. When lightning
is coming, bend forward and wait the lightning go away. It is safe to go into the car. It is important tocollect the information about lightning from the weather news on the radio. Working environment
sometimes influences your health. For example, you become heat stroke under the scorching sun, andbecome hypothermia in the cold area. Do not drink spring water without permission. Pay attention to
the disease such as Japanese fever river and echinococcus, and be careful of the dangerous wild
animals such as wild bears and feral dogs. In case of unforeseen accidents, you have to be committed toyour safety primarily, and all participants, and contact a responsible person and your college. When
getting lost, you should go up to a mountain ridge you can look around and try to find a way out. Use acell phone. If you get lost completely, refrain from moving around. It is better to stay there. Do not
move at night. There are many cases where the mountain stream is shut by dam or waterfalls. By
walking a lot, you are needlessly exhausting yourself. Do not go down along a mountain stream
without reason.
Refer to the following website.
“Field Research Safety Manual”
Edited by Ecological Society of Japan, Field Safety Management Committee
http://www.esj.ne.jp/safety/manual/

10.5. Overseas Research
10.5.1. Introduction
In the overseas research and field working, it is possible to be killed by accidents or diseases
because there are dangerous animals, diseases such as endemic, and public health problem due to poor
hygiene. Collect information of target areas or regions as much as you can and be cautious about healthmanagement, hygiene control, and accident prevention. Instructors have responsibility for the student
safety management and accident prevention, and students should observe general regulations even89though the teacher does not attend your overseas research. In case of an accident, you stay calm andmake the best effort for the rescue and contact your instructor or institute. The instructor and instituteshould give support appropriately.
Notes on safety management and accidents prevention-Make an appropriate plan and notification has to be done before going abroad.
-Change your plan depending on the conditions.
-Ensure an emergency contact number.
-In case of an accident, you stay calm and require for help in accordance with the emergency contact
system.
10.5.2. Plan and notification(1) Plan
Consult your instructor for your plan and write “oversea plan” before making your plan by
yourself. Writing “oversea plan” makes you collect information, research the condition of your area,and predict dangerous matters. There is a case that overseas research can not progress as you think bychange of the condition. Make preparations in good time and add some more research plans according
to the situation. Moreover, ensure your emergency contact number in the field beforehand. Beginners
should work with experienced people and learn the method of overseas research from them. To go to adangerous research area, you work in a team centered on experience people.
(2) Notification
When students go abroad, they have an obligation to inform the Student Section of School of
Agriculture. Teachers need to submit “Overseas travel notification” and its schedule to the General
Affairs Section. “Overseas travel plan” and “Schedule” should be provided to your instructor, related
offices and your parents beforehand.
(3) Preparation
Disease prevention
Collect information about disease prevention one month before going abroad. Receive avaccination if necessary after consulting a doctor. There are quarantine disease vaccine, basic
vaccination, and voluntary vaccination. When you go to an area where malaria is present, you can get a
medicine for malaria prevention but it is necessary to confirm a period after injection whether there areside-effects or not. If you have previous disease, fill out your health condition on “Overseas travel plan.”
Avoid lack of sleep and go abroad in good health. You carry personal medicine.
Insurance
It is the responsibility of everyone to have “personal accident insurance for student pursuing
education and research,” however, the expense for medical treatment and hospital, rescue at the time of90

accidents may not be covered enough. It is better to have “overseas travel accident insurance.” For thearea of insurance coverage, make sure you include the expense for a disease, rescue, changing the planof travel, emergency temporary return.
Other preparedness
Consult an experienced person about the necessity materials for research, equipments, andprocedure. Adequate confirmation of operation capability is needed beforehand especially for carrying
equipments required preliminary check. There is different danger lurking in overseas research from indomestic research. Contact a counterpart closely, and collect information about the field from thewebsite (Foreign Ministry overseas security HP: http://www.pubanzen.mofa.go.jp/). In preparation for
emergency, carry the phone number or address at your local embassy or consular office provided byForeign Ministry HP. Moreover the copy of overseas plan, airplane ticket, and overseas travel accident
insurance should be left to your family and laboratory.
10.5.3. Stay
(1) Secure security
Confirm the contact number to Japan or counterparts, and know the place, facilities, the contact
number of local ambulance, police and hospital. Prepare for having local information by internet,
newspaper and TV. Understand a foreseen risk and have a meeting about the safety beforehand.
Change or reschedule research plan due to the weather condition. When you are caught in thunderstorm during outdoors research, evacuate to a safer place immediately. Do not drive in a research area.
If any, rent a car with a driver. Do not use a bike or bus. If you have no other choice but to use a bike,wear a helmet and be aware of a car accident and crime. Observe the local low and pay attention to the
local religion. In an accident, get a document required by the insurance company. In case of a diseaseor injury, a medical certificate and receipt are needed. In case of a theft, a police certificate is needed.
(2) Health management
Time differences and different dietary life may damage your life and you may be easily infectedwith a disease. The management of dietary life and sleeping becomes very important. In some areas,
there are many causes to damage people’s health such as hygienic condition, dangerous animals, andendemic disease so be careful of health damage. Pay attention to food, water, ice pool, and river
thoroughly.
In poor physical condition, stop a research immediately. Oversea research requires a lot of energy
and needs a rest and health management more than domestic research. In poor physical condition,
contact a teacher or counterpart and go to hospital. At the same time, contact your laboratory
immediately in Japan. A disease transmitted by a mosquito such as malaria, yellow fever, and dengue
fever is spreading, so please be aware not to be bitten by a mosquito. Canine madness is also spreading
all over the world except a part of area, so pay attention to wild dogs.

