4-aminoazobenzene

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: Acceptable publication which meets basic scientific principles

Data source

Reference

Reference 1

Reference Type:

publication

Title:

The bacterial mutation test. Six tests for carcinogenicity

Author:

Anderson, D. and Styles, J.A.

Year:

1978

Bibliographic source:

Br J Cancer 37: pp. 924-30

Reference 2

Reference Type:

secondary source

Title:

4-Aminoazobenzene. BUA-Stoffbericht 217

Author:

Beratergremium für Altstoffe (BUA)

Year:

1998

Bibliographic source:

BUA der Gesellschaft dt. Chemiker, S. Hirzel Wissenschaftliche Verlagsgesellschaft, Stuttgart

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

only 4 strains used

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-aminoazobenzene

EC Number:

200-453-6

EC Name:

4-aminoazobenzene

Cas Number:

60-09-3

Molecular formula:

C12H11N3

IUPAC Name:

4-[(E)-2-phenyldiazen-1-yl]aniline

Details on test material:

no data

Method

Target gene:

his

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium TA98, TA100, TA1535, TA1538

Details on mammalian cell type (if applicable):

- Type and identity of media: Minimal Glucose Agar Medium (1.5% Difco Bacto agar in Vogel-Bonner medium with 2% glucose)
- Properly maintained: yes
- Periodically checked for: histidine requirements, deep rough, DNA excision-repair deficiency, R-factor plasmid, rate of reversion and purity

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

4, 20, 100, 500, 2500 µg/plate

Vehicle / solvent:

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

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

with metabolic activation

Migrated to IUCLID6: for strain TA1538 and TA98

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-(1-chloro-2-isopropylaminoethyl)naphthalene for strains TA1535 and TA100

Remarks:

with metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 2 to 3 days
- Expression time (cells in growth medium): 2 to 3 days

Evaluation criteria:

The results were accepted as positve if:
- there was a 2-fold increase over the negative control count for any strain;
- the negative-control cultures had counts within about 50% of the mean value;
- the positive-control cultures had counts greater than twice the negative-control values;
- the correct strains responded to the appropriate positive-control compounds;
- there was a background lawn indicating at least 10% survival

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

3fold at 500µg AAB/plate

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

5fold at 100µg AAB/plate

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

6fold at 500µg AAB/plate

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

5fold at 20µg AAB/plate

Cytotoxicity / choice of top concentrations:

not specified

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'.

Any other information on results incl. tables

Mean Number of S. Typhimurium Revertant Colonies in Each Strain with No Treatment (C) or with 0.1ml/Plate Water or DMSO

	TA1535			TA1538			TA98			TA100
	C	water	DMSO	C	water	DMSO	C	water	DMSO	C	water	DMSO
mean	30.4	30.33	25.87	28.83	31.47	26.4	47.11	48.74	42.06	63.57	60.90	57.92
s.d.	17.15	21.82	14.82	15.00	15.58	11.12	31.27	33.27	30.48	43.44	31.4	25.46
s.e.	3.13	4.0	2.70	2.74	2.84	2.03	5.28	5.62	5.08	9.48	6.85	4.89
n	30	30	30	30	30	30	35	35	36	21	21	27

Induction of Revertants in 4 Test Strains of S. Typhimurium

	TA1535	TA1538	TA98	TA100	Test result
maximum fold increase in Salmonella revertants above the concurrent negativ-control values	3	5	6	5	+
concentration of test substance (µg/plate)	500	100	500	20