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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Oct. 18, 2000 - Dec. 01, 2000
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose:
reference to same study
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
Oct. 18, 2000 - Dec. 01, 2000
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
See read-across justification attached in section 13.2
Reason / purpose:
read-across source
Related information:
Composition 1
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay
Test material information:
Composition 1
Specific details on test material used for the study:
(if 0,0 then see remarks)
6-Octen-1-ol, 3,7-dimethyl-, acetate
EC-No: Chemical Name:
205-775-0
Remark Rel. 2: according to OECD 471 and GLP, but the plate incorporation method was used
for both experiments; AMES test using strains TA98, TA100, TA102, TA1535, TA1537
up to 5000 μg/plate in DMSO;
Target gene:
histidine
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
0, 5, 15, 50, 150, 500, 1500 µg/Plate
Negative controls:
yes
Solvent controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
mitomycin C
other: 2-Aminoanthracene
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding (if applicable):
0.1mL of bacteria

DURATION
- Preincubation period: 11-12 hours
- Exposure duration: 48 to 72 h
at 37°C in the dark

NUMBER OF REPLICATIONS:
3
Rationale for test conditions:
This test uses several specially constructed histidine-requiring (his-) mutants of Salmonella typhimurium and measures his- to his+ reversion induced by chemicals which cause base changes or frameshift mutations in the genome of this organism.
Evaluation criteria:
A reduction in the number of revertant colonies and/or a diminution of the background lawn was taken as an indication of bacteriotoxicity.
Statistics:
X²-test
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Additional information on results:
In the absence of S9-mix the test substance was bacteriotoxic towards the strains TA102 at 150µg/plate, towards the strains TA1537, and TA100 at 500µg/plate, towards the strain TA98 at 1500μg/plate, and towards the strain TA1535 at 5000μg/plate. In the presence of S9-mix the test substance was bacteriotoxic towards the strains TA102 at 500µg/plate and towards the strains TA1535, TA1537, TA100, and TA98 at 1500µg/plate. Precipitation of the test compound on the plates was not observed.

see attachment

Conclusions:
In conclusion, these results indicate that the test substance under the experimental conditions described, was not mutagenic to Salmonella typhimurium strains TA1535, TA1537, TA98, TA100, and TA102 in the presence and absence of a metabolizing system.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
Designation: CITRONELLYL ACETATE EXTRA
Appearance: colourless to pale yellowish liquid
Specific details on test material used for the study:
Chemical Name: 6-Octen-1-ol, 3,7-dimethyl-, acetate
EC-No: 205-775-0

Method

Target gene:
histidine
Species / strain
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
0, 5, 15, 50, 150, 500, 1500 µg/Plate
Controls
Negative controls:
yes
Solvent controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
mitomycin C
other: 2-Aminoanthracene
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding (if applicable):
0.1mL of bacteria

DURATION
- Preincubation period: 11-12 hours
- Exposure duration: 48 to 72 h
at 37°C in the dark

NUMBER OF REPLICATIONS:
3
Rationale for test conditions:
This test uses several specially constructed histidine-requiring (his-) mutants of Salmonella typhimurium and measures his- to his+ reversion induced by chemicals which cause base changes or frameshift mutations in the genome of this organism.
Evaluation criteria:
A reduction in the number of revertant colonies and/or a diminution of the background lawn was taken as an indication of bacteriotoxicity.
Statistics:
X²-test

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Additional information on results:
In the absence of S9-mix the test substance was bacteriotoxic towards the strains TA102 at 150µg/plate, towards the strains TA1537, and TA100 at 500µg/plate, towards the strain TA98 at 1500μg/plate, and towards the strain TA1535 at 5000μg/plate. In the presence of S9-mix the test substance was bacteriotoxic towards the strains TA102 at 500µg/plate and towards the strains TA1535, TA1537, TA100, and TA98 at 1500µg/plate. Precipitation of the test compound on the plates was not observed.

Any other information on results incl. tables

See attachment

Applicant's summary and conclusion

Conclusions:
In conclusion, these results indicate that the test substance under the experimental conditions described, was not mutagenic to Salmonella typhimurium strains TA1535, TA1537, TA98, TA100, and TA102 in the presence and absence of a metabolizing system.