Toluidine Blue O Chemical Properties

Melting point:

>200℃

storage temp. 

Inert atmosphere,Room Temperature

solubility 

H2O: soluble1mg/mL

form 

Solid

pka

2.4, 11.6(at 25℃)

Colour Index 

52040

color 

Dark green to black

PH

2.8 (H2O, 25℃)

Water Solubility 

3.82 g/100 mL

λmax

626 nm, 630 nm

Merck 

14,9520

Stability:

Stable. Incompatible with strong oxidizing agents.

Biological Applications

Bound hyaluronan assay; DNA assay; bioelectronic applications; biofuel cells; microbial fuel cells; detecting bacteria,body fluid,nucleic acids,45,proteins,viruses; identifying Gram-negative bacteria,b-amyloid reducing agents

EPA Substance Registry System

Tolonium chloride (92-31-9)

Safety Information

Risk Statements 

36/37/38

Safety Statements 

22-24/25

WGK Germany 

3

RTECS 

SP5670000

TSCA 

Yes

HS Code 

32041300

Toxicity

LD50 in mice, rats, rabbits (mg/kg): 27.56, 28.93, 13.44 i.v. (Haley, Storlarsky)

MSDS

• Language:English Provider:3-Amino-7-dimethylamino-2-methylphenazathionium chloride
• Language:English Provider:SigmaAldrich
• Language:English Provider:ACROS

Toluidine Blue O Usage And Synthesis

Chemical Properties

Toluidine Blue O is a dark green or black-gray powder with bronze luster. It is soluble in water (3.82g/100ml), producing a blue-violet solution, slightly soluble in ethanol (0.57g/100ml) to give a blue solution, very slightly soluble in chloroform, and almost insoluble in ether.

Originator

Blutene,Abbott,US,1953

Uses

Toluidine Blue O is a metachromatic dye suitable for a wide variety of histological staining procedures. It is used for direct dyeing and printing of wool and silk, as well as a biological stain and hemostatic agent.

Definition

ChEBI: Toluidine Blue O is an organic chloride salt having 3-amino-7-(dimethylamino)-2-methylphenothiazin-5-ium (tolonium) as the counterion. It is a blue nuclear counterstain that can be used to demonstrate Nissl substance and is also useful for staining mast cell granules, both in metachromatic and orthochromatic techniques. It has a role as a fluorochrome, a histological dye and a diagnostic agent. It contains a tolonium.

Preparation

by heavy chromate salt in the presence of sodium content and deal with N1,N1-dimethylbenzene-1,4-diamine, transforming it into the content and sulfonic acid, this product then through heavy chromate salt and O-Methylaniline processing, into a unique of sulfur acid amine generation, then use heavy chromate salt and copper sulfate closed loop and oxidation, Finally,?the?zinc chloride?salt?precipitation

Manufacturing Process

As taken from US Patent 416,055 (probably the oldest patent on the manufacture of a currently-used drug): In carrying out this process about 6 pbw of dimethyl-p-phenylenediamine was dissolved in about 18 pbw of hydrochloric acid of about 1.16 specific gravity and then a solution of about 3.8 pbw of nitrite of soda in about 6 pbw of water was gradually added. The hydrochlorate of nitroso-dimethylaniline thus produced in the well-known manner is then submitted to the reducing action of zinc-dust by adding, first about 30 pbw of hydrochloric acid of about 1.16 specific gravity and then (in small portions at a time) about 10 pbw of zinc-dust as is well understood by chemists. The solution of hydrochlorate of paramido-dimethylaniline thus obtained is afterwards diluted with about 250 pbw of water and then the uncombined hydrochloric acid contained in the solution is, if any, neutralized by the addition of an alkali. There are then added about 16 pbw of sulfate of alumina and about 13 pbw of thiosulfate of sodium, (hyposulfite of soda) and immediately afterwards a solution of about 5 pbw of bichromate of potash in about 60 pbw of water is quickly run in.
In this stage of the process the formation of an acid sulfureted compound of paramidodimethylaniline takes place, possessing the formula C8H11N2SSO3H(paramido-dimethylaniline-thiosulfonic acid). Without previous separation of this intermediate compound a solution of about 5.3 pbw of orthotoluidine, in the requisite amount of dilute hydrochloric acid (about 6 pbw of hydrochloric acid, SG about 1.16, diluted with about 6 pbw water) and shortly afterwards a solution of about 14 pbw of bichromate of potash in about 160 parts by weight of water is then added under constant agitation, when a precipitate will be formed chiefly consisting of a green indamine possessing in its dry condition the formula C15H17N3S2O3. In order to transform the same into toluidine-blue, about 50 pbw of a solution of chloride of zinc of about 1.5 specific gravity are added and the mixture thus obtained is boiled during about half an hour, when, after cooling, the toluidine-blue thus formed will separate out and may then be filtered and purified, if necessary, by repeated solution in water and precipitation by means of chloride of sodium and chloride of zinc.
In the above described process the sulfate of alumina may be dispensed with and replaced by as much hydrochloric, sulfuric, or acetic acid as will be required to liberate the thiosulfuric acid from the thiosulfate of sodium employed.
Toluidine-blue prepared as above described presents the following characteristic properties: It consists principally of the hydrochlorate of dimethyltoluthionine, the composition of which corresponds to the formula C15H15N3SHCl.

Therapeutic Function

Coagulant

Staining Procedures

The staining solution is prepared by dissolving 0.05% toluidine blue O in buffer, pH 4.4-6.8. Sections or whole tissues are stained for 5-10 min followed by differentiation using the same buffer until excess stain is removed. Cellulose and other glucans become red-violet depending on pH, lignified elements and other polyphenols become bluish-green. Fungal hyphae are often distinguished from the surrounding host tissue because of the difference in cytoplasmic pH.

Properties and Applications

Soluble in water for blue purple, soluble in ethanol to blue. The strong sulfuric acid for yellow green, diluted to blue. The dye solution to join sodium hydroxide have dark purple precipitation.

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