(1S,4R,4aS,9aR)-4,4a,9,9a-tetrahydro-1H-1,4-methanofluorene

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:

04 July 2002 - 21 February 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to OECD, EU, US FDA and Japanese (EPA, MHLW and METI) test guidelines, and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

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:

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

Version / remarks:

(Commission Directive 2000/32/EC; L136, 57-64, Annex 4D)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: US FDA Administration Center for Food Safey & Applied Nutrition Office of Premarket Approval, Redbook 2000, Toxicological Principles for the Safety of Food Ingredients, IV.C.1.a Bacterial Reverse Mutation Test, July 07, 2000.

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Version / remarks:

"Kanpoan 287 - EPA", "Eisei 127 - MHLW", and "Heisei 09/10/31 Kikyoku 2 - METI"

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(1R,2S,10R,11S)-tetracyclo[9.2.1.0²,¹⁰.0³,⁸]tetradeca-3(8),4,6,12-tetraene

EC Number:

700-736-9

Cas Number:

2149571-40-2

Molecular formula:

C14H14

IUPAC Name:

(1R,2S,10R,11S)-tetracyclo[9.2.1.0²,¹⁰.0³,⁸]tetradeca-3(8),4,6,12-tetraene

Test material form:

other: liquid

Details on test material:

- Name of test material (as cited in study report): MTF
- Substance type: organic
- Physical state: liquid
- Analytical purity: 99.1%
- Lot/batch No.: 010920
- Stability under test conditions: > 3 days in water
- Storage condition of test material: Approx. 4°C. The container has to be tightly sealed with nitrogen. Each time, the test item is used, the container has to be refilled with nitrogen.

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 102

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

rat-liver post mitochondrial supernatant (S9 fraction)

Test concentrations with justification for top dose:

Range finding test:
strains S. typhimurium TA 100 and E. coli WP2 uvrA (with and without methabolic activation): 20.6, 61.8, 185.2, 555.6, 1666.7 and 5000 µg/plate


without metabolic activation:
strains TA 100, TA 102: 1.0, 3.3, 10, 33 and 100 µg/plate
strain TA 1537: 10, 33, 100, 333 and 1000 µg/plate
strains TA 1535, TA 98, WP2 uvrA: 33, 100, 333, 1000 and 2500 µg/plate

with metabolic activation (pre-incubation assay):
strains TA 100, TA 1537, TA 102: 10, 33, 100, 333 and 1000 µg/plate
strains TA 1535, TA 98, WP2 uvrA: 33, 100, 333, 1000 and 2500 µg/plate

Due to a wide range of toxicity in the pre-incubation assay with microsomal activation, additional concentrations were tested with metabolic activation:
strains TA 100, TA 1537: 0.3, 1.0, 3.3, 10 and 33 µg/plate
strains T A 1535, T A 98, T A 102: 3.3, 10, 33, 100 and 333 µg/plate.

Controls

Controls 1

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Without metabolic activation

Controls 2

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

Without metabolic activation

Controls 3

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Without metabolic activation

Controls 4

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

Without metabolic activation

Controls 5

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

mitomycin C

Remarks:

Without metabolic activation

Controls 6

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

With metabolic activation

Controls 7

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

other: 2-AA

Remarks:

With metabolic activation

Results and discussion

Test results

Test results 1

Key result

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

During the described mutagenicity test and under the experimental conditions reported, the test item did not induced gene mutations by base-pair changes or frameshifts in the genome of the strains used.


Therefore, MTF is considered to be non-mutagenic in this Salmonella typhimurium and Escherichia coli reverse mutation assay.