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.
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.
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.
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
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.
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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Standard( Cotton )
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Light Fastness
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Persperation Fastness
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Ironing Fastness
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Soaping
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Fading
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Stain
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Fading
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Stain
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Fading
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Stain
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C
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2-3
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1-2
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