New research on ozonated olive oil with a high peroxide value

The International Ozone Association has recently published a paper on the use of ozonated olive oil, specifically as it relates to in-vitro cytotoxicity for topical applications.  In recent years, Ozonated olive oil has become a popular application for ozone, however, very little research is published on this application.  Therefore, we are excited to make you aware of this new research.

Ozonated Olive Oil with a High Peroxide Value for Topical Applications: In-Vitro Cytotoxicity Analysis with L929 Cells

IOA logo

Received 17 Jan 2017,  Accepted 07 Jun 2017, Accepted author version posted online: 13 Jun 2017, Published online: 27 Jul 2017

Previous studies have shown that ozonated vegetable oils have been used topically for healing of cutenous wounds. The aim of this study is to evaluate the dose dependent use of ozonated olive oil with high peroxide value (OZ) on the viability of cells for preventing side effects in topical applications. To the best of our knowledge, there are no reports investigaing the effect of peroxide value of ozonated olive oil associated with its cytotoxic activity on mouse non-neoplastc fibroblast cell lines (L929). Therefore, the present study was carried out by using OZ alone and/or in combination with glycerol and olive oil. In our study OZ was prepared by using pure olive oil. Both olive oil and glycerol are non-toxic and can be mixed with OZ uniformly. The cytotoxic activity of samples against L929 fibroblasts was assessed using the tetrazolium salt 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay. The peroxide value of synthesized OZ was found to be in the range of 2700–2900 mEq O2/kg oil. The OZ/olive oil group did not show any cell death at all concentrations tested (p > 0.05) however OZ/glycerol group showed statistically significant reductions in viability at higher concentrations (p = 0.004–0.006) compared to the control group. Conclusively, using OZ/olive oil with a peroxide value of 2700–2900 mEq O2/kg oil for short-term incubation was non-cytotoxic to the L929 fibroblast cell line.

Access this entire paper and download HERE

Tech tips on creating your own ozonated olive oil HERE

Want more information on the IOA and how to join?  Click HERE

How to make your own Ozonated Olive Oil – tech tip

Ozonated olive oil is all the craze these days.  I won’t get into the Why of ozonated olive oil here, only the HOW.

Image result for olive oil

Ozonated olive oil is very simple (in theory) it is only olive oil + ozone.  Take any normal bottle of olive oil, or other similar oil and bubble ozone into that water.  After time this will create ozonated olive oil!  Simple right?   Keep reading…

Ozone is a gas that is only partially soluble into water.  Therefore it will require proper work to get ozone to dissolve into a liquid.  This is why when you bubble ozone into liquid you smell so much ozone above that liquid, most of the ozone is escaping.  For more information about ozone solubility, follow this link.

Other great tips on how to more efficiently create ozonated olive oil.

  • Always use ozone produced from oxygen.
    • Air fed ozone generators can introduce impurities
    • oxygen is more soluble into liquid that air
    • ozone produced from oxygen will be higher concentration (increased solubility)
  • Use an ozone generator that produces the highest concentration of ozone
  • Keep your ozonated olive oil cool, but not too cold
  • Use the tallest column possible to bubble ozone into your oil
  • Use the smallest pore diffuser possible to bubble ozone into your oil
    • smaller bubbles have greater surface area and will dissolve easier
  • Keep it running 24/7
    • Keep ozone concentration as high as possible, don’t give it a break, that only slows things down
  • Use glass containers to reduce impurities

See sample set-up below:

ozone olive oil in production
How to create ozonated olive oil

Necessary components:

  • Oxygen Source – we show an oxygen bottle, however an oxygen concentrator will be more cost effective long term.  And much more convenient.
  • Ozone Generator – Choose an ozone generator capable of producing high concentrations of ozone from oxygen.  Our pictures shows the smallest ozone generator we would recommend.
  • Reaction Vessel – use a reaction vessel made of glass that is as tall as possible.  Ensure it is sealed and vents excess gas to a safe location.  Use a bubble diffuser at the bottom with fine pores to create small bubbles.

A successful batch of ozonated olive oil should take only 24 – 72 hours to produce.  Ideally 24 – 48 hours.  More time allows more impurities to enter the system.

Should you have any questions about this ozone application or any others feel free to contact our staff today, we would be glad to discuss this with you.


Ozone Chamber for Safe Ozone Use

Introducing the Ozone Chamber for safe ozone use.  This is a great device that can be used at home, or in your lab to harness the power of ozone without ozone escaping into the ambient air.

Ozone chamber with scrubbers and diffusers
Ozone Chamber – $99.50

The Ozone Chamber comes with an ozone scrubber to place on the off-gas vent that will destroy all off-gassed ozone from your application.  Tubing and a diffuser are used to bubble ozone in the bottom of the liquid or other substance.  We provide the Ozone Chamber with a spare scrubber and diffuser.


The Ozone Chamber can be used with the OXS-300 Ozone Generator to easily apply ozone to fruits, vegetables, and produce you may have.  Using ozone in food products will prolong  the shelf life of foods, and reduce bacteria, pathogens and pesticides.

Treat fruit with Ozone using the Ozone Chamber and Ozone Generator

How to use:

Open the top of the Ozone Chamber using the handles.  Place produce and water into the chamber.  Ozone will dissolve into the water and help kill all bacteria that the water touches.  This activated water is a powerful pathogen killer.


The Ozone Chamber can also be used to safely ozonate water for drinking or for washing other items.

Image shows the larger 1KNT Ozone Generator used with oxygen to create high levels of dissolved ozone in water safely.

Create your Own Ozonated Olive Oil using the Ozone Chamber and an ozone generator.  Place the olive oil in the chamber and ozonate for as long as you need, without the risk of excess ozone off-gassing into room or ambient air!

Common Uses for the Ozone Chamber:

  • Sanitize Fruits and Vegetables
  • Remove mold from Fruits and Vegetables
  • Ozonate water for drinking
  • Treat water with high levels of ozone for sanitation
  • Production of Ozonated Olive Oil
  • Material compatibility testing with ozone
  • Chemical Synthisis
  • Laboratory Experiments with ozone