91

(3) Contact between teacher and student
Teacher and student should exchange the contact numbers and student contact teacher in thefollowing cases.
-When disaster or serious accident happened.
-When a student is in an accident or disease.
-When changing a schedule.
-When arriving at the area.
-Around the time of leaving the area.
-Before the time of leaving the area.
Teachers should contact students constantly. If there is an emergency call from a student, theteacher contacts a department head immediately.
10.5.4. After returning to Japan
After returning to Japan, take care of your health. There are diseases you found developed one
month after you returned such as malaria and type A hepatitis.
When you feel physical damage, fever, and digestive trouble, you should consult a doctor
immediately. When you have a fever or diarrhea in airport, you go to the quarantine station to consult adoctor. Submit necessary documents immediately after returning to Japan.
10.5.5. Others
It is effective to use a cell-phone for long and short stay. Use a cell-phone rental service fortravelers or carry a GSM mobile phone and buy a SIM card at the site. Moreover there is a SIM cardhaving a universal number. Select an appropriate way depending on the term of research or the case of
the field. In the case of long-stay or visiting the field many times, it is better to buy a cell-phone at thesite. Contact an information center about traveler’s insurance. Some credit card companies offer theoverseas traveler’s insurance, so it is recommended to contact a customer service in advance.
Refer to the following website.
http://www.anzen.mofa.go.jp/manual/index.html （ Safety guide for Japanese living overseas ）http://www.anzen.mofa.go.jp/index.html（Foreign minister overseas safety HP）
http://www.forth.go.jp/（Ministry of Health For Traveler’s Health）
http://www.cgiar.org/（CGLAR: Consultative Group on International Agriculture）
http://www.mobistar.jp/（cell-phone rental service for travelers）
http://www.mobell.co.jp/simcard（World SIM card）

9211. Disposal of wastes
We use various chemicals in the University. When you conduct an experiment using chemicals in
the laboratory, you must handle them and the wastes properly. Release of harmful waste solution to theenvironment causes pollution to groundwater and their accumulation.
As the Japanese law requires everyone to perform the proper waste disposal, you need to know theproperty of the wastes well. It is the responsibility of the person who generates the waste to dispose
his/her wastes properly.
In Kyushu University, rules regarding effluent standards and ways of waste disposal have been
established
in
the
"Rule
for
effluentandwaste
managements.”(http://www.kyushu-u.ac.jp/university/rule/zenbun/2004kisoku104.pdf)

11.1. Classification of the laboratory wastes
Except for some special substances such as PCB, the laboratory wastes can be processed using any
method classified in Table 11-1. The liquid waste is classified into inorganic and organic wastes (Table
11-1, 11-2 and Fig. 11-1). Especially, inorganic liquid waste that is collected by Special Liquid Waste
Processing Facility, Kyushu University once a month is called "special liquid wastes". As described
below, other inorganic liquid wastes except for the special liquid wastes, or some organic liquid wastes
and harmful solids that cannot entrust to the agents must be processed as "Chemicals being discarded"
once a year.
Table 11-1. Classification of the laboratory wastes
Category

Particularwaste

liquid(inorganic)

Organic liquid waste

Regular pickup

Separated garbage inthe lab.

Group

Code

Collecting method,

inorganic mercury

A

２０ L Specified plastic container

cyanide and arsenicBfluoric

C

heavy metals

D—aheavy metals
substancesandCollecting day

First Tuesday in every monthorganic D — bphotographic fixer

E

Halogenated organic solvents

H—a

Other organics

H—b

Drum can in the collecting place
The day 10 in every month

Chemicals being discarded (dangerous andharmful solid and liquid)

Novembersummer)Pseudo-medical wastes

June and December

burnable garbage (include the plastic wastes)

carry out to the garbage yard
Entrusted agent collects them as
neededharmful chemical-adhered matter (both burnable
and nonburnable)bottle (cleaned chemical bottles)
no burnable garbage

