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Genetic toxicity in vitro

Description of key information

In vitro gene mutation study in Bacteria was conducted by U. S. National Library of Medicine(CCRIS ;, 2017) to evaluate the mutagenic potential of target chemical((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5), Other Name; Magenta and Basic Fuchsin.In the study Magenta was assessed for its possible mutagenic potential. For this purpose the reverse mutation assay was performed on Salmonella typhimurium TA98, TA 100, TA 1535, 1537, TA 1538 and E.Coli WP2UVRA.t material was exposed at the concentration of 5-1000 µg/plate for TA98, TA 100, E.Coli WP2UVRA and 1-200 µg/plate for TA 1535, 1537,TA 1538 in the absence of S9 metabolic activation. While all strains were exposed to the test material at the concentration of 20-5000µg/plate in the presence of S9.Cytotoxicity was also observed. No mutagenic effects were observed. Therefore Magenta was considered to be non mutagenic with and without S9. Hence the substance cannot be classified as gene mutant in vitro.

Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from CCRIS
Qualifier:
according to guideline
Guideline:
other: As mention below
Principles of method if other than guideline:
To evaluate the mutagenic potential of Magenta in Salmonella typhimurium TA98, TA 100, TA 1535, 1537,TA 1538 and E.Coli WP2UVRA.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
IUPAC name:Magenta
commen name : Basic violet 14
Molecular formula :C20H19N3.ClH
molecular weight: 337.8 g/mol
InChI:1S/C20H19N3.ClH/c1-13-12-16(6-11-19(13)23)20(14-2-7-17(21)8-3-14)15-4-9-18(22)10-5-15;/h2-12,21H,22-23H2,1H3;1H/b20-14-,21-17?
Smiles:C(\c1cc(c(N)cc1)C)(c1ccc(N)cc1)=C1/C=CC(=N)C=C1.Cl
Target gene:
Histidine for Salmonella typhimurium and tryptophan for E.Coli.
Species / strain / cell type:
bacteria, other: Salmonella typhimurium TA98, TA 100, TA 1535, 1537, TA 1538and E.Coli WP2UVRA.
Details on mammalian cell type (if applicable):
Not apllicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
not specified
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S-9, Phenobarbital and Beta-naphthoflavone
Test concentrations with justification for top dose:
-S9; 5-1000 µg/plate for TA98, TA 100and E.Coli WP2UVRA.
-S9; 1-200 µg/plate for TA 1535, 1537 and TA 1538
+S9;20-5000µg/plate ,For all strain
Vehicle / solvent:
Vehicle
- Vehicle(s)/solvent(s) used: DMSO
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
not specified
Details on test system and experimental conditions:
Details on test system and conditions
METHOD OF APPLICATION: Preincubation
Rationale for test conditions:
Not specified
Evaluation criteria:
Evaluation was done considering a dose dependent increase in the number of revertants/plate.
Statistics:
Not specified
Key result
Species / strain:
bacteria, other: Salmonella typhimurium TA98, TA 100, TA 1535, 1537,TA 1538 and E.Coli WP2UVRA.
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Remarks on result:
other: No mutagenic effect were observed
Conclusions:
Magenta (632-99-5) was evaluated for its mutagenic potential in Salmonella typhimurium TA98, TA 100, TA 1535, 1537, TA 1538 and E.Coli WP2UVRA. The test results were considered to be negative in the presence and absence of S9.
Executive summary:

In the study Magenta was assessed for its possible mutagenic potential. For this purpose the reverse mutation assay was performed on Salmonella typhimurium TA98, TA 100, TA 1535, 1537, TA 1538 and E.Coli WP2UVRA.t material was exposed at the concentration of 5-1000 µg/plate for TA98, TA 100, E.Coli WP2UVRA and 1-200 µg/plate for TA 1535, 1537,TA 1538 in the absence of S9 metabolic activation. While all strains were exposed to the test material at the concentration of 20-5000µg/plate in the presence of S9.Cytotoxicity was also observed. No mutagenic effects were observed. Therefore Magenta was considered to be non mutagenic with and without S9. Hence the substance cannot be classified as gene mutant in vitro.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Genetic toxicity in Vitro

In different studies, ((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5), Other Name; Magenta, Para- Rosaniline HCL and Basic Fuchsin has been investigated for potential of mutagenic response with and without metabolic activation. The studies are based on in vitro gene mutation study in mammalian cells and bacteria for target chemical. Different studies based on these experiments are concluded below

In vitro gene mutation study in Bacteria was conducted by U. S. National Library of Medicine(CCRIS ;, 2017) to evaluate the mutagenic potential of target chemical((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5), Other Name; Magenta and Basic Fuchsin.In the study Magenta was assessed for its possible mutagenic potential. For this purpose the reverse mutation assay was performed on Salmonella typhimurium TA98, TA 100, TA 1535, 1537, TA 1538 and E.Coli WP2UVRA.t material was exposed at the concentration of 5-1000 µg/plate for TA98, TA 100, E.Coli WP2UVRA and 1-200 µg/plate for TA 1535, 1537,TA 1538 in the absence of S9 metabolic activation. While all strains were exposed to the test material at the concentration of 20-5000µg/plate in the presence of S9.Cytotoxicity was also observed. No mutagenic effects were observed. Therefore Magenta was considered to be non mutagenic with and without S9. Hence the substance cannot be classified as gene mutant in vitro.

