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EC number: 248-792-9 | CAS number: 28043-10-9
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2015
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study performed under GLP.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 015
- Report date:
- 2015
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
- Version / remarks:
- Guideline stipulated by the Japanese Ministry of Health, Labour and Welfare, Ministry of Economy, Trade and Industry and Ministry of the Environment (revised March 31st, 2011)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- inspected March 2013; signature: May 2013
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Methyl 2,6,6-trimethylcyclohex-2-ene-1-carboxylate
- EC Number:
- 248-792-9
- EC Name:
- Methyl 2,6,6-trimethylcyclohex-2-ene-1-carboxylate
- Cas Number:
- 28043-10-9
- Molecular formula:
- C11H18O2
- IUPAC Name:
- methyl 2,6,6-trimethylcyclohex-2-ene-1-carboxylate
- Details on test material:
- - Physical state: Liquid
- Storage condition of test material: At room temperature protected from light
- Other: Colourless liquid
Constituent 1
Method
Species / strainopen allclose all
- Species / strain / cell type:
- E. coli WP2 uvr A
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat liver microsomal enzymes (S9 homogenate)
- Test concentrations with justification for top dose:
- Dose range finding study: 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 ug/plate
Experiment 1: 52, 164, 512, 1600, 5000 ug/plate
Experiment 2: 275, 492, 878, 1568, 2800, 5000 ug/plate
Experiment 3 (strains TA1537, TA100): 17, 52, 164, 512, 1600, 5000 ug/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Dimethyl sulfoxide (DMSO)
- Justification for choice of solvent/vehicle: A solubility test was performed. The test substance could not be dissolved in water. The test substance was soluble in dimethyl sulfoxide. Dimethyl sulfoxide was used in the assay.
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 2-nitrofluorene
- sodium azide
- methylmethanesulfonate
- other: ICR-191; 2-aminoanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: The plates were inverted and incubated in the dark at 37.0 ± 1.0 °C for 48 ± 4 h. After this period revertant colonies (histidine independent (His+) for Salmonella typhimurium bacteria and tryptophan independent (Trp+) for Escherichia coli were counted. The revertant colonies were counted automatically with a Colony Counter.
NUMBER OF REPLICATIONS: 3
DETERMINATION OF CYTOTOXICITY
- Method: Relative total growth (bacterial background lawn) and reduction in the number of revertants
OTHER:
Dose range finding test on TA100 and WP2urvA with and without 5% (v/v) S9-mix; First mutation assay on TA1535, TA1537 and TA98 with and without 5% (v/v) S9-mix. To obtain more information about the possible mutagenicity of the test substance, a second mutation experiment was performed on all strains, in the absence of S9-mix and in the presence of 10% (v/v) S9-mix. Based on the results of the first mutation assay, the test substance was tested up to the dose level of 5000 μg/plate in strains TA1535, TA1537, TA98, TA100 and WP2uvrA. - Evaluation criteria:
- See 'Any other information on materials and methods' for details on evaluation of the assay and positive criteria.
- Statistics:
- No formal hypothesis testing was done. See 'Any other information on materials and methods' for details on the acceptability and evaluation criteria of the assay.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- In strains TA100, TA1535 and TA1357, cytotoxicity was seen at the 1600 µg/plate or above (with or without S9-mix); TA98 strain at 2800 µg/plate (with S9-mix) or was tested to the limit dose.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of test substance on the plates was observed at the start of the incubation period and at 5000 µg/plate at the end of the incubation period. Except for the tester strains TA1535 (presence of S9-mix, end of the incubation period). The precipitation at the start of the incubation period was observed as oily droplets of the test substance. In experiment 3 on strains TA1537 and TA100 only: Precipitation of the test substance on the plates was observed at the start of the incubation period at concentrations of 1600 and 5000 μg/plate. At the end of the incubation period precipitate was only observed in tester strain TA100 at the top dose of 5000 μg/plate.
