library.onepin.app/methylene-blue Peptide Education Library
Education only, not medical advice 21+ Every dose is an example to finalize with a licensed provider

OnePin Peptide Library · Cognitive & Mood

Cognitive & Mood Peptide Not FDA-approved · investigational

Methylene Blue

Methylthioninium Chloride · MB · Tetramethylthionine

Methylene blue (methylthioninium chloride) is a synthetic phenothiazine dye first produced in 1876, making it one of the oldest synthetic drugs in medicine. It is FDA-approved for the treatment of acquired methemoglobinemia (where it acts as an electron carrier to reduce methemoglobin back to functional hemoglobin) and is on the WHO List of Essential Medicines. It has a long history of use in diagnostic procedures, as a surgical dye, and as an antiseptic.,At the mitochondrial level, methylene blue functions as an alternative electron carrier in the electron transport chain, accepting electrons from NADH and transferring them to cytochrome c. This bypass mechanism can sustain ATP production even when Complex I or Complex III are impaired, which has generated significant interest in its potential for neurodegenerative diseases, cognitive enhancement, and aging. At low doses (0.5-2mg/kg), it acts as an electron donor (antioxidant), while at higher doses it paradoxically becomes a pro-oxidant.,Research interest in methylene blue has surged in recent years, particularly for cognitive enhancement, neuroprotection, and mitochondrial support. Preclinical studies show improvements in memory, mitochondrial function, and neuroprotection in various disease models. A modified form (LMTM/TRx0237) has been tested in Phase III Alzheimer's trials with mixed results. Low-dose oral methylene blue is used off-label in nootropic and longevity communities, though human clinical data for cognitive enhancement in healthy individuals remains limited.

Quick Start
Route
Oral
Start low
0.5-1mg/kg
Frequency
1x daily
Timing
Can be taken with food to reduce GI discomfort

Oral or IV. Morning preferred (energizing). Low doses (0.5-2mg/kg). Can stain urine blue.

Start here. Every protocol below is ordered lowest-first. Begin at the smallest effective dose, hold about a week to assess tolerance, and step up only if needed.

Tiered Protocols · lowest first
Quick Start
Conservative start
0.5-1mg/kg · Oral · 1x daily
Lowest starting point. Hold about a week to assess tolerance before stepping up.
0.5-1mg/kg
1x daily
Intermediate
Mitochondrial Respiration
0.5-1 mg/kg (approx 10-50mg) · Oral · Daily
human study · Rojas, Bruchey & Gonzalez-Lima 2011 (Prog Neurobiol 96:32-45)
0.5-1 mg/kg (approx 10-50mg)
Daily
Reconstitution CalculatorU-100
050100u
Draw to
units
01

How it works

Acts as an alternative electron carrier in the mitochondrial electron transport chain, cycling between oxidized (blue) and reduced (colorless leuco-methylene blue) forms. It can accept electrons from NADH and donate them directly to cytochrome c, bypassing Complex I-III blockades. This sustains ATP production and reduces electron leak/ROS generation from dysfunctional mitochondria.,In methemoglobinemia, methylene blue is reduced by NADPH-methemoglobin reductase to leucomethylene blue, which then donates electrons to reduce methemoglobin (Fe3+) back to functional hemoglobin (Fe2+), restoring oxygen-carrying capacity.,Exhibits dose-dependent hormetic behavior: at low concentrations (0.5-2mg/kg), it acts as an antioxidant by improving electron flow and reducing ROS. At high concentrations, it overwhelms reduction capacity and generates superoxide, becoming pro-oxidant. This hormetic window is critical for therapeutic use.

