Methylene Blue

Methylene Blue structural formula

CAS Number61-73-4
Molecular FormulaC16H18ClN3S
Molecular Weight319.851
InChI KeyCXKWCBBOMKCUKX-UHFFFAOYSA-M
LogP2.72
Synonyms
  • Methylene blue
  • 3,7-Bis(dimethylamino)phenothiazin-5-ium chloride
  • Phenothiazinium, 3,7-bis(dimethylamino)-, chloride (1:1)
  • 61-73-4
  • Phenothiazin-5-ium, 3,7-bis(dimethylamino)-, chloride
  • 3,7-BIS(DIMETHYLAMINO)PHENAZATHIONIUM CHLORIDE
  • 3,7-Bis(dimethylamino)phenothiazin-5-ium chloride
  • C.I. basic blue 009
  • Chlorure de methylthioninium
  • Chromosmon
  • cloruro de metiltioninio
  • Izit Crystal Dye
  • METHYLENBLAU
  • Methylene blue polychrome
  • Methylenium ceruleum
  • METHYLTHIONINE CHLORIDE
  • methylthioninium chloride
  • Methylthioniniumchlorid
  • NSC 617593
  • Phenothiazin-5-ium, 3,7-bis(dimethylamino)-, chloride (1:1)
  • Schultz 1038
  • Swiss blue
  • Tetramethylthionine
  • Tetramethylthionine chloride
  • D and C Blue No. 1
  • D And C Blue Number 1
  • C.I. Basic Blue 9
  • M-B Tabs
  • Aizen Methylene Blue BH
  • Aizen Methylene Blue FZ
  • Calcozine Blue ZF
  • Caswell No. 567
  • Ceruleum methylenum
  • CI Basic Blue 9
  • EINECS 200-515-2
  • EPA Pesticide Chemical Code 039505
  • Ext D and C Blue No. 1
  • External Blue 1
  • Hidaco Methylene Blue Salt Free
  • Leather Pure Blue HB
  • Methylene Blue A
  • Methylene Blue B
  • Methylene Blue D
  • Methylene Blue G
  • Methylene Blue N
  • Methylene Blue 2B
  • Methylene Blue 2BF
  • Methylene Blue 2BN
  • Methylene Blue 2BP
  • Methylene Blue BB
  • Methylene Blue BBA
  • Methylene Blue BD
  • Methylene Blue BP
  • Methylene Blue BPC
  • Methylene Blue BX
  • Methylene Blue BZ
  • Methylene Blue chloride
  • Methylene Blue FZ
  • Methylene Blue GZ
  • Methylene Blue HGG
  • Methylene Blue IAD
  • Methylene Blue JFA
  • Methylene Blue NZ
  • Methylene Blue SG
  • Methylene Blue SP
  • Methylene Blue ZF
  • Methylene Blue Zinc Free
  • Methylene Blue ZX
  • Methylenum coeruleum
  • Methylthionini chloridum
  • Methylthionium chloride
  • Mitsui Methylene Blue
  • NSC 215213
  • Sandocryl Blue BRL
  • Schultz No. 1038
  • Tetramethylene Blue
  • Urolene Blue
  • Yamamoto Methylene Blue B
  • Yamamoto Methylene Blue ZF
  • Methylthioninii chloridum
  • Metiltioninio cloruro
  • Modr Methylenova
  • Modr Rozpoustedlova 8
  • Modr Zasadita 9
  • UNII-8NAP7826UB
  • Basic Blue 9
  • C.I. 52015
  • Methylene Blue anhydrous
  • Solvent blue 8
  • azul de metileno
  • bleu de methylene
  • 152071-32-4
  • 1001913-24-1
  • 1001914-35-7
  • 105504-42-5
  • 121067-62-7
  • 1219638-11-5
  • 12262-49-6
  • 1341-90-8
  • 167498-52-4
  • 6476-03-5
  • 923927-02-0
  • 97130-83-1

Applications:

UV-Vis Spectrum of Methylene Blue

July 16, 2024
UV-Vis Spectrum of Methylene Blue. Absorption Maxima: 246 nm, 292 nm.

For optimal results in HPLC analysis, it is recommended to measure absorbance at a wavelength that matches the absorption maximum of the compound(s) being analyzed. The UV spectrum shown can assist in selecting an appropriate wavelength for your analysis. Please note that certain mobile phases and buffers may block wavelengths below 230 nm, rendering absorbance measurement at these wavelengths ineffective. If detection below 230 nm is required, it is recommended to use acetonitrile and water as low UV-transparent mobile phases, with phosphoric acid and its salts, sulfuric acid, and TFA as buffers.
For some compounds, the UV-Vis Spectrum is affected by the pH of the mobile phase. The spectra presented here are measured with an acidic mobile phase that has a pH of 3 or lower.

Application Analytes:
Methylene Blue
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.

HPLC Method for Analysis of Methylene blue on Primesep 100 Column

December 7, 2022

High Performance Liquid Chromatography (HPLC) Method for Analysis of Methylene blue Primesep 100 by SIELC Technologies.