RANGE-FINDING/SCREENING STUDIES:
Test substance was tested in the tester strains TA100 and WP2uvrA with concentrations of 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 μg/plate in the absence and presence of S9-mix. Based on the results of the dose range finding test, the following dose range was selected for the mutation assay with the tester strains, TA1535, TA1537 and TA98 in the absence and presence of S9-mix: 52, 164, 512, 1600 and 5000 μg/plate. To obtain more information on TA1537 and TA100 a third mutagenicity experiment was conducted at a concentration range of 17 to 5000 μg/plate in the absence and presence of 10% (v/v) S9-mix. This was based on the results of the preceding tests.
COMPARISON WITH HISTORICAL CONTROL DATA:
The negative control values were within the laboratory historical control data ranges. The positive control values were not all within the laboratory historical control data ranges, specifically TA98 (first experiment, absence of S9-mix) and TA1537 (second experiment, absence of S9-mix). However, these responses were judged not to impact the reliability of the study. The strain-specific positive control values were at least three times the concurrent vehicle control group mean indicating that the test conditions were adequate and that the metabolic activation system functioned properly. Historical control data from experiments was presented within the study report. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
Table 1 Dose range finding test: Mutagenic response of test substance in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay
Dose (µg/plate) |
Mean number of revertant colonies/3 replicate plates (± S.D.) with one strain of Salmonella typhimurium and one Escherichia coli strain
|
|||
|
TA100 |
|
WP2uvrA
|
|
Without S9-mix
|
|
|
|
|
Positive control |
632 |
± 10 |
1450 |
± 32 |
Solvent control |
104 |
± 15 |
32 |
± 6 |
1.7 |
116 |
± 14 |
35 |
± 9 |
5.4 |
112 |
± 5 |
36 |
± 12 |
17 |
118 |
± 13 |
34 |
± 5 |
52 |
118 |
± 15 |
39 |
± 6 |
164 |
103 |
± 4 |
38 |
± 10 |
512 |
110 |
± 14 n |
28 |
± 4 |
1600 |
80 |
± 12 s NP |
24 |
± 5 NP |
5000 |
78 |
± 21 s SP |
23 |
± 5 n SP |
With S9-mix #1
|
|
|
|
|
Positive control |
1553 |
± 167 |
196 |
± 18 |
Solvent control |
112 |
± 5 |
34 |
± 13 |
1.7 |
115 |
± 14 |
38 |
± 4 |
5.4 |
103 |
± 11 |
39 |
± 11 |
17 |
109 |
± 14 |
43 |
± 7 |
52 |
101 |
± 13 |
42 |
± 15 |
164 |
92 |
± 7 |
39 |
± 4 |
512 |
101 |
± 7 n |
37 |
± 3 |
1600 |
85 |
± 6 s NP |
33 |
± 8 NP |
5000 |
57 |
± 2 m SP |
28 |
± 9 n SP |
#1: Plate incorporation assay (5% S9)
NP: No precipitate
SP: Slight Precipitate
m: Bacterial background lawn moderately reduced
n: Normal bacterial background lawn
s: Bacterial background lawn slightly reduced
Table 2 Experiment 1: Mutagenic response of test substance in the Salmonella typhimurium reverse mutation assay
Dose (µg/plate) |
Mean number of revertant colonies/3 replicate plates (± S.D.) with different strains of Salmonella typhimurium
|
|||||
|
TA1535 |
|
TA1537
|
|
TA98 |
|
Without S9-mix
|
|
|
|
|
|
|
Positive control |
857 |
± 44 |
586 |
± 27 |
1379 |
± 128 |
Solvent control |
17 |
± 2 |
6 |
± 2 |
19 |
± 8 |
52 |
21 |
± 2 |
15 |
± 8 |
17 |
± 6 |
164 |
27 |
± 9 |
9 |
± 1 |
16 |
± 3 |
512 |
23 |
± 7 |
7 |
± 1 |
16 |
± 2 n |
1600 |
15 |
± 7 n NP |
10 |
± 7 n NP |
15 |
± 6 s NP |
5000 |
- |
e SP MC |
10 |
± 8 m SP |
16 |
± 6 m SP |
With S9-mix #1
|
|
|
|
|
|
|
Positive control |
273 |
± 19 |
424 |
± 41 |
