Reaction mass of bis[3-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-) and bis[5-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

May 11, 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

not specified

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Bis[5-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

EC Number:

281-692-3

EC Name:

Bis[5-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

Cas Number:

84012-52-2

Molecular formula:

C36H42Cl4N12Zn

IUPAC Name:

bis[5-({4-[benzyl(methyl)amino]phenyl}diazenyl)-1,4-dimethyl-4,5-dihydro-1H-1,2,4-triazol-1-ium] tetrachlorozincate(2-)

Constituent 2

Reference substance name:

Bis[3-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

EC Number:

281-650-4

EC Name:

Bis[3-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

Cas Number:

84000-94-2

Molecular formula:

C36H42Cl4N12Zn

IUPAC Name:

bis(3-[(E)-2-{4-[benzyl(methyl)amino]phenyl}diazen-1-yl]-1,4-dimethyl-1H-1,2,4-triazol-4-ium); tetrachlorozincdiuide

Test material form:

solid: particulate/powder

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

0.2, 2, 20, 200 and 2000 µg per Petri dish.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

no

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

N-ethyl-N-nitro-N-nitrosoguanidine

other: daunomycine

Remarks:

without S9

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

no

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-anthramine

Remarks:

with S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar.
The bacterial strains were kept as frozen broth cultures, in aliquots of 0.5 ml at -70 °C with 8.0 % dimethylsulfoxide (DMSO). Fresh cultures were prepared by adding 0.1 ml of a thawed stock culture to 15 ml of nutrient broth (normal Difco nutrient broth for strains TA 1535 and TA 100, and double strength Difco nutrient broth for TA 1537 and TA 98, in 0.5 % NaCI). A nutrient agar (2.5% Difco nutrient broth, 1.2 % Difco agar) was also streaked. The broth was incubated in the dark in a shaking water bath at 37 °C for 16 hours, while the plate was incubated at 37 °C overnight. Broths and plates were then transferred to the refrigerator, for up to one week.

NUMBER OF REPLICATIONS:3

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

at 200 µg only

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium, other: TA 100 and TA 1535

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium, other: TA 1535, TA 1537 and TA 100

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium, other: TA 1537 and TA 98

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Under the experimental conditions defined in the protocol, and employing a doubling of the spontaneous reversion rate and dose-effect relationship as criteria of mutagenicity, product was found to be mutagenic for S. typhimurium strain TA 98 in the presence of a liver microsomal enzyme preparation (S9-mix) from male rats pretreated with Aroclor 1254. The evidence for mutagenicity existed at a a concentration level of 200 µg of product per Petri dish, at 2000 µg the decrease of the number of revertants is due to a toxic effect as well as the absence of the background "lawn" in strains TA 1535 and TA 100 at the highest concentration (2000 µg) without metabolic activation.

Applicant's summary and conclusion

Conclusions:

The test substance was found to be mutagenic for S. typhimurium strain TA 98 in the presence of a liver microsomal enzyme preparation (S9-mix) at cytotoxic concentratons.
A toxic effect in strains TA 1535 and TA 100 at the highest concentration (2000 µg) without metabolic activation was observed.

Executive summary:

The substance was tested according to the OECD Guideline 471 with the Salmonella typhimurium strains TA I535, TA I537, TA 98 and TA l00 at concentrations from 0.2 to 2000 µg per Petri dish both in the presence and absence of metabolic activation.

Under the test condition the test substance was found to be mutagenic for S. typhimurium strain TA 98 in the presence of a liver microsomal enzyme preparation (S9-mix) at cytotoxic concentrations. Moreover a toxic effect in strains TA 1535 and TA 100 at the highest concentration (2000 µg) without metabolic activation was observed. Based on the results observed the substance, under the test conditions, is considered as mutagenic.