Ozone Saline IV Drip: Protocol, Uses and Safety

At a Glance:

• An ozone saline drip involves bubbling medical-grade ozone-oxygen through 200 to 250 mL of sterile 0.9% NaCl, then giving it as an IV drip. The Madrid Declaration sets the dose by body weight (1 to 5 µg/kg) and says the patient must not receive more than 2 µg/mL of dissolved ozone, because going higher is dangerous and can cause phlebitis [15]. Some published studies list higher concentrations, but they report them in different ways, so they cannot be compared directly.
• Ozone saline drip is a systemic oxidative therapy option that avoids the complexities and risk of blood handling associated with major autohemotherapy, since ozone saline drips use pre-saturated saline rather than withdrawn blood [1].
• When ozone reacts with chloride ions in saline, chlorine byproducts can form. In the one lab study that measured them, only chlorate was found (chlorite and perchlorate were not detected), and far less formed in an ozone-resistant bag than in a plastic saline bottle [2]. This is still a debated point in the safety of ozone saline drip [1].
• The ozone saline drip became popular in busy Russian hospitals because it eliminates the need for blood withdrawal and reinfusion, making it a practical option for busy hospitals and situations where blood removal may be impractical. 
• Major proponents of ozone therapy, like Dr. Bocci and the World Federation of Ozone Therapy, consider ozone saline drip to be higher risk due to chlorite formation and of lower benefit than autohemotherapy due to lower ozone doses. 

Ozone saline drip (O3SS) is a controversial ozone treatment. While easier to administer and avoiding difficulties related to blood removal and handling, saline ozonation runs the risk of generating chlorine byproducts such as chlorate, which can be harmful at sufficient doses. This article covers the pros and cons in comparison with other ozone IV treatments, safety limitations, and the latest clinical evidence. Please note that this article is to provide evidence-based information, not to encourage any specific treatments. 

How an Ozone Saline Drip Works

Biochemical Dynamics of Ozonated Saline Infusion

The ozone saline drip begins with medical-grade ozone being bubbled through sterile 0.9% sodium chloride (NaCl) in an ozone-resistant glass container. Madrid gives 5 minutes with a microbubbling device or 15 minutes with conventional bubbling, both at 200 mL/min [15].

Unlike major autohemotherapy and other blood ozone treatments, where blood's natural antioxidant buffering capacity moderates the reaction, saline lacks these protective proteins. As a result, the drip delivers dissolved ozone (O₃), hydrogen peroxide (H₂O₂), and reactive oxygen and chloride species directly into your vein [2,1]. This distinction is what gives the ozone saline drip its unique pharmacological profile compared to blood-based ozone methods.

Safety testing of the ozone-saline interaction is a critical consideration. The dissolved ozone the patient receives is capped at 2 µg/mL under Madrid [15], which is much lower than what you’d use in any blood ozone treatments. 

Also, ozonated saline can contain potentially harmful chlorinated byproducts. Bocci and others have raised concerns about chlorite [1]. When researchers measured them, they found chlorate, which rose with higher ozone concentrations and longer storage, but no chlorite or perchlorate. Chlorate levels were 0.9 to 7.7 µg/L in an ozone-resistant blood bag versus 45 to 208 µg/L in a polypropylene saline bottle, which is one more reason to use glass or ozone-resistant containers [2]. The Madrid Declaration holds that little or no chlorine byproduct forms under therapeutic conditions (0.9% saline, low ozone concentrations), and that the lab findings of concern used much higher ozone concentrations or plastic bags [15]. 

In addition, the ozone concentration in the prepared solution falls once bubbling stops [1]. Madrid says a solution made by microbubbling holds its concentration for more than 30 minutes and can be given without bubbling, while with conventional bubbling the gas keeps running during the drip [15].

Differences Between Ozonated Saline Infusion and Autohemotherapy

Understanding the key biochemical differences between intravenous (IV) ozonated saline and ozone autohemotherapy (O3-AHT) requires examining how ozone interacts with biological substrates in each method, as the reaction environment fundamentally shapes the downstream molecular effects.

Autohemotherapy

In major autohemotherapy (MAH), a volume of blood (typically 100–200 mL) is drawn into an anticoagulated container and mixed with a precise ozone/oxygen gas mixture outside the body. This is the most extensively studied method in the clinical literature.

Upon direct contact with whole blood components, ozone and oxygen react within seconds to the full spectrum of blood substrates, including:

• Plasma lipids
• Albumin
• Antioxidants (glutathione, ascorbic acid, uric acid)
• Red blood cell membranes, white blood cells, and platelets

Ozone reacts with polyunsaturated fatty acids in plasma and cell membranes, generating reactive oxygen species as early, short-lived messengers and lipid oxidation products (LOPs), including lipid peroxides, 4-hydroxynonenal (4-HNE), and ozonides. These LOPs act as controlled, low-level oxidative stress signals.

With MAH, the clinician can precisely control the ozone concentration per mL of blood, and the reaction is completed before the treated blood re-enters circulation. This makes the dose-response relationship more predictable.

IV Ozonated Saline

In IV ozonated saline, ozone gas is bubbled through sterile normal saline (typically 0.9% NaCl) until a target dissolved ozone concentration is reached. This ozonated saline is then infused intravenously.

Normal saline lacks the biological substrates present in whole blood. There are no lipids, no albumin, no antioxidants, no blood cells in the saline itself. Ozone dissolved in saline exists primarily as molecular ozone (O₃) and its aqueous decomposition products, including reactive oxygen species such as hydroxyl radicals (·OH), superoxide anion (O₂·⁻), and hydrogen peroxide (H₂O₂). Each of these species is naturally occurring in the human body and performs a specific role [3]. Among them, hydrogen peroxide is the most stable one, which is most likely to last long enough to affect your physiology.

While MAH generates LOPs as key secondary messengers through lipid ozonation, IV ozonated saline delivers primarily dissolved ozone and H₂O₂ directly into the venous blood. The ozone and ROS then react with blood components in vivo (inside the body) rather than ex vivo, generating a smaller amount of LOPs and activating Nrf2.

This makes H₂O₂ the dominant mediator in ozone saline drip. Given the rapid decomposition of ozone and other oxidative species in aqueous solution, hydrogen peroxide (H₂O₂) is likely the predominant reactive species. 

Theoretically, hydrogen peroxide and other oxidative molecules in ozonated saline can theoretically influence other aspects of your physiology, including [3]:

• Temporarily improving insulin function (this is typically observed across most biooxidative therapies)
• Initiating wound healing signals
• Supporting protein folding
• Influencing inflammatory balance

Ozone Saline Drip: The Clinical Evidence

Ozone saline drip studies have been published as part of various retrospective studies and case series, especially during the pandemic, especially in India and Europe. Most prepandemic mechanistic and randomized controlled trials are Russia-based studies. Overall, the evidence is early: mostly small, single-centre or uncontrolled studies, plus a few small randomized trials from Russia. The results are encouraging, especially in combination with standard care.

A retrospective study by Sharma et al. evaluated IV ozonized saline as COVID-19 prophylaxis for healthcare workers (N=64) in a dedicated hospital setting. When compared to health care workers who just received standard prophylaxis, the combination with ozone resulted in significantly lower positive tests [4]. PRELIMINARY (retrospective)

In a single-arm pilot clinical study, Schwartz et al. administered IV ozonized saline solution (200 mL at 3–5 µg/NmL of gas, which is about 0.75 to 1.25 µg/mL dissolved using Madrid's 25% rule [15], daily for 10 days) as a complement to standard care in 25 hospitalized adult patients with mild to severe COVID-19 at Virgen De La Paloma Hospital, Madrid. The authors reported a tendency toward clinical improvement (reduced dyspnea, weakness, and body temperature), improvements in laboratory markers (D-dimer, fibrinogen, LDH, CRP), zero mortality, and no treatment-related adverse events[7]. PRELIMINARY (no control group)

