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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
Remarks:
Experimenta data from various test chemicals
Justification for type of information:
Data for the target chemical is summarized based on data from various test chemicals
Cross-referenceopen allclose all
Reason / purpose:
read-across source
Reference
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
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 to determine the mutagenic nature of the test chemical
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
- Name of test material: Cineol (eucalyptol)
- IUPAC name: 1,3,3-Trimethyl-2-oxabicyclo[2.2.2]octane
- Molecular formula: C10H18O
- Molecular weight: 154.2512 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data
Target gene:
Histidine
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
Liver S-9 fractions were routinely prepared from male Sprague-Dawley rats and male Syrian hamsters that were injected, ip, with Aroclor 1254 (200 mg/ml in corn oil) at 500 mg/kg.
Test concentrations with justification for top dose:
0, 3.3, 10, 33, 100, 333, 1000 or 3333 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 2-Aminoanthracene (2-AA), 4-Nitro-o-phenylenediamine (NOPD)
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- 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: Triplicate

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:
A dose related increase in the number of revertants was noted whether it be twofold over background or not
Statistics:
No data
Species / strain:
other: TA1535, TA1537, TA98, and TA100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
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: To select the dose range for the mutagenesis assay, the test chemical was checked for toxicity to TA100 up to a concentration of 10 mg/plate or the limit of solubility, both in the presence and absence of S-9 mix. One or more parameters were used as an indication of toxicity: viability on complete medium and reduced numbers of revertant colonies per plate and/or thinning or absence of the bacterial lawn. If toxicity was not apparent in the preliminary toxicity determination, the highest dose tested was 10 mg/plate; otherwise the upper limit of solubility was used. If toxicity was observed, the doses of test chemical were chosen so that the high dose exhibited some degree of toxicity. Occasionally, in the earlier tests, the high dose was greater than 10 mg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA: No data

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data
Remarks on result:
other: No mutagenic potential

Table: Mutagenicity of the test chemical

Dose (µg/plate)

TA100

TA1535

NA

RLI

HLI

NA

RLI

HLI

0

94±7.1

120±8.2

91±5.5

4±1.2

7±1.3

8±3.3

3.3

85±6.0

 

 

 

 

 

10

76±6.0

 

 

 

 

 

33

79±7.9

127±20.4

105±15.0

4± 1.2

6±0.3

6±0.9

100

95±11.2

113± 16.2

100±6.4

4±0.9

8±2.0

8±1.0

333

97 ±3.2

92 ±6.5

88±8.0

4±0.7

4±0.7

4±1.9

1000

 

104±6.4

93±4.0

3±0.6

t

5±1.3

3333

 

t

3±3.0

T

T

T

Pos

773±61.2

2096±89.8

1834±29.2

1260±85.5

157±27.

180±39.4

 

Dose (µg/plate)

TA1537

TA98

NA

RLI

HLI

NA

RLI

HLI

0

4±1.8

10±2.6

7±1.0

19±2.6

27±4.4

24±3.8

3.3

7±1.2

 

 

 

 

 

10

8±3.6

6±1.7

9±0.6

19±2.1

 

 

33

6±1.3

7±1.5

9±2.1

14±2.3

27±3.7

22±1.5

100

4±0.5

5±0.7

8±3.4

15±3.2

23±6.2

19±9.5

333

T

6±2.5

7±1.9

21±0.9

14±2.2

17±4.1

T: complete clearing of background lawn

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

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed by the preincubation protocol using Salmonella typhimurium strains TA1535, TA1537, TA98, and TA100 both in the presence and absence of S9 metabolic activation system. Preincubation was carried at 37°C for 20 mins followed by exposure period of 48 hrs at dose levels of 0, 3.3, 10, 33, 100, 333, 1000 or 3333µg/plate. DMSO was used as solvent control and concurrent positive control chemicals were included in the study. A dose related increase in the number of revertants was noted whether it be twofold over background or not.

 

The test chemical did not induce mutation in the Salmonella typhimurium strains TA1535, TA1537, TA98, and TA100 both in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Reason / purpose:
read-across source
Reference
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
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
Salmonella/microsome reverse mutation assay was performed to determine the mutagenic nature of the test chemical
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Target gene:
Histidine
Species / strain / cell type:
other: TA98 and TA100
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
S9 metabolic activation system was obtained from liver of Aroclor 1254 induced male Sprague Dawley rats
Test concentrations with justification for top dose:
0, 9.85, 19.7, 39.3, 78.5, 157, 313, 625, 1250, 2500 or 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: 100% ethanol
- Justification for choice of solvent/vehicle: The test chemical was soluble in 100% ethanol
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
100% ethanol
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
2-nitrofluorene
sodium azide
other: 2- aminoanthracene (with S9, TA98 and TA100)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- 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: Triplicate

