Caryophyllene, Reaction product with Acetic anhydride and acetic acid

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:

September 05-23 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

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:

other: "Comission Directive 2000/32/EC, L1362000, Aneexe 4D," dated May 19, 2000.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

[1R-(1α,2α,5β,8β)]-4,4,8-trimethyltricyclo[6.3.1.02,5]dodecan-1-yl acetate

EC Number:

260-555-1

EC Name:

[1R-(1α,2α,5β,8β)]-4,4,8-trimethyltricyclo[6.3.1.02,5]dodecan-1-yl acetate

Cas Number:

57082-24-3

Molecular formula:

C17H28O2

IUPAC Name:

4,4,8-trimethyltricyclo[6.3.1.0~2,5~]dodec-1-yl acetate

Constituent 2

Reference substance name:

1-((1S,2S,5R)-1,4,4-trimethyltricyclo[6.3.1.02,5]dodec-8-en-9-yl)ethan-1-one

Molecular formula:

C17H26O

IUPAC Name:

1-((1S,2S,5R)-1,4,4-trimethyltricyclo[6.3.1.02,5]dodec-8-en-9-yl)ethan-1-one

Constituent 3

Reference substance name:

(3S,3aS,7R,9aS)-1,1,7-trimethyldecahydro-3a,7-methanocyclopenta[8]annulen-3-yl acetate

Cas Number:

77346-43-1

Molecular formula:

C17H28O2

IUPAC Name:

(3S,3aS,7R,9aS)-1,1,7-trimethyldecahydro-3a,7-methanocyclopenta[8]annulen-3-yl acetate

Constituent 4

Reference substance name:

1-((1S,2S,5R,9S)-1,4,4-trimethyltricyclo[6.3.1.02,5]dodec-7-en-9-yl)ethan-1-one

Cas Number:

Not applicable

Molecular formula:

C17H26O

IUPAC Name:

1-((1S,2S,5R,9S)-1,4,4-trimethyltricyclo[6.3.1.02,5]dodec-7-en-9-yl)ethan-1-one

Constituent 5

Reference substance name:

4,4,8-trimethyltricyclo[6.3.1.02,5]dodecan-1-ol

EC Number:

207-458-2

EC Name:

4,4,8-trimethyltricyclo[6.3.1.02,5]dodecan-1-ol

Cas Number:

472-97-9

Molecular formula:

C15H26O

IUPAC Name:

4,4,8-trimethyltricyclo[6.3.1.0~2,5~]dodecan-1-ol

Constituent 6

Reference substance name:

(3S,3aS,7R,9aS)-1,1,7-trimethyldecahydro-3a,7-methanocyclopenta[8]annulen-3-ol

Cas Number:

62560-57-0

Molecular formula:

C15H26O

IUPAC Name:

(3S,3aS,7R,9aS)-1,1,7-trimethyldecahydro-3a,7-methanocyclopenta[8]annulen-3-ol

Constituent 7

Reference substance name:

2,2,4,7a-tetramethyl-1,2,3,6,7,7a-hexahydro-1,3a-ethanoindene

Cas Number:

4545-68-0

Molecular formula:

C15H24

IUPAC Name:

2,2,4,7a-tetramethyl-1,2,3,6,7,7a-hexahydro-1,3a-ethanoindene

Constituent 8

Reference substance name:

not applicable

Cas Number:

not applicable

Molecular formula:

not applicable

IUPAC Name:

not applicable

Test material form:

liquid

Specific details on test material used for the study:

Name (as stated in the report): VETYNAL EXTRA
Batch: 9000474504
Expiration date: June 15, 2004

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Test concentrations with justification for top dose:

All Salmonella strains with and without S9: 33; 100; 333; 1000; 2500; and 5000 µg/plate

Vehicle / solvent:

ethanol

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-o-phenylene-diamine, 4-NOPD; 2-aminoanthracene, 2-AA

Results and discussion

Test results

Key result

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

In conclusion, it can be stated that during the distributes mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes of frameshifts in the genome of the strains used.

Therefore, VETYNAL EXTRA is considered to be non-mutagenic in the Salmonella thyphimurium reverse mutation assay.

Executive summary:

This study was performed to investigate the potential of VETYNAL EXTRA to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100, and TA 102.

 

The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations:

33; 100; 333; 1000; 2500; and 5000 µg/plate

 

Slight toxic effects, evident as a reduction in the number of revertants, were observed in strain TA 100 with S9 mix in experiment I, and without S9 mix in the experiment II.

 

The plate incubated with the test item showed normal background growth up to 5000 µg/plate with and without S9 mix in all strains used.

 

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with VETYNAL EXTRA at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.

 

Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies.