1. Methylene blue trihydrate

Methylene blue trihydrate  (Synonyms: C.I. Basic Blue 9 trihydrate)

Cat. No.: HY-B1359 Purity: ≥98.0%
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Methylene blue trihydrate (C.I. Basic Blue 9 trihydrate) is a guanylyl cyclase (sGC), monoamine oxidase A (MAO-A) and NO synthase (NOS) inhibitor. Methylene blue trihydrate is a vasopressor and is often used as a dye in several medical procedures. Methylene blue trihydrate has antinociception, antimalarial, antidepressant and anxiolytic activity effects. Methylene Blue trihydrate has the potential for methemoglobinemias, neurodegenerative disorders and ifosfamide-induced encephalopathytreatment.

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Methylene blue trihydrate Chemical Structure

Methylene blue trihydrate Chemical Structure

CAS No. : 7220-79-3

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Solid + Solvent
10 mM * 1 mL in DMSO
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Customer Review

Based on 13 publication(s) in Google Scholar

Other Forms of Methylene blue trihydrate:

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  • Biological Activity

  • Purity & Documentation

  • Customer Review

Description

Methylene blue trihydrate (C.I. Basic Blue 9 trihydrate) is a guanylyl cyclase (sGC), monoamine oxidase A (MAO-A) and NO synthase (NOS) inhibitor. Methylene blue trihydrate is a vasopressor and is often used as a dye in several medical procedures. Methylene blue trihydrate has antinociception, antimalarial, antidepressant and anxiolytic activity effects. Methylene Blue trihydrate has the potential for methemoglobinemias, neurodegenerative disorders and ifosfamide-induced encephalopathytreatment[1][2][3].

IC50 & Target

Guanylyl cyclase (sGC)[1].
Monoamine oxidase A (MAO-A)[1].
NO synthase (NOS)[1]

In Vitro

By acting as an alternative electron acceptor/donor Methylene blue restores mitochondrial function, improves neuronal energy production and inhibits the formation of superoxide[1].
Methylene blue inhibits cytochrome P450 (CYP) isozymes. Methylene blue is an odorless, water soluble, dark blue-green crystalline powder, which turns blue whenever mixed in solution. Methylene blue is a vasopressor that impacts the NO synthetic pathway by inhibiting inducible NOS and inhibiting the subsequent activation of sGC. In addition, by binding to the iron heme moiety of sGC and causing enzyme inhibition, Methylene blue blocks accumulation of cyclic GMP (cGMP), competing directly with NO in its ability to activate soluble guanylyl cyclase[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

In Vivo

Methylene Blue (1, 5, and 25 μg/rat) significantly decreases sevoflurane minimum alveolar anesthetic concentration (MAC) and brain cyclic guanosine monophosphate (cGMP) content in a dose-dependent manner in male Sprague-Dawley rats (7-week-old, 200-250 g)[2].
Methylene blue is used as a dye in chromoendoscopy, and is sprayed onto the mucosa of the gastrointestinal tract in order to identify dysplasia, or pre-cancerous lesions[2].
Methylene blue is able to restore vascular tone, normalize mean arterial pressures (MAP), and reduce vasopressor usage[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Clinical Trial
Molecular Weight

373.90

Formula

C16H24ClN3O3S

CAS No.
Appearance

Solid

Color

Light brown to black

SMILES

CN(C1=CC(SC2=C/C(C=CC2=N3)=[N+](C)\C)=C3C=C1)C.O.[Cl-]

Shipping

Room temperature in continental US; may vary elsewhere.

Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 6 months
-20°C 1 month
Solvent & Solubility
In Vitro: 

DMSO : 100 mg/mL (267.45 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.6745 mL 13.3726 mL 26.7451 mL
5 mM 0.5349 mL 2.6745 mL 5.3490 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (6.69 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.5 mg/mL (6.69 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: ≥98.0%

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 2.6745 mL 13.3726 mL 26.7451 mL 66.8628 mL
5 mM 0.5349 mL 2.6745 mL 5.3490 mL 13.3726 mL
10 mM 0.2675 mL 1.3373 mL 2.6745 mL 6.6863 mL
15 mM 0.1783 mL 0.8915 mL 1.7830 mL 4.4575 mL
20 mM 0.1337 mL 0.6686 mL 1.3373 mL 3.3431 mL
25 mM 0.1070 mL 0.5349 mL 1.0698 mL 2.6745 mL
30 mM 0.0892 mL 0.4458 mL 0.8915 mL 2.2288 mL
40 mM 0.0669 mL 0.3343 mL 0.6686 mL 1.6716 mL
50 mM 0.0535 mL 0.2675 mL 0.5349 mL 1.3373 mL
60 mM 0.0446 mL 0.2229 mL 0.4458 mL 1.1144 mL
80 mM 0.0334 mL 0.1672 mL 0.3343 mL 0.8358 mL
100 mM 0.0267 mL 0.1337 mL 0.2675 mL 0.6686 mL
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  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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Methylene blue trihydrate
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