Methylene Blue

• Mitochondrial Function Enhancement
• Neuroprotection
• Cellular Energy Production
• Antioxidant Activity

• Cognitive Enhancement
• Memory Support
• Mood Regulation
• Antimicrobial Properties
• Methemoglobinemia Treatment

Methylene Blue (MB) exerts its biological effects through multiple mechanisms, with its primary action centered on its role as an electron cycler in mitochondrial respiration. MB acts as an alternative electron carrier in the electron transport chain (ETC), accepting electrons from NADH and transferring them to cytochrome c, effectively bypassing complexes I-III. This unique property allows MB to enhance mitochondrial respiration and ATP production, particularly when the normal ETC function is compromised. At low doses (0.5-4 mg/kg), MB increases cytochrome c oxidase (complex IV) activity, enhancing oxygen consumption and ATP production.

This mitochondrial enhancement is particularly significant in tissues with high energy demands, such as the brain and heart. MB’s redox cycling properties also contribute to its antioxidant effects. In its reduced form (leucomethylene blue), MB can scavenge reactive oxygen species (ROS) and reactive nitrogen species (RNS), including superoxide, hydrogen peroxide, and peroxynitrite. Unlike conventional antioxidants that become depleted after neutralizing free radicals, MB can be regenerated through the ETC, allowing for sustained antioxidant activity.

In the central nervous system, MB exhibits neuroprotective properties through multiple mechanisms. It inhibits monoamine oxidase (MAO), increasing levels of neurotransmitters like serotonin, dopamine, and norepinephrine, which may contribute to its effects on mood and cognition. MB also inhibits nitric oxide synthase (NOS), reducing nitric oxide production and subsequent peroxynitrite formation, which can damage neurons. Additionally, MB inhibits the aggregation of tau protein and prevents the formation of amyloid plaques, two pathological hallmarks of Alzheimer’s disease.

This may explain its potential benefits in neurodegenerative conditions. MB influences cellular signaling pathways, including the activation of the Nrf2-Keap1 pathway, which upregulates antioxidant defense systems and phase II detoxification enzymes. It also modulates the mTOR pathway, potentially influencing cellular metabolism, autophagy, and protein synthesis. In the context of methemoglobinemia (a condition where hemoglobin is oxidized and cannot transport oxygen effectively), MB acts as an electron donor to reduce methemoglobin back to functional hemoglobin, restoring oxygen-carrying capacity.

This is its most well-established medical use. MB also possesses antimicrobial properties through multiple mechanisms. It can intercalate with DNA, disrupting microbial replication. When photoactivated, MB can generate singlet oxygen, which damages microbial cell membranes and proteins.

It can also interfere with electron transport in microbial cells, disrupting their energy metabolism. At the cellular level, MB can modulate autophagy, the process by which cells recycle damaged components. This may contribute to its potential anti-aging effects by promoting the clearance of dysfunctional mitochondria and protein aggregates. MB’s diverse mechanisms of action explain its wide range of potential applications, from treating methemoglobinemia and septic shock to potential benefits in neurodegenerative diseases, psychiatric conditions, and antimicrobial applications.

For cognitive and mitochondrial support (non-medical use): 0.5-2 mg/kg body weight, typically translating to 30-100 mg for an average adult. Medical applications may use different dosages under professional supervision.

Methylene Blue has moderate oral bioavailability, estimated at approximately 40-70%. It is rapidly absorbed from the gastrointestinal tract, with peak plasma concentrations typically occurring within 1-2 hours after ingestion. The compound readily crosses the blood-brain barrier, particularly in its reduced form (leucomethylene blue), allowing it to exert effects in the central nervous system.

