Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 269-522-6 | CAS number: 68259-31-4
- 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
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2000-05-17 to 2000-06-02
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 000
- Report date:
- 2000
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
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- 8-isopropyl-6-methylbicyclo[2.2.2]oct-5-ene-2-carbaldehyde
- EC Number:
- 267-308-7
- EC Name:
- 8-isopropyl-6-methylbicyclo[2.2.2]oct-5-ene-2-carbaldehyde
- Cas Number:
- 67845-30-1
- Molecular formula:
- C13H20O
- IUPAC Name:
- 8-isopropyl-6-methylbicyclo[2.2.2]oct-5-ene-2-carbaldehyde
- Reference substance name:
- 7-isopropyl-5-methylbicyclo[2.2.2]oct-5-ene-2-carbaldehyde
- EC Number:
- 299-795-7
- EC Name:
- 7-isopropyl-5-methylbicyclo[2.2.2]oct-5-ene-2-carbaldehyde
- Cas Number:
- 93904-56-4
- Molecular formula:
- C13H20O
- IUPAC Name:
- 7-isopropyl-5-methylbicyclo[2.2.2]oct-5-ene-2-carbaldehyde
- Reference substance name:
- rel-(1S,2R,4R)-1-isopropyl-4-methylbicyclo[2.2.2]oct-5-ene-2-carbaldehyde
- Cas Number:
- 36208-34-1
- Molecular formula:
- C13H20O
- IUPAC Name:
- rel-(1S,2R,4R)-1-isopropyl-4-methylbicyclo[2.2.2]oct-5-ene-2-carbaldehyde
- Test material form:
- liquid
Constituent 1
Constituent 2
impurity 1
- Specific details on test material used for the study:
- Name: Lierral
Batch Number: 9000365675
Aggregate state at room temperature: liquid
Colour: colourless to pale yellow
Purity: 99.5% (sum of three peaks)
Storage: room temperature
Expiration date: December 20, 2000
Method
- Target gene:
- Histidine operon
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Bacterial suspensions were transferred into flasks containing nutrient medium. A solution of 20 µL ampicillin (25 µg/mL) was added to the strains TA98, TA100, and TA102. Additionally 20 µL tetracycline (2 µg/mL) was added to strain TA102. The nutrient medium contains per litre: 8 g Merck Nutrient Broth and 5 g NaCl.
- Properly maintained: Yes. The strain cultures were stored as stock cultures in ampoules with nutrient broth + 5 % DMSO in liquid nitrogen. Regular checking of the properties of the strains regarding the membrane permeability, ampicillin- and tetracycline-resistance as well as spontaneous mutation rates is performed in the testing laboratory. - Additional strain / cell type characteristics:
- other: All strains possess the rfa- mutation. TA1537, TA98, TA1535 and TA100 possess the uvrB- mutation; TA102 possess the uvrB+ mutation. TA98, TA100 and TA 102 all have the R-factor plasmid pKM101.
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- - Experiment 1: 3, 10, 33, 100, 333 and 1000 µg/plate
- Experiment 2: 1, 3, 13, 33, 100 and 333 µg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Dimethyl sulphoxide (DMSO)
- Justification for choice of solvent/vehicle: The solvent was chosen because of its solubility properties and its relative non-toxicity to the bacteria.
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- methylmethanesulfonate
- other: 4-nitro-o-phenylene-diamine, 2-aminoanthracene
- Details on test system and experimental conditions:
- - EXPERIMENT 1
METHOD OF APPLICATION: in agar (plate incorporation)
The following materials were mixed in a test tube and poured onto selective agar plates:
- 100 µL of test solution at each dose level, solvent or positive control;
- 500 µL S9 mix (for test with metabolic activation) or S9 mix substitution buffer (for test without metabolic activation);
- 100 µL bacteria suspension (prior to use, bacterial cultures were incubated in a shaking water bath for 4 hours at 37 °C); and
- 2000 µL overlay agar (containing per litre: 6.0 g Agar, 6.0 g NaCl, 10.5 mg L-Histidine x HCI x H₂O and 12.2 mg Biotin).
