1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

other: read across from analogue substance

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted following official guidelines performed on analogue susbtance 1

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1981

Report date:

1986

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,3-bis(2,3-diaminophenylazo)benzene hydrochloride

EC Number:

233-314-3

EC Name:

1,3-bis(2,3-diaminophenylazo)benzene hydrochloride

Cas Number:

10114-58-6

Molecular formula:

C18H18N8.2ClH

IUPAC Name:

Basic Brown 1 HCl

Test material form:

semi-solid (amorphous): gel

Remarks:

migrated information: paste

Method

Target gene:

his

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

S-9 mix

Test concentrations with justification for top dose:

20ug-5000 ug/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: complete solubility

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

other: without S-9 mix:N-methyl-N'-nitro-N-nitroso-guanidine (MNNC) (in DMS0) for the strains TA 100, TA 98 and TA 1535/ 4-nitro-o-phenylenediamine (in DMSO) for the strain TA 1538/ 9-aminoacridine chloride monohydrate (in DMOS) for the strain TA1537

Remarks:

with S9-mix: aminoanthracene for the strains TA100,Ta98,TA1538,TA1537 and TA1535

Details on test system and experimental conditions:

METHOD OF APPLICATION: Test tubes containing 2 ml portions of soft agar which consists of 100 ml agar (0.6% agar + 0.6% NaCl) and 10 ml aminoacid solution (minimal aminoacid solution for the determination of mutants: 0.5 mM histidine + 0.5 mM biotin) are kept in a water bath at 450C, and the remaining components are added in the following order: 0.1 ml test solution /0.1 ml bacterial suspension/0.5 ml S-9 mix (in tests with metabolic activation) or 0.5 ml phosphate buffer (in tests without metabolic activation). After mixing, the samples are poured onto the Vogel-Bonner agar plates (minimal glucose agar plates) within approx. 30 seconds.

DURATION
After incubation for 48 hours at 370C in the dark, the bacterial colonies (his+ revertante) are counted.

Test system:
Ist Experiments
Strains: TA 1535, TA 100, TA 1537, TA 1538, TA 98
Doses: 0, 20, 100, 500, 2500 and 5000 uig/plate
Solvent: DNSO
Type of test, standard plate test with and without
test condition: S-9 Mix
Number o f plates: 4 test plates per dose or per control

2nd Experiment
Strain: TA 1538
Doses: 0, 20, 100, 500, 2500 and 5000 Šug/plate
Solvent: DIVSO
Type of test,
test condition: standard plate test with S-9 mix
Number of plates: 4 test plates per dose or per control

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

Weakly positive reaction only with S-9 mix at 2500 >ig - 5000 ,ug/plate (factor 2.5)

Cytotoxicity / choice of top concentrations:

not specified

Test results 3

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

from about 20ug to 100 ug

Cytotoxicity / choice of top concentrations:

not specified

Test results 4

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

from about 500 ug/plate (TA 1537) onward with an increase in the number >of his+ >revertants by a factor of - 20 at - 500 >ug/plate

Cytotoxicity / choice of top concentrations:

not specified

Test results 5

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

Without S-9 mix only slight increase in the rate at 2500 uig/plate (factor 2.0).After the addition of S-9mix mutagenicity was observed from ca. 100 ug/plate onward with an increase in the number of revertant colonies by a factor >20 at 2500-5000 ug/plate

Cytotoxicity / choice of top concentrations:

not specified

Additional information on results:

no bacteriotoxic effect was observed

Applicant's summary and conclusion

Conclusions:

The substance was tested for in vitro gene mutation in bacteria following OECD 471. Under the experimental conditions the substance shows positive results with metabolic activation.

Executive summary:

The analogue substance 1 was tested for mutagenicity to bacteria with strains TA1535, TA100, TA1537, TA1538 and TA98 of Salmonella Typhimurium with and without metabolic activation. Positive reactions were detected with metabolic activation for all strains at low doses except for strain TA1535.