Trypan blue

Structural formula
General
Surname	Trypan blue
other names	Tetrasodium 3,3 '- [(3,3'-dimethyl-4,4'-biphenyldiyl) di-2,1-diazenediyl] bis (5-amino-4-hydroxy-2,7-naphthalene disulfonate) ( IUPAC ); CI Direct Blue 14; CI 23850; Benzamine blue;
Molecular formula	C 34 H 24 N 6 Na 4 O 14 S 4
Brief description	dark blue solid
External identifiers / databases
EC number	200-786-7
ECHA InfoCard	100,000,715
PubChem	6296
ChemSpider	21159397
Wikidata	Q61482373
properties
Molar mass	960.8 g mol −1
Physical state	firmly
Melting point	> 300 ° C (decomposition)
solubility	moderate in water (10 g l −1 at 25 ° C, free acid); insoluble in ethanol (free acid);
safety instructions
GHS labeling of hazardous substances danger
H and P phrases	H: 350
	P: 201-308 + 313
Toxicological data	6200 mg kg −1 ( LD 50 , rat , oral )
	As far as possible and customary, SI units are used. Unless otherwise noted, the data given apply to standard conditions .

Trypan Blue ( CI Direct Blue 14) is an anionic bisazo dye . The dye is made from o -tolidine as the diazo component and H-acid as the coupling component.

Use and properties

The dark blue stained with trypan blue cytoplasm of Hyaloperonospora parasitica

Trypan blue is mainly sold and used as the tetrasodium salt, in which the four sulfonic acid groups are dissociated .

Trypan blue is used in microbiology and in cell culture to determine the cell viability of cells . The substance is not absorbed by living cells, but dead and perforated cells absorb the dye and are colored dark blue. If the dye is not absorbed, one can draw conclusions about the condition of the cell membrane and not about the functionality of the cell interior.

Trypan blue binds to proteins. Because of the cytotoxicity of trypan blue, the evaluation must be carried out immediately after the addition of the dye.

In the flow cytometry, trypan blue is used as it is the intracellular autofluorescence, inter alia, by NADH and riboflavin caused reduced.

Trypan blue is teratogenic , which was first recognized in 1948. The carcinogenic properties of trypan blue were later determined, which are mainly due to the metabolized product tolidine.

Trypan blue is widely used in eye surgery . With the help of the dye, eye surgeons identify transparent or pathologically altered tissue that should be removed from the eye. This can be the lens capsule or epiretinal membranes ( gliosis ). Another area of application is the staining of the Descemet membrane for performing a DMEK ( Descemet Membrane Endothelial Keratoplasty ).

The name is derived from the ability of trypan blue to kill trypanosomatida .

history

Trypan blue was first synthesized together with trypan red in 1904 by Paul Ehrlich . Edwin Goldmann injected the dye into the cerebrospinal fluid ( liquor cerebrospinalis ) of dogs in 1913 . He realized that the entire central nervous system (brain and spinal cord) was stained, but no other organ. This was important evidence for the existence of the blood-brain barrier .

literature

ME Farah et al: Current concepts of trypan blue in chromovitrectomy. In: Dev Ophthalmol 42, 2008, pp. 91-100. PMID 18535383 (review article)
BJ Vote among others: Trypan blue-assisted vitrectomy. In: Retina 24, 2004, pp. 736-738. PMID 15492627
RJ Ford and FF Becker: The characterization of trypan blue-induced tumors in Wistar rats. In: Am J Pathol 106, 1982, pp. 326-331. PMID 7065117
AR Beaudoin and JM Roberts: Teratogenic action of the thyroid stimulating hormone and its interaction with trypan blue. In: J Embryol Exp Morphol 15, 1966, pp. 281-289. PMID 5964278

Web links

Commons : Trypan Blue - collection of images, videos, and audio files

Individual evidence

↑ ^a ^b ^c ^d ^e ^f ^g Data sheet Trypan blue (CI 23850) (PDF) from Merck , accessed on February 5, 2019.
↑ External identifiers or database links for trypan blue (free acid) : CAS number: 2538-83-2, PubChem : 6297 , ChemSpider : 14833963 , DrugBank : DB09158 , Wikidata : Q419642 .
^ VL Mosiman et al.: Reduction of cellular autofluorescence in the "flow cytometry": an in situ method. In: Cytometry 30, 1997, pp. 151-156. PMID 9222101
↑ J. Gillman et al .: A preliminary report on hydrocephalus, spina bifida and other congenital anomalies in the rat produced by trypan blue. In: S Afr J Med Sci 13, 1948, pp. 47-90.
↑ MM Turbow: Trypan blue induced teratogenesis of rat embryos cultivated in vitro. In: J Embryol Exp Morphol 15, 1966, pp. 387-395. PMID 5964283
^ T. Gillman and RC Hallowes: Ultrastructural changes in rat livers induced by repeated injections of trypan blue. In: Cancer Res , 32, 1972, pp. 2393-2399. PMID 4563139
↑ EE Goldmann: vital staining on the central nervous system. In: Abh. Preuss. Akd. Knowledge Phys. - Math. Kl. I 1, 1913, pp. 1-13.

[merck-1] ↑ ^a ^b ^c ^d ^e ^f ^g Data sheet Trypan blue (CI 23850) (PDF) from Merck , accessed on February 5, 2019.

[2] External identifiers or database links for trypan blue (free acid) : CAS number: 2538-83-2, PubChem : 6297 , ChemSpider : 14833963 , DrugBank : DB09158 , Wikidata : Q419642 .

[3] VL Mosiman et al.: Reduction of cellular autofluorescence in the "flow cytometry": an in situ method. In: Cytometry 30, 1997, pp. 151-156. PMID 9222101

[4] J. Gillman et al .: A preliminary report on hydrocephalus, spina bifida and other congenital anomalies in the rat produced by trypan blue. In: S Afr J Med Sci 13, 1948, pp. 47-90.

[5] MM Turbow: Trypan blue induced teratogenesis of rat embryos cultivated in vitro. In: J Embryol Exp Morphol 15, 1966, pp. 387-395. PMID 5964283

[6] T. Gillman and RC Hallowes: Ultrastructural changes in rat livers induced by repeated injections of trypan blue. In: Cancer Res , 32, 1972, pp. 2393-2399. PMID 4563139

[7] EE Goldmann: vital staining on the central nervous system. In: Abh. Preuss. Akd. Knowledge Phys. - Math. Kl. I 1, 1913, pp. 1-13.