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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 500-007-3 | CAS number: 9003-50-3
- 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:
- 29 March 2016 to 03 May 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- Study performed in accordance with OECD test guideline in compliance with GLP.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
- Report date:
- 2016
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 490 (In Vitro Mammalian Cell Gene Mutation Tests Using the Thymidine Kinase Gene)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian cell transformation assay
Test material
- Reference substance name:
- Heptanal, oligomeric reaction products with aniline
- EC Number:
- 500-007-3
- EC Name:
- Heptanal, oligomeric reaction products with aniline
- Cas Number:
- 9003-50-3
- Molecular formula:
- (C7H14O)n.(C6H7N)n
- IUPAC Name:
- Heptanal, oligomeric reaction products with aniline
- Test material form:
- liquid
- Details on test material:
- Test item: 207368/A
Identification: Hepteen Base®
Appearance: Clear amber liquid (determined by Charles River Den Bosch)
Batch: LT5C30Y170
Purity/Composition: 99.7%
Test item storage: At room temperature
Stable under storage conditions until: 30 November 2016 (retest date)
1
Method
- Target gene:
- thymidine-kinase locus (TK-locus)
Species / strain
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- Source: American Type Culture Collection, (ATCC, Manassas, USA) (2001).
Stock cultures of the cells were stored in liquid nitrogen (-196°C). The cultures were checked for mycoplasma contamination. Cell density was kept below 1 x 106 cells/ml.
Cell culture
Horse serum
Horse serum (Life Technologies) was inactivated by incubation at 56°C for at least 30 minutes.
Basic medium
RPMI 1640 Hepes buffered medium (Dutch modification) (Life Technologies, Bleiswijk, The Netherlands) containing penicillin/streptomycin (50 U/ml and 50 μg/ml, respectively) (Life Technologies), 1 mM sodium pyruvate (Sigma, Zwijndrecht, The Netherlands) and 2 mM L-glutamin (Life Technologies).
Growth medium
Basic medium, supplemented with 10% (v/v) heat-inactivated horse serum (=R10 medium).
Exposure medium
For 3 hour exposure:
Cells were exposed to the test item in basic medium supplemented with 5% (v/v) heat-inactivated horse serum (R5-medium).
For 24 hour exposure:
Cells were exposed to the test item in basic medium supplemented with 10% (v/v) heat-inactivated horse serum (R10-medium).
Selective medium
Selective medium consisted of basic medium supplemented with 20% (v/v) heat-inactivated horse serum (total amount of serum = 20%, R20) and 5 μg/ml trifluorothymidine (TFT) (Sigma).
Non-selective medium
Non-selective medium consisted of basic medium supplemented with 20% (v/v) heat-inactivated horse serum (total amount of serum = 20%, R20). - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mixture
- Test concentrations with justification for top dose:
- Hepteen Base® was poorly soluble in the exposure medium, the highest tested concentration was 164 μg/ml exposure medium.
- Vehicle / solvent:
- Ethanol
Controls
- Untreated negative controls:
- yes
- Remarks:
- Ethanol (Merck, Darmstadt, Germany).
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- methylmethanesulfonate
- Details on test system and experimental conditions:
- Test System: L5178Y/TK+/--3.7.2C mouse lymphoma cells.
Source: American Type Culture Collection, (ATCC, Manassas, USA) (2001).
Stock cultures of the cells were stored in liquid nitrogen (-196°C). The cultures were checked for mycoplasma contamination. Cell density was kept below 1 x 106 cells/ml.
Cell culture
Horse serum
Horse serum (Life Technologies) was inactivated by incubation at 56°C for at least 30 minutes.
Basic medium
RPMI 1640 Hepes buffered medium (Dutch modification) (Life Technologies, Bleiswijk, The Netherlands) containing penicillin/streptomycin (50 U/ml and 50 μg/ml, respectively) (Life Technologies), 1 mM sodium pyruvate (Sigma, Zwijndrecht, The Netherlands) and 2 mM L-glutamin (Life Technologies).
Growth medium
Basic medium, supplemented with 10% (v/v) heat-inactivated horse serum (=R10 medium).
Exposure medium
For 3 hour exposure:
Cells were exposed to the test item in basic medium supplemented with 5% (v/v) heat-inactivated horse serum (R5-medium).
