GOLDEN YELLOW SCJ 1006

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:

1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD compliant GLP study without deviations

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Version / remarks:

92/69/EEC

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Identification: FAT 40543/A
Description: Red powder
Batch: TV RN 196-200
Purity: 90%
Test substance storage: At room temperature in the dark
Stability under storage conditions: Stable
Expiry date: November 01, 2000
Stability in vehicle: Dimethylsulfoxide: not indicated

Method

Target gene:

gene of histidine-requiring Salmonella typhimurium bacterial strains resulting in histidine-independent strains, and in a gene of tryptophan-requiring Escherichia coli bacterial strain resulting in a tryptophan-independent strain

Species / strain

Species / strain / cell type:

bacteria, other: Salmonella typhimurium TA 1535, TA 1537, TA 98 , TA 100 Escherichia coli WP2 uvrA

Additional strain / cell type characteristics:

other: histidine-requiring and tryptophan-requiring

Metabolic activation:

with and without

Metabolic activation system:

Aroclor-induced rat liver S9

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 3, 10, 33, 100, 333, 1000, 3330, 5000 µg/plate (all tested in triplicates)
Concentration range in the main test (without metabolic activation): 3, 10, 33, 100, 333, 1000, 3330, 5000 µg/plate (all tested in triplicates)

Vehicle / solvent:

Solvent: Dimethylsulfoxide

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

for TA1535 without metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

for TA1537 without metabolic activation

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: duanomycin

Remarks:

for TA98 without metabolic activation

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

for TA100 without metabolic activation

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

for WP2uvrA without metabolic activation

Controls 6

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

all strains with metabolic activation

Details on test system and experimental conditions:

Concentration of the test substance resulting in precipitation: 100 µg/plate

Evaluation criteria:

No formal hypothesis testing was done. A test substance is considered negative (not mutagenic) in the test if:
a) The total number of revertants in any tester strain at any concentration is not greater than two times the solvent control value, with or without metabolic activation.
b) The negative response should be reproducible in at least one independently repeated experiment.
A test substance is considered positive (mutagenic) in the test if:
a) It induces 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. However, any mean plate count of less than 20 is considered to be not significant.
b) The positive response should be reproducible in at least one independently repeated experiment.
The preceding criteria were not absolute and other modifying factors might
enter into the final evaluation decision.

Results and discussion

Test results

Test results 1

Species / strain:

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

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

TA1537 only at concentrations with precipitation

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation as of 100 µg/plate and above

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation as of 100 µg/plate and above

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

without

Genotoxicity:

positive

Remarks:

only at concentrations with precipitation

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation as of 100 µg/plate and above

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 98

Metabolic activation:

without

Genotoxicity:

positive

Remarks:

only at concentrations with precipitation

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation as of 100 µg/plate and above

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 100

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation as of 100 µg/plate and above

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 6

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitation as of 100 µg/plate and above

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Observations:
Significant increases in revertants were observed in the precipitating range only (>=100 micrograms/plate) in strains TA 1537, TA 98, in the absence of S9 and strain TA 1537 in the presence of S9.
At non-precipitating concentrations (<100 micrograms/plate), significant increases in revertants were observed in strains TA 1535, TA 98, TA 100 and WP2 uvrA in the presence of S9.

No toxicity was observed.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

In the absence of S9-mix, FAT40543/A induced in strain TA1537 5- and 4-fold dose-related increases in the number of revertant (His+) colonies, in experiment 1 and 2 respectively. In strain TA98 the test substance induced 7-and 6-fold dose-related increases, in experiment 1 and 2 respectively. These increases were observed only at precipitating concentrations. In the tester strains TA1535, TA100 and WP 2 uvrA, FAT40543/A did not induce a dose-related increase in the number of revertant colonies in both independent experiments. In the presence of S9-mix, FAT40543/A induced in strain TA1535 14- and 7-fold dose-related increases in the number of revertant (His+) colonies, in experiment 1 and 2 respectively. In strain TA1537 the test substance induced 6- and 9-fold dose-related increases, in experiment 1 and 2 respectively. In. TA98 the test substance induced dose related 4-fold increases in both experiments. FAT 40543/A induced in strain TA100 12- and 8-fold dose-related increases, in experiment 1 and 2 respectively. In WP 2 uvrA the test substance induced dose related 2-fold increases in the number of revertant (Trp+) colonies in both experiments. The increases in tester strain TA1537 were observed only at precipitating concentrations. The increases in the four other strains were observed at non-precipitating concentrations also.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive with metabolic activation
positive without metabolic activation

Based on the results of this study it is concluded that FAT 40543/A is mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

Executive summary:

FAT 40543/A was tested in the Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of Salmonella typhimurium (TA1535, TA1537, TA100 and TA98) and in the Escherichia coli reverse mutation assay with the tryptophan-requiring strain WP2 uvrA in two independent experiments. FAT 40543/A was tested up to and including concentrations of 5000 µg/plate in the absence and presence of S9-mix. In the absence of S9-mix, FAT 40543/A induced in strain TA1537 5- and 4-fold dose-related increases in the number of revertant (His+) colonies, in experiment 1 and 2 respectively. In strain TA98 the test substance induced 7-and 6-fold dose-related increases, in experiment 1 and 2 respectively. These increases were observed only at precipitating concentrations. In the tester strains TA1535, TA100 and WP2 uvrA, FAT 40543/A did not induce a dose-related increase in the number of revertant colonies in both independent experiments. In the presence of S9-mix, FAT40543/A induced in strain TA1535 14- and 7-fold dose-related increases in the number of revertant (His+) colonies, in experiment 1 and 2 respectively. In strain TA1537 the test substance induced 6- and 9-fold dose-related increases, in experiment 1 and 2 respectively. In. TA98 the test substance induced dose related 4-fold increases in both experiments. FAT 40543/A induced in strain TA100 12- and 8-fold dose-related increases, in experiment 1 and 2 respectively. In WP2 uvrA the test substance induced dose related 2-fold increases in the number of revertant (Trp+) colonies in both experiments. The increases in tester strain TA1537 were observed only at precipitating concentrations. The increases in the four other strains were observed at non-precipitating concentrations also.

Based on the results of this study it is concluded that FAT 40543/A is mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.