Methylrosanilinium chloride exerts its antimicrobial effects primarily through interaction with bacterial cell walls and membranes. As a triphenylmethane dye, it binds to bacterial peptidoglycan and disrupts cell wall integrity, leading to cell death. It also has antifungal activity through similar membrane-disrupting mechanisms. Its anthelmintic activity is related to its ability to interfere with parasite metabolism and motility. It is effective against some Gram-positive bacteria but has limited activity against Gram-negative bacteria. Its mechanism as a dye involves binding to cellular components for histological staining.

In vitro, methylrosanilinium chloride demonstrates antibacterial and antifungal activity. It is effective against some Gram-positive bacteria and fungi. Its activity has been characterized in standard antimicrobial susceptibility tests, showing concentration-dependent inhibition of microbial growth. It is also used as a biological stain in various histological and microbiological applications. Its anthelmintic activity has been demonstrated in vitro against nematodes. Its antimicrobial activity is related to its ability to bind to and disrupt cell membranes.

In vivo, methylrosanilinium chloride is used topically for the treatment of bacterial and fungal infections of the skin and mouth. It is also used as a blood additive to prevent transmission of Chagas disease. When applied topically, it is effective against localized infections, including thrush and skin infections. Its systemic absorption is minimal, making it suitable for topical use. It has been used as an anthelmintic for nematode infections. Its in vivo efficacy is well-established for these indications.

In vitro enzyme/receptor binding studies are not applicable to methylrosanilinium chloride, as its mechanism is based on physicochemical interactions with cell walls and membranes rather than binding to specific enzymes or receptors. Its activity as a dye involves binding to cellular components such as DNA, proteins, and peptidoglycan. Binding studies may involve spectrophotometric or fluorometric measurement of dye binding to bacterial cells or purified components. Antimicrobial activity is assessed by standard susceptibility testing methods.

In vitro cellular assays for methylrosanilinium chloride involve culturing bacteria, fungi, or parasites in the presence of serial dilutions of the compound. Antimicrobial susceptibility is assessed by broth microdilution or agar diffusion methods to determine MIC values. For antifungal activity, similar methods are used with fungal cultures. Cytotoxicity in mammalian cells is evaluated using standard viability assays to assess safety for topical use. For anthelmintic activity, nematodes are incubated with the compound, and motility and viability are assessed microscopically.

In vivo animal studies for methylrosanilinium chloride are limited, as it is primarily used topically in humans. Preclinical studies may involve topical application to animal models of skin or oral infections to assess efficacy and safety. For its use as a blood additive to prevent Chagas disease transmission, animal studies have evaluated its ability to kill Trypanosoma cruzi in blood. Standard protocols involve administration of the compound, followed by assessment of infection rates or parasite clearance. Toxicity studies evaluate local irritation and systemic effects.

Absorption, Distribution and Excretion
This study investigated tissue residues of gentian violet using nine male and nine female Hubbard adult broiler breeders. Each broiler was orally administered 14C-labeled gentian violet. The half-life (T_1/2) of 14C-labeled gentian violet in blood differed between males and females (1.43 hours and 1.68 hours, respectively). Radioactivity was detected in the muscle of only one broiler eight hours after administration. Radioactivity was detected in the liver 120 hours after administration and in the kidneys 432 hours after administration. All eggs collected within the first 144 hours contained very low but detectable radioactivity. Metabolism/Metabolites Direct electron spin resonance spectroscopy revealed that gentian violet undergoes single-electron reduction in the cytochrome P450 monooxygenase system, generating a carbon-centered radical.
Pharmacokinetic properties of methylrosanilinium chloride indicate minimal systemic absorption after topical application. When used as a blood additive, it is present in the blood and is metabolized and cleared. Its systemic bioavailability is low, which contributes to its favorable safety profile for topical use. It is excreted primarily via urine and bile. Its use as a dye and topical antimicrobial agent does not typically require detailed pharmacokinetic characterization.

Non-Human Toxicity Values
Oral LD50 in rats: 420 mg/kg
Intraperitoneal LD50 in rats: 17 mg/kg for both juvenile and adult rats
Intraperitoneal LD50 in rats: 8900 μg/kg
Oral LD50 in mice: 96 mg/kg
Intraperitoneal LD50 in mice: 5100 μg/kg
Toxicological data for methylrosanilinium chloride indicate that it is generally safe for topical use at recommended concentrations. It may cause skin irritation or staining in some individuals. It is listed as a substance of very high concern (SVHC) by the European Chemicals Agency due to its potential carcinogenicity and environmental persistence. Systemic toxicity is rare with topical use, but ingestion can cause gastrointestinal irritation and systemic effects. It should not be used on open wounds or mucous membranes in large amounts. Its use as a food additive is not permitted in many countries.