Taking on an empty stomach may improve absorption, though it may increase gastrointestinal side effects, Sublingual administration potentially bypasses first-pass metabolism, Liposomal formulations may enhance cellular uptake and bioavailability, Enteric-coated preparations can protect from stomach acid degradation, Combining with a small amount of vitamin C may enhance reduction to the leucomethylene blue form, which more readily crosses the blood-brain barrier, Nanoparticle delivery systems are being investigated to improve bioavailability, Avoiding high-fat meals during administration, as they may delay absorption

For cognitive enhancement or mitochondrial support, Methylene Blue is typically taken in the morning or early afternoon to avoid potential sleep disruption due to its energizing effects. Taking on an empty stomach approximately 30 minutes before meals may optimize absorption, though some individuals may need to take it with a light meal to reduce gastrointestinal discomfort. For those using MB for cognitive performance, taking 30-60 minutes before mentally demanding tasks may be beneficial. Avoid taking within 4 hours of bedtime to prevent potential sleep disturbances.

When used for antimicrobial purposes (under medical supervision), timing may vary based on the specific protocol. For medical applications such as methemoglobinemia treatment, timing is determined by medical necessity and administered by healthcare professionals. Consistent daily timing is recommended for regular use to maintain stable blood levels.

• Blue/green discoloration of urine and feces (harmless but expected)
• Temporary blue discoloration of skin with higher doses
• Gastrointestinal discomfort, nausea
• Headache
• Dizziness
• Anxiety or restlessness
• Increased blood pressure (dose-dependent)
• Photosensitivity (increased sensitivity to sunlight)
• Hemolytic anemia (rare, primarily in G6PD-deficient individuals)
• Serotonin syndrome (when combined with serotonergic medications)

• Glucose-6-phosphate dehydrogenase (G6PD) deficiency – can cause severe hemolytic anemia
• Current use of serotonergic medications (SSRIs, SNRIs, MAOIs, triptans, certain opioids) – risk of serotonin syndrome
• Pregnancy and breastfeeding – insufficient safety data, potential risks to fetus/infant
• Severe renal impairment – altered clearance and potential toxicity
• Known hypersensitivity to methylene blue or similar dyes
• Intraspinal injection – neurotoxicity risk
• Patients undergoing serotonergic anesthesia
• Children (for non-medical use)

• Serotonergic medications (SSRIs, SNRIs, MAOIs, triptans, tramadol, etc.) – risk of potentially fatal serotonin syndrome
• Dapsone – may increase risk of hemolytic reactions
• Primaquine and other oxidative medications – increased risk of hemolysis
• Acetaminophen (paracetamol) – MB may reduce its effectiveness
• Aniline dyes – potential additive toxicity
• Calcium channel blockers – potential interaction affecting blood pressure
• Antipsychotic medications – potential for CNS effects
• Hypoglycemic agents – may affect blood glucose levels

For non-medical use, doses above 4 mg/kg are generally not recommended due to increased risk of side effects and potential paradoxical inhibition of mitochondrial function. Medical applications may use higher doses under professional supervision for specific conditions. The FDA-approved dose for methemoglobinemia is 1-2 mg/kg, with cumulative doses not to exceed 7 mg/kg due to risk of hemolytic anemia. Long-term safety of regular use has not been well-established in clinical trials.

In the United States, Methylene Blue has a complex regulatory status. It is FDA-approved as a prescription medication for several specific medical conditions: 1) Treatment of methemoglobinemia (acquired and congenital forms); 2) Visualization aid during surgical procedures; 3) Treatment of cyanide poisoning (as part of the cyanide antidote kit). The FDA has issued specific warnings about the risk of serotonin syndrome when Methylene Blue is used in patients taking serotonergic psychiatric medications. As a prescription medication, it is not legally marketed as a dietary supplement under DSHEA regulations.

However, some compounding pharmacies may prepare Methylene Blue formulations under prescription for off-label uses. The FDA has not evaluated or approved Methylene Blue for cognitive enhancement, anti-aging, or other non-medical uses that have gained popularity in biohacking communities.

