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Ozonated Saline IV Drip (O3SS) Protocol
Weight-based dosing, preparation, concentration calculation and safety for the ozonated saline (O3SS) IV drip.
Quick answer: The ozonated saline (O3SS) IV drip uses 0.4–2 µg/mL in 200–250 mL, daily or every other day. MediSkill does not advise this protocol; standard IV ozone (MaH) is the method it teaches.
Warning
MediSkill does not advise this protocol
- It is hard to do correctly. It is easy to overdo it and burn (irritate and damage) veins.
- If too much is given, ozonated saline stays active longer than ozone gas. That raises the risks for people with contraindications and the risk of hemolysis (breaking down red blood cells).
- It cannot be done in a standard saline bag. It needs special equipment and sterile glassware.
- We are not aware of any company that supplies sterile, single-use glassware for ozonated saline infusion. The systems on the market are designed to reuse the same glassware between sessions without sterilizing it. This should never be done without proper sterility.
We present Madrid's method below for reference only. Standard IV ozone (major autohemotherapy) is the method we teach: see standard IV ozone.
What are the basics of the ozonated saline IV drip?
- Ozonated saline (O3SS) is sterile 0.9% saline bubbled with ozone and given as a slow IV drip. No blood is drawn.
- The dose is set by body weight, not by feel. This is the one method where Madrid calculates the dose from weight instead of starting low and stepping up. Low is 1 µg/kg, medium is 2 to 3 µg/kg, and high is 4 to 5 µg/kg.
- Volume is 200 to 250 mL. A cycle is 6 to 10 sessions, given daily or every other day, adjusted to the case.
- The ozone that dissolves in the saline is about 25% of the gas concentration bubbled in. To reach a target, set the generator to 4 times the dissolved concentration.
- Upper limit: 2 µg/mL dissolved in the saline. Madrid says going above it is dangerous and can cause phlebitis.
Source: Madrid Declaration on Ozone Therapy, 4th edition (ISCO3, 2025), section 3.3 and Table 1B. Paraphrased.
Preparation
Madrid compares four ways to prepare the solution:
- Three-needle method. 250 mL of 0.9% saline in a glass bottle, bubbled constantly before and during the drip so it stays saturated. Madrid calls it reliable.
- Indirect method. Saturate for about 15 minutes, then infuse quickly because the ozone decays. Madrid calls it unreliable.
- Closed single-use kit. A phthalate-free kit that serves for both MAH and O3SS. Madrid calls it reliable.
- Microbubbling in reusable borosilicate glass. Saturate for 5 minutes at 200 mL/min. The concentration then holds for more than 30 minutes without bubbling, so the drip runs with the generator disconnected.
With conventional bubbling, saturate for 15 minutes at 200 mL/min, then give the solution at 120 drops per minute with the same bubbling running. Madrid notes that microbubbling removes the risk of ozone gas reaching the vein when no one is watching the end of the drip.
What dose and schedule are used for the ozonated saline IV drip?
| Level | Dose | Dissolved concentration | Generator gas concentration (4x) |
|---|---|---|---|
| Low | 1 µg/kg | 0.4 µg/mL | 1.6 µg/mL |
| Medium | 2 µg/kg | 0.8 µg/mL | 3.2 µg/mL |
| Medium | 3 µg/kg | 1.2 µg/mL | 4.8 µg/mL |
| High | 4 µg/kg | 1.6 µg/mL | 6.4 µg/mL |
| High | 5 µg/kg | 2.0 µg/mL | 8.0 µg/mL |
Values are Madrid's worked example: an 80 kg patient and 200 mL of saline, at a flow of 200 mL/min. Dose in µg is dose per kg times body weight. Dissolved concentration is that dose divided by the saline volume in mL.
Madrid's uses by dose level
| Level | Madrid lists it for |
|---|---|
| Low (1 µg/kg) | Immune support and cardiovascular conditions. |
| Medium (2 to 3 µg/kg) | Detoxification in endotoxemia and chronic inflammatory conditions, viral infections, and neurodegenerative conditions. |
| High (4 to 5 µg/kg) | Highly infectious bacterial and viral illness such as sepsis, acute skin infections and burns, and autoimmune conditions where an immunosuppressive effect is wanted. |
Madrid's treatment examples
| For | Dose | Method | How often | Sessions |
|---|---|---|---|---|
| Chronic degenerative inflammatory, cardiovascular and obstetric conditions | 1–2 µg/kg | Microbubbling, 5 minutes | Every other day | 2–3 |
| Same | 1–2 µg/kg | Conventional bubbling, 15 minutes | Daily | 5 |
| Acute bacterial or viral infection, sepsis, burns, skin infection, autoimmune disease | 3–5 µg/kg | Microbubbling, 5 minutes | Daily | 3–4 |
| Same | 3–5 µg/kg | Conventional bubbling, 15 minutes | Daily | 5–6 |
Source: Madrid sections 14.9 and 14.10, at 200 mL/min. These courses are shorter than the general 6 to 10 sessions above. For each, Madrid says to switch to conventional intravenous administration as soon as possible. The 3 to 5 is µg per kg of body weight, not µg/mL.