For 24 hour exposure:
Cells were exposed to the test item in basic medium supplemented with 10% (v/v) heat-inactivated horse serum (R10-medium).
Selective medium
Selective medium consisted of basic medium supplemented with 20% (v/v) heat-inactivated horse serum (total amount of serum = 20%, R20) and 5 μg/ml trifluorothymidine (TFT) (Sigma).
Non-selective medium
Non-selective medium consisted of basic medium supplemented with 20% (v/v) heat-inactivated horse serum (total amount of serum = 20%, R20).
Environmental conditions
All incubations were carried out in a humid atmosphere (80 - 100%, actual range 57 – 91%) containing 5.0 ± 0.5% CO2 in air in the dark at 37.0 ± 1.0°C (actual range 35.0 – 37.3°C). Temperature and humidity were continuously monitored throughout the experiment. The CO2 percentage was monitored once on each working day. Temporary deviations from the temperature, humidity and CO2 percentage may occur due to opening and closing of the incubator door. Any variation to these conditions were evaluated and maintained in the raw data. - Rationale for test conditions:
- Rationale: Recommended test system in international guidelines (e.g. OECD).
- Evaluation criteria:
- Data evaluation and statistical procedures
In addition to the criteria stated below, any increase of the mutation frequency should be evaluated for its biological relevance including comparison of the results with the historical control data range.
The global evaluation factor (GEF) has been defined by the IWGT as the mean of the negative/solvent MF distribution plus one standard deviation. For the micro well version of the assay the GEF is 126.
A test item is considered positive (mutagenic) in the mutation assay if it induces a MF of more than MF(controls) + 126 in a dose-dependent manner. An observed increase should be biologically relevant and will be compared with the historical control data range.
A test item is considered equivocal (questionable) in the mutation assay if no clear conclusion for positive or negative result can be made after an additional confirmation study.
A test item is considered negative (not mutagenic) in the mutation assay if:none of the tested concentrations reaches a mutation frequency of MF(controls) + 126.
Results and discussion
Test results
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Solubility
Hepteen Base® precipitated in the exposure medium at concentrations of 52 μg/ml and above. The concentration used as the highest test item concentration for the dose range finding test was 164 μg/ml.
Dose range finding test
In the dose range finding test, L5178Y mouse lymphoma cells were treated with a test item concentration range of 1.7 to 164 μg/ml in the absence of S9-mix with 3- and 24-hour treatment periods and in the presence of S9-mix with a 3-hour treatment period.
In the absence of S9-mix, the relative suspension growth was 49% at the test item concentration of 164 μg/ml compared to the relative suspension growth of the solvent control.
In the presence of S9-mix, no toxicity in the relative suspension growth was observed up to test item concentrations of 164 μg/ml compared to the solvent control.
The relative suspension growth was 30% at the test item concentration of 17 μg/ml compared to the relative suspension growth of the solvent control. Hardly any cell survival was observed at the test item concentration of 52 μg/ml and upwards after 24 hours treatment.
Mutation experiment
The percentages of cell survival and the mutation frequencies for various concentrations of Hepteen Base®. Individual colony counts of cloning and selective plates and cell counts during subculturing are tabulated and listed in “Any other information on results incl. tables).
First mutagenicity test
Based on the results of the dose range finding test, the following dose range was selected for the first mutagenicity test:
Without S9-mix: 0.17, 0.54, 1.7, 5.4, 17, 30, 52, 85 and 164 μg/ml exposure medium.
With S9-mix: 0.05, 0.17, 0.54, 1.7, 5.4, 17, 52 and 164 μg/ml exposure medium.
Evaluation of toxicity
In the absence of S9-mix, the dose level of 52 μg/ml was not used for mutation frequency measurement, since this dose level showed an inconsistent RSG.
In the presence of S9-mix, no toxicity was observed and all dose levels were evaluated.
The dose levels selected to measure mutation frequencies at the TK-locus were:
Without S9-mix: 0.17, 0.54, 1.7, 5.4, 17, 30, 85 and 164 μg/ml exposure medium.
With S9-mix: 0.05, 0.17, 0.54, 1.7, 5.4, 17, 52 and 164 μg/ml exposure medium.
In the absence of S9-mix, the relative total growth of the highest test item concentration was 32% compared to the total growth of the solvent controls.