Separation type: Liquid Chromatography Mixed-mode

HPLC Method for Analysis of Methylene blue on Primesep 100 Column
HPLC Method for Analysis of Methylene blue on Primesep 100 Column by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analyses of Methylene blue

Methylene blue can be retained and analyzed on a Primesep 100 mixed-mode stationary phase column using an isocratic analytical method with a simple isocratic mobile phase of water, Acetonitrile (MeCN), and a sulfuric acid (H2SO4) buffer.  This analysis method can be UV detected at 600 nm with high resolution and peak symmetry.

Condition

ColumnPrimesep 100, 4.6×150 mm, 5 µm, 100A
Mobile PhaseMeCN/H2O – 80/20%
BufferH3PO4 – 0.2%
Flow Rate1.0 ml/min
DetectionUV, 600 nm
Peak Retention Time5.92 min

Description

Class of CompoundsDyes
Analyzing CompoundsMethylene blue

Application Column

Primesep 100

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
Methylene Blue
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.

Analysis of Methylene Blue Purity in Reversed-Phase Cation-Exchange and Reversed-Phase Cation-Exclusion Modes

December 5, 2013

Methylene blue is a heterocyclic dye with a wide range of use in biology and chemistry. Purity of dyes rarely exceed 70% and separation of methylene blue and related impurities is required. Two alternative methods for purity analysis were developed on Primesep C and Primesep D mixed-mode HPLC columns. Primesep C is a reversed-phase cation-exchange (anion-exclusion column) column and Primesep D is reversed-phase anion-exchange (cation-exclusion) column. Various buffers can be used for the method. Depending on the buffer, method can be compatible with UV, LC/MS and prep chromatography. Method can be used for prep separation of methylene blue and related impurities.

Condition 1

Column Primesep D, 4.6×150 mm, 5 µm, 100A
Mobile Phase Gradient MeCN – 5-20%
Buffer Gradient NaH2PO4 pH 2.5 70-20mM 15 min
Flow Rate 1.0 ml/min
Detection UV, 300 nm

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Condition 2

Column Primesep C, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN – 35%
Buffer AmFm pH 3.5 – 35 mM
Flow Rate 1.0 ml/min
Detection UV, 300 nm

 

Description

Class of Compounds
 Duy, Hydrophobic, Ionizable
Analyzing Compounds  Methylene Blue

 

Application Column

Primesep D

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Primesep C

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
Methylene Blue

Application Detection:
ELSD Detection
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.

HPLC Dye Separation on SHARC1 Column

July 10, 2013

 

Condition

Column Sharc 1, 3.2×100 mm, 3 µm, 100A
Mobile Phase MeCN/EtOH – 99/1%
Buffer H2SO4 – 0.5%
Flow Rate 1.5 ml/min
Detection UV, 500, 650 nm

 

Description

Class of Compounds
Dye, Hydrophilic, Ionizable
Analyzing Compounds Methyl Red, Methyl Orange, Methylene Blue, Nile Blue A

 

Application Column

SHARC 1

The SHARC™ family of innovative columns represents the first commercially available columns primarily utilizing separation based on hydrogen bonding. SHARC stands for Specific Hydrogen-bond Adsorption Resolution Column. Hydrogen bonding involves an interaction or attraction between a bound hydrogen atom and molecules containing electronegative atoms, such as oxygen, nitrogen, and fluorine.

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Application Analytes:
Methyl Orange
Methyl Red
Methylene Blue
Nile Blue A

Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.

HPLC Separation of Dyes

November 21, 2010


HPLC.cloud
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Condition 1

Column Newcrom B, 3.2×50 mm, 3 µm, 100A
Mobile Phase MeCN/H2O – 30/70%
Buffer H3PO4 – 0.2%
Flow Rate 0.5 ml/min
Detection UV, 275 nm

 


Various dyes are widely used in food, textile, and pharmaceutical industries. Dies are aromatic compounds containing ionizable groups. Methylene blue, crystal violet lactone, and crystal violet were separated on a Primesep D HPLC column. Method can be used for separation of other dyes and pigments that are basic and hydrophobic in nature.

Condition  2

Column Primesep D, 4.6×50 mm, 3 µm, 100A
Mobile Phase MeCN
Buffer H3PO4 – 0.2%
Flow Rate 0.5 ml/min
Detection UV, 300 nm

 

Description

Class of Compounds
Dye, Hydrophilic, Ionizable
Analyzing Compounds Methylene Blue, Crystal violet lactone, Crystal violet

 

Application Column

Newcrom B

The Newcrom columns are a family of reverse-phase-based columns. Newcrom A, AH, B, and BH are all mixed-mode columns with either positive or negative ion-pairing groups attached to either short (25 Å) or long (100 Å) ligand chains. Newcrom R1 is a special reverse-phase column with low silanol activity.

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Primesep D

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
Crystal Violet
Crystal Violet Lactone
Methylene Blue

Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.