Tetramethylbenzidine Chemical Properties

Melting point:

168-171 °C(lit.)

Boiling point:

100 °C

Density 

1

refractive index 

1.5519 (estimate)

storage temp. 

2-8°C

solubility 

Slightly soluble. <0.1 g/100 mL at 20°C.

pka

4.49±0.10(Predicted)

form 

tablet

color 

Cream

Water Solubility 

Slightly soluble. <0.1 g/100 mL at 20 ºC

Sensitive 

Light Sensitive

BRN 

2808541

Stability:

Stable, but moisture sensitive and may be light sensitive. Incompatible with water, strong oxidizing agents.

CAS DataBase Reference

54827-17-7(CAS DataBase Reference)

EPA Substance Registry System

3,3',5,5'-Tetramethylbenzidine (54827-17-7)

Safety Information

Hazard Codes 

Xi,Xn

Risk Statements 

36/38-36/37/38-22-41

Safety Statements 

26-36-36/37-39

RIDADR 

UN 2796 8/PG 2

WGK Germany 

3

RTECS 

DV2300000

F 

8-10

TSCA 

Yes

HazardClass 

6.1

HS Code 

29215990

Hazardous Substances Data

54827-17-7(Hazardous Substances Data)

MSDS

• Language:English Provider:3,3',5,5'-Tetramethylbenzidine
• Language:English Provider:SigmaAldrich
• Language:English Provider:ACROS
• Language:English Provider:ALFA

Tetramethylbenzidine Usage And Synthesis

Description

Tetramethylbenzidine(TMB) is a chromogenic substrate used in staining procedures in immunohistochemistry as well as being a visualising reagent used in enzyme-linked immunosorbent assays (ELISA). It is a white solid that forms a pale blue-green liquid in solution with ethyl acetate. TMB is degraded by sunlight and by fluorescent lights. It is a soluble chromogen substrate for horseradish peroxidase detection systems. Tetramethylbenzidine is recommended for ELISA procedures.

Chemical Properties

Tetramethylbenzidine is a white or light yellow solid, odorless, tasteless, insoluble in water, easily soluble in acetone, ether, dimethyl sulfoxide, dimethylformamide and other organic solvents. It forms a pale blue-green liquid in solution with ethyl acetate. TMB is degraded by sunlight and by fluorescent lights.

Uses

3,3,5,5-Tetramethyl benzidine is used as a reagent in a sensitive staining procedure for the detection of low levels of heme-associated peroxidase activity of cytochrome P-450 on SDS-polyacrylamide or agarose gel; non-carcinogenic substitute for benzidine as reagent for the detection of blood and determination of hemoglobin content.

Preparation

The synthesis of 3,3',5,5'-Tetramethylbenzidine involved using 2,6-dimethylaniline as the raw material. The process included activation, oxidative coupling, and purification, resulting in the production of pure material. The yield of 3,3,5,5-tetramethylbenzidine was 65%.

Reactions

TMB can act as a hydrogen donor for the reduction of hydrogen peroxide to water by peroxidase enzymes such as horseradish peroxidase.

Shows the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to 3,3',5,5'-tetramethylbenzidine diimine
The resulting diimine causes the solution to take on a blue colour, and this colour change can be read on a spectrophotometer at the wavelengths of 370 and 650 nm.

General Description

3,3',5,5'-tetramethylbenzidine appears as pale yellow crystals or off-white powder. (NTP, 1992)

Air & Water Reactions

Insoluble in water.

Reactivity Profile

Tetramethylbenzidine is sensitive to prolonged exposure to light . Neutralizes acids in exothermic reactions to form salts plus water. May be incompatible with isocyanates, halogenated organics, peroxides, phenols (acidic), epoxides, anhydrides, and acid halides. Flammable gaseous hydrogen may be generated in combination with strong reducing agents, such as hydrides.

Fire Hazard

Flash point data for Tetramethylbenzidine are not available, however, Tetramethylbenzidine is probably combustible.

Biochem/physiol Actions

3,3′,5,5′-Tetramethylbenzidine/TMB can be used as a chromogen to increase the progression of product obtained in peroxidase reaction. In food and environmental decontamination procedures, TMB can be used in in?situ free available chlorine (FAC) monitoring of chlorite-based sanitizers.

Carcinogenicity

The carcinogenic potential of Tetramethylbenzidine is uncertain due to conflicting evidence. It is not mutagenic by the Ames test, and did not induce formation of tumors in a single-arm study of 24 rats. On that evidence, it has been used as a replacement for carcinogenic compounds such as benzidine and o-phenylenediamine.
DOI: 10.1177/26.2.24068

Tetramethylbenzidine(54827-17-7)Related Product Information

• Octamethylcyclotetrasiloxane
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• N,N,N',N'-Tetramethylethylenediamine
• Pigment Yellow 12
• Tetramethylguanidine
• Tetramethylpyrazine
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• 3,3',5,5'-Tetramethylbenzidine dihydrochloride
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• N-(3-Sulfopropyl)-3,3',5,5'-tetramethylbenzidine sodium salt
• 3,3'',5,5-TETRAMETHYLBENZIDINESULFATE(TMB-S)
• N,N,N',N'-Tetramethylbenzidine, 97.50%,N,N,N',N'-TETRAMETHYLBENZIDINE
• TMB 124