N,N'-di-sec-butyl-p-phenylenediamine

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:

key study

Study period:

1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Although the test protocol was not described in the publication (in stead, reference to another publication was made), the results described in the paper combine the results of Ames testing of 300 substances by a renowed Research institute (National Institute of Environmental Health Sciences, Genetic toxicity department) in the framework of the National Toxicology Program.

Data source

Reference

Reference Type:

publication

Title:

Salmonella Mutagenicity Tests: IV. Results from the testing of 300 chemicals.

Author:

Zeiger E, Anderson B, Haworth S, Lawler T and Mortelmans K

Year:

1988

Bibliographic source:

Environmental and Molecular Mutagenesis 11 (Suppl. 12), 1-158

Materials and methods

Principles of method if other than guideline:

Ames test

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-di-sec-butyl-p-phenylenediamine

EC Number:

202-992-2

EC Name:

N,N'-di-sec-butyl-p-phenylenediamine

Cas Number:

101-96-2

Molecular formula:

C14H24N2

IUPAC Name:

N,N'-di-sec-butyl-p-phenylenediamine

Details on test material:

- Name: N,N'-di-sec-butyl-p-phenylenediamine
- Analytical purity: > 82%
- Source: Pfaltz & Bauer

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 97

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 98

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 100

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 1535

Metabolic activation:

with and without

Metabolic activation system:

rat and hamster S-9 fractions

Test concentrations with justification for top dose:

0.000 - 0.330 - 1.000 - 3.300 - 10.000 - 33.000 - 100.000 - 200.000 - 333.000 µg/plate (as tested by lab. MIC)
0.000 - 0.300 - 1.000 - 3.000 - 10.000 - 16.000 - 33.000 - 66.000 - 100.000 - 166.000 - 333.000 µg/plate (as tested by lab. SRI)

Vehicle / solvent:

DMSO

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 4-nitro-o-phenylenediamine, 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation assay

DURATION
- Preincubation period: 20 minutes
- Expression time (cells in growth medium): 2 days

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
The toxicity assay was performed using TA100 or the system developed by Waleh et al (Waleh, N.S., Rapport, S.J., Mortelmans, K. 1982. Development of a toxicity test to be coupled to the Ames Salmonella assay and the method of construction of the required strains. Mutat. Res. 97:247-256.). Toxic concentrations were those that produced a decrease in the number of his+ colonies, or a clearing in the density of the background lawn, or both.

Evaluation criteria:

Evaluations were made at both the individual trial and overall chemicals. Individual trials were judged mutagenic (+), weakly mutagenic (+W), questionable (?) or non-mutagenic (-), depending on the magnitude of the increase of his+ revertants, and the shape of the dose-response. A trial was considered questionable (?) if the dose-response was judged insufficiently high to support a call of "+W", if only a single dose was elevated over the control, or if the increase seen was not dose related. It was not necessary for a response to reach twofold background for a chemical to be judged mutagenic.
A chemical was judged mutagenic (+) or weakly mutagenic (+W) if it produced a reproducible dose-related response over the solvent control in replicate trials. A chemical was judged questionable (?) if the results of individual trials were not reproducible, if increases in his+ revertants did not meet the criteria for a "+W" response, or if only single doses produced increases in his+ revertants in repeat trials. Chemicals were judged non-mutagenic (-) if they did not meet the criteria for a mutagenic or questionable response.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The test substance was not mutagenic in the Ames/Salmonella assay to strains TA97, TA98, TA100 or TA1535 in the presence or absence of metabolic activation.

Executive summary:

The substance was tested in the framework of the National Toxicology Program and coordinated by the National Institute of Environmental Health Sciences, Genetic Toxicology Department, Cellular and Genetic Toxicology Branch. The test was performed in two independent laboratories (Microbiological Associates Inc. (MIC) and SRI International), which both concluded the test substance to be non mutagenic in this assay. As such, a good interlaboratory reproducibility of the assay was shown.