DURATION
- Exposure duration: After solidification the plates were incubated upside down for at least 48 hours at 37 °C in the dark.
NUMBER OF REPLICATIONS: For each strain and dose level, including the controls, three plates were used.
DETERMINATION OF CYTOTOXICITY
- Method: Evaluation of background bacterial lawn
- EXPERIMENT 2
METHOD OF APPLICATION: pre-incubation
The following materials were mixed in a test tube and incubated:
- 100 µL of test solution at each dose level, solvent or positive control;
- 500 µL S9 mix (for test with metabolic activation) or S9 mix substitution buffer (for test without metabolic activation); and
- 100 µL bacteria suspension (prior to use, bacterial cultures were incubated in a shaking water bath for 4 hours at 37 °C).
- After pre-incubation, 2.0 mL overlay agar (as above, 45 °C) was added to each tube. The mixture was poured on minimal agar plates.
DURATION
- Pre-incubation period: 37 °C for 60 minutes.
- Exposure duration: after solidification the plates were incubated upside down for at least 48 hours at 37 °C in the dark.
NUMBER OF REPLICATIONS: For each strain and dose level, including the controls, three plates were used.
DETERMINATION OF CYTOTOXICITY
- Method: Evaluation of background bacterial lawn - Evaluation criteria:
- ACCEPTABILITY OF THE ASSAY
The assay is considered acceptable if it meets the following criteria:
- Regular background growth in the negative and solvent control;
- The spontaneous reversion rates in the negative and solvent control are in the range of historical data; and
- The positive control substances should produce a significant increase in mutant colony frequencies.
EVALUATION OF RESULTS
- A test material is considered positive if either a dose related increase in the number of revertants or a biologically relevant increase for at least one test concentration is induced.
- A test material producing neither a dose related increase in the number of revertants nor a biologically relevant positive response at any one of the test points is considered non-mutagenic in this system.
- A biologically relevant response is described as follows: a test material is considered mutagenic if the number of reversions is at least twice the spontaneous reversion rate in strains TA98, TA100, and TA102 or thrice in strains TA1535 and TA1537.
Also, a dose-dependent and reproducible increase in the number of revertants is regarded as an indication of possibly existing mutagenic potential of the test material regardless whether the highest dose induced the criteria described above or not. - Statistics:
- A statistical analysis of the data is not required.
Results and discussion
Test results
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Distinct toxic effects, evident as a reduction in the number of revertants, were observed as summarised in Table 1.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Reduced background growth was observed in strains TA1535, TA1537, TA98 and TA100 at 1000 µg/plate in the first experiment and in TA1537 and TA98 in the second experiment.
No substantial or reproducible increase in revertant colony numbers of any of the five tester strains was observed following treatment with the test material at any concentration level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no reproducible tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.
Positive controls showed a distinct increase in induced revertant colonies.
