Reaction products of Sodium 1-amino-4-bromo-9,10-dioxoanthracene-2-sulphonate coupled with Aniline, subsequently coupled with p-(1,1-dimethylpropyl)phenol, subsequently coupled with Ethyl benzoylacetate, subsequently coupled with Fuming Sulfuric acid, sodium salts.

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

other company data

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Acid Red 143

IUPAC Name:

Acid Red 143

Method

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538

Metabolic activation:

with and without

Metabolic activation system:

rat liver metabolic activation system (S-9)

Test concentrations with justification for top dose:

Range Finder Experiment and Experiment 1: 1.6, 8, 40, 200, 1000 and 5000 µg/plate, with and without metabolic activation

Vehicle / solvent:

- Vehicle: sterile purified water.

Controls

Untreated negative controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: 2-Aminoanthracene

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

When the data from the Range-Finder Experiment were analysed at the 1 % level using Dunnett's test a statistically significant increase in revertant numbers was observed following TA 100 treatments in the presence of S-9. As this increase was small in magnitude and was observed at the lowest dose treatment, this was not considered to be indicative of test item mutagenic activity in this strain under this treatment condition. No other test item treatments induced statistically significant increases in revertant numbers, when the data were analysed at the 1 % level using Dunnett's test.

TOXICITY
No clear evidence of toxicity, as manifest by a marked decrease in revertant numbers or a thinning of the background bacterial lawn was observed following the any of the dose treatments in any of the strains in the presence or the absence of S-9.

PRECIPITATION
No precipitation of test agent was observed following any of the treatments in either the presence or absence of S-9.

Any other information on results incl. tables

Summary of mean revertant colonies (-S9) - Experiment 1

Dose Level µg/plate	TA 98	TA 100	TA 1535	TA 1537	TA 1538
Water	36± 5	117± 12	13± 1	15± 2	19± 6
Test item 1.6	41± 9	118± 7	10± 5	13± 4	18± 5
Test item 8	32± 10	132± 12	9± 1	16± 2	17± 7
Test item 40	39± 4	122± 1	13± 1	9± 1	17± 2
Test item 200	42± 7 (M)	116± 3	13± 3 (M)	12± 5 (M)	14± 4 (M)
Test item 1000	43± 7 (M)	110± 11 (M)	8± 2 (M)	12± 4 (M)	20± 3 (M)
Test item 5000	36± 2 (M)	94± 9 (M)	11± 6 (M)	12± 2 (M)	18± 7 (M)
2-Nitrofluorene 5	664± 35				338± 25
Sodium azide 2		763± 22	443± 18
9-Aminoacridine 50				136± 23

(M) Plates counted manually

Summary of mean revertant colonies (+S9) - Experiment 1

Dose Level µg/plate	TA 98	TA 100	TA 1535	TA 1537	TA 1538
Water	40± 5	171± 15	14± 3	13± 4	27± 4
Test item 1.6	39± 5	209± 17	10± 2	15± 2	32± 3
Test item 8	49± 6	168± 20	18± 1	17± 6	22± 6
Test item 40	49± 7	185± 4	22± 7	12± 2	24± 8
Test item 200	40± 6	156± 8	17± 2	15± 2 (M)	25± 6 (M)
Test item 1000	44± 2 (M)	118± 12 (M)	16± 4 (M)	13± 6 (M)	23± 6 (M)
Test item 5000	37± 4 (M)	107± 14 (M)	9± 5 (M)	17± 1 (M)	26± 1 (M)
Benzo[a]pyrene 10	247± 35
2-Aminoanthracene 5		1831± 55	164± 17	358± 44	961± 46

(M) Plates counted manually

Applicant's summary and conclusion

Conclusions:

It was concluded that test item did not induce mutation in Salmonella typhimurium strains TA 98, TA l00, TA l535, TA 1537 and TA 1538 under the conditions employed in the screening study.

Executive summary:

A screening study was conducted on test substance to determine reverse mutation in five histidine-requiring strains of Salmonella typhimuium. The strains tested were TA 98, TA 100, TA 1535, TA 1537 and TA 1538. Both the range finder experiment and experiment 1 were conducted at the concentrations of 1.6, 8, 40, 200, 1000 and 5000 µg/plate, with and without metabolic activation.

When the data from the Range-Finder Experiment were analysed at the 1 % level using Dunnett's test a statistically significant increase in revertant numbers was observed following TA 100 treatments in the presence of S9. As this increase was small in magnitude and was observed at the lowest dose treatment, this was not considered to be indicative of test item mutagenic activity in this strain under this treatment condition. No other test item treatments induced statistically significant increases in revertant numbers, when the data were analysed at the 1 % level using Dunnett's test.

No clear evidence of toxicity, as manifest by a marked decrease in revertant numbers or a thinning of the background bacterial lawn was observed following the any of the dose treatments in any of the strains in the presence or the absence of S9.

No precipitation of test agent was observed following any of the treatments in either the presence or absence of S9.

Conclusion

It was concluded that test item did not induce mutation in Salmonella typhimurium strains TA 98, TA l00, TA l535, TA 1537 and TA 1538 under the conditions employed in the screening study.