What are the best tips when using the ozonated saline IV drip?
- Use ozone-resistant glass, silicone or PTFE for everything that touches the gas or the solution. Avoid plastic bags for bubbling: they release phthalates, which are toxic and raise inflammation.
- Use medical-grade oxygen from a cylinder. Madrid does not accept industrial oxygen or oxygen concentrators for O3SS.
- If the generator does not give decimals, Madrid rounds up (for example 1.6 to 2.0 µg/mL). The patient then receives about 0.5 µg/mL, still under the 2 µg/mL limit.
- Start the drip right after saturation. Any delay lowers the dose delivered, except after microbubbling, where the concentration holds for more than 30 minutes (Madrid).
- Per our saline drip article, a healthy patient should eat beforehand: the drip can lower blood sugar, so have a plan to treat low blood sugar. Patients on insulin or other glucose-lowering medicine should check with their physician.
What are the precautions for the ozonated saline IV drip, and who should avoid it?
- Phlebitis happens when very high, non-therapeutic doses are used. Watch the IV site throughout.
- Accidental gas embolism happens with conventional bubbling when bubbling is not stopped at 50 mL and the drip is unsupervised. Stay with the patient.
- Never give ozone gas by direct IV injection. Madrid classes it as a route that should not be used under any circumstances.
- If a side effect occurs, Madrid points to the ISCO3 first-aid guidance and asks that it be reported on the ISCO3 adverse-event form.
Contraindications
The Madrid Declaration lists these contraindications for systemic ozone:
- Direct inhalation of ozone gas. The fluid lining the lungs has no antioxidants to buffer it, so inhaled ozone is toxic.
- Glucose-6-phosphate dehydrogenase (G6PD) deficiency (favism, acute hemolytic anemia). Madrid lists it as a contraindication and recommends testing, but whether it is absolute is debated. MediSkill protocols start low and go slow; see the contraindications article.
- Toxic hyperthyroidism (severe Graves' disease)
- Thrombocytopenia (platelets under 50,000) and severe coagulation disorders
- Severe cardiovascular instability
- Acute alcohol intoxication
- Recent acute myocardial infarction
- Acute massive hemorrhage
- Convulsive states
- Hemochromatosis
- Patients receiving intravenous copper or iron therapy
Pregnancy. Madrid advises avoiding ozone therapy in the first trimester (weeks 0 to 13) and reserves its use in pregnancy for specialists in gynecology and obstetrics.
More detail
Equipment
- Medical ozone generator and medical-grade oxygen cylinder with regulator
- Sterile 0.9% saline, 200 to 250 mL
- Ozone-resistant glass bottle or a phthalate-free closed kit, with ozone-resistant tubing
- Standard IV start supplies and IV pole
- Ozone destructor for gas that is not dissolved
Worked example
Patient 80 kg, low dose of 1 µg/kg, 200 mL of saline. Dose is 80 µg. Divided by 200 mL, the dissolved concentration is 0.4 µg/mL. Multiplied by 4, the generator is set to 1.6 µg/mL.
What the studies used
Most O3SS research is in Russian-language journals. Study doses are listed as printed and are not Madrid dosing. Gas concentrations were given for making the solution, not the dissolved dose.
- In a randomized trial of 140 people with post-COVID fatigue, 200 mL daily for 10 days (gas at 2.0 mg/L, then 3.0 to 4.0 mg/L) lowered a blood oxidative-stress marker more than the comparison group, with no adverse events reported (Soldatenko 2025).
- In 139 people with leg artery disease, 8 to 10 infusions of 200 mL over 20 minutes improved the ankle-brachial index (0.64 vs 0.50) over standard therapy alone (Makarov 2015).
- In 100 people having colorectal surgery, 5 infusions at 1 to 6 µg/mL in solution went with fewer complications afterward (14% vs 22%) (Gataullin 2014, Zhurnal MediAl).
Learn more
Background, benefits and research: Ozone saline IV drip, full article. IV by blood: IV ozone (MaH) protocol. Comparing routes: Comparing home ozone therapies.
Related protocols: IV Ozone Protocol: Major Autohemotherapy (MaH), Minor Autohemotherapy (MiAH) Protocol, EBOO Protocol.
Other protocols
For licensed practitioners. Educational reference only, not medical advice. Follow your scope of practice and local regulations.