In a single-subject study, Thorp et al. administered IV ozonated normal saline (500 mL at 100 µg/mL) to one subject across seven paired hypoxia-chamber experiments (four with ozone, three without). They found that ozonated saline runs showed a 31% increase in hypoxia tolerance time (p<0.0001) and significantly longer time to oxygen saturation nadir (p<0.05), while control runs without ozone showed a 43% decrease in tolerance (p<0.0001). No infusion-related adverse events were reported [8]. PRELIMINARY

In a non-randomized comparative clinical study, Lebed et al. analyzed treatment outcomes in 98 post-operative brain tumor patients (51 receiving standard intensive therapy alone, 47 receiving standard therapy plus daily IV ozonated saline at 1.2–1.5 mg/L) alongside 30 blood donors as baseline controls. They found that ozone-treated patients demonstrated enhanced anti-edematous effects with periorbital edema resolution beginning by day 3 in most patients and in 100% by day 4. No specific adverse events were reported per group[9]. PRELIMINARY (not randomized)

Pregnancy is a contraindication for ozone therapy in MediSkill's clinic guide. These researchers chose to study it anyway, and we report the study here for completeness, not as a recommendation. In a comparative study, Gretchkanev administered IV ozonated physiological saline (400 mL NaCl saturated at 400 mcg/L, five daily sessions) to 90 patients with threatened abortion as part of complex treatment, compared with 40 patients receiving conventional treatment only (total n=130, divided by trimester). Results indicated that the ozone group showed an approximately 22.1% increase in total plasma antioxidative activity, approximately 3.3x higher progesterone levels, 2.4–3.6x higher placental lactogen, a 20–50% decrease in circulating immune complexes, and clinically faster pain relief with reduced need for spasmolytics. No specific adverse clinical events were reported, and the authors noted that higher ozone concentrations (800 and 1200 mcg/L) produced adverse biochemical effects in vitro, supporting their dose selection[10]. PRELIMINARY (not randomized)

In a randomized clinical trial, 118 patients with widespread progressive psoriasis (ages 18–66) were randomized into three groups: Group I (n=42, traditional therapy), Group II (n=42,  traditional therapy + IV O3SS [200 mL at 2.5 mg/L, mean 7.5 sessions]), and Group III (n=34, plasmapheresis + autotransfusion of ozone-modified erythrocyte suspension plus 200 mL ozonated saline as a bolus, mean 3.6 sessions). Results showed that ozone groups showed more pronounced and faster skin improvement, reductions in seromucoid and C-reactive protein, correction of coagulogram abnormalities, and normalization of lipid peroxidation. Group III was most effective, and ozone was well tolerated across groups[11]. MODERATE (small single-centre RCT)

In a randomized clinical trial, 40 patients with obliterating atherosclerosis of the lower limb arteries (Fontaine stages IIA–IIB) were randomly divided into two equal groups: Group 1 (n=20) receiving standard conservative vascular therapy alone, and Group 2 (n=20) receiving the same baseline therapy plus daily IV infusions of ozonized physiological saline (400 mL at 4–7 mg/L). Results showed that ozone increased fibrinolytic activity and activation of the anticoagulant pathway, with a sustained hypocoagulative effect lasting up to 6 months[12]. MODERATE (small single-centre RCT)

A randomized controlled animal study by Ma et al. studied 90 rabbits with VX2 tumors. Note that this tested ozonated saline injected into the tumor, not an IV drip. Rabbits  were divided into three groups receiving intratumoral injections of normal saline (control), 20 μg/mL ozonated saline, or 40 μg/mL ozonated saline. Both ozonated saline groups showed significantly lower tumor growth rates and increased intratumoral necrosis compared to control, with the mechanism potentially partially mediated by elevated IL-6 and TNF-α[13]. PRELIMINARY (animal study)

In a study focused on erythrocyte rheology, Katiukhin gave a course of IV ozonated saline to patients with complex pathology (dosing as listed in the table) and measured red blood cell aggregation, deformability and membrane permeability after each session and at 2 weeks, 1 month and 2 months. Rheological properties improved after the sessions and stayed improved at follow-up. There was no separate control group[5]. PRELIMINARY

A note on units: the Russian studies report ozone in mg/L, which is the same as µg/mL. Thorp's 100 µg/mL most likely refers to the gas going into the saline, not the ozone dissolved in it. Madrid's limit of 2 µg/mL is for the ozone dissolved in the solution the patient receives, which is only about 25% of the gas concentration bubbled in. Several studies in the table list higher concentrations, many as the gas concentration used to make the solution, so they cannot be compared directly with Madrid's limit. They are what the studies reported, not MediSkill dosing.