93(submit

the

list

in

94

11.2. Disposal of inorganic liquid wastes (special wastes)
To handle various kinds of liquid and solid wastes generated in the experiment in the laboratory
safely, you need to prepare their separate containers appropriately according to their property, toxicity
and hazardous natures. In addition, you must hand them over to the Faculty personnel of the Special
Liquid Waste Processing Facility, Kyushu University or to an outsourced personnel.
11.2.1. Acidic and alkaline liquid wastes
You can release acidic and alkaline wastes that are free from a heavy metal or organic substance,by neutralizing them to pH 5-9. If it is strong acid or strong alkali, you can classify them as "heavy
metal including waste solution (special waste solution)" when it is dangerous to do neutralization.
When the acidic or alkaline waste includes any organic substance, you can classify them as "otherorganic liquid waste" and neutralize it to pH 5-9, or "Liquid waste with heavy metals and organic
substances" even if it includes no heavy metals when it is difficult to do neutralization.
11.2.2. Liquid waste with inorganic mercury (special liquid waste)
When liquid waste includes any heavy metal, it must be classified as "inorganic mercury liquid
waste".
The mercury liquid waste with cyanogen or arsenic compound is classified into "Chemicals being
discarded" when it is alkaline. Don't neutralize because of the danger of the hydrogen cyanide (cyanidefume).
The amount of soluble organic substances does not matter.
11.2.3. Organic mercury
The liquid waste with organic mercuric compound such as alkyl mercury and phenyl mercury must
be commissioned to the outsourced processor as "chemical waste, etc" or classified into "inorganic
mercury liquid waste (special liquid waste)" by pre-treatment as follows;
Alkyl mercury --> After adding HCl to change to acidic waste, reduce it to metallic mercury byadding aluminum powder or iron powder, and leaving it overnight.
Phenyl mercury --> Make an oxidative degradation by adding diluted sulfuric acid and potassium
permanganate solution, and heating for 2 to 3 hours at 70 to 90°C.
11.2.4. Liquid waste with cyanogen and arsenic (special liquid waste)
The liquid waste with free cyanide and arsenic compound such as cyanide, cyanic acid, thiocyanicacid, thiocyanate, hydrogen arsenide and arsenic acid must be stocked after you make it more than95

pH10. Heavy metal and organic substance can be included.
When heavy metal liquid waste includes cyanogen or arsenic, you must classify and store thewaste as "Liquid waste with cyanogen and arsenic". On this occasion, the amount of mercury must be
1mg/L or less and that of fluorine must be 15 mg/L or less.
Do not mix the waste with acidic liquid waste.
11.2.5. Liquid waste with fluorine (special liquid waste)
The liquid waste of fluorine compound such as fluoride or silicofluoride must be stocked to makeit pH 6 or more.
The amount of mercury, cyanogen and arsenic in the waste must be 1 mg/L or less, respectively.

11.2.6. Liquid waste with heavy metals (special liquid waste)
Toxic liquid waste with heavy metals such as berylium, osmium, thallium and selenium is treatedas "waste chemicals for outsourced processor". When it can be determined as low concentration andlow risk, you can classify it as "Liquid waste with heavy metals and organic substances".
The content of the following: mercury, cyanogen and arsenic must be 1mg/L or less, that offluorine 15 mg/L or less and that of organic substance 4 g/L or less.
11.2.7. Liquid waste with heavy metals and organic substances (special liquid
waste)The liquid waste with more than 4 g/L organic metal compound, metal carbonyl, chelating agent
such as EDTA and polyamine or organic substances is classified into this category.
The contents of mercury, cyanogen and arsenic must be 1mg/L or less, respectively, and that offluorine 15 mg/L or less.
The organic substance is restricted to water-soluble. Separated oil layer in it must be removed
before handing over to the Special Liquid Waste Processing Facility, Kyushu University.
11.2.8. Photographic effluent
Fixing solution (special liquid waste) --> Argentine must be recovered from it. Set it aside from thedevelopment waste.
Photographic developer --> Store it as "other organic liquid waste".

96

11.3. Disposal of the special liquid wastes
11.3.1. Method for fractionation and pool
The special liquid wastes must be pooled in the rectangular 20 L polyethylene-made container(270×270×385 mm, over 1.7 mm in thickness). Contact a special liquid wastes facility regarding
purchase of the container.
Attach the 15cm or more black tape here for organic substances 4 g
or more and liquid wastes including chelate agent.
Yellow tape for liquid wastes*Write the laboratory name,numberofaffiliation(medicalmanagement system login number),container number on the upper
aspect.
example：○○laboratory 940401 No.3
*Write the name of department on the side of container.

(1) Each liquid waste is distinguished by the colored tape wrapped around theupper part of the container.
red: mercury
blue: cyanogen and arsenic
brown: fluorine
yellow: heavy metal
black: fixer
yellow and black: heavy metals with organic substances(2) The volume of the waste in the container
The level of the liquid waste in the container must be less than the level of the colored tapedescribed above.
(3) The storing of the container
Uncapped or uncovered container or broken container can not be used for the processing. You must
make a check the packing and the inner cap to prevent spilling. In addition, you must always clean theoutside of the container.
Deteriorated container should not be used and must be processed as "experimental burnablegarbage" or "Harmful chemical-adhered material". Keep the container out of direct sunlight.
97

11.3.2. Method of disposal processing(1) Inspection before the request
Check the following points in the laboratory before requesting for processing.
1. Whether or not the liquid waste undergone appropriate processing such as pH adjustment?
2. Is the volume of waste less than the level of attached colored tape on the container?
3. No leak of the waste?
4. Is the outside of the container clean?
5. Is the colored tape for the identification wrapped around the container?
6. Are the organic solvent, oils and fats separated form the liquid waste on the surface or bottom layer?
7. Is there any solid substance floating or precipitated in the container?
If there is any solid substance in the container, you must clear it off with net or other appropriate
tools.
(2) Preparing the forms for the request of special liquid waste processing andsubmission to the Facilities Affairs Section, Faculty of Agriculture (Nogakubu
Hozen-kakari)
When faculty staff in the laboratory requests for waste processing, fill in the form and submit it to
the Facilities Affairs Section, Faculty of Agriculture (Hozen-kakari). The form can be downloaded fromthe following webpage（http://kan-an.jimu.kyushu-u.ac.jp/download-muki-denpyo.htm）.
You have to fill in the login ID (6 digit number) of reagent management system since 2008.
The form must be written in Japanese. Usually, laboratory staff prepares the document. If you
cannot use Japanese language and you need to prepare it, ask a Japanese staff in your laboratory forhelp.
無機系廃液 処理依頼伝票 [センター保管用]
月
部局・研究院