Acts as an alternative electron carrier in the mitochondrial electron transport chain, cycling between oxidized (blue) and reduced (colorless leuco-methylene blue) forms. It can accept electrons from NADH and donate them directly to cytochrome c, bypassing Complex I-III blockades. This sustains ATP production and reduces electron leak/ROS generation from dysfunctional mitochondria.,In methemoglobinemia, methylene blue is reduced by NADPH-methemoglobin reductase to leucomethylene blue, which then donates electrons to reduce methemoglobin (Fe3+) back to functional hemoglobin (Fe2+), restoring oxygen-carrying capacity.,Exhibits dose-dependent hormetic behavior: at low concentrations (0.5-2mg/kg), it acts as an antioxidant by improving electron flow and reducing ROS. At high concentrations, it overwhelms reduction capacity and generates superoxide, becoming pro-oxidant. This hormetic window is critical for therapeutic use.

02

What to expect

Early
Days 1–7

Days 1-7: Blue urine (normal and expected). Possible subtle cognitive effects. Mild GI adjustment period.

03

Evidence

HumanStrong
AnimalStrong
In-vitroPresent
2016Multimodal Randomized Functional MR Imaging of the Effects of Methylene Blue in the Human Brain · Rodriguez P et al, Radiology ↗peer reviewed
2016Methylene blue modulates functional connectivity in the human brain · Rodriguez P et al, Brain Imaging and Behavior ↗peer reviewed
2014Therapeutic benefits of methylene blue on cognitive impairment during chronic cerebral hypoperfusion · Auchter A et al, Journal of Alzheimer's Disease ↗peer reviewed
2018Potential of Low Dose Leuco-Methylthioninium Bis(Hydromethanesulphonate) (LMTM) Monotherapy for Treatment of Mild Alzheimer's Disease · Wilcock GK et al, Journal of Alzheimer's Disease ↗peer reviewed
04

Interactions & stacking

What's safe to combine, and what belongs in a separate pin.

Safe & synergistic

RapamycinmTOR inhibition and mitochondrial electron transport support address aging through independent mechanisms
EpicatechinBoth support mitochondrial function - epicatechin via PGC-1alpha biogenesis, methylene blue via electron transport bypass
EpithalonNo direct interaction - telomerase activation and mitochondrial support are complementary
BPC-157No pharmacological interaction - BPC-157 tissue repair works via distinct pathways (NO, GH)

Keep separate

SSRIs and SNRIsMethylene blue is a potent MAO-A inhibitor - combining with serotonergic medications causes potentially fatal serotonin syndrome
Injectable Vitamin CVitamin C can reduce methylene blue to leucomethylene blue, disrupting its electron carrier cycling and therapeutic mechanism
MelatoninMethylene blue inhibits MAO-A which metabolizes serotonin and melatonin - may cause unpredictable melatonin accumulation
Dasatinib+QuercetinQuercetin has redox activity that may interfere with methylene blue's precise hormetic dose-response - unpredictable oxidative interactions
05

Pharmacokinetics

Illustrative plasma concentration · 24 h
Half-life
~5-6 hours (terminal half-life ~24 hours due to tissue distribution)
Peak · Tmax
~1-2 hours oral, immediate IV
Elimination
~4-5 days (due to extensive tissue binding and long terminal half-life)
Activity
12-24 hours (mitochondrial electron carrier effects persist due to tissue retention)

Storage. Store at room temperature (15-25C). Protect from light. Aqueous solutions are stable but light-sensitive. Pharmaceutical grade (USP) required for medical use.

06

Regulatory status

FDA-approved (ProvayBlue, generic) for treatment of acquired methemoglobinemia. Investigational for Alzheimer's disease (LMTM failed Phase III primary endpoints). Off-label use for cognitive enhancement, vasoplegic syndrome, and ifosfamide encephalopathy. WHO Essential Medicine.

Put it to work

Calculate, log & track

Open Methylene Blue straight into OnePin with your vial and dose pre-filled, then let it handle the syringe math and remind you before the vial runs out.

Open Methylene Blue in the app →

Go deeper

The guide & community

The complete Methylene Blue walkthrough, real protocol discussion and coaching live inside the BlessUp community on Skool.

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