In the presence of S9-mix, no toxicity was observed up to and including the highest tested dose level tested. .
Evaluation of the mutagenicity
No significant increase in the mutation frequency at the TK locus was observed after treatment with Hepteen Base® either in the absence or in the presence of S9-mix. The numbers of small and large colonies in the Hepteen Base® treated cultures were comparable to the numbers of small and large colonies of the solvent controls.
Second mutagenicity test
To obtain more information about the possible mutagenicity of Hepteen Base®, a second mutation experiment was performed in the absence of S9-mix with a 24-hour treatment period.
Based on the results of the dose range finding test, the following dose levels were selected for mutagenicity testing: 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40 and 45 μg/ml exposure medium.
Evaluation of toxicity
The dose levels of 25 to 45 μg/ml were not used for mutation frequency measurement, since these dose levels were too toxic for further testing. The dose levels selected to measure mutation frequencies at the TK-locus were: 0.1, 0.5, 1, 5, 10, 15 and 20 μg/ml exposure medium.
The relative total growth of the highest test item was 7% compared to the total growth of the solvent controls;
Evaluation of mutagenicity
No significant increase in the mutation frequency at the TK locus was observed after treatment with Hepteen Base®. The numbers of small and large colonies in the Hepteen Base® treated
Applicant's summary and conclusion
- Conclusions:
- The mutation frequency found in the solvent control cultures was within the acceptability criteria of this assay and within the 95% control limits of the distribution of the historical negative control database.
Positive control chemicals, methyl methanesulfonate and cyclophosphamide, both produced significant increases in the mutation frequency. In addition, the mutation frequency found in the positive control cultures was within the 95% control limits of the distribution of the historical positive control database. It was therefore concluded that the test conditions were adequate and that the metabolic activation system (S9-mix) functioned properly.
The suspension growth (SG) over the two-day expression period for cultures treated with ethanol was between 16 and 22 (3 hours treatment) and 136 and 150 (24 hours treatment).
In the absence of S9-mix, Hepteen Base® did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in a repeat experiment with modification in the duration of treatment.
In the presence of S9-mix, Hepteen Base® did not induce a significant increase in the mutation frequency.
In conclusion, Hepteen Base® is not mutagenic in the TK mutation mouse lymphoma test system under the experimental conditions described in this report. - Executive summary:
Evaluation of the mutagenic activity of Hepteen Base® in an in vitro mammalian cell gene mutation test with L5178Y mouse lymphoma cells.
This report describes the effects of Hepteen Base® on the induction of forward mutations at the thymidine-kinase locus (TK-locus) in L5178Y mouse lymphoma cells. The test was performed in the absence of S9-mix with 3 and 24-hour treatment periods and in the presence of S9-mix with a 3 hours treatment period (rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone).
The study procedures described in this report were based on the most recent OECD guideline no. 490.
Batch LT5C30Y170 of Hepteen Base® was a clear amber liquid with a purity of 99.7%. The test item was dissolved in ethanol.
In the first experiment, Hepteen Base® was tested up to concentrations of 164 μg/ml in the absence and presence of S9-mix. The incubation time was 3 hours. No toxicity was observed at this dose level in the absence and presence of S9-mix. Hepteen Base® precipitated in the culture medium at this dose level.
In the second experiment, Hepteen Base® was tested up to concentrations of 20 μg/ml in the absence of S9-mix. The incubation time was 24 hours. The RTG was 7%.
The mutation frequency found in the solvent control cultures was within the acceptability criteria of this assay and within the 95% control limits of the distribution of the historical negative control database.
Positive control chemicals, methyl methanesulfonate and cyclophosphamide, both produced significant increases in the mutation frequency. In addition, the mutation frequency found in the positive control cultures was within the 95% control limits of the distribution of the historical positive control database. It was therefore concluded that the test conditions were adequate and that the metabolic activation system (S9-mix) functioned properly.
In the absence of S9-mix, Hepteen Base® did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in an independent experiment with modification in the duration of treatment.
In the presence of S9-mix, Hepteen Base® did not induce a significant increase in the mutation frequency.
It is concluded that Hepteen Base® is not mutagenic in the mouse lymphoma L5178Y test system under the experimental conditions.
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.