学科・部門

日

【内 線】

講座名

所属等整理番号

下記の廃液の排出責任を負いますので、処理方お願い致します。
【職

名】

分別記号

【取扱担当者】

廃液名

容器番号

含 有 物 質 名 と そ の 重 量 (g 、 mg)
又は 濃度(％、g/L、ppm)

98

ｐＨ

缶数

計
農学系保全係へ、処理依頼伝票及び受入伝票とも提出して下さい
所属等整理番号：通常は薬品管理システムの「所属 Login ＩＤ（６桁の数値）」です。

(3) Collection of the special liquid wastes, pickup and return of the empty
containers
The special liquid wastes form in your laboratory will be carried to the collecting place (P.108)
according to the instruction by the Facilities Affairs Section, Faculty of Agriculture (Nogakukei
Hozen-kakari). The collection is carried out morning of the first Tuesday every month. If the day isholiday, it is the next day. The empty container will be returned to the collecting place the next month,
you must pick it up according to the Faculty personnel.
(4) Return of the liquid wastes that cannot be processed
The liquid wastes that cannot be processed, and did not meet conditions for disposal or haveincorrect information in the form will be returned to you with the form along with the reason. You musttake them back and request again after appropriate pre-processing.
Table 11-2.
CodeA

Special liquid wastes
Liquid waste nameCategorized tape

Conditions

Inorganic mercury liquid

Inorganic mercury liquid wastes

wastesInclude heavy metal or water soluble organic matter.
Alkali liquid wastes including cyanide is categorized “wastes”

Red tape
Cyanide or arsenic liquidBwastesCyanide or arsenic liquid wastes
Heavy metal other than mercury and water soluble organic matterand cyanide.

Blue tapeconditions： pH > 10、Hg < 1 mg/L

Fluorine liquid waste

Inorganic fluoride such as fluorine、silicofluoride

C

Heavy metal other than mercury, water soluble organic matter.
Brown tapeconditions：pH > 6、Hg < 1 mg/L、cyanide・arsenic < 1 mg/L

Heavy metal

Heavy metal including cadmium, lead, chrome, copper, zinc, iron,

Liquid wastes

mangan, nickelic, cobalt, tin.

D—aconditions：organic substances < 4 g/L、Hg < 1 mg/L, cyanide,
Yellow tapearsenic < 1 mg/L
fluorine < 15 mg/L

99

Organic substances
D—b

E

Heavy

metal

Organic metal, metal carbonyl compound, EDTA, chelate such as
liquidpolyamine or heavy liquid wastes including organic substances 4 g

wastesor more.

Yellow＋black tapeconditions：Hg < 1 mg/L

Photographic fixer wastes

Notes: Photographic developing wastes, ammonia water, diluted

Black tapesolution for slide processor “other organic liquid wastes”［Notes］
1. Elimination of solid material: When there is a solid material or sediment in the liquid wastes, filter it with amesh, like nets for catching insects. The sediment should be handled as wastes chemicals.
2. Removed oil: When special liquid wastes are separated organic solvent and lubricating oil, please remove
the oil. Removed oil is processed as other organic liquid wastes (H-b).
3. Contaminant prohibited materials: nuclear fuel material, radioactive ingredient, and contaminated materials
by pathogenic organism. Chemical materials caused odor or health disorder are handled as “wastes
chemicals.” If it is diluted and probably not dangerous, it is handled as “Heavy metal liquid wastes
including organic substances.

11.4. Disposal of organic liquid wastes
Organic solvent waste, hazardous organic solution, dense detergent waste and oil waste areclassified as organic liquid wastes. Especially, you must handle the volatile organic compound such asdichloromethane and benzene because of the strict sewer standards by Fukuoka-city. The watercontaining the organic solvent must be processed as "organic liquid wastes, others".
11.4.1. Notes about the separated storage of the organic liquid wastes
When you store the organic liquid wastes in the same place, you must check whether it conforms
to the Fire Service Law or not and follow the instructions by a dangerous object handler. Keep themfrom places where there might be sudden rise of temperature.
You need to use appropriate and safe container for the contents and do not use thepolyethylene-made container for the inorganic liquid wastes or mix them.
When the waste is polymerizable compound, high polymer compound or tarry substance and it
may form precipitate during the storage or by mixing with other wastes, do not mix other substanceswith the organic liquid wastes.
11.4.2. How to request the disposal of organic liquid wastes
Regarding the organic liquid wastes described below, bring them to the collecting place for organic
liquid wastes and transfer them into the drum can according to the instruction of the Facilities Affairs
Section, Faculty of Agriculture (Nogakukei Hozen-kakari) after the submission of the "application form
for the disposal of organic liquid waste" by the end of the month. The form can be downloaded from100the website. （http://www.agr.kyushu-u.ac.jp/noujimu/）
Usually, the form is written by laboratory staff.
the 10th day of every month.

The collection is conducted at 10:00 to 10:30 on

11.4.3. Notes at the request-The acidic waste should be neutralized to avoid the corrosion of the drum can.
The organic solvent containing heavy metals whose concentration is more than the level of effluent
standards should be classified as "Chemicals being discarded".
-Heavy metal solution with 4 g/L or more of organic substances should be classified into "liquid
waste with heavy metals and organic substances (D-b)" in the special liquid wastes category.
-Special flammable materials that have ignition temperature of 100°C or less, burning point of-20°C or less or boiling point of 40°C are classified as "Chemicals being discarded".
有機系廃液 処理依頼書
提出日付

平成

年

月

日

保全係担当者 殿
分野名

分 野 責 任 者
取

扱者

連絡電話番号
下記の有機系廃液の処理を依頼します。
記
有機系廃液の種類

処理依頼 予定数量 （㍑）

H-a

ハロゲン含有機溶剤

H-b

その他の有機廃液

計※ 依頼条件 ： 重金属濃度は排水基準以下
有機系廃液の分別
クロロホルム、四塩化炭素、クロロベンゼン等の脂肪族並びに
ハロゲン系 有機溶剤＝

芳香族ハロゲン化物で、希釈されていない廃ハロゲン化有機
溶剤

101

非ハロゲン系有機溶剤、有機溶剤と接触した水、写真現像廃その他の有機廃液＝

液、有害有機物質の水溶液、洗剤濃厚廃液、アンモニア水、油類（第３、４石油類）、エステル類、フェノール類、アルコール類、ポンプのドレン等

11.5. Chemicals being discarded
Concerning about the disposal of chemicals in the original container, if there is no danger of heatgeneration or explosion, you can reduce the processing costs by the following methods. The chemicals
that are difficult to pre-process in the laboratory must be classified and store using the usual method in
the laboratory until the day for disposal of "chemicals being discarded".
11.5.1. Chemicals being discarded (Class A)-Organic solvent and water soluble organic chemicals: Take out them from the bottle or container
and classify as "organic liquid wastes.”-Water-soluble heavy metal inorganic salt: If the amount of the substance is small, make a solution
and classify as "heavy metal wastes". If there are several bottles of the same chemicals and they can bemixed together, and put them in a same bottle to reduce the number of bottles.
-Acidic and alkaline chemicals: Neutralize them and dispose. If not possible, classify as "heavy
metal wastes".
11.5.2. Solid wastes with hazardous substances (Class B)
Muddy materials, precipitated and catalytic substances with mercury, cadmium, cyanogen, lead or
chrome, or broken mercury thermometer are classified into "chemicals being discarded" and keep themsafely until the day for the collection.
Notes regarding storage and moving:
Prevent the spillage, runoff and infiltration to the ground.
Pack carefully to prevent the leak when carrying the container.
Broken mercury thermometer should be put in a heavy-duty container that can be sealed.
11.5.3. How to request the outsourcing disposal of “chemicals being discarded”
The chemicals being discarded are processed by an outsourced company once a year, in November.
The form of the list can be downloaded from the web site of the Special Liquid Waste Processing
102

Facility, Kyushu University (http://kan-an.jimu.kyushu-u.ac.jp/). The form is usually prepared by alaboratory staff.

Ask staff in your laboratory as needed.

廃薬品等 処理依頼申込書[H○年度学外委託処理用]
Ｈ○年

○

月

申

部局･部門

○○○・○○○○○○

込

講座･分野

○○○○○○○○

ﾀﾞｲﾔﾙｲﾝ123-4567

者

職

○ ○ ○

氏 名

○ ○ ○ ○

名日○

下記リストの廃薬品について、排出責任を負いますので、処理方お願い致します。
排出責任者

○○ ○○

Ａ．廃 薬 品（購入時の容器に入ったもの）
TEL 下 4 桁

分別

No.

薬

4567

A

1

4567

A

4567

品

名

容器容量

単位

本数

性状

無水酢酸

500

mL

3

L

2

内容物不明

25

mL

1

S

A

3

フェノール

1000

mL

1

L+S

A

4

mL

A

5

mL

A

6

mL
計

5

Ｂ．廃液、汚泥、固形廃棄物、小分けした薬品等
TEL 下 4 桁

分別

No.

容量･重量

単位

個数

性状

15

kg

1

L

2

廃 液 ・ 固 形 物 等 の 品 名
水 銀 入 り 実 験 器 具(15 30 25cm)
重金属含有汚泥(ビニール袋入り)

4567

B

1

4567

B

5

kg

2

S

4567

B

3

不明廃液(ポリ容器入り)

20

L

1

L+S

4567

B

4

水銀温度計(5 本、ﾋﾞﾆｰﾙ袋)

0.1

kg

1

S

B

5
計

5

容器容量

単位

個数

性状

Ｃ．アンプル瓶、スプレー缶 (注 １)
TEL 下 4 桁

分別

No.

