Bicyclo[3.1.1]hept-2-ene, 2,6,6-trimethyl-, phosphosulfurized

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The in vitro gene mutation study in bacteria was conducted on the registered substance according to OECD Testing Guideline 471. The registered substance was considered to be non-mutagenic in bacteria under the conditions of the test, for both plate incorporation and pre-incubation methods, with and without metabolic activation.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 20 April 2003 to 21 July 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Deviations:

no

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMHW Genotoxicity Testing Guideline, PAB Notification No. 1604

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

- Type and identity of media: Nutrient broth
- Properly maintained: yes

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9

Test concentrations with justification for top dose:

5, 15, 50, 150, 500, 1500, 5000 ug/plate

Vehicle / solvent:

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

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

2-nitrofluorene

sodium azide

N-ethyl-N-nitro-N-nitrosoguanidine

benzo(a)pyrene

other: 2-Aminoanthracene

Remarks:

2-aminoanthracene and benzo(a)pyrene done in the presence of S9. The other substances done in the absence of S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: 10 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 72 hours

Evaluation criteria:

To count the number of bacterial colonies present to look for a reproducible increase in revertant colony number of at least twice the concurrent solvent/vehicle control, with evidence of a positive dose-response relationship.

Statistics:

None performed

Key result

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:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

See attached background material.

Conclusions:

Interpretation of results (migrated information): negative

There was no significant increase in revertant colony numbers over control counts in any of the tester strains regardless of the presence of S9 or the concentration of the test substance. The substance was considered as non-mutagenic under the conditions of the test.

Executive summary:

The in vitro genotoxicity in bacteria of the test substance was determined in accordance with the OECD Guideline for Testing of Chemicals 471. Range finding (Experiment 1) and pre-incubation (Experiment 2) methods were performed at up to seven dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolizing system (10 % v/v liver S9 in standard cofactors).

The dose for Experiment 1 was 5000 µg/plate and led to the range for Experiment 2 being 5 to 5000 µg/plate. The experiment was repeated on a separate day (pre-incubation method) using fresh cultures of the bacterial strains and fresh test item formulations.

There were no significant increases in the frequency of revertant colonies recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation (S9-mix) in Experiment 1. Similarly, no toxicologically significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation (S9-mix) in Experiment 2. The test substance was therefore considered to be non-mutagenic under the conditions of the test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

The in vitro genotoxicity in bacteria of the registered substance was determined in accordance with the OECD Guideline for Testing of Chemicals 471. Ames plate incorporation (Experiment 1) and pre-incubation (Experiment 2) methods were performed both with and without the addition of a rat liver homogenate metabolizing system (10% liver S9 in standard co-factors). There were no significant increases in the frequency of revertant colonies recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation (S9-mix) in Experiment 1. Similarly, no toxicologically significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation (S9-mix) in Experiment 2. The registered substance was therefore considered to be non-mutagenic under the conditions of the test.

Justification for selection of genetic toxicity endpoint
Study was conducted by a GLP accredited laboratory using OECD Testing Guideline 471. The study was conducted on the registered substance. In accordance with Annex VII of REACH, no further testing was performed since the in vitro gene mutation study in bacteria returned a negative result.

Justification for classification or non-classification

An in vitro gene mutation study in bacteria was conducted on the registered substance and returned a negative result. In accordance with Annex VII of REACH, no further testing was performed considering this result. Therefore, the registered substance did not meet the criteria for classification according to Regulation (EC) No.1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures.