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Practitioner protocol
EBOO Protocol (Cautioned)
Madrid's ozone concentration, gas volume, blood flow and equipment notes for extracorporeal blood oxygenation-ozonation, an advanced hospital procedure.
Quick answer: EBOO (extracorporeal blood oxygenation-ozonation) is an advanced hospital procedure. Madrid lists 0.1–0.4 µg/mL, 18 L of gas and a blood flow of 30–35 mL/min, treating about 4 L of blood per hour. MediSkill recommends starting with standard IV ozone (MAH).
Warning
Requires hands-on training from an experienced physician
EBOO should only be performed by a practitioner trained in it by an experienced physician.
Start with standard IV ozone. Major autohemotherapy (MAH) is the method MediSkill teaches and the one we recommend practitioners start with. It has far more published research behind it, needs simpler equipment and does not require a hospital. See the IV ozone (MaH) protocol. Come back to EBOO once MAH is routine in your practice. For background, benefits and cost, read EBOO therapy: benefits, side effects, cost and comparing IV ozone therapies.
Practitioner-only procedure. EBOO is a blood procedure. Madrid says it must be performed in a hospital. This sheet is a reference for trained, licensed clinicians. It is not a self-treatment guide and not medical advice.
What are the basics of EBOO?
- EBOO stands for extracorporeal blood oxygenation-ozonation. Blood leaves a vein, passes through a gas exchange device where it meets the ozone-oxygen mixture, and returns to another vein. Madrid compares it to hemodialysis and calls it an advanced variant of MAH.
- Compared with MAH, it treats a larger volume of blood (Madrid gives 4 L per hour) at a lower ozone concentration (under 1 µg/mL).
- Madrid lists the main uses as severe peripheral arterial disease, coronary artery disease, severe dyslipidemia, Madelung's disease, vascular deafness, necrotizing fasciitis, antibiotic-resistant sepsis or infections, ischemic stroke, chronic heart failure and viral hepatitis C. That is where the method is used. It is not a promise of results.
- Madrid says EBOO is used in the United States, Italy, Russia, Ukraine and Malaysia, and rarely in some Latin American countries.
- Madrid gives no session count or schedule for EBOO, so none is listed here.
Source: Madrid Declaration on Ozone Therapy, 4th edition (ISCO3, 2025), section 3.4 and its table of systemic routes. Paraphrased.
What settings does EBOO use?
| Level | Concentration | Gas volume | Ozone dose |
|---|---|---|---|
| Low | 0.1 µg/mL | 18 L | 1.8 mg |
| Medium | 0.25 µg/mL | 18 L | 4.5 mg |
| High | 0.4 µg/mL | 18 L | 7.2 mg |
Madrid's note on this row: the optimal blood flow is 30 to 35 mL/min. Madrid's method text says the procedure treats 4 L of blood per hour. Madrid does not reconcile the two numbers, so confirm the flow and run time against your device's instructions.
What are the best tips when using EBOO?
- Use an ozone-resistant gas exchanger. Madrid warns that most hemodialysis filters are made of polysulfone, cuprophane or other materials that ozone attacks, and that using them for EBOO risks producing toxic products. It adds that many people use them anyway.
- Madrid describes two basic set-ups. One uses microporous hollow-fiber exchangers made of ozone-resistant polypropylene, with blood in the fibers and the gas mixture outside them, at about 99% oxygen and 1% ozone. The other uses a rotor and a glass bottle that turns horizontally on a device with three pieces of ozone-resistant polypropylene.
- Madrid's text (about 1% ozone, which is roughly 20 µg/mL) and its table (0.1–0.4 µg/mL) disagree. Confirm the setting with your device maker and start at the table's low level.
- Madrid says that if the procedure runs longer than an hour, the patient needs an extra dose of heparin (1 mL, 5,000 IU) every hour.
- At the end, the blood is pushed out of the lines and the exchanger with saline, then the cannulas are removed.
- Use the device's bubble trap and air detector, and never let air reach the vein. Check the lines for air before you start and during the run.
- Use medical oxygen from a tank and regulator. An oxygen concentrator is not for blood work.
- Madrid's base rules apply: first do no harm, and titrate from the low level.
Equipment (generic)
- Ozone generator with an ozone-resistant gas path (quartz glass, titanium, silicone) and an ozone monitor
- Medical oxygen tank and regulator
- Ozone-resistant gas exchange device (for example hollow-fiber polypropylene)
- Blood pump, sterile tubing and venous access for two lines
- Heparin and saline
What are the precautions for EBOO, and who should avoid it?
- Never breathe ozone gas. Keep the room ventilated.
- Sterile technique for both venous lines. Madrid says most adverse events come from malpractice, not from the method.
- Blood contacts the whole circuit, so every part that touches blood or gas must be ozone-resistant and single-use or properly sterilized.
- Heparin carries its own bleeding risk. Check platelets and coagulation first, as the contraindications below say.
- Report any side effect with the ISCO3 adverse event form, as Madrid asks.
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. MediSkill treats it as an absolute contraindication for high-dose methods (10-Pass, EBOO, HiDose).
- 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
What the studies used
EBOO has little controlled evidence: one small randomized trial, plus case reports and equipment measurements. Study settings are listed as printed and are not Madrid dosing.
- In 28 people with peripheral artery disease, EBOO produced more regression of skin lesions than intravenous prostacyclin, with better pain, itching and well-being scores. Limb blood supply did not differ, and the authors saw no side effects over 210 EBOO sessions (Di Paolo 2005). Madrid cites this trial.
- A clinic took 85 measurements of gas leaving the circuit during one-hour EBOO runs in 12 patients. Blood took up on average 37% of the ozone supplied, and uptake rose by up to 30% when blood flow went from 30 to 40 mL/min. It used gas at 10 to 60 µg/mL, far above Madrid's range, and a hemodialysis filter. It measured ozone uptake, not patient outcomes, and reported no untoward effects in at least 400 sessions (Rowen 2023).
- Case reports describe 4 to 6 sessions: four weekly sessions of 1 to 1.2 L of blood in three people with advanced Parkinson disease, with G6PD checked first and ozone at 90 µg/mL (far above Madrid's range) (Cureus 2026), and two series of three sessions in one woman with chronic anemia (Bennett 2025). Neither had a comparison group.
Where sources differ: Madrid's concentration (0.1 to 0.4 µg/mL) is much lower than the figures in the clinic reports above. This sheet gives Madrid's numbers.
Learn more
Where to start: IV ozone (MaH) protocol and the MAH article. EBOO background: EBOO therapy: benefits, side effects, cost. Equipment and cost: EBOO machine price. Comparison: Comparing IV ozone therapies.
Related protocols: Bladder (Vesicourethral) Ozone Instillation Protocol, Prostate Ozone Injection Protocol, Ozonated Saline IV Drip (O3SS) Protocol.
Other protocols
For licensed practitioners. Educational reference only, not medical advice. Follow your scope of practice and local regulations.