Preparation and Administration

Despite requiring no blood draw, ozone saline drip comes with its own risk. The Madrid Declaration says the patient must not receive more than 2 µg/mL of dissolved ozone: going above it is dangerous and can cause phlebitis [15]. The World Federation of Ozone Therapy also warns that higher concentrations may create dangerous levels of chlorine-containing oxidative species [14].

Bubbling and Saturation Protocol

Medical-grade ozone is bubbled through 200 to 250 mL of sterile 0.9% NaCl in an ozone-resistant container. Madrid calls glass the best material and warns that plastic bags release phthalates, which are toxic and raise inflammation [15]. It gives two ways to saturate the solution, both at 200 mL/min: 5 minutes with a microbubbling device, or 15 minutes of conventional bubbling. The dose is set by body weight: 1 µg/kg is low, 2 to 3 µg/kg is medium and 4 to 5 µg/kg is high. Only about 25% of the ozone in the bubbled gas dissolves, so the generator is set to 4 times the dissolved target, and the patient may not receive more than 2 µg/mL dissolved [15]. For an 80 kg person given 200 mL at 1 µg/kg, the dose is 80 µg, or 0.4 µg/mL dissolved (1.6 µg/mL on the generator); the worked table is on the protocol sheet. This range is notably lower than concentrations used in other ozone delivery methods, reflecting the fact that saline cannot buffer reactive species the way blood components can. Bocci et al. reviewed the preparation of ozonated saline and emphasized that ozone-resistant containers and tubings (glass, silicone, or PTFE) are even more important, as standard plastics degrade on contact with ozone and may leach harmful byproducts [1]. 

Infusion Timing

Madrid describes two ways to give the drip [15]. With microbubbling, the solution is saturated for 5 minutes and holds its concentration for more than 30 minutes, so it can be given without bubbling, which removes the risk of gas reaching the vein when no one is watching the end of the drip. With conventional bubbling, the solution is saturated for 15 minutes and given while the gas keeps bubbling, at about 120 drops per minute. Some studies drip the solution over an hour. Where the solution is not kept saturated, any delay between saturation and infusion reduces the dose the patient receives.

How Often

Madrid gives a cycle of 6 to 10 procedures, done every day or every other day depending on the severity of the case and always customized. Its own treatment examples are shorter, and each says to switch to conventional IV administration as soon as possible: for chronic inflammatory and cardiovascular conditions, 1 to 2 µg/kg for 2 to 3 sessions every other day with microbubbling, or 5 daily sessions with conventional bubbling; for acute infections, 3 to 5 µg/kg daily for 3 to 4 sessions with microbubbling, or 5 to 6 with conventional bubbling [15]. The 3 to 5 here is µg per kg of body weight, not µg/mL.

Safety Monitoring and Concentration Limits

Clinicians must monitor the infusion site throughout the procedure for signs of local vein irritation or phlebitis. With conventional bubbling, someone must stay with the patient and stop the bubbling when about 50 mL of solution is left, or gas can reach the vein [15]. Because the ozone saline drip functions through a mechanism described as oxidative preconditioning, strict adherence to concentration limits is essential. 

Since H₂O₂ can potentiate insulin action, ozone saline drip can lower blood sugar. So, healthy patients should be advised to eat prior to the treatment and clinics should be ready to treat hypoglycemia. Patients on insulin or other glucose-lowering medicines should monitor their blood sugar and involve their physician, since doses may need adjusting. 

Contraindications to the ozone saline drip include pregnancy (an absolute contraindication in MediSkill's clinic guide), toxic hyperthyroidism (severe Graves' disease), thrombocytopenia under 50,000 and severe coagulation disorders. Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a consideration (lower antioxidant capacity, so start low and monitor), and an absolute contraindication only for high-dose treatments (10-Pass, EBOO and HiDose). The full contraindication list for IV ozone is on the IV ozone protocol page. Patients with a history of thrombophlebitis should be assessed carefully before intravenous access is established.