1161 |
± 58 |
Solvent control |
22 |
± 14 |
6 |
± 2 |
27 |
± 11 |
52 |
45 |
± 16 |
10 |
± 4 |
37 |
± 2 |
164 |
36 |
± 6 |
12 |
± 6 |
31 |
± 5 |
512 |
31 |
± 13 n |
11 |
± 4 |
29 |
± 8 |
1600 |
32 |
± 8 m |
7 |
± 4 n NP |
19 |
± 6 n NP |
5000 |
e NP MC |
5 |
± 2 m SP |
18 |
± 2 s SP |
#1: Plate incorporation assay (5% S9)
MC: Microcolonies
NP: No precipitate
SP: Slight Precipitate
e: Bacterial background lawn extremely reduced
m: Bacterial background lawn moderately reduced
n: Normal bacterial background lawn
s: Bacterial background lawn slightly reduced
Table 3 Experiment 2: Mutagenic response of test substance in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay
Dose (µg/plate) |
Mean number of revertant colonies/3 replicate plates (± S.D.) with different strains of Salmonella typhimurium and one Escherichia coli strain |
|||||||||
|
TA1535 |
|
TA1537 |
|
TA98 |
|
TA100 |
|
WP2uvrA |
|
Without S9-mix
|
|
|
|
|
|
|
|
|
|
|
Positive control |
939 |
± 74 |
1565 |
± 137 |
793 |
± 268 |
800 |
± 42 |
1242 |
± 62 |
Solvent control |
16 |
± 3 |
7 |
± 6 |
27 |
± 2 |
99 |
± 7 |
28 |
± 10 |
275 |
13 |
± 5 |
- |
- |
- |
- |
- |
- |
- |
- |
492 |
10 |
± 1 |
3 |
± 2 n |
20 |
± 3 |
73 |
± 13 s |
21 |
± 2 |
878 |
7 |
± 2 |
3 |
± 2 s |
16 |
± 10 |
51 |
± 14 m |
23 |
± 2 |
1568 |
5 |
± 1 n |
3 |
± 3 m |
11 |
± 1 n |
47 |
± 12 m |
20 |
± 6 |
2800 |
7 |
± 2 s NP |
|
e NP MC |
15 |
± 1 s NP |
52 |
± 6 m NP |
25 |
± 10 NP |
5000 |
5 |
± 2 m SP |
|
e NP MC |
9 |
± 1 s SP |
|
e SP MC |
27 |
± 4 n SP |
|
|
|
|
|
|
|
|
|
|
|
With S9-mix #1
|
|
|
|
|
|
|
|
|
|
|
Positive control |
295 |
± 30 |
342 |
± 40 |
406 |
± 54 |
1279 |
± 105 |
175 |
± 23 |
Solvent control |
12 |
± 3 |
10 |
± 2 |
32 |
± 2 |
110 |
± 18 |
31 |
± 4 |
275 |
14 |
± 5 |
- |
- |
- |
- |
- |
- |
- |
- |
492 |
11 |
± 1 |
12 |
± 7 n |
30 |
± 6 |
91 |
± 19 n |
28 |
± 3 |
878 |
9 |
± 4 n |
11 |
± 4 s |
23 |
± 3 n |
73 |
± 9 s |
30 |
± 7 |
1568 |
6 |
± 4 m |
3 |
± 2 m |
17 |
± 3 s |
60 |
± 6 m |
31 |
± 13 |
2800 |
|
e MC |
2 |
± 1 m NP |
15 |
± 6 m NP |
60 |
± 4 m NP |
29 |
± 3 NP |
5000 |
|
e SP MC |
|
e NP MC |
11 |
± 1 m SP |
|
e SP MC |
26 |
± 4 n SP |
#1: Plate incorporation assay (10% S9)
MC: Microcolonies
MP: Moderate Precipitate
NP: No precipitate
SP: Slight Precipitate
e: Bacterial background lawn extremely reduced
m: Bacterial background lawn moderately reduced
n: Normal bacterial background lawn
s: Bacterial background lawn slightly reduced
- : Not tested
Table 4 Experiment 3: Mutagenic response of test substance in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay
Dose (µg/plate) |
Mean number of revertant colonies/3 replicate plates (± S.D.) with different strains of Salmonella typhimurium and one Escherichia coli strain |
|||
|
TA1537 |
|
TA100 |
|
Without S9-mix
|
|
|
|
|
Positive control |
1070 |
± 57 |
848 |
± 61 |
Solvent control |
9 |
± 3 |
91 |
± 6 |
17 |
11 |
± 4 |
106 |
± 12 |
52 |
10 |
± 6 |
105 |
± 12 |
164 |
13 |
± 4 |
95 |
± 8 |
512 |
9 |
± 4 |
87 |
± 7 n |
1600 |
4 |
± 1 n |
|
e NP MC |
5000 |
|
e SP MC |
|
e SP MC |
|
|
|
|
|
With S9-mix #1
|
|
|
|
|
Positive control |
368 |
± 17 |
1287 |
± 52 |
Solvent control |
11 |
± 4 |
107 |
± 8 |
17 |
15 |
± 8 |
102 |
± 12 |
52 |
14 |
± 3 |
98 |
± 6 #2 |
164 |
10 |
± 3 |
91 |
± 22 |
512 |
11 |
± 1 |
81 |
± 11 n |
1600 |
6 |
± 4 n |
85 |
± 14 s NP |
5000 |
|
e NP MC |
|
e SP MC |
#1: Plate incorporation assay (10% S9)
#2: Mean of two plates, one plate could not be determined
