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EC number: 263-134-0 | CAS number: 61790-41-8
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Based on the results of the read across Ames study, the test substance is not considered to be genotoxic.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 12 May 2017 - 26 July 2017
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 21 July 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- 31 May 2008
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- d.d. 3 November 2015
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- - S. typhimurium: Histidine gene
- E. coli: Tryptophan gene - Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat liver S9-mix induced with Aroclor 1254
Rat liver microsomal enzymes (S9 homogenate) were obtained from Trinova Biochem GmbH, Giessen, Germany and were prepared from male Sprague Dawley rats that had been injected intraperitoneally with Aroclor 1254 (500 mg/kg body weight). Each S9 batch is characterized with the mutagens benzo-(a)-pyrene and 2-aminoanthracene, which require metabolic activation, in tester strain TA100 at concentrations of 5 μg/plate and 2.5 μg/plate, respectively.
Preparation of S9-Mix
S9-mix was prepared immediately before use and kept on ice. S9-mix contained per 10 ml: 30 mg NADP (Randox Laboratories Ltd., Crumlin, United Kingdom) and 15.2 mg glucose-6-phosphate (Roche Diagnostics, Mannheim, Germany) in 5.5 ml or 5.0 ml Milli-Q water (first or second experiment respectively) (Millipore Corp., Bedford, MA., USA); 2 ml 0.5 M sodium phosphate buffer pH 7.4; 1 ml 0.08 M MgCl2 solution (Merck); 1 ml 0.33 M KCl solution (Merck). The above solution was filter (0.22 μm)-sterilized. To 9.5 ml of S9-mix components 0.5 ml S9-fraction was added (5% (v/v) S9-fraction) to complete the S9-mix in the first experiment and to 9.0 ml of S9-mix components 1.0 ml S9-fraction was added (10% (v/v) S9-fraction) to complete the S9-mix in the second experiment. - Test concentrations with justification for top dose:
- Experiment 1
Preliminary test (without and with 5% S9-mix) TA100 and WP2uvrA: 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate
Main study:
TA1535, TA1537 and TA98: without and with 5% S9-mix: 0.18, 0.55, 1.7, 5.4, 17 and 52 µg/plate
Additional:
TA1535 and TA98: with 5% S9-mix: 17, 52, 164 and 512 µg/plate
Experiment 2:
All strains: without S9-mix: 4.7, 8.5, 15, 27, 48 and 86 µg/plate
All strains: with 10% S9-mix: 8.5, 15, 27, 48, 86 and 154 µg/plate
Additional:
TA1535, TA1537, TA98 and TA100: without S9-mix: 0.83, 1.5, 2.6, 4.7, 8.5 and 15 µg/plate
WP2uvrA: with 10% S9-mix: 86, 154, 275 and 492 µg/plate - Vehicle / solvent:
- - Vehicle used: Milli-Q Water
- Justification for choice of solvent/vehicle: Test compound was soluble in water and water has been accepted and approved by authorities and international guidelines - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- without S9; 5 µg/plate in saline for TA1535
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: ICR-191
- Remarks:
- without S9; 2.5 µg/plate in DMSO for TA1537
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 2-nitrofluorene
- Remarks:
- without S9; 10 µg/plate in DMSO for TA98
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- without S9; 650 µg/plate in DMSO for TA100
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- Remarks:
- without S9; 10 µg/plate in DMSO for WP2uvrA
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- in DMSO; 2.5 μg/plate for TA1535 (5% + 10% S9-mix) and TA1537 (5% S9-mix); 5 μg/plate for TA1537 (10% S9-mix); 1 μg/plate for TA98 (5% + 10% S9-mix) and TA100 (5% S9-mix); 2 μg/plate for TA100 (10% S9-mix); 15 μg/plate for WP2uvrA (5% + 10% S9-mix).
- Details on test system and experimental conditions:
- Method of application: in agar (plate incorporation)
Duration
- Exposure duration: 48 ± 4 hour
Number of replications:
- Doses of the test substance were tested in triplicate in each strain. Two independent experiments were conducted.
Number of cells evaluated: 10E8 per plate
Determination of cytotoxicity
- Method: The reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies.
Other examinations:
- The presence of precipitation of the test compound on the plates was determined.