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: Yes, bacterial growth was observed

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:
To be considered mutagenic, a test article treatment had to induce at least twice the number of revertants/plate in atleast 1 tester strain compared to those induced in the appropriate vehicle control and exhibit an increasing number of revertants/plate with increasing test article dosage
Statistics:
Mean and standard deviation
Species / strain:
other: TA98 and TA100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at and above 1250 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
No data
Remarks on result:
other: No mutagenic potential

Table: Mutagenicity of the test chemical

Doses (µg/plate)

Revertants/plate

Background lawn evaluation code

TA98

TA100

With S9

 

 

 

0

38±2

81±7

1

9.85

34±6

90±2

1

19.7

39±4

78±9

1

39.3

36±10

89±11

1

78.5

31±4

77±13

1

157

29±6

82±13

1

313

31±2

76±10

1

625

29±3

76±11

1

1250

26±4

62±10

2

2500

16±4

7±6

3

5000

0±0

0±0

5

Positive control

1189±66

1259±53

1

0

17±3

70±9

1

9.85

19±2

68±3

1

19.7

16±2

63±3

1

39.3

23±5

71±10

1

78.5

22±5

65± 5

1

157

19±4

68±12

1

313

21±5

63±6

1

625

19± 4

61±5

1

1250

8±4

36±7

2

2500

2±2

0±0

3

5000

0±0

0±0

5

Positive control

116±7

479±35

1

Evaluation Codes: I = normal; 2 = slightly reduced; 3 = moderately reduced; 4 = extremely reduced; 5 = absent.

Conclusions:
The test chemical did not induce mutation in the Salmonella typhimurium strains TA98 and TA100 both in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.
Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed by the plate incorporation method using Salmonella typhimurium strains TA100 both in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in 100% ethanol and doses selected for the study were 0, 9.85, 19.7, 39.3, 78.5, 157, 313, 625, 1250, 2500 or 5000 µg/plate. Concurrent solvent control and positive chemical was also included in the study. The test chemical did not induce mutation in the Salmonella typhimurium strains TA98 and TA100 both in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Reason / purpose:
read-across source
Reference
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
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 to determine the mutagenic nature of the test chemical
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Target gene:
Histidine
Species / strain / cell type:
S. typhimurium TA 100
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
without
Metabolic activation system:
S9 metabolic activation system
Test concentrations with justification for top dose:
0, 0.01, 0.1, 1.0, 2.5, 5.0 or 10.0 µmoles/plate
Vehicle / solvent:
No data
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Details are not specified
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data
- Exposure duration: No data
- Expression time (cells in growth medium): No data
- 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: Triplicate

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:
The plates were observed for revertant colonies
Statistics:
No data
Species / strain:
S. typhimurium TA 100
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
doses at and above 2.5 µmoles/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
No data
Remarks on result:
other: No mutagenic potential

Table: Mutagenicity of the test chemical

Doses

Revertantsa

0

125±10 (99±11)

0.01

125±25

0.1

113±17

1.0

138±10c

2.5

-

(74±12)

5.0

-

(0)c

10.0

0d,e

a: The mean of revertants for 3 plates + S.D.

b: The values in parentheses are for the concurrent comparisons at 2.5 and 5.0 #moles in which the bacteria were from the same

overnight culture.

c Reduction in background lawn.

d No background lawn.

e Dose showed slight precipitation in agar.

               

Conclusions:
The test chemical did not induce mutation in the Salmonella typhimurium strains TA100 both in the absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.
Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed by the plate incorporation method using Salmonella typhimurium strains TA100 both in the absence of S9 metabolic activation system. The doses selected for the study were 0, 0.01, 0.1, 1.0, 2.5, 5.0 or 10.0µmoles/plate. Concurrent solvent control chemical was also included in the study. The test chemical was toxic to the bacterial strain but it did not induce mutation in the Salmonella typhimurium strains TA100 both in the absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Data source

Reference
Reference Type:
review article or handbook
Title:
WoE of gene mutation in vitro toxicity study for CAS no 1135-66-6
Author:
Sustainability Support Services (Europe) AB
Year:
2018
Bibliographic source:
WoE report, Sustainability Support Services (Europe) AB, 2018

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:
WoE for the target CAS is summarized based on data from various test chemicals
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of the test chemical: (2S)-1,3,4,5,6,7-hexahydro-1,1,5,5-tetramethyl-2H-2,4a-methanonaphthalene
- Molecular weight: 204.3546 g/mol
- Molecular formula: C15H24

