Esterification product of castor oil and tetrahydromethyl-1,3-isobenzofuranedione

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: guideline study and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Esterification product of castor oil and tetrahydromethyl-1,3-isobenzofuranedione

EC Number:

700-064-6

Cas Number:

2105830-60-0

Molecular formula:

Not applicable as this is a UVCB substance

IUPAC Name:

Esterification product of castor oil and tetrahydromethyl-1,3-isobenzofuranedione

Constituent 2

Reference substance name:

[TN]Multiester P97-463[/TN][SPEC][/SPEC][AM]100 %[/AM]

IUPAC Name:

[TN]Multiester P97-463[/TN][SPEC][/SPEC][AM]100 %[/AM]

Details on test material:

Multiester P97-463
Batch No: NB2890-61

Method

Target gene:

hisC3076 (frameshift): S. typhimurium TA1537
hisD3052/R-factor (frameshift): S. typhimurium TA98
hisG46 (base-pair substitutions): S. typhimurium TA1535
hisG46/R-factor (base-pair substitutions): S. typhimurium TA100
WP2uvrA (base-pair substitutions): E.coli

R-factor: plasmid pKM101 (increases error-prone DNA repair)

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium TA698, TA100, TA1535, TA 1537, Escherichia coli WP2uvrA

Additional strain / cell type characteristics:

other: each Salmonella strain, additional mutations: rfa, gal, chl, bio, uvrB

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9-mix (prepared from adult male Wistar rats, liver induced by phenobarbital and ß-naphthoflavone)

Test concentrations with justification for top dose:

1st experiment (dose range finding): 100, 333, 1000, 3330 and 5000 μg/plate; for TA100 and WP2uvrA additionally 3, 10, 33 μg/plate
2nd experiment: 100, 333, 1000, 3330 and 5000 μg/plate

Vehicle / solvent:

The test substance was dissolved in dimethyl sulfoxide. The stock solutions were treated with ultrasonic waves until the test substance had completely dissolved. Test substance concentrations were used within 15 minutes to 2 hours after preparation.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: various (see remarks)

Remarks:

Without metabolic activation: sodium azide, 9-aminoacridine, 2.nitrofluorene, methylmethanesulfonate, 4-nitroquinoline N-oxide; with metabolic activation: 2-aminoanthracene

Results and discussion

Test results

Species / strain:

other: Salmonella typhimurium TA698, TA100, TA1535, TA 1537, Escherichia coli WP2uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

Precipitation of Multiester P97-463 on the plates was observed at the start of the incubation period at concentrations of 1000 μg/plate and upwards and at 5000 μg/plate at the end of the incubation period.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

All bacterial strains showed negative responses over the entire dose range, i.e. no significant dose-related increase in the number of revertants in two independently repeated experiments. The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Based on the results of this study it is concluded that Multiester P97-463 is not mutagenic in the Salmonella typhimurium reverse mutation assay an in the Escherichia coli reverse mutation assay.

Executive summary:

Multiester P97-463 was tested in the Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of Salmonella typhimurium (TA1535, TA1537, TA98 and TA100) and in the Escherichia coli reverse mutation assay with a tryptophan-requiring strain of Escherichia coli (WP2uvrA). The test was performed in two independent experiments in the presence and absence of S9-mix (rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone). The study procedures described in this report were based on the most recent OECD and EEC guidelines.

Batch NB2890-61 of Multiester P97-463 was a very viscous amber liquid. The test substance was dissolved in dimethyl sulfoxide. In the dose range finding test, Multiester P97-463 was tested up to concentrations of 5000 μg/plate in the absence and presence of S9-mix in the strains TA100 and WP2uvrA.

Multiester P97-463 precipitated on the plates at the dose level of 5000 μg/plate. The bacterial background lawn was not reduced at any of the concentrations tested and no decrease in the number of revertants was observed. Results of this dose range finding test were reported as part of the first experiment of the mutation assay. Based on the results of the dose range finding test, Multiester P97-463 was tested in the first mutation assay at a concentration range of 100 to 5000 μg/plate in the absence and presence of 5% (v/v) S9-mix in tester strains TA1535, TA1537 and TA98. In an independent repeat of the assay with additional parameters, Multiester P97-463 was tested at the same concentration range as the first assay in the absence and presence of 10% (v/v) S9-mix in tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA. Multiester P97-463 precipitated on the plates at the top dose of 5000 μg/plate. The bacterial background lawn was not reduced at any of the concentrations tested and no biologically relevant decrease in the number of revertants was observed. Multiester P97-463 did not induce a significant dose-related increase in the number of revertant (His+) colonies in each of the four tester strains (TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trp+) colonies in tester strain WP2uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed in an independently repeated experiment. In this study, the negative and strain-specific positive control values were within our laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

Based on the results of this study it is concluded that Multiester P97-463 is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.