品

名

4567

C

1

メチルメルカプタン

50

mL

1

L

4567

C

2

スプレー缶（テフロン）

300

mL

1

L

C

3
計

2

ｱﾝﾌﾟﾙ
長(cm)
15
7Φ 2
0Ｌ

注１) 金属製容器 に密 封 され たもので、容 器の構造によっては処理不能のため集荷できない場合があります 。
ガスボンベ 類 は本 処 理 では該 当 外 ですので申し込まないで下さい。

11.6. Separated refuse generated in the laboratory and daily life
The solid wastes with hazardous materials such as mercury and cadmium should be handledproperly. Clean empty bottles for chemicals and harmless wastes in the laboratory are considered as103industrial wastes or recycling materials and must be distinguished from daily refuse.
11.6.1. General wastes from business activities
The wastes from Kyushu University are divided into general waste from business activities andindustrial waste. The latter is further classified into the industrial wastes subject to special control andthe other industrial wastes.
When you process the wastes in the laboratory by yourself, you must ask the person in charge inthe Faculty to confirm that the processing route is authorized. In Kyushu University, all of the wastes
except for the burnable daily refuse are treated as the industrial wastes. This includes the laboratory
wastes such as empty bottles for chemicals and glassware, empty beverage cans and bottles that areclassified into "scrap metals" and "glass and ceramic garbage" according to the related law.
If the separation of the wastes is not done properly, the collecting agency’s sorting takes time andcan become dangerous. You must observe the precautions indicated in the poster or in following notes;
1. Do not mix different kinds of wastes in the same garbage bag.
2. Needles or sharp materials are dangerous. Put them in a container to avoid injury or accident.
3. Remove caps of beverage bottles, cans and plastic bottles and empty them.
11.6.2. Separated refuse in the laboratory(1) Bottles (recyclable) [The backyards of 3rd, 4th and 6th buildings]
Washed chemical bottles of 500 mL or more (soft glass) are recyclable. The bottles that are notcleansed well will be classified as "non burnable", and those with heavy smudge are classified into
"harmful chemical-adhered matter".
The bottles with frosted interface to the cap (hard glass, thermal glass) and glassware such as
beaker and measuring cylinder are classified into "glass or ceramic garbage".
(2) Harmful chemical-adhered matter (both burnable and unburnable) [Thebackyard of 3rd, 4th and 6th buildings]
Harmful chemical-adhered in laboratory instruments (glass bottles, glass wastes, plastic wastes)
and used silica gel must be put in the drum can with lid. Powdered or granular material such as silica
gel must be enclosed in a bag or case with a label written on it.
(3) Laboratory burnable refuse [The backyards of 3rd, 4th and 6th buildings]
This includes plastic-made lab wares and empty bottles for chemicals. If possible, wash andreuse. Use a transparent garbage bag for them.
(4) Pseudo-medical wastes [West entrance of 2nd building]
Syringes and needles used outside of the University hospital and medical school are treated as104

pseudo-medical wastes and are collected twice a year (June and December) with dry cells and spraycans. The date will be announced by the Facilities Affairs Section, Faculty of Agriculture(Hozen-kakari).
(5) Unburnable refuse [The backyards of 3rd, 4th and 6th buildings]
Non-recyclable bottles of chemicals, glass wares such as beaker and aluminum foil are processed
as industrial wastes similar to daily unburnable refuse.
(6) Animal carcass [South side of 7th building, Practical room for anatomy]
The animal carcass except for large animals such as bovine, swine or goat is collected as infectious
medical wastes once a month. You must freeze and store the carcasses and put them into a container
designed for the carcass. When you bring the container to the room, you must prepare the request sheet
for processing. Concerning about the carcasses of the large animals, you must contact with professional
agency and do the processing individually.
(7) Floor mat for laboratory rodents and animal excrement [The backyard of 3rd,
4th and 6th buildings, University Farm in Hara-machi town]
The floor mat and excrement of rodents can be processed as non-infectious medical wastes if you
can certify that they are non-infectious wastes. For the certification, you must perform a microbial
monitoring paid by your research fund once a year, at least, and submit the certificate to Head Officialof Faculty of Agriculture until 5th in every month. Paste the distributed "seal" on the garbage bag(double thick transparent, 10 kg or less), then take it out to the backyard of the 3rd, 4th, or 6th building
on a twice-weekly basis: only in the afternoon on designated collection days. (The schedule is subject
to change after April 2011.) The cost is beneficiary’s liability. In the case of infectious medical wastes,
which do not have a certificate, laboratory staff should ask assistance from a special industrial waste
dealer individually.
Floor mat for laboratory rodents and animal excrement can be also processed for the composting in
the University Farm in Hara-machi town according to the "Agreement on the disposal of floor mat andexcrement". At first, submit the “ Application form of floor mat and excrement” to the Strategic
Planning Section. Then, bring floor mat and excrement to the composting shed after writing in the sheetof the "Disposal record of floor mat and excrement" in the Farm office. Do not mix plastic materials or
harmful substances in the wastes and bring back the plastic bag or container.
11.6.3. Daily separated refuse(1) Used paper (newspaper, cardboard, magazine, miscellaneous paper, pieces of
paper) [The backyard of 7th building]-The materials that can not mix: tissue paper, thermal paper, coated paper, oiled paper, carbon paper,
coated paper, photos, compound paper, cellophane, tape, fabric, pair, metal, etc.
-Pieces of paper: the amount of the pieces of paper mixed in burnable garbage bags is checked every
year. These papers (scratch paper, cut paper shredder, envelopes, postcards, etc.)" are put into the clear
105