Method

Target gene:
Histidine
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Remarks:
2
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
other: TA98 and TA100
Remarks:
3
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 100
Remarks:
4
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
Liver S-9 fractions were routinely prepared from male Sprague-Dawley rats and male Syrian hamsters that were injected, ip, with Aroclor 1254 (200 mg/ml in corn oil) at 500 mg/kg.
Test concentrations with justification for top dose:
2. 0, 3.3, 10, 33, 100, 333, 1000 or 3333 µg/plate
3. 0, 9.85, 19.7, 39.3, 78.5, 157, 313, 625, 1250, 2500 or 5000 µmoles/plate
4. 0, 0.01, 0.1, 1.0, 2.5, 5.0 or 10.0µmoles/plate
Vehicle / solvent:
2. - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO

3. - Vehicle(s)/solvent(s) used: 100% ethanol
- Justification for choice of solvent/vehicle:The test chemical was soluble in 100% ethanol

4. No data
Controlsopen allclose all
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 2-Aminoanthracene (2-AA), 4-Nitro-o-phenylenediamine (NOPD)
Remarks:
2
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
100% ethanol
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
2-nitrofluorene
sodium azide
other: 2- aminoanthracene (with S9)
Remarks:
3
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Not specified
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Remarks:
4
Details on test system and experimental conditions:
2. METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- 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: Triplicate

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

3. METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period:No data
- Exposure duration:48 hrs
- Expression time (cells in growth medium):48 hrs
- 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:Triplicate

NUMBER OF CELLS EVALUATED:No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:Yes, bacterial growth was observed

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

OTHER: No data

4. METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period:No data
- Exposure duration:No data
- Expression time (cells in growth medium):No data
- 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:Triplicate

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:
2. A dose related increase in the number of revertants was noted whether it be twofold over background or not

3. To be considered mutagenic, a test article treatment had to induce at least twice the number of revertants/plate in atleast 1 tester strain compared to those induced in the appropriate vehicle control and exhibit an increasing number of revertants/plate with increasing test article dosage

4. A dose related increase in the number of revertants was noted whether it be twofold over background or not
Statistics:
No data

Results and discussion

Test resultsopen allclose all
Species / strain:
other: TA1535, TA1537, TA98, and TA100
Remarks:
2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
other: TA98 and TA100
Remarks:
3
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at and above 1250 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Remarks:
4
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
2. 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: To select the dose range for the mutagenesis assay, the test chemical was checked for toxicity to TA100 up to a concentration of 10 mg/plate or the limit of solubility, both in the presence and absence of S-9 mix. One or more parameters were used as an indication of toxicity: viability on complete medium and reduced numbers of revertant colonies per plate and/or thinning or absence of the bacterial lawn. If toxicity was not apparent in the preliminary toxicity determination, the highest dose tested was 10 mg/plate; otherwise the upper limit of solubility was used. If toxicity was observed, the doses of test chemical were chosen so that the high dose exhibited some degree of toxicity. Occasionally, in the earlier tests, the high dose was greater than 10 mg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA: No data

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data

3. No data

4. No data
Remarks on result:
other: No mutagenic potential

Applicant's summary and conclusion

Conclusions:
The test chemical did not induce mutation in the Salmonella typhimurium strains both in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.
Executive summary:

Data available for the various test chemicals was reviewed to determine the mutagenic nature of the test chemical. The studies are as mentioned below:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed by the preincubation protocol using Salmonella typhimurium strains TA1535, TA1537, TA98, and TA100 both in the presence and absence of S9 metabolic activation system. Preincubation was carried at 37°C for 20 mins followed by exposure period of 48 hrs at dose levels of 0, 3.3, 10, 33, 100, 333, 1000 or 3333µg/plate. DMSO was used as solvent control and concurrent positive control chemicals were included in the study. A dose related increase in the number of revertants was noted whether it be twofold over background or not. The test chemical did not induce mutation in the Salmonella typhimurium strains TA1535, TA1537, TA98, and TA100 both in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

In anothet study, gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed by the plate incorporation method using Salmonella typhimurium strains TA100 both in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in 100% ethanol and doses selected for the study were 0, 9.85, 19.7, 39.3, 78.5, 157, 313, 625, 1250, 2500 or 5000 µg/plate. Concurrent solvent control and positive chemical was also included in the study. 2-Methylisoborneol did not induce mutation in the Salmonella typhimurium strains TA98 and TA100 both in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

In yet another study, gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed by the plate incorporation method using Salmonella typhimurium strains TA100 both in the absence of S9 metabolic activation system. The doses selected for the study were 0, 0.01, 0.1, 1.0, 2.5, 5.0 or 10.0 µmoles/plate. Concurrent solvent control chemical was also included in the study. The test chemical was toxic to the bacterial strain but it did not induce mutation in the Salmonella typhimurium strains TA100 both in the absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Based on the data available, the test chemical did not induce mutation in the Salmonella typhimurium strains both in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.