1,4-bis(mesitylamino)anthraquinone

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:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well performed GLP compliant OECD guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,4-bis(mesitylamino)anthraquinone

EC Number:

204-155-7

EC Name:

1,4-bis(mesitylamino)anthraquinone

Cas Number:

116-75-6

Molecular formula:

C32H30N2O2

IUPAC Name:

1,4-bis(mesitylamino)anthraquinone

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

rat S 9 mix

Test concentrations with justification for top dose:

plate incorporation test (experiment I) and pre-incubation test (experiment II)
33; 100; 333; 1000; 2500; and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solvent was chosen because of its solubility properties and its relative non-toxicity to the bacteria

Controls

Untreated negative controls:

yes

Remarks:

untreated

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Without metabolic activation: sodium azide, NaN3 (TA 1535, TA 100) / 4-nitro-o-phenylene-diamine, 4-NOPD (TA 1537, TA 98) / methyl methane sulfonate, MMS (WP2 uvrA) /// With metabolic activation: 2-aminoanthracene, 2-AA (all tester strains)

Details on test system and experimental conditions:

- Experiment I: Plate Incorporation Test
- Experiment II: Pre-Incubation Test
- number of replicates: 3
- preincubation period: 1 hour at 37°C
- exposure duration: at least 48 hours at 37°C

Evaluation criteria:

A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.

A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration.

An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.

A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant.

Statistics:

No statistical evaluation of the data is required according to OECD guideline 471

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

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

without S9 mix

Concentration µg/plate	Revertants/plate mean from three plates
Strain	TA 1535		TA 1537		TA 98		TA 100		WP2 uvrA
Exp	I	II	I	II	I	II	I	II	I	II
Negative control	10	13	7	7	25	28	78	130	38	34
Solvent control	11	19	6	10	21	29	81	102	30	34
Positive control #	1016	927	53	69	167	199	1175	1044	295	177
33	12	14	5	9	23	33	66	102	30	29
100	13	15	8	7	21	34	74	114	43	31
333	8	14	4	6	23	25	61	110	30	36
1000	8	16	7	5	24	28	61	101	34	24
2500	12	15	5	7	21	28	67	104	37	25
5000	9	13	4	5	23	26	52	101	29	32

with S9 Mix

Concentration µg/plate	Revertants/plate mean from three plates
Strain	TA 1535		TA 1537		TA 98		TA 100		WP2 uvrA
Exp	I	II	I	II	I	II	I	II	I	II
Negative control	12	13	5	13	29	38	87	123	37	33
Solvent control	19	14	7	12	34	34	87	116	30	33
Positive control ##	125	92	59	44	442	513	482	477	169	186
33	14	20	9	13	37	37	72	125	39	40
100	13	17	7	16	31	37	67	131	42	34
333	16	16	10	13	32	37	80	132	29	36
1000	13	15	9	14	34	35	75	114	31	32
2500	15	15	11	15	31	38	88	118	31	36
5000	10	16	9	12	28	32	67	122	35	36

# Sodium azide (10.0 µg/plate) strains TA 1535 and TA 100 4-nitro-o-phenylene-diamine strains TA 1537 (50 µg/plate) and TA 98 (10.0 µg/plate) Methyl methane sulfonate (4 µg/plate) strain WP2 uvrA

## 2-aminoanthracene (2.5 µg/plate) strains TA 1535, TA 1537, TA 98, and TA 100 2-aminoanthracene (10.0 µg/plate) strain WP2 uvrA

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

During the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair
changes or frameshifts in the genome of the strains used.

Executive summary:

The test item was assessed for its potential to induce gene mutations in the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using Salmonella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100, and the Escherichia coli strain WP2 uvrA according to OECD guideline 471.

The assay was performed in two independent experiments both with and without rat liver microsomal activation. Each concentration and the controls were tested in triplicate. The test item was tested at the following concentrations:

33; 100; 333; 1000; 2500; and 5000 µg/plate

The plates incubated with the test item showed normal background growth up to 5000 µg/plate with and without S9 mix in both experiments.

No toxic effects, evident as a reduction in the number of revertants, occurred in the test groups with and without metabolic activation.

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with the test item at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.

Appropriate reference mutagens were used as positive controls. They showed a distinct increase of induced revertant colonies.