Eu: In the European Union, Methylene Blue is approved as a medicinal product for treating methemoglobinemia and as a diagnostic aid. The European Medicines Agency (EMA) has issued warnings similar to the FDA’s regarding the risk of serotonin syndrome. A derivative of Methylene Blue (LMTX/TRx0237) has been in clinical trials for Alzheimer’s disease and frontotemporal dementia in Europe.

Canada: Health Canada regulates Methylene Blue as a prescription drug for methemoglobinemia and as a medical dye. It is not approved as a natural health product or supplement.

Australia: The Therapeutic Goods Administration (TGA) classifies Methylene Blue as a prescription-only medicine for treating methemoglobinemia and as a diagnostic agent. It is not approved as a complementary medicine.

Japan: In Japan, Methylene Blue is approved as a pharmaceutical for methemoglobinemia and as a medical dye. It is not approved as a food supplement or functional food.

Uk: The Medicines and Healthcare products Regulatory Agency (MHRA) regulates Methylene Blue as a prescription-only medicine for treating methemoglobinemia and as a diagnostic aid.

International Organizations: The World Health Organization (WHO) includes Methylene Blue on its List of Essential Medicines for the treatment of methemoglobinemia.

medium to high

$1.00-$5.00 per day for typical doses (30-100 mg of pharmaceutical-grade)

Methylene Blue presents a complex value proposition that varies significantly based on source, quality, and intended use. Pharmaceutical-grade (USP) Methylene Blue is moderately expensive compared to many common supplements, with monthly costs ranging from $30-150 depending on dosage and source. This higher cost reflects the need for pharmaceutical-grade purity and specialized manufacturing processes. The value proposition is strongest for specific medical conditions where Methylene Blue has established efficacy, such as methemoglobinemia, where it is often the treatment of choice and highly cost-effective compared to alternatives.

For cognitive enhancement and neuroprotection, the cost-efficiency is less clear due to limited clinical evidence, though some users report significant benefits that may justify the cost. When compared to other cognitive enhancers or mitochondrial support supplements, Methylene Blue is generally more expensive than options like CoQ10 or alpha-lipoic acid, but potentially more effective for some users, particularly at enhancing mitochondrial function. The cost-efficiency is significantly reduced when considering non-pharmaceutical grade sources (which should be avoided due to safety concerns) or when used without proper medical guidance. Compounded formulations from pharmacies tend to be more expensive than commercial pharmaceutical preparations but may offer specific dosing or delivery advantages.

For research purposes, the cost is generally justified by Methylene Blue’s unique mechanism of action and well-documented effects on mitochondrial function. The long history of medical use and established safety profile (when used appropriately) adds value compared to newer, less-studied compounds. Overall, Methylene Blue represents a moderate to high-cost intervention with potentially significant benefits for specific applications, but requires careful sourcing and ideally medical supervision to maximize its value and safety profile.

Pharmaceutical-grade methylene blue solutions: 2-3 years when properly stored; Tablets/capsules: 2-3 years; Compounded formulations: typically 6 months to 1 year depending on preparation method and storage conditions

Store in tightly closed, light-resistant containers (amber or opaque bottles). Keep at room temperature (15-30°C or 59-86°F), away from direct light, which can degrade the compound. Protect from excessive heat and freezing temperatures. Some liquid formulations may require refrigeration after opening – follow specific product guidelines.

Avoid exposure to strong oxidizing or reducing agents that can alter methylene blue’s redox state. Keep away from moisture, as methylene blue is hygroscopic and can absorb water from the environment. Once a solution is prepared or a container is opened, the shelf life may be reduced – follow specific product recommendations for use after opening.