[1].The response of selected members of the archaea to the gram stain. Microbiology, 1996. 142 ( Pt 10): p. 2887-95.

[2].Curr Protoc Microbiol, 2005. Appendix 3: p. Appendix 3C.

[3].Statins potentiate cytostatic/cytotoxic activity of sorafenib but not sunitinib against tumor cell lines in vitro. Cancer Lett, 2010. 288(1): p. 57-67.

According to an independent committee of scientific and health experts, gentian violet may be carcinogenic. Hexamethyl-p-rosaniline chloride is a green to dark green powder. (NTP, 1992) Crystal violet is an organochloride, a monochloride of the crystal violet cation. It was once used in creams for topical treatment of bacterial and fungal infections, effective against some Gram-positive bacteria (especially Staphylococcus) and some pathogenic fungi (including Candida), but its use declined after reports of carcinogenicity in animals. It was also used for staining wood, silk, and paper, as well as for histological staining. It can be used as a histological dye, preservative, antibacterial agent, antifungal agent, and insect repellent. It contains the crystal violet cation. Gentian violet is a blue aniline derivative dye with antifungal and antimitotic properties. Gentian violet (GV) can dissociate into positive ions (GV+) and negative ions (Cl-), which can penetrate Gram-positive and Gram-negative bacterial cells. GV+ ions interact with negatively charged components of the bacterial cell wall, including lipopolysaccharides, peptidoglycans, and DNA. This substance is also a mutagen and mitotic toxin. GV induces photodynamic activity through a free radical mechanism. Furthermore, this agent dissipates action potentials on prokaryotic or eukaryotic cell membranes by inducing membrane permeability, leading to respiratory depression and subsequent cell death.
A mixture of violet dyes with antibacterial, antifungal, and anthelmintic properties.
See also: Gentian Violet Cationic (containing the active moiety); Brilliant Green; Cod Liver Oil; Gentian Violet (component); Acridine Yellow; Gentian Violet; Sodium Propionate (component)...See more...

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Therapeutic Uses
Topical anti-infective; anti-nematode; Rosaniline dye
Gentian violet has been used in medicine for nearly a century: as a topical disinfectant, an oral anthelmintic, and in recent years as a blood additive to prevent the spread of Chagas disease. ...
Therapeutic Category: Anti-infective (Topical). It has been used as an anthelmintic (for nematodes) and a blood additive to prevent the transmission of Chagas disease through blood transfusions.
Therapeutic Classification (Veterinary): Anti-infective (topical); antimicrobial agent in poultry feed.
For more complete data on the therapeutic uses of gentian violet (7 types in total), please visit the HSDB record page.
Drug Warnings
Contact with granulation tissue can cause permanent pigmentation of the skin. This dye should not be used on ulcerative lesions of the face. Its staining properties are a significant drawback.
Six newborns developed oral ulcers after treatment with gentian violet (crystal violet) for oral candidiasis.Use a 0.5% or 1% aqueous solution twice daily.
Methylrosanilinium chloride (crystal violet, gentian violet) is a widely used dye and topical antimicrobial agent. It is available over-the-counter for the treatment of minor skin and mouth infections. It is also used as a biological stain in histology and microbiology. Its use as a blood additive to prevent Chagas disease transmission is an important public health application in endemic regions. It is regulated as a pharmaceutical ingredient in some jurisdictions and as a dye in others. Its safety and efficacy have been established over many years of use.

C25H30CLN3

407.9788

407.212

548-62-9

11057

Light green to green solid powder

1.19 g/cm3 (20ºC)

215ºC

40ºC

1.463

0

3

4

29

542

0

ZXJXZNDDNMQXFV-UHFFFAOYSA-M

InChI=1S/C25H30N3.ClH/c1-26(2)22-13-7-19(8-14-22)25(20-9-15-23(16-10-20)27(3)4)21-11-17-24(18-12-21)28(5)6;/h7-18H,1-6H3;1H/q+1;/p-1

[4-[bis[4-(dimethylamino)phenyl]methylidene]cyclohexa-2,5-dien-1-ylidene]-dimethylazanium;chloride

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ≥ 100 mg/mL (~245.11 mM)
H2O : ~5 mg/mL (~12.26 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)