Registration Dossier
Registration Dossier
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EC number: 214-494-2 | CAS number: 1135-66-6
Genetic toxicity: in vitro
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 for cross-reference:
- 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
- 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:
- S. typhimurium, 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
- 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.
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
- Reason / purpose for cross-reference:
- 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
- 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:
- S. typhimurium, 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:
- S. typhimurium, 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
- 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.
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.
- Reason / purpose for cross-reference:
- 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
- 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
- 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.
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.
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:
- 2 018
- Bibliographic source:
- WoE report, Sustainability Support Services (Europe) AB, 2018
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- 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 substance name:
- (2S)-1,3,4,5,6,7-hexahydro-1,1,5,5-tetramethyl-2H-2,4a-methanonaphthalene
- EC Number:
- 214-494-2
- EC Name:
- (2S)-1,3,4,5,6,7-hexahydro-1,1,5,5-tetramethyl-2H-2,4a-methanonaphthalene
- Cas Number:
- 1135-66-6
- Molecular formula:
- C15H24
- IUPAC Name:
- (2S)-1,3,4,5,6,7-hexahydro-1,1,5,5-tetramethyl-2H-2,4a-methanonaphthalene
- 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
Constituent 1
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:
- S. typhimurium, 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:
- S. typhimurium, 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:
- S. typhimurium, 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.
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