Department of Biochemistry Yong Loo Lin School of Medicine, NUS Standard Operating Procedure

Title: Safe

Doc. No: Version No: Issue date: Page:

SOP-BCH-WVS-007 001 8 Nov 2010 1 of 3

Handling of Liquid Nitrogen

Dr. Takao Inoue A/P Caroline Lee
Approved By

Mr. Wong Veen Senn Dr. Lim Cheh Peng

Prepared by

Review Date

1.

OBJECTIVE The objective of this SOP is to provide guidance on the safe handling of liquid nitrogen.

2.

SCOPE This SOP is applicable to all labs in the Department of Biochemistry, NUS.

3.

RESPONSIBILITY 3.1 Principal Investigators The PI has the primary responsibility for the implementation of this SOP in their work area. This involves: (a) Preparation, proper storage, transfer and use of liquid nitrogen. (b) Providing appropriate PPE to employees and students where applicable. (c) Training of employees/users. 3.2 Users Users of cryogenic liquids must: (a) Adhere to safe handling procedures (b) Wear appropriate PPE when handling cryogenic liquids (c) Attend required training sessions.

4.

PROCEDURES 4.1 Personal Protective Equipment (PPE) (a) A face shield, cryogloves and a lab coat must be worn whenever you are removing or keeping samples in a liquid nitrogen Dewar. (b) Tuck the sleeves of your lab coat into the cryoglove so that your arm is fully covered. (c) Trousers should be worn over shoes/boot tops to prevent shoes from filling with liquid nitrogen in the event of a spillage. In general, before handling a liquid nitrogen Dewar, ensure that your bare skin will not come into contact with the liquid. (d) Watches, rings, and other jewelry should NOT be worn. (e) If liquid nitrogen spills onto your cryogloves or shoes, remove them IMMEDIATELY. The low temperatures numb everything so fast that no pain is felt while frostbite is incurred. The damage, which can be equivalent to second or third degree burns, is only noticed after gloves or shoes are removed, and requires treatment in a hospital. (f) Unprotected body parts should not come in contact with vessels or pipes that contain cryogenic liquids because extremely cold material may bond firmly to the skin and tear flesh if separation is attempted.

Department of Biochemistry Yong Loo Lin School of Medicine, NUS Standard Operating Procedure
Title: Safe

Doc. No: Version No: Issue date: Page:

SOP-BCH-WVS-007 001 8 Nov 2010 2 of 3

Handling of Liquid Nitrogen

4.2 Storage (a) Cryogenic liquids should be handled and stored in containers that are designed for the pressure and temperature to which they may be subjected. The most common container for cryogenic liquids is a double-walled, evacuated container known as a Dewar flask. (b) Containers and systems containing cryogenic liquids should have pressure relief mechanisms. (c) Cylinders and other pressure vessels such as Dewar flasks used for the storage of cryogenic liquids should not be filled more than 80% of capacity, to protect against possible thermal expansion of the contents and bursting of the vessel by hydrostatic pressure. If the possibility exists that the temperature of the cylinder may increase to above 30C, a lower percentage (i.e., 60 percent capacity) should be the fill limit. (d) This SOP must be attached to every liquid nitrogen Dewar.

4.3 Handling and Transport (a) Escape of nitrogen from a Dewar can cause localized oxygen depletion leading to asphyxiation. (b) Hence, opening a liquid nitrogen Dewar should be done only in a wellventilated area. If this is done in a cold room, its door must be left ajar. Alternatively, the Dewar tank is carted out from the cold room to a wellventilated area when in use. (c) Refilling a liquid nitrogen Dewar should also be done in an open, wellventilated area. (d) Objects that are in contact with cryogenic liquid should be handled with tongs or cryogloves. (e) When transporting liquid nitrogen in a lift, the Dewar must not be accompanied. Instead, two people should be assigned to transport the Dewars, one to load and one to receive at the destination floor. To prevent others from entering the lift, a Do Not Enter sign should be displayed prominently on the Dewar.

4.4 Training All staff using or handling cryogenic liquids must receive training which includes hazards associated with its use, care, selection and use of protective equipment and emergency procedures. New users of liquid nitrogen should receive instruction in its use from experienced members of the academic or technical staff.

Department of Biochemistry Yong Loo Lin School of Medicine, NUS Standard Operating Procedure
Title: Safe

Doc. No: Version No: Issue date: Page:

SOP-BCH-WVS-007 001 8 Nov 2010 3 of 3

Handling of Liquid Nitrogen

4.5 First Aid (a) Warm the affected area of the body rapidly by immersion in water not exceeding 40C, with body heat, or by exposure to warm air. In the event of massive exposure, the emergency shower should be used to warm the body. All clothing must be removed prior to showering. Maintain the affected area of the victim at normal body temperature until medical help arrives. (b) Calm the victim and prevent aggravation of the injury. In the event of frostbite on the feet, the affected person should not attempt to walk. Do not rub or massage the affected parts of the body. (c) Use a mild soap to clean the affected area to prevent infection. Dressings need to be applied if the skin is intact. (d) Flush eyes, if affected, with warm water for 15 minutes.

5. RECORDS Nil 6. REFERENCES Nil