Light exposure (particularly UV light) – methylene blue is highly photosensitive and can degrade when exposed to light, Oxidizing agents – can alter methylene blue’s redox state, Reducing agents – can convert methylene blue to leucomethylene blue, changing its properties, Extreme pH conditions – stability is optimal at neutral to slightly acidic pH, Heat – accelerates degradation reactions, Moisture – can promote hydrolysis and degradation, Microbial contamination – particularly in liquid formulations, Metal ions – can catalyze degradation reactions, Prolonged storage in solution form – gradual degradation occurs even under optimal conditions

Synthesis Methods

Natural Sources

Quality Considerations

Methylene Blue has a rich and diverse history spanning nearly 150 years, evolving from a textile dye to an important pharmaceutical agent with multiple medical applications. The compound was first synthesized in 1876 by Heinrich Caro at BASF in Germany, initially developed as a textile dye for cotton. Its journey into medicine began in 1891 when Paul Ehrlich, a pioneer of chemotherapy, discovered that Methylene Blue could selectively stain malaria parasites. This led to its first medical application when Ehrlich and Paul Guttmann used it to treat malaria in 1891, making it one of the first synthetic drugs used in modern medicine.

During the early 20th century, Methylene Blue’s applications expanded. In 1933, Bernhard Heinz discovered its effectiveness in treating methemoglobinemia, a condition where hemoglobin is unable to effectively release oxygen to tissues. This remains one of its primary FDA-approved medical uses today. During World War II, Methylene Blue was included in military first aid kits as a treatment for cyanide poisoning, though this application has since been superseded by more effective antidotes.

In the 1950s, Methylene Blue’s properties as a monoamine oxidase inhibitor (MAOI) were discovered, revealing its potential effects on neurotransmitter systems. This finding would later become relevant to its psychiatric and neurological applications. The 1960s and 1970s saw Methylene Blue used in surgical contexts as a dye for identifying tissues and tracing lymphatic drainage. It became an important tool in sentinel lymph node biopsies and for identifying parathyroid glands during surgery.

In the 1980s, researchers began investigating Methylene Blue’s potential in treating vasoplegic syndrome, a form of severe hypotension that can occur during cardiac surgery. This application has become standard practice in many cardiac surgical centers. The 1990s marked the beginning of serious investigation into Methylene Blue’s neuroprotective properties. Researchers discovered its ability to inhibit nitric oxide synthase and its effects on the electron transport chain in mitochondria, suggesting potential benefits for neurodegenerative conditions.

In the early 2000s, interest in Methylene Blue’s cognitive-enhancing properties grew, with studies demonstrating improvements in memory and attention in both animal models and human subjects. This led to increased interest in its potential for treating conditions like Alzheimer’s disease and other forms of dementia. A significant development came in 2008 when researchers at the University of Aberdeen developed a derivative of Methylene Blue called Rember (methylthioninium chloride), which showed promise in clinical trials for Alzheimer’s disease by inhibiting tau aggregation. This was later reformulated as LMTX (TRx0237), which continues to be investigated in clinical trials.

In recent years, Methylene Blue has gained attention in the biohacking and anti-aging communities due to its effects on mitochondrial function and potential neuroprotective properties. While not approved for these uses, some individuals use low-dose Methylene Blue as a cognitive enhancer or for potential anti-aging benefits. Throughout its history, Methylene Blue has demonstrated remarkable versatility, transitioning from an industrial dye to a vital medical treatment and now to a compound of interest for its potential cognitive and neuroprotective properties. This evolution reflects both the serendipitous nature of many medical discoveries and the value of continuing to investigate established compounds for new therapeutic applications.

Tucker D, Lu Y, Zhang Q. (2018) From Mitochondrial Function to Neuroprotection—an Emerging Role for Methylene Blue. Molecular Neurobiology, Rojas JC, Bruchey AK, Gonzalez-Lima F. (2012) Neurometabolic mechanisms for memory enhancement and neuroprotection of methylene blue. Progress in Neurobiology

Clinical trials evaluating methylene blue derivatives (e.g., TRx0237/LMTX) for Alzheimer’s disease and frontotemporal dementia, Studies investigating methylene blue for traumatic brain injury and post-traumatic stress disorder, Research on methylene blue’s potential in treating COVID-19-related complications, Investigations into methylene blue’s effects on mitochondrial function in aging and neurodegenerative diseases