Other confirmatory test as In vitro gene mutation study in mammalian cells was conducted by William Au and T. C. Hsu.(Environmental Mutagenesis,1979) the mutagenic potential of target chemical ((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5), Other Name; Magenta and Basic Fuchsin in mammalian cell line. In Vitro Assay System Basic Fuchsin(632-99-5)was assessed for its mutagenic potential. For this purpose chromosome aberration test was performed on Chinese hamster ovary cell line (CHO). Chinese hamster ovary cell line (CHO) in McCoy 5a medium was used for this study. Exponentially growing cultures in 10 ml medium were treated with 1-20 µM of chemical. A culture treated with the same amount (0.1 ml) of water was used as controls. All cultures were incubated for five hours after the introduction of the test chemical.Colcemid (0.04 µg / d final concentration) was added to each culture during thelast hour of incubation and all cultures were harvested for conventional cytogenetic preparations, stained with Giemsa and coded. The average number of breaks per metaphase was calculated and was used for comparing the clastogenic activities of different compounds.The number of chromosome breaks per metaphase is 0.18. This was not a significant increase. The significant value for the number of chromosome breaks per metaphase by some water-soluble dyes was 0.32. Therefore Basic Fuchsin was considered to be non mutagenic in Chinese hamster ovary cell line (CHO) by chromosome aberration test. Hence test substance cannot be classified as gene mutant in vitro.

Further supported by other experimental study conducted by J. Mirsalis et.al (Abstracts of the fifteenth annual meeting of the environmental mutagen society held at San Antonio, Texas March 3–6, 1983) ,Environmental and Molecular Mutagenesis,1983) to evaluate the mutagenic potential of target chemical ((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5).Genetic toxicity in vitro for Para- Rosaniline HCL was assessed for its possible mutagenic potential. For this purpose Unscheduled DNA assay was performed.Hepatocytes from male Fischer-344 rats were incubated with3H-TdR along with test substance at the test concentration of2.2ug/ml. Positive and negative controls were used .Unscheduled DNA synthesiswas measured by quantitative autoradiography as net grains/nucleus (NG). No mutagenic effects were observed in test substance as compared to the value of positive and negative control. Therefore Para- Rosaniline HCL was considered to be non mutagenic for Unscheduled DNA assay. Hence the substance cannot be classified as gene mutant in vitro.

Another supporting in vitro gene mutation study in bacteria was conducted by Mortelmans K et al.(Environmental Mutagenesis) to evaluate the mutagenic potential of target chemical ((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5), Other Name; Magenta, Para- Rosaniline HCL and Basic Fuchsin.In vitro Genetic toxicity study was assessed for Basic Fuchsin to find its possible mutagenic potential. For this purpose Bacterial reverse mutation assay was performed on Salmonella typhimurium TA1535, TA1537, TA98, and TA100. The test material was exposed in the presence and absence of metabolic activation at the concentration of 0,0.030,0.1,0.160,0.3 ,0.33 ,0.670,1,3.3,10,33,100,333 and 1000µg/plate. Mutagenic effects were observed. Therefore Basic Fuchsin was considered to be non mutagenic in Salmonella typhimurium TA98, TA100, TA 1535, TA 1537 and TA97 Bacterial reverse mutation assay. Hence the substance canbe classified as gene mutant in vitro.

Another supporting in vitro gene mutation study in bacteria was conducted by Tsutomu Yamaguchi (Agric. Biol. Chem.,1988) to evaluate the mutagenic potential of target chemical ((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5), Other Name; Magenta, Para- Rosaniline HCL and Basic Fuchsin.In vitro Genetic toxicity study was assessed for Basic Fuchsin to find its possible mutagenic potential. For this purpose Bacterial reverse mutation assay was performed on Salmonella typhimurium TA100. The test material was exposed in the presence and absence of metabolic activation at the concentration of 5 µg/plate. Mutagenic effects were observed. Therefore Fuchsin was considered to be non mutagenic in Salmonella typhimurium TA100 Bacterial reverse mutation assay. Hence the substance canbe classified as gene mutant in vitro.

 

Bacterial reverse mutation assay is a screening test for mutagenic response, while mammalian cell gene mutation assay is a confirmatory test for mutagenic response. As per the above studies the target chemical ((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5), Other Name; Magenta, Para- Rosaniline HCL and Basic Fuchsin is considered to be non mutagenic because the result for in vitro chromosome abbreviation as well as Bacterial reverse mutation assay are negative. Thus basedon the majority data for the target chemical((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5)does notexhibits gene mutation toxicity in vitro. Hence, the chemical isnot likely to classify as a gene mutant in vitro.

Justification for classification or non-classification

Thus based on the majority data for the target chemical((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5)does not exhibits gene mutation toxicity in vitro.

Bacterial reverse mutation assay is a screening test for mutagenic response, while mammalian cell gene mutation assay is a confirmatory test for mutagenic response. As per the above studies the target chemical ((4-(4-aminophenyl)(4-iminocyclohexa-2,5-dienylidene)methyl)-2-methylaniline hydrochloride)(632-99-5), Other Name; Magenta, Para- Rosaniline HCL and Basic Fuchsin is considered to be non mutagenic because the result for in vitro chromosome abbreviation as well as Bacterial reverse mutation assay are negative. Hence, the chemical isnot likely to classify as a gene mutant in vitro.