Handling and storage of liquid nitrogen

May 2021

This information sheet provides advice for developing, maintaining and implementing local safe work practices for all workers at the university who are required to use liquid nitrogen. Liquid nitrogen is commonly used across Monash University for the purposes of snap-freezing and long-term storage of biological samples and in cold traps on vacuum lines/equipment

1. What is liquid nitrogen?

LN2 (liquid nitrogen) is a cryogenic liquid and is the liquefied form of nitrogen gas at atmospheric pressure and subzero temperature. Its properties are listed in the following table.

Dangerous goods class

liquid nitrogen symbol

Boiling point (°C)


Colour of gas


Odour of gas


Taste of gas


Expansion ratio


Gas density (27°C air = 1·18g/L)


2. Hazards associated with Liquid Nitrogen


When LN2 boils it produces gaseous nitrogen, which displaces oxygen from the air. This results in an oxygen-deficient atmosphere, particularly if vented into a closed space, and asphyxia can result.

The health effects of diminished oxygen in the environment can be acute and the impact severe as detailed below.

O2 > 19·5%

Normal function

14% < O2 ≤ 19·5%

Diminution of physical and intellectual performance without person's knowledge

O2 ≤ 14%

Fainting, unconsciousness, death depending on level and duration without adequate oxygen

Cold burns

Due to its extremely low temperature, the cold vapour and gas that are produced when LN2 boils can give rise to cold burns or frostbite upon contact with unprotected skin. Exposure may also produce discomfort in breathing and can provoke an asthma attack in susceptible individuals.

Fire in oxygen-enriched atmosphere

LN2 can condense oxygen from the atmosphere. Liquid oxygen creates potentially flammable or explosive conditions by greatly increasing the combustibility of many materials (e.g. solvents, hydrocarbons).

Pressure build-up/Ice plug formation

Due to the large expansion ratio (liquid to gas), if LN2 is evaporating within a closed system, the pressure can build up with a high risk of explosion. Ice plugs can form on the neck of narrow storage vessels if there is no relief vent, as water is readily condensed by LN2 to form ice. This will lead to pressure build up in the storage vessel.

3. First aid

In no circumstances is it considered safe to enter an oxygen-deprived environment for the purposes of patient rescue.


  1. Immediate assistance should be sought from emergency services by dialling 333 or 0-000 from any internal telephone.
  2. Once in attendance, emergency personnel will manage the situation.

Cold burns

  1. Flush affected area with tepid or unheated tap water.
  2. Apply non-stick dressing and seek medical attention.
  3. If skin is adhered to a cold metal surface, run cold water between the skin and the metal surface, rather than attempting to pull skin away from the surface.

4. Risk management & safe work practices

Risk assessments must be completed by each area where LN2 is handled and stored using the information detailed in the Safety Data Sheet (SDS) and that which is detailed below. It is important to consider the task being performed and the quantities of LN2 used in order to select the most appropriate control measures. Safe work instructions must then be developed and control measures implemented and followed.

Storage facilities

The control measures required in facilities used for using and/or storing LN2 differ depending on the lowest likely oxygen level.

Lowest likely oxygen level
(as a result of room dimension
& volume of LN2 stored)

Risk control measures
(engineering & administrative)

O2 > 19·5%

Basic requirements:

  • signage
  • written procedures for emergencies displayed
  • windows/grilles partly open at all times
  • water for burns available
  • no cold rooms or unventilated rooms
  • training for all users
  • no need to evacuate immediate area.

14% < O2 ≤ 19·5%

In addition to the basic requirements:

  • room mechanically ventilated for heating/cooling
  • access controlled
  • glass panel in upper half of door
  • single, audible, visible low oxygen detector - 1m off floor
  • in room emergency stop button for reticulated systems
  • evacuate immediately if incident occurs.

O2 ≤ 14%

All of the above controls required, plus:

  • CCTV cameras
  • no after-hours access
  • evacuate immediately if incident occurs.
Dewars and storage vessels
  • Storage vessels are classed as pressure vessels and must meet the requirements of AS2030.4 They have an internal volume capacity of ≥ 50L and are used for bulk storage. In most cases, these are leased from the supplier and it is their responsibility to maintain the storage vessels in accordance with the standard.
  • Dewars are portable, double walled containers that are open-necked, free-venting and non-pressurised. Only dewars that meet AS 1894 should be used and a regular physical inspection should be carried out to check the external surface integrity.
Transport and filling operations
  • Containers that meet the requirements of AS 1894 must be used when handling small volumes of LN2 for use in the laboratory, e.g. for snap freezing of samples. Domestic thermo flasks are not appropriate for this purpose, as they are not designed to be free-venting and can result in pressure build-up and a subsequent explosion.
  • Only appropriately trained personnel should perform LN2 filling and transferring tasks. Controls to minimise the likelihood of splashes as well as exposure to escaping liquid/gases must be in place, e.g. using LN2 pumps and ensuring there is adequate ventilation.
  • Appropriate trolleys must be used for transporting dewars between buildings to ensure that the dewar is secure at all times.
  • Dewars must never be accompanied in lifts due to the risk of asphyxiation. In areas where dewars need to be transported between floors via lift, a clear procedure must be in place and available to all staff. Where possible, access to intervening floors should be locked out or the lift must be clearly sign-posted whilst LN2 dewars are in transit with a trained member of staff receiving the dewar at the final destination.
  • For transport between different locations, dewars must be firmly secured in an open truck, utility or trailer. Alternatively, a cryoporter or dry shipper can be used and these are also suitable when samples need to be mailed.

In addition to the engineering and administrative controls outlined above, appropriate Personal Protective Equipment (PPE) must be selected and used, based on the principle that it should prevent LN2 from touching the skin and eyes.

  • Thermo-insulated gloves (elbow length and loose fitting).
  • Safety glasses/full face shield.
  • Long sleeve lab coat.
  • Apron/Long pants.
  • Fully enclosed footwear.
Induction and training

All users working with LN2 must complete the online "Cryogenic Material Basic Safety Principles" training in myDevelopment. Users who wish to access LN2 storage and/or dispensing facilities will need to complete a local area induction. The induction must be recorded using the Local Area Cryogenics Facility Induction checklist and the records maintained locally. The induction must be delivered by the relevant facility manager or their authorised delegate.

5. References

6. Further information

For further information, contact your local OHS Consultant or Occupational Health and Safety: