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Liquid Nitrogen Safety In The Lab: 10 Tips To Avoid LN2 Dangers

Posted in: Lab Safety
Liquid nitrogen safety - using liquid nitrogen to make icecream highlights potential safety issues

The image of liquid nitrogen has been glamorized recently in TV cookery programs by chefs such as Heston Blumenthal. However, whether you are going to use it to make homemade ice cream or to snap-freeze samples in the lab, liquid nitrogen safety is critical, as this reagent is not something to fool around with.

In this article, we highlight the dangers of liquid nitrogen, discuss the necessity of appropriate liquid nitrogen safety training, and share our top tips for avoiding liquid nitrogen dangers in the lab.

Liquid Nitrogen Safety: Why is it Important in the Lab?

Liquid nitrogen (also referred to as LN2) is a cryogenic liquid commonly used in labs. In biology research settings, its primary use is for preserving biological materials such as tissue samples and cells. This is a critical application in many research settings, which means if you work in a biology lab, you will probably have to handle liquid nitrogen at some point.

Unfortunately, researchers have had life-changing injuries and even lost their lives because of accidents involving liquid nitrogen, making it critical that only working with this liquid fully appreciates the dangers. [1,2]

Liquid Nitrogen Dangers

It’s Extremely Cold

The boiling point of liquid nitrogen is -196ºC/-321ºF, and it freezes at -210ºC/-346ºF.

Between these values, liquid nitrogen is in the liquid state. The extreme cold of this liquid means that contact with bare skin can result in cold burns, cell damage, and life-changing injuries such as retinal detachment in a person’s eye.

Liquid nitrogen can also cause common materials such as plastic, carbon steel, and rubber to fatigue and eventually fracture.

It’s an Asphyxiant

One of the most dangerous properties of liquid nitrogen is its ability to boil into large volumes of gas.

The liquid-to-gas expansion ratio is 1:694 at 20ºC/68ºF.

This means 1 L of liquid nitrogen can expand to 694 L of gas. [3] If enough liquid boils in a confined space, the air can be displaced, resulting in an oxygen deficiency.

The reduced oxygen concentration can quickly result in feeling dizzy and render you unconscious with little warning, as nitrogen gas is colorless, odorless, and tasteless.

How to Calculate Percentage Oxygen Depletion Incase of Dewar Failure

Wondering how much oxygen would get displaced if the liquid nitrogen Dewar in your lab failed while full? Need to know this calculation for your risk assessments? Here’s how to work it out.

Air is 21% oxygen and 78% nitrogen.

Generally speaking, we shouldn’t enter rooms containing less than 20% oxygen. Oxygen depletion alarms usually sound at 19.5%, and some two-stage alarms sound again at 18%.

We can use the expansion ratio of LN2 and the volume of a lab to calculate how much the atmospheric oxygen percentage would remain after a spillage.

The equation we need is:

%O2 remaining = 100 x (21 x [ Volumeroom – Volumereleased gas ]) / [ Volumeroom ]

And since it’s useful to keep all units in meters, m:

1 L = 0.001 m3

Let’s imagine a scenario in which a 25 L Dewar of liquid nitrogen fails in a 300 m3 lab. How much oxygen would remain? If 1L of liquid nitrogen produces 694 L of gas, 25 L of liquid produces 17,350 L of gas. In meters cubed, that’s 17,350/1000 = 17.35 m3.

Subbing into our equitation, we get:

100 x (21 x [ 300 – 17.35 ]) / 300 = 19.79%

So it wouldn’t trip the oxygen depletion alarm, but it’s close. Now imagine how small a lift is. Never travel in a lift with liquid nitrogen. If you need to transport it between floors, send the Dewar in the lift and take the stairs.

Also, this is why areas where liquid nitrogen is stored and handled should have appropriate oxygen sensors that alarm when the oxygen concentration falls.

It Can Cause Explosions

The large expansion ratio also means that if liquid nitrogen boils in a confined space with no escape, the pressure buildup can result in an explosion. Therefore, liquid nitrogen storage equipment should have appropriate pressure-relief devices.


Liquid nitrogen safety training should include correct PPE and handling proceedures as shown in image.
Figure 1. Image of researcher safely using liquid nitrogen with correct safety equipment.

Top Tips for Safety Handling Liquid Nitrogen

Now you know the dangers of liquid nitrogen, let’s look at how you can handle it safely in the lab.

1. Get Proper Liquid Nitrogen Safety Training

Before you start work with liquid nitrogen in the lab, whether it is using a Dewar flask for flash-freezing or placing your cells in for cryo-storage, you need to get proper safety training.

Speak to your supervisor or lab safety manager—they should provide appropriate training, including on PPE, safe handling, and what to do in an emergency.

The potential dangers of liquid nitrogen mean you should get some basic safety training if used in the same area you work in, even if you aren’t directly working with it.