plastic bags. Cellophane tape, cellophane film or packing tape should be removed from envelopes andwrapping paper if it is put on the paper.
(2) Beverage bottles [The backyards of 3rd, 4th and 6th buildings]
Remove lids of bottles, empty them and put into the basket in the recycling yard waste. Reusablebottles (beer bottles, 1.8 L bottles) should be returned to the shop.
(3) Beverage cans [The backyards of 3rd, 4th and 6th buildings]
The cans should be made empty and put in a clear plastic bag.
(4) PET bottles [The backyards of 3rd, 4th and 6th buildings]
PET bottles should be made empty, squashed and put in a clear plastic bag.
(5) Styrofoam [The backyard of 3rd, 4th and 6th buildings]
Styrofoam that can be recycled is box- or board-form polystyrene only. Styrofoam has a mark of"PS" or "Recycle" on it. Other foamed materials such as polypropylene, tips-type, sponge-type andsmall pieces of packing materials are classified into "burnable wastes".
(6) Fluorescent tube [West entrance of 2nd building]Fluorescent tubes contain mercury. They will be collected together with cells, battery andpseudo-medical wastes on June and December. Wait an announcement from the Facilities Affairs
Section, Faculty of Agriculture (Hozen kakari).
(7) Dry-cell battery [West entrance of 2nd building]Dry cells, lithium cells and batteries will be collected together with pseudo-medical wastes on Juneand December. Keep them in the laboratory and wait an announcement from the Facilities Affairs
Section, Faculty of Agriculture (Hozen kakari).
(8) Spray cans [The backyards of 3rd, 4th and 6th buildings]
Spray cans must be punched a hole to make empty the gas. The empty can is classified into a scrap
metal.
(9) Scrap metals [The backyards of 3rd, 4th and 6th buildings]
Small scrap metals can be recycled. The scrap metals equipped with plastic piece or others areclassified into unburnable garbage (glass and ceramic garbage).
Needles must be put into a sealed can.
Empty 18 L can (oil tin) should be pressed if possible.
Aluminum foil is classified into unburnable garbage.
(10) Daily burnable garbage [The backyards of 3rd, 4th and 6th buildings]
106

Do not mix plastic wastes, food scraps in daily life, and the pieces of paper that can be recycled.
(11) Bulk trash or large garbage [The backyard of 4th building]
Large valuable metals such as steel rack or desk should be carried out according to the monthly
instruction from the Supplies Section (Youdo kakari).
(12) Wood wastes [The backyard of 6th building]
The large wooden desk or bookshelf that is difficult to destroy will be classified into the bulk trash.
(13) Useless computer, Consumer electronics recycling [The backyard of 2nd
building annex]
They should be carried out according to the monthly instruction from the Supplies Section (Youdokakari).

107

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12. VDT operation
network security

andcomputersafetymanagement,

12.1. VDT operation
VDT operation indicates that works used Visual Display Terminals such as computer
display and keyboard. At the VDT operation, notes the following points.

12.1.1. Operation environment(1) Adjust the room lightning to remove the distinction between light and dark.
(2) Set up a blind or curtain at the window if necessary to shed out the direct sunlight into
the display.
(3) About the display, adjust the position of screen, tilt of back and forth, and aspect ofright and left and try to prevent the glare.
(4) When there is an unpleasant noise from the VDT or peripheral equipment, takemeasures to reduce the noise.
12.1.2. Working hours
Do not exceed one hour in series for working hours. Set up the break from 10 to 15
minutes to the next work. Moreover set up ones or twice small break for the successivework.
12.1.3. VDT equipment(1) Sit upright in a chair with your spine pressed against the back of the chair and with all
the sole of your foot touched on the floor while at work.
(2) Keep your visual distance 40 cm or more from the display. If necessary, use glasses tocorrect your vision.
(3) To reduce the distance between a display, key board, and documents, keep all things
you see within an appropriate sight.

12.2. Computer safety management and network security
Computer is one of very important tools for the research and has precise mechanism
inside. Therefore it is necessary to handle it carefully. According to the prevalence of
network system, many cases of infections caused by the viruses or worms are being109

