Reaction mass of 2-(1,1-dimethylpropyl)anthraquinone and 2-(1,2-dimethylpropyl)anthraquinone

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study. However, the updated and currently applicable guideline states use of 5 strains instead of 4, which are used here.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

a fifth strain was not included (requested in the updated and currently applicable OECD guideline)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Test material form:

liquid: viscous

Details on test material:

- Substance type: Technical product.
- Storage condition of test material: No data.

Method

Target gene:

histidine gene

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

Liver S9-mix prepared from Aroclor 1254-induced Wistar or Sprague Dawley rats

Test concentrations with justification for top dose:

100, 333, 1000, 3330 and 5000 µg/plate.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Sodium azide (TA1535 -S9), 9-aminoacridine (TA1537 -S9), daunomycine (TA98 -S9), methylmethanesulfonate (TA100 -S9), 2-aminoanthracene (TA1535, TA1537, TA100, TA98 +S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation);

DURATION
- Exposure duration: 48 hours.

NUMBER OF REPLICATIONS: two separate experiments were conducted with three replicates per dose level.

Evaluation criteria:

A test substance is considered positive in the Ames test if:
1) It induced at least a 2-fold, dose related increase in the number of revertants with respect to the number induced by the solvent control in any of the tester strains, either with or without metabolic activation.
2) The positive response should be reproducible in at least one independently repeated experiment.

Statistics:

The revertant colonies have been counted automatically with a colony counter or manually, if less than 40 colonies per plate were present.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA98, TA1537

Metabolic activation:

without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium, other: TA98, TA1537

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium, other: TA100, TA1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

The strains TA1537 and TA98, in the presence of S9-mix, and the strains TA1535 and TA100, in the presence and absence of S9-mix showed
negative responses over the entire dose range of the test substance, in two independently repeated experiments. However, in the absence of S9-mix the test substance did induce a reproducible 3- to 21-fold, dose-related increase in the number of revertant colonies in tester strains TA1537 and
TA98.

Any other information on results incl. tables

Table 1: Summary of the results

Parameter	Strain
	TA1535		TA1537		TA98		TA100
Metabolic activation	with	without	with	without	with	without	with	without
Genotoxicity	no	no	no	yes	no	yes	no	no
Cytotoxicity	no	no	no	no	no	no	no	no
Vehicle control valid	yes	yes	yes	yes	yes	yes	yes	yes
Positive control valid	yes	yes	yes	yes	yes	yes	yes	yes

Applicant's summary and conclusion

Conclusions:

2-Amylanthraquinone was tested in the Ames test for its ability to induce mutations in five histidine dependent Salmonella typhimurium strains. Two independent mutation tests were performed, each in presence and absence of a metabolic activation system (S9-mix). The bacterial strains were exposed to 0-5000 μg/plate 2-amylanthraquinone. In the absence of S9-mix the test substance induced a reproducible 3- to 21-fold, dose-related increase in the number of revertant colonies in tester strains TA1537 and TA98. The test substance is considered to be mutagenic under the experimental conditions employed.