2. Know How to Store and Transport Liquid Nitrogen

The storage of liquid nitrogen containers is highly regulated.  There are specific health and safety requirements that require good ventilation and limit the number of storage containers in one room. Make sure you are familiar with your institute’s policies.

When aliquoting a smaller amount for use in a laboratory procedure, only use a container specified for liquid nitrogen. Do not use a domestic vacuum flask or any old container hanging around the lab. Uninsulated containers can freeze to skin and tear it when removed. Unsuitable containers can also suddenly crack and spill liquid nitrogen everywhere.

3. Wear Appropriate Personal Protective Equipment

When using liquid nitrogen, you want to protect your skin, particularly your face and eyes, from splashes. Therefore, you need to wear personal protective equipment (PPE), including a lab coat, specialized gloves, and goggles.

Note that depending on the type of gloves you use, you may only be protected from the extremely low temperatures of stored items, not from direct exposure to liquid nitrogen, so don’t submerge your hands.

While small splashes of liquid nitrogen may not cause a burn, thanks to the Leidenfrost effect, [3] if allowed to pool, liquid nitrogen can cause severe burns.

So be aware of liquid nitrogen getting down the cuffs of your gloves!

For added protection, you should also consider wearing goggles under a full-face shield. I’ve heard of cases where people have been splashed in the eye when organizing cryotubes in a Dewar, despite having a full-face shield. The stuff can shoot up from all angles!

Also, you should ensure that your footwear doesn’t leave any exposed areas, or else you could end up with frostbitten feet.

4. Work in a Well-Ventilated Area and Avoid Confined Spaces

As discussed in the above section on liquid nitrogen dangers, asphyxiation is a real risk. Ensure any area you are working in with liquid nitrogen is well-ventilated, avoid confined spaces such as elevators and monitor oxygen levels.

If this can’t be avoided, speak to your lab safety manager about the necessary steps to ensure that risks are appropriately managed, such as not allowing others to ride in the elevator and ensuring a colleague is aware.

5. Do Not Work with Liquid Nitrogen When You are Alone in the Lab or After Hours

When working with liquid nitrogen, particularly when dispensing from a large to a small container, make sure a colleague is nearby and is aware of what you are doing. The importance of this is highlighted by the tragic death of a researcher working alone. [2]

6. Use Only the Necessary Amount of Liquid Nitrogen

The large expansion volume of liquid nitrogen means that you should limit the amount you use and handle to the minimum you need. Using excessive amounts can dramatically increase the risk of asphyxiation.

7. Don’t Go Diving into a Storage Container to Retrieve a Dropped Sample

No sample is worth asphyxiating yourself from the vapors or burning your face and hands when the liquid bubbles up.

8. Never Use Liquid Nitrogen in a Closed Container

Liquid nitrogen boils rapidly and can cause ice dams to form or pressure to build up, resulting in explosions. Ensure storage containers are properly vented, and never use liquid nitrogen in a closed system.

9. Never Put Liquid Nitrogen Down the Sink

It can cause the pipes to crack, and you will have an expensive repair bill. Furthermore, liquid nitrogen poured down one drain can come up another, which could be a nasty surprise for a colleague in the lab next door.

10. Watch Out for Exploding Cryotubes

Cryotubes should be safe for liquid nitrogen, right? Not always. Liquid nitrogen can unknowingly enter cryotubes through minute cracks and expand quickly when the tube thaws. An exploding cryotube can be like a mini-missile, and a former colleague of mine has a mark on her forehead to prove it!

Liquid Nitrogen Safety in the Lab Summarized

Liquid nitrogen safety is important when working in the lab. If you or others in your lab are working with liquid nitrogen, ensure you have the appropriate liquid nitrogen safety training.

Some key considerations to avoid the potential dangers of liquid nitrogen include working in a well-ventilated area and monitoring oxygen levels if working with large amounts.

Never work with liquid nitrogen alone or out of hours in case of emergencies. Wear appropriate PPE to minimize the risk of cold burns and other liquid nitrogen-related injuries. Use appropriate storage containers to prevent explosions from pressure build-up or leaks from damaged containers.

Do you have any other tips for liquid nitrogen safety? Leave a comment below if so.

Originally published 6 January 2014. Reviewed and updated November 2021. Reviewed and updated again December 2022.

References

  1. Tristan Kirk. Imperial College London fined £70,000 after scientist suffocated in lab filled with nitrogen. Evening Standard. 13 December 2017
  2. Safety problems led to lab death. BBC News. 20 June 2000
  3. Liquid nitrogen. Wikipedia. Accessed 16 November 2021
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1 Comment

  1. Zia on June 13, 2019 at 9:48 am

    Hi sir
    Can i put liquid nitrogen in stainless steel bottle and depth into water.
    I need your prestigious time

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