reported. To avoid the network failure caused by infection, appropriate knowledge isneeded. For the safety of computer, indicate the following points.
12.2.1. Notes on handling the hardware of the computer(1) As computer is very weak against shake and impact, please handle it carefully.
(2) If you do not shut down a computer in a predetermined manner, you may cause fatal computer error.
When shutting down a computer, please complete a shut down procedure. If you have a plan toknock out power, please shut down your computer beforehand.
(3) When changing the memory or hardware of the computer or opening the hardware,
there is a fear of being gotten a shock. Pull out power plug beforehand.
(4) Computer is weak against static electricity. Do not wear a sweater or touch the door
knob and window frame to avoid the static electricity.
(5) Computer is weak against water and dust. Keep the laboratory clean.
(6) Dispose of used printer ink cartridge or toner in an appropriate way.
12.2.2. Precaution in using computer(1) When setting up new computer and connecting with internal network, submit terminal
attachment application form to the LAN administrator and connect a computer with adesignated IP address by the administrator.
Moreover, when connecting local network in the laboratory, observe the instructionof the administrator in the laboratory.
(2) If you connect a computer without permission, there is a possibility that othercomputer may not connect the network.
(3) It is legally prohibited to use unauthorized computer with other’s IP address or attack
vulnerability on the security.
(4) When the computer failure happened, report it to the administrator immediately. At
that time, report about the condition of failure briefly and simply (5W1H)to theadministrator.
(5) When finding the vulnerability in OS (Windows, Mac) or software, there is possibilityof attack by malicious programs such as worm. Suspend using the program
immediately and use modification program.
(6) It is highly possible that Windows is infected by the virus. Install virus software andcheck a virus regularly.
(7) Make a back up copy of important file to the storage medium (hardware, CD-R)
regularly.
(8) Do not install file swapping software (WinMX, Winny, BitTorrent) because it ispossible to infringe a copyright. The virus (Antiny) infected by file swapping software
is found. There is a fear that internal data may be leaked.
110(9) When using wireless LAN, restrict access to your web pages to keep out the hackers.
(10) Learn to improve your ability of knowledge about hardware and software.
12.2.3. Precaution in using e-mail(1) E-mail account received in school should be used for the purpose of research. If you
want to use e-mail personally, make another account out of school.
(2) When sending e-mail, avoid using half size of character receiver may not read or
machine dependence character such as ①,②,Ⅰ,Ⅱ,㍉,kg,㈱.
(3) Avoid using HTML mail may become the route of infection of virus, send e-mail by text.
Set up the mail software not to create HTML mail.
(4) Set up the mail software not to open a mail automatically because there is a mailinfected with the virus.
(5) Avoid sending mails with attached large file. If you need to send a large file, you should
send it separately or using a file transfer protocol. There is no problem in sending filesof 1 MB or less. Be careful not to be over the limit of mail server.
(6) If you receive a suspicious e-mail attachment, you should not open it because there ishigh possibility that it is infected with the virus.
(7) Contact the administrator and terminate your account if you are not using your mailaccount anymore.
(8) There are many incidents of taking personal information such as card number, ID,
Password by people posing as a credit company or bank official page. Do not click the
URL in the mail and give your personal information easily even though the mailseems reliable.
12.2.4. Precaution in using Web browser
As a web browser, Internet Explorer, Netscape, Safari, Firefox, Opera are known.
They sometimes cause computer failures. Please note the following points.
(1) When sending important information such as personal data with a web browser, you
should confirm whether the site is reliable or not or if you need to send it or not. It is
important to confirm whether it is SSL compatible site or not.
(2) Do not use Web browser personally. Do not use if for net shopping or seeing unrelatedsite to the research.
(3) As there is a malevolent program being run when you view the website, nullify theeffect of Web browser such as Java-script, Java, ActiveX or set up the permission
every time the computer starts.

111

12.2.5. Precaution in using server
When using supercomputer and servers used by more than one person, please note thefollowing points.
(1) Avoid using easy password imagined from the user. Create a password with more than
8 characters and mix the capital and small letter, number and code. A part of softwareor OS have a function to record ID and Password, but it is better not to use it because
there is a fear for others to log in by your password or ID.
(2) To report the problem immediately, you should confirm who the computer
administrator is.
(3) Read the computer instruction published by the administrator carefully and observe it.
(4) When using charged machines such as supercomputer or database, contact theadministrator beforehand and use it appropriately. While using it, take note of charged
money, connection time and hard disk utilization.
12.2.6. Precaution as an administrator
When you become an administrator of the server used by more than one person, you hold aposition of great responsibility if something does happen. Be careful of the following
points and you should manage the servers very carefully.
(1) Confirm the condition of the computer and related networks on regular basis.
-User account (delete unnecessary account)-The condition of CPU, memory, disk, peripheral device.
-Types of application installed and OS, version, other setting status.
-When connecting with network, confirm the setting items such as hostname, IP
address, DNS address, gateway address.
-The constitution of network connected with computer, Firewall, its setting status(2) Confirm who is administrator (Network, mail server, computer center etc) and contact
them immediately if something happens.
(3) When leaving university for a long time, inform your contact number to the person
involved in the laboratory, just in case.
(4) Turn off the power of the computer that will not be used for the next 24 hours.
(5) When using a server, do not run unnecessary service.

112

Contributors
Honjoh, Ken-ichi
Hasegawa, Masumi
Nishimura, Shotaro
Yamada, Naotaka
Doi, Katsumi
Sakuta, Kotaro
Onikura, Norio
Kuwano, Eiichi
Matsuoka, Ken
Sato, Masao
Mitsuoka, Muneshi
Nakamura, Mako
Nakao, Miki
Westover, Layne
Chowdhury, Vishwajit Sur
Eljamal, Osama
Hizon, Amelia
Nomura, Hisako
Sato, Mizuho

Department of Bioscience and Biotechnology
Department of Agro-environmental Sciences
Department of Bioresource Sciences
Department of Bioscience and Biotechnology
Department of Bioscience and Biotechnology
Department of Agro-environmental Sciences
Department of Bioresource Sciences
Professor Emeritus
Department of Bioscience and Biotechnology
Department of Bioscience and Biotechnology
Department of Agro-environmental Sciences
International Affairs and Foreign Student Support of Faculty ofAgriculture
Department of Bioscience and Biotechnology
International Undergraduate Program in Agriculture
International Undergraduate Program in Agriculture
International Undergraduate Program in Agriculture
International Undergraduate Program in Agriculture
International Undergraduate Program in Agriculture
Student Section of School of Agriculture

Kasuya, Yuriko

English translator

113