MC: Microcolonies
NP: No precipitate
SP: Slight Precipitate
a: Bacterial background lawn absent
e: Bacterial background lawn extremely reduced
m: Bacterial background lawn moderately reduced
n: Normal bacterial background lawn
s: Bacterial background lawn slightly reducedApplicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
Under the conditions of this study the test substance is not considered to be mutagenic. Negative control values were within laboratory historical control data. The strain-specific positive control values were at least three times the concurrent vehicle control group mean indicating that the test conditions were adequate and that the metabolic activation system functioned properly. - Executive summary:
The study was performed to OECD TG 471, EU Method B.13/14, EPA OPPTS 870.5100 and the Japan Guidelines for Screening Mutagenicity of Chemicals in accordance with GLP; to evaluate the mutagenic activity of the test substance in the Salmonella typhimurium and the Escherichia coli in a reverse mutation assay (with independent repeat). In the dose range finding test, the test substance was tested up to concentrations of 5000 µg/plate in the absence and presence of S9-mix in the strains TA100 and WP2uvrA. The test substance precipitated on the plates at the top dose of 5000 μg/plate. In tester strain TA100, toxicity was observed at dose levels of 1600 and 5000 μg/plate in the absence and presence of S9-mix. In tester strain WP2uvrA, no toxicity was observed at any of the dose levels tested. Results of this dose range finding test were reported as part of the first mutation assay. Based on the results of the dose range finding test, the test substance was tested in the first mutation assay at a concentration range of 52 to 5000 µg/plate in the absence and presence of 5% (v/v) S9-mix in the tester strains TA1535, TA1537 and TA98. The test substance precipitated on the plates at the top dose of 5000 μg/plate. The test substance was tested up to or beyond a precipitating dose level. Toxicity was observed in all three tester strains in the absence and presence of S9-mix. In a follow-up experiment of the assay with additional parameters, the test substance was tested at a concentration range of 275 to 5000 µg/plate in tester strain TA1535 and at a concentration range of 492 to 5000 µg/plate in tester strains TA1537, TA98, TA100 and WP2uvrA in the absence and presence of 10% (v/v) S9-mix. The test substance precipitated on the plates at the top dose of 5000 μg/plate. Toxicity was observed in all tester strains in the absence and presence of S9-mix. Except tester strain WP2uvrA, where no toxicity is observed. The negative control values were within the laboratory historical control data ranges. The positive control values were not all within the laboratory historical control data ranges, specifically TA98 (first experiment, absence of S9-mix) and TA1537 (second experiment, absence of S9-mix). However, these responses were judged not to impact the reliability of the study. The strain-specific positive control values were at least three times the concurrent vehicle control group mean indicating that the test conditions were adequate and that the metabolic activation system functioned properly. Under the conditions of this study, it is concluded that that the test substance was not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.
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