p,p',p''-tris(diethylamino)trityl alcohol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from NTP report

Data source

Reference

Reference Type:

publication

Title:

Toxicity Studies of Malachite Green Chloride and Leucomalachite Green Administered in Feed to F344/N Rats and B6C3F1 Mice

Author:

U.S. Department of Health and Human Services

Year:

2004

Bibliographic source:

National Toxicology Program Toxicity Report Series Number 71

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Similar to OECD 471

Principles of method if other than guideline:

Gene mutation toxicity study was performed to evaluate the mutagenic nture of Malachite Green

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

[4-[α-[4-(dimethylamino)phenyl]benzylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride

EC Number:

209-322-8

EC Name:

[4-[α-[4-(dimethylamino)phenyl]benzylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride

Cas Number:

569-64-2

IUPAC Name:

N-(4-{[4-(dimethylamino)phenyl](phenyl)methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride

Constituent 2

Reference substance name:

Malachite Green

IUPAC Name:

Malachite Green

Test material form:

other: Solid

Details on test material:

- Name of test material: Malachite Green
- Molecular formula: C23H25ClN2
- Molecular weight: 364.9175 g/mol
- Smiles notation: C(\c1ccc(N(C)C)cc1)(=C1/C=C\C(=[N+](\C)C)C=C1)c1ccccc1.[ClH-]
- Substance type: Organic
- Physical state: Green crystal with metallic lustre
- Analytical purity: 95%

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA97, TA98, TA100, TA102, TA104, and TA1535

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix (metabolic activation enzymes and cofactors from Aroclor 1254-induced male Sprague Dawley rat or Syrian hamster liver)

Test concentrations with justification for top dose:

0, 0.1, 0.3, 1.0, 3.3 or 10.0 µg/plate

Vehicle / solvent:

No data

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

methylmethanesulfonate

mitomycin C

other: 4-nitroo-phenylenediamine, 2-aminoanthracene

Remarks:

No data

Details on test system and experimental conditions:

METHOD OF APPLICATION:preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: Each trial consisted of triplicate plates

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data

Evaluation criteria:

y, a positive response is defined as a reproducible, dose-related increase in histidine-independent (revertant) colonies in any one strain/activation combination. An equivocal response is defined as an increase in revertants that is not dose related, is not reproducible, or is not of sufficient magnitude to support a determination of mutagenicity. A negative response is obtained when no increase in revertant colonies is observed following chemical treatment. There is no minimum percentage or fold-increase required for a chemical to be judged positive or weakly positive.

Statistics:

Revertants are presented as mean ± standard error from three plates

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA97, TA98, TA100, TA102, TA104, or TA1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

No data

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without

Malachite green failed to induce mutation in Salmonella typhimurium strains TA97, TA98, TA100, TA102, TA104, or TA1535 with or without S9 metabolic activation and hence is not likely to be mutagenic in vitro.

Executive summary:

Gene mutation toxicity study was performed to evaluate the mutagenic nature of Malachite Green. The study was performed as per the protocol given by Zeiger et al. The material was used at dose levels of 0, 0.1, 0.3, 1.0, 3.3 or 10.0 µg/plate using Salmonella typhimurium strains TA97, TA98, TA100, TA102, TA104, or TA1535 with or without S9 metabolic activation. Preincubation assay was performed and the plates were incubated for 48 hrs before evaluation. The plates were evaluated for a dose dependent increase in the number of revertants. Each trial consisted of triplicate plates of concurrent positive and negative controls and five doses of malachite green chloride. The high dose was limited by toxicity. Malachite green failed to induce mutation in Salmonella typhimurium strains TA97, TA98, TA100, TA102, TA104, or TA1535 with or without S9 metabolic activation and hence is not likely to be mutagenic in vitro.