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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:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
Salmonella Mutagenicity Tests: II. Results From The Testing Of 270 Chemicals
Author:
Mortelmans,K, Haworth,S, Lawlor,T, Speck,W, Tainer,B And Zeiger,E
Year:
1986
Bibliographic source:
Environ. Mutagen. 8(Suppl. 7):1 -119, 1986

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
Gene mutation toxicity study was performed for Geranyl acetate to evaluate its mutagenic nature
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
- Name of test material (as cited in study report): Geranyl acetate - Molecular formula (if other than submission substance): C12H20O2
- Molecular weight (if other than submission substance): 196.288 g/mol
- Smiles notation (if other than submission substance): C(\CC\C=C(\C)C)(=C\COC(C)=O)C
- InChl (if other than submission substance): 1S/C12H20O2/c1-10(2)6-5-7-11(3)8-9-14-12(4)13/h6,8H,5,7,9H2,1-4H3/b11-8+
- Substance type: Organic
- Physical state: Liquid
Specific details on test material used for the study:
- Name of test material: Geranyl acetate
- IUPAC name: 3,7-dimethylocta-2,6-dien-1-yl acetate
- Molecular formula: C12H20O2
- Molecular weight (if other than submission substance): 196.288 g/mol
- Substance type: Organic
- Physical state: No data available
- Purity: 69.6 %
- Impurities (identity and concentrations): No data available

Method

Target gene:
Histidine
Species / strain
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
Male Sprague-Dawley rats and male Syrian hamsters were routinely used for the S9 preparation of the liver fractions
Test concentrations with justification for top dose:
0, 1, 3, 10, 33, 100, 333, 1000, 3333 µg/plate
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO
Controls
Negative controls:
not specified
Solvent controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 4-nitro-o-phenylenediamine (TA98; -S9), 2-aminoanthracene (all strains, +S9)
Details on test system and conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hr
- Expression time (cells in growth medium): 48 hr
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: At least five dose levels of the chemicals were tested, with three plates per dose level.

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data
Rationale for test conditions:
No data
Evaluation criteria:
1) mutagenic response: a dose-related, reproducible increase in the number of revertants over background, even if the increase was less than twofold;
2) nomutagenic response: when no increase in the number of revertants was elicited by the chemical;
3) questionable response: when there was an absence of a clear-cut dose-related increase in revertants; when the dose-related increases in the number of revertants were not reproducible;
or when the response was of insufficient magnitude to support a determination of mutagenicity
Statistics:
Mean and Standard error of mean

Results and discussion

Test results
Species / strain:
other: TA100, TA1535, TA1537, TA98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
Toxic in the absence of S9 at and above 100 µg/plate
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: No mutagenic potential
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: No data
- Other confounding effects: No data

RANGE-FINDING/SCREENING STUDIES: The chemical was initially tested with strain TA100 in the presence and the absence of the metabolic activation systems, over a wide dose range with an upper limit of 10 mg/plate, or less when solubility problems were encountered. Toxicity was evidenced by one or more of the following phenomena: appearance of his+ pinpoint colonies, reduced numbers of revertant colonies per plate, or thinning or absence of the bacterial lawn. Nontoxic chemicals were tested in the initial experiment up to the 10 mg/plate dose level, or to a level determined by their solubility. Toxic chemicals were tested up to a high dose which exhibited some degree of toxicity.

COMPARISON WITH HISTORICAL CONTROL DATA: No data

Any other information on results incl. tables

Table: Mutation data for the test chemical Geranyl acetate

Dose (µg/plate)

TA100

NA

10% HLI

10% RLI

Mean

SEM

Mean

SEM

Mean

SEM

0

81

3.8

116

6.6

100

9.9

1

76

2.6

93

1.5

 

 

3

84

4.1

118

4.2

104

3.5

10

75

2.6

97

3.2

129

6.6

33

69

6.7

99

9.2

98

7.4

100

T

 

103

3.2

91

6.5

333

 

 

 

 

94

8.6

1000

 

 

 

 

 

 

3333

 

 

 

 

 

 

Positive control

671

49.2

2091

79.8

442

39.4

 

Dose (µg/plate)

TA1535

NA

10% HLI

10% RLI

Mean

SEM

Mean

SEM

Mean

SEM

0

3

0.0

5

0.9

6

1.2

1

5

1.2

 

 

 

 

3

6

0.3

 

 

 

 

10

4

0.9

 

 

 

 

33

3

0.3

4

1.2

6

1.8

100

T

 

10

1.2

8

0.7

333

 

 

5

0.9

6

2.3

1000

 

 

4

0.3

7

0.7

3333

 

 

2

0.9

3

1.2

Positive control

657

40.5

109

6.4

49

3.1

 

Dose (µg/plate)

TA1537

NA

10% HLI

10% RLI

Mean

SEM

Mean

SEM

Mean

SEM

0

4

0.9

5

1.0

5

1.8

1

5

1.5

 

 

 

 

3

2

1.2

 

 

 

 

10

4

0.9

 

 

 

 

33

2

0.9

3

1.0

9

1.0

100

T

 

6

1.9

3

0.3

333

 

 

4

0.7

4

0.7

1000

 

 

4

0.3

4

0.3

3333

 

 

0

0.0

T

0.3

Positive control

58

9.0

122

4.8

39

8.3

 

Dose (µg/plate)

TA98

NA

10% HLI

10% RLI

Mean

SEM

Mean

SEM

Mean

SEM

0

9

1.3

12

3.6

12

1.8

1

7

0.9

 

 

 

 

3

8

0.9

 

 

 

 

10

6

1.0

 

 

 

 

33

4

1.0

13

0.9

11

0.3

100

T

 

13

4.1

12

2.6

333

 

 

13

4.2

16

2.2

1000

 

 

13

1.0

11

2.9

3333

 

 

7

2.6

7

0.0

Positive control

267

99.6

543

19.6

301

15.0

T: toxic

Applicant's summary and conclusion

Conclusions:
Geranyl acetate did not induce mutation in the Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.
Executive summary:

Gene mutation toxicity study was performed for geranyl acetate to evaluate its mutagenic nature. The study was performed as per the preincubation protocol using Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system at doses of 0, 1, 3, 10, 33, 100, 333, 1000, 3333 µg/plate. DMSO was used at the vehicle. The plates were incubated for 48 hrs after 20 mins preincubation before the evaluation of the revertant colonies could be made. Geranyl acetate did notinduce mutation in the Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.