Additional experiments:
Since in the first experiment in the presence of S9-mix, no toxicity and no precipitate was observed at the highest dose levels tested in the tester strains TA1535 and TA98, an additional experiment was performed. In the second mutation experiment, due to the cytotoxicity, only two to three analyzable dose levels were left for the determination of the mutagenicity of the test substance in the tester strains TA1535, TA1537, TA98 and TA100 in the absence of S9-mix. Furthermore, in tester strain WP2uvrA no dose level with toxicity or precipitate on the plates was observed in the presence of S9-mix. Therefore an additional mutation experiment was performed. - Rationale for test conditions:
- A dose-range finding test was performed and the highest concentration of the test item used in the subsequent mutation assay was 5000 μg/plate or the level at which the test item inhibited bacterial growth.
- Evaluation criteria:
- ACCEPTABILITY CRITERIA
A Salmonella typhimurium reverse mutation assay and/or Escherichia coli reverse mutation assay is considered acceptable if it meets the following criteria:
a) The vehicle control and positive control plates from each tester strain (with or without S9-mix) must exhibit a characteristic number of revertant colonies when compared against relevant historical control data generated at Charles River Den Bosch.
b) The selected dose-range should include a clearly toxic concentration or should exhibit limited solubility as demonstrated by the preliminary toxicity range-finding test or should extend to 5 mg/plate.
c) No more than 5% of the plates are lost through contamination or some other unforeseen event. If the results are considered invalid due to contamination, the experiment will be repeated.
INTERPRETATION
A test substance is considered negative (not mutagenic) in the test if:
a) The total number of revertants in the tester strain TA100 or WP2uvrA is not greater than two (2) times the concurrent vehicle control, and the total number of revertants in tester strains TA1535, TA1537 or TA98 is not greater than three (3) times the concurrent vehicle control.
b) The negative response should be reproducible in at least one follow-up experiment.
A test substance is considered positive if:
a) The total number of revertants in the tester strain TA100 or WP2uvrA is greater than two (2) times the concurrent vehicle control, or the total number of revertants in tester strains TA1535, TA1537, TA98 is greater than three (3) times the concurrent vehicle control.
b) In case a follow up experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one follow up experiment. - Statistics:
- No formal hypothesis testing was done.
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Under the study conditions, the test substance was not mutagenic in the Salmonella typhimurium reverse mutation assay and Escherichia coli reverse mutation assay, with or without metabolic activation.
- Executive summary:
An in vitro study was conducted to determine the mutagenic potential of test substance, TMAC O (91.9% acive), according to OECD Guideline 471 and EU Method B.13/B.14, in compliance with GLP. 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. 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 0.18 to 52 μg/plate in the absence and presence of 5% (v/v) S9-mix in the tester strains TA1535, TA1537 and TA98. The test substance did not precipitate on the plates at this dose level.Cytotoxicity, as evidenced by a decrease in the number of revertants, reduction of the bacterial background lawn and/or the presence of microcolonies, was observed in all tester strains in the absence and presence of S9-mix, except for the tester strains TA1535 and TA98 in the presence of S9. So, an additional experiment was performed with these two tester strains TA1535 and TA98, using concentration range of 17 to 512 μg/plate of the test substance. No precipitation on the plates was observed at this dose level, however, cytotoxicity was observed in both tester strains. In a follow-up experiment of the assay with additional parameters, the test substance was tested at concentration ranges of 4.7 to 86 μg/plate and 8.5 to 154 μg/plate in the absence and presence of 10% (v/v) S9-mix, respectively, in the tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA. The test substance did not precipitate on the plates at this dose level. Cytotoxicity, was observed in all tester strains in the absence and presence of S9-mix. Except for tester strain WP2uvrA, where no cytotoxicity was observed in the presence of S9 mix. In the second mutation experiment, due to the cytotoxicity, only two to three analyzable dose levels were left for the determination of the mutagenicity of the test substance in the tester strains TA1535, TA1537, TA98 and TA100 in the absence of S9-mix. Furthermore, in tester strain WP2uvrA no dose level with toxicity or precipitate on the plates was observed in the presence of S9-mix. Therefore an additional mutation experiment was performed. In this additional experiment, the test substance was tested at concentration ranges of 0.83 to 15 μg/plate in strains TA1535, TA1537, TA98 and TA100 in the absence of S9 mix and at 86 to 492 μg/plate in tester strain WP2uvrA in the presence of S9 mix. The test substance seemed to show slight precipitation at the highest tested concentration of 15 μg/plate in the tester strains TA98 and TA100 in the presence of a reduced bacterial background. Cytotoxicity, was observed in all tester strains in the absence or presence of S9-mix. Except for tester strain TA1535, where no cytotoxicity was observed in the absence of S9 mix. The test substance did not induce a significant dose-related increase in the number of revertant (His+) colonies in each of the four tester strains (TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trp+) colonies in the tester strain WP2uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed in a follow-up experiment. In this study, acceptable responses were obtained for the negative and strain-specific positive control substances, indicating that the test conditions were adequate and that the metabolic activation system functioned properly. Under the study conditions, the test substance was not mutagenic in the Salmonella typhimurium reverse mutation assay and Escherichia coli reverse mutation assay, with or without metabolic activation (Gijsbrechts, 2017).
Reference
Results
Dose-range Finding Test/First Mutation Experiment
Test substance (dried) was tested in the tester strains TA100 and WP2uvrA at 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 first mutation experiment with the tester strains, TA1535, TA1537 and TA98 in the absence and presence of S9-mix: 0.18, 0.55, 1.7, 5.4, 17 and 52 μg/plate.
Precipitate
Precipitation of the test substance on the plates was not observed at the start or at the end of the incubation period in any tester strain.
Toxicity
To determine the toxicity of the test substance, the reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies were examined. Cytotoxicity, as evidenced by a decrease in the number of revertants, reduction of the bacterial background lawn and/or the presence of microcolonies, was observed in all tester strains in the absence and presence of S9-mix, except for the tester strains TA1535 and TA98 in the presence of S9.
Mutagenicity
No increase in the number of revertants was observed upon treatment with the test substance under all conditions tested.
First Mutation Experiment (additional)
Since in the first experiment in the presence of S9-mix, no toxicity and no precipitate was observed at the highest dose levels tested in the tester strains TA1535 and TA98, an additional experiment was performed. Based on the results of the first mutation experiment, the following dose-range was selected for the additional mutation experiment with the tester strains, TA1535 and TA98 in the presence of S9-mix: 17, 52, 164 and 512 μg/plate. Precipitation of the test substance on the plates was not observed at the start or at the end of the incubation period.
Toxicity
Cytotoxicity, as evidenced by a decrease in the number of revertants, reduction of the bacterial background lawn and/or the presence of microcolonies, was observed in both tester strains.
Mutagenicity
In the additional mutation experiment, no increase in the number of revertants was observed upon treatment with Test substance (dried) under all conditions tested.
Second Mutation Experiment
To obtain more information about the possible mutagenicity of the test substance, a second mutation experiment was performed in the absence and 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 levels of 86 and 154 µg/plate in strains TA1535, TA1537, TA98, TA100 and WP2uvrA in the absence and presence of S9 mix, respectively. Precipitation of the test substance on the plates was not observed at the start or at the end of the incubation period.
Toxicity
Cytotoxicity, as evidenced by a decrease in the number of revertants, reduction of the bacterial background lawn and/or the presence of microcolonies, was observed in all tester strains in the absence and presence of S9-mix. Except for tester strain WP2uvrA, where no cytotoxicity was observed in the presence of S9 mix.
Mutagenicity
In the second mutation assay, no increase in the number of revertants was observed upon treatment with Test substance (dried) under all conditions tested.
Second Mutation Experiment (additional)
In the second mutation experiment, due to the cytotoxicity, only two to three analyzable dose levels were left for the determination of the mutagenicity of the test substance in the tester strains TA1535, TA1537, TA98 and TA100 in the absence of S9-mix. Furthermore, in tester strain WP2uvrA no dose level with toxicity or precipitate on the plates was observed in the presence of S9-mix. Therefore an additional mutation experiment was performed. In this additional experiment, the test substance was tested at concentration ranges of 0.83 to 15 µg/plate in strains TA1535, TA1537, TA98 and TA100 in the absence of S9 mix and at 86 to 492 µg/plate in tester strain WP2uvrA in the presence of S9 mix.
Precipitate
No precipitate was observed at the start of the incubation period. Although in the tester strains TA1535 and TA1537, as well as in the second mutation experiment, no precipitation was observed, the test substance seemed to show slight precipitation at the highest tested concentration of 15 µg/plate in the tester strains TA98 and TA100 in the presence of a reduced bacterial background.
Toxicity
Cytotoxicity, as evidenced by a decrease in the number of revertants, reduction of the bacterial background lawn and/or the presence of microcolonies, was observed in all tester strains in the absence or presence of S9-mix. Except for tester strain TA1535, where no cytotoxicity was observed in the absence of S9 mix.
Mutagenicity
In the second mutation assay, no increase in the number of revertants was observed upon treatment with Test substance (dried) under all conditions tested.
Discussion
All bacterial strains showed negative responses over the entire dose-range, i.e. no significant dose-related increase in the number of revertants in two experiments. The negative and strain-specific positive control values were within the laboratory historical control data. The strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly, except the response for TA98 in the presence of S9-mix in the second experiment. The purpose of the positive control is as a reference for the test system, where a positive response is required to check if the test system functions correctly. Since the value was more than 8 times greater than the concurrent solvent control values, this deviation in the mean plate count of the positive control had no effect on the results of the study.
Conclusion
In conclusion, based on the results of this study it is concluded that Test substance (dried) is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
An in vitro study was conducted to determine the mutagenic potential of test substance, TMAC O (91.9% acive), according to OECD Guideline 471 and EU Method B.13/B.14, in compliance with GLP.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.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 0.18 to 52 μg/plate in the absence and presence of 5% (v/v) S9-mix in the tester strains TA1535, TA1537 and TA98. The test substance did not precipitate on the plates at this dose level.Cytotoxicity, as evidenced by a decrease in the number of revertants, reduction of the bacterial background lawn and/or the presence of microcolonies, was observed in all tester strains in the absence and presence of S9-mix, except for the tester strains TA1535 and TA98 in the presence of S9. So, an additional experiment was performed with these two tester strains TA1535 and TA98, using concentration range of 17 to 512 μg/plate of the test substance. No precipitation on the plates was observed at this dose level, however, cytotoxicity was observed in both tester strains. In a follow-up experiment of the assay with additional parameters, the test substance was tested at concentration ranges of 4.7 to 86 μg/plate and 8.5 to 154 μg/plate in the absence and presence of 10% (v/v) S9-mix, respectively, in the tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA. The test substance did not precipitate on the plates at this dose level. Cytotoxicity, was observed in all tester strains in the absence and presence of S9-mix. Except for tester strain WP2uvrA, where no cytotoxicity was observed in the presence of S9 mix.In the second mutation experiment, due to the cytotoxicity, only two to three analyzable dose levels were left for the determination of the mutagenicity of the test substance in the tester strains TA1535, TA1537, TA98 and TA100 in the absence of S9-mix. Furthermore, in tester strain WP2uvrA no dose level with toxicity or precipitate on the plates was observed in the presence of S9-mix. Therefore an additional mutation experiment was performed. In this additional experiment, the test substance was tested at concentration ranges of 0.83 to 15 μg/plate in strains TA1535, TA1537, TA98 and TA100 in the absence of S9 mix and at 86 to 492 μg/plate in tester strain WP2uvrA in the presence of S9 mix. The test substance seemed to show slight precipitation at the highest tested concentration of 15 μg/plate in the tester strains TA98 and TA100 in the presence of a reduced bacterial background. Cytotoxicity, was observed in all tester strains in the absence or presence of S9-mix. Except for tester strain TA1535, where no cytotoxicity was observed in the absence of S9 mix.The test substance did not induce a significant dose-related increase in the number of revertant (His+) colonies in each of the four tester strains (TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trp+) colonies in the tester strain WP2uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed in a follow-up experiment.In this study, acceptable responses were obtained for the negative and strain-specific positive control substances, indicating that the test conditions were adequate and that the metabolic activation system functioned properly.Under the study conditions,it was concluded that the read acrosssubstance was not mutagenic in Salmonella typhimurium strain TA98, TA100, TA1535 or TA 1537 and E.Coli WP2uvrA, with or without metabolic activation(Gijsbrechts, 2017).
Based on the results of the read across study, similar non-mutagenic potential can be expected for the test substance.
Justification for classification or non-classification
Based on the results of the read across Ames study, the test substance does not warrant aclassification for genotoxicity according to the EU CLP criteria (Regulation 1272/2008/EC).
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.
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