Aniline

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment.

Data source

Reference

Reference Type:

publication

Title:

Reproducibility of microbial mutagenicity assays: I. Tests with Salmonella typhimurium and Escherichia coli using a standardized protocol.

Author:

Dunkel VC, Zeiger E, Brusick D, McCoy E, McGregor D, Mortelmans K, Rosenkranz HS, Simmon VF

Year:

1984

Bibliographic source:

Environ Mutagen. 6, Suppl 2, 1-251.

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

maximum test concentration < 5000 µg/plate

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Anilinium chloride

EC Number:

205-519-8

EC Name:

Anilinium chloride

Cas Number:

142-04-1

IUPAC Name:

anilinium chloride

Constituent 2

Reference substance name:

aniline, hydrochloride

IUPAC Name:

aniline, hydrochloride

Details on test material:

- Name of test material (as cited in study report): aniline HCl
- Analytical purity: 98.5 + 1.2 %

Method

Species / strain

Species / strain / cell type:

other: S. typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538, E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 mix from rats, mice or hamsters treated with Aroclor 1254 (500 mg/kg)

Test concentrations with justification for top dose:

0.3, 1.0, 3.3, 10, 33.3, 100, and 333.3 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol; distilled water
- Justification for choice of solvent/vehicle:

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

distilled water or ethanol

True negative controls:

other: a total of 59 substance were evaluated, some of which were positive

Positive controls:

yes

Positive control substance:

other: with S9: 2-AA (2-aminoanthracene); without S9: TA1537: 9-AA (9-aminoacridine), TA100 and TA1535: Na azide (sodium azide), TA98 and TA1538: 2-NF (2-nitrofluorene): E. coli: AF-2 (2-2(-furyl)-3-(5-nitro-2-furyl)acrylarnide), MNNG (N-methyl-NI-nitro-N-nitros

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
The bacterial strain, test chemical, and S-9 mix, when required, were added to 2 ml of molten top agar at 45°C. The contents were then mixed and poured onto minimal agar plates. All plates were prepared in triplicate, and concurrent positive and negative controls were run at all times. Plates were incubated at 37°C for 48 hr prior to counting.

METHOD OF APPLICATION: preincubation

The bacterial strain, test chemical, and S-9 mix, when required, were added to tubes. The contents were mixed and incubated at 37°C for 20-30 min. At the end of the incubation period 2 ml of top agar was added to each tube, and the contents were mixed and poured onto the minimal agar plates. Plates were incubated at 37°C for 48 hr.
Since the prime objective of this study was to evaluate inter-laboratory reproducibility, the laboratories were only required to do a single assay with each coded chemical.

DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency

Evaluation criteria:

A positive response was defined as a dose-related increase with at least two doses being greater than or equal to twofold background, unless the background was less than ten, in which case a threefold increase was required. A question mark (?) was defined as a test in which only a single dose was equal to or greater than twofold (or threefold) background. Low-level, non-dose-related increases were scored as negative, as were single doses whose increase was the result of widely divergent replicate plate Counts.
An experiment was designated "no test" (NT) if the majority of plates were contaminated or showed a toxic response, controls were not run, or, in the case of TA100, if the mean background Count plus the standard error of the mean was equal to or greater than 300. It must be emphasized that these rules were selected in order to facilitate comparison of data and are generally not used in any of the authors' laboratories. There were numerous instances in which the laboratory's or the authors' judgment disagreed with the results obtained from using these fixed criteria. However, these rules were necessary for comparing the results; they are not recommended for use beyond this comparison study.
Although the protocol only called for one experiment, laboratories occasionally performed additional experiments. In these cases only the initial experiment was selected for analysis and presentation to maintain comparability across laboratories. In order to obtain analyzable data for each laboratory, exceptions to this rule were made when (1) the testing laboratory repeated the experiment at the Same or lower doses because of high levels of toxicity or contamination or (2) the positive or negative control values were outside the laboratory's acceptable range. Otherwise the data are presented as reported to the study organizers. All data and supporting experimental design information were entered on standard forms.

Statistics:

not specified

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538, E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity observed up to 333.3 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.