The test material did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
Any other information on results incl. tables
Table 1: Summary of Toxic Effects (µg/plate)
Strain |
Experiment 1 |
Experiment 2 |
||
-S9 |
+S9 |
-S9 |
+S9 |
|
TA 1535 |
333 to 1000 |
1000 |
33 to 333 |
333 |
TA 1537 |
333 to 1000 |
333 to 1000 |
333 |
333 |
TA 98 |
100 to 1000 |
1000 |
333 |
333 |
TA 100 |
100 to 1000 |
333 to 1000 |
100 to 333 |
333 |
TA 102 |
/ |
/ |
333 |
/ |
/ = no relevant toxic effects observed
Table 2: Experiment 1
+/- S9 Mix |
Concentration (µg/plate) |
Mean number of colonies/plate |
||||
Base-pair Substitution Type |
Frameshift Type |
|||||
TA100 |
TA1535 |
TA102 |
TA98 |
TA1537 |
||
- |
Solvent 0 3 10 33 100 333 1000 |
132 183 113 110 87 48 0 0 |
12 10 12 9 9 10 4 0 |
256 279 251 265 202 152 153 149 |
32 26 31 23 25 10 11 3 |
8 7 8 7 4 4 3 1 |
+ |
Solvent 0 3 10 33 100 333 1000 |
159 167 100 129 82 90 0 0 |
11 14 14 12 14 15 9 2 |
279 281 276 300 299 255 219 239 |
31 32 25 32 29 26 17 9 |
13 10 14 11 13 11 6 0 |
Positive Controls |
||||||
- |
Name |
SA |
SA |
MMS |
4-NOPD |
4-NOPD |
Concentration (µg/plate) |
10 |
10 |
5 (µl/plate) |
10 |
50 |
|
Mean no. colonies/plate |
1002 |
1200 |
1156 |
253 |
49 |
|
+ |
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
Concentration (µg/plate) |
2.5 |
2.5 |
10 |
2.5 |
2.5 |
|
Mean no. colonies/plate |
1132 |
238 |
1088 |
800 |
130 |
SA = Sodium azide
2AA = 2-Aminoanthracene
4-NOPD = 4-Nitro-o-phenylene-diamine
MMS = Methyl methane sulfonate
Table 3: Experiment 2
+/- S9 Mix |
Concentration (µg/plate) |
Mean number of colonies/plate |
||||
Base-pair Substitution Type |
Frameshift Type |
|||||
TA100 |
TA1535 |
TA102 |
TA98 |
TA1537 |
||
- |
Solvent 0 1 3 10 33 100 333 |
117 159 131 176 137 159 56 18 |
13 9 13 11 14 6 5 7 |
244 219 208 282 257 193 166 104 |
21 20 21 17 23 9 14 10 |
7 7 9 5 7 5 4 0 |
+ |
Solvent 0 1 3 10 33 100 333 |
152 169 162 212 203 200 166 53 |
11 10 5 7 7 11 10 1 |
220 277 176 203 243 194 144 131 |
36 40 28 35 37 36 25 0 |
10 8 8 14 13 11 8 0 |
Positive Controls |
||||||
- |
Name |
SA |
SA |
MMS |
4-NOPD |
4-NOPD |
Concentration (µg/plate) |
10 |
10 |
5 (µl/plate) |
10 |
50 |
|
Mean no. colonies/plate |
527 |
926 |
1095 |
155 |
62 |
|
+ |
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
Concentration (µg/plate) |
2.5 |
2.5 |
10 |
2.5 |
2.5 |
|
Mean no. colonies/plate |
981 |
207 |
1101 |
677 |
128 |
SA = Sodium azide
2AA = 2-Aminoanthracene
4-NOPD = 4-Nitro-o-phenylene-diamine
MMS = Methyl methane sulfonate
Applicant's summary and conclusion
- Conclusions:
- The test material is considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay in both the presence and absence of metabolic activation.
- Executive summary:
The mutagenic potential of the test material was assessed in a bacterial reverse mutation assay conducted under GLP conditions and in accordance with the standardised guidelines OECD 471 and EU Method B.14.
The study was performed according to the plate incorporation test (Experiment 1) and the pre-incubation test (Experiment 2) using the Salmonella typhimurium strains TA1535, TA1537, TA98, TA100, and TA102.
The assay was performed in two independent experiments both with and without liver microsomal activation. Negative, solvent and positive controls were conducted concurrently. Each concentration, including the controls, was tested in triplicate. The test material was tested at the following concentrations in DMSO:
- Experiment 1:3, 10, 33, 100, 333 and 1000 µg/plate
- Experiment 2:1, 3, 13, 33, 100 and 333 µg/plate
Distinct toxic effects, evident as a reduction in the number of revertants, occurred in almost all strains at higher concentrations. Reduced background growth was observed in strains TA1535, TA1537, TA98 and TA100 at 1000 µg/plate in the first experiment and in TA1537 and TA98 in the second experiment.
No substantial or reproducible increase in revertant colony numbers of any of the five tester strains was observed following treatment with the test material at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no reproducible tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.
The positive controls showed a distinct increase in induced revertant colonies.
In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test material did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
Therefore, the test material is considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay in both the presence and absence of metabolic activation.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.