For most clinics, major autohemotherapy is the better-studied place to start. Practitioners can learn it in the free IV Ozone 101 training.

This content is for educational purposes and does not constitute medical advice.

References: 

1 Bocci, V. (2010) The actual six therapeutic modalities. In OZONE, pp 35–74, Springer Netherlands, Dordrecht

2 Ma, L., Wen, S., Yuan, J., Zhang, D., Lu, Y.-L., Zhang, Y., et al. (2020) Detection of chlorite, chlorate and perchlorate in ozonated saline. Exp. Ther. Med., Spandidos Publications 20, 2569–2576

3 Sies, H., Berndt, C. and Jones, D. P. (2017) Oxidative stress. Annu. Rev. Biochem., Annual Reviews 86, 715–748

4 Sharma, A., Shah, M., Sane, H., Gokulchandran, N., Paranjape, A., Khubchandani, P., et al. (2021) Intravenous ozonized saline therapy as prophylaxis for healthcare workers (HCWs) in a dedicated COVID-19 hospital in India - A retrospective study. Eur. Rev. Med. Pharmacol. Sci., Eur Rev Med Pharmacol Sci 25, 3632–3639

5 Katiukhin, L. N. (2016) Influence the course of treatment injections of ozonated saline on the rheological properties of erythrocytes in patients with complex pathology. Fiziol. Cheloveka, Pleiades Publishing Ltd 42, 100–105

6 Sharma, A., Shah, M., Lakshmi, S., Sane, H., Captain, J., Gokulchandran, N., et al. (2021) A pilot study for treatment of COVID-19 patients in moderate stage using intravenous administration of ozonized saline as an adjuvant treatment-registered clinical trial. Int. Immunopharmacol., Elsevier BV 96, 107743

7 Ozone in COVID-19 https://jppres.com/jppres/ozone-in-covid-19/

8 Thorp, J. A., Hollonbeck, S. A., Viglione, D. D., Green, P. C., Hodge, J. R., Tamburro, J. A., et al. (2020) Novel therapy for COVID-19 does intravenous ozonated-saline affect blood and tissue oxygenation? J. Gynecol. Res. Obstet., Peertechz Publications Private Limited 6, 046–050

9 Лебедь, С. Л. (2005) Патофизиологическое обоснование применения озонированного физиологического раствора в послеоперационном периоде у больных с новообразованиями головного мозга, кандидат медицинских наук, Нижний Новгород

10 Gretchkanev, G. O. (2001) Ozonetherapy as the main component of the complex treatment of threatened abortion

11 Викторович, Б. В. (2011) Эффективность озонотерапии в комплексном лечении псориаза. Вестник Российского университета дружбы народов. Серия: Медицина 0, 97–102

12 Винник, Ю. С., Дунаевская, С. С. and Антюфриева, Д. А. (2016) ВЛИЯНИЕ ОЗОНИРОВАННОГО ФИЗИОЛОГИЧЕСКОГО РАСТВОРА НА СИСТЕМУ ГЕМОСТАЗА ПРИ ОБЛИТЕРИРУЮЩЕМ АТЕРОСКЛЕРОЗЕ СОСУДОВ НИЖНИХ КОНЕЧНОСТЕЙ. Vestn. St. Petersbg. Univ. Med. 11, 30–35

13 Ma, Q., Yang, C., Jiang, X., Liu, J., Shi, Y., Li, H., et al. (2018) Effectiveness of ozonated saline in the treatment of VX2 tumors in rabbits. J. Interv. Med., J Interv Med 1, 143–149

14 Feng, H. X. and Baeza-Noci, J. (2017) Ozonated saline solution and ozone therapy, World Federation of Ozone Therapy, Bologne

15 Madrid Declaration on Ozone Therapy, 4th edition (ISCO3, 2025), sections 3.3 and 14.9 to 14.10.