Registration Dossier
Registration Dossier
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: 224-320-7 | CAS number: 4306-88-1
- 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:
- 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
- Report date:
- 2016
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- adopted 21st July 1997
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- dated May 30, 2008
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Version / remarks:
- EPA 712-C-98-247, August 1998
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- 2,6-di-tert-butyl-4-nonylphenol
- EC Number:
- 224-320-7
- EC Name:
- 2,6-di-tert-butyl-4-nonylphenol
- Cas Number:
- 4306-88-1
- Molecular formula:
- C23H40O
- IUPAC Name:
- 2,6-di-tert-butyl-4-nonylphenol
- Test material form:
- liquid
- Details on test material:
- - Density: 0.895 g/cm3
Constituent 1
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No. of test material: Batch: 222375101
- Expiration date of the lot/batch: 02 May 2018
- Purity test date: 02 September 2016
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- Stability under test conditions:
Method
- Target gene:
- histidine
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Cytokinesis block (if used):
- histidine
- Metabolic activation:
- with and without
- Metabolic activation system:
- The S9 liver microsomal fraction was prepared at Eurofins Munich. Male Wistar rats were induced with phenobarbital (80 mg/kg bw) and (3-naphthoflavone (100 mg/kg bw) for three consecutive days by oral route.
- Test concentrations with justification for top dose:
- Pre-experiment for toxicity: 3.16, 10.0, 31.6, 100, 316, 1000, 2500 and 5000 µg/plate
Experiment I and II: 31.6,100, 316, 1000, 2500 and 5000 µg/plate
The test item concentrations to be applied in the main experiments were chosen according to the results of the pre-experiment. 5000 µg/plate was selected as the maximum concentration. The concentration range covered two logarithmic decades. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solvent was compatible with the survival of the bacteria and the S9 activity.
Controlsopen allclose all
- Untreated negative controls:
- yes
- Remarks:
- A. dest
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- methylmethanesulfonate
- other: 4-nitro-o-phenylene-diamine
- Positive control substance:
- other: 2-aminoanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION:
Experiment I in agar (plate incorporation)
Experiment II preincubation
DURATION Experiment I (plate incorporation)
- Preincubation period: No
- Exposure duration: at 37 °C for at least 48 h in the dark
- Selection time (if incubation with a selection agent): at least 48 h
DURATION Experiment II (preincubation)
- Preincubation period: for 60 min at 37 °C
- Exposure duration: 60 min plus at least 48 h
- Selection time (if incubation with a selection agent): 60 min plus at least 48 h
SELECTION AGENT (mutation assays): 10.5 mg L-histidine x HCI x H2O
NUMBER OF REPLICATIONS: triplicate
DETERMINATION OF CYTOTOXICITY
- Method: Cytotoxicity can be detected by a clearing or rather diminution of the background lawn (indicated as "N" or "B", respectively in the result tables) or a reduction in the number of revertants down to a mutation factor of approximately ≤ 0.5 in relation to the solvent control.
- Any supplementary information relevant to cytotoxicity: - Evaluation criteria:
- The Mutation Factor is calculated by dividing the mean value of the revertant counts by the mean values of the solvent controI (the exact and not the rounded values are used for calculation).
A test item is considered as mutagenic if:
-a clear and dose-related increase in the number of revertants occurs and/or
-a biologically relevant positive response for at least one of the dose groups occurs
-in at least one tester strain with or without metabolic activation.
A biologically relevant increase is described as follows:
-if in tester strains TA 98, TA 100 and TA 102 the number of reversions is at least twice as high
-if in tester strains TA 1535 and TA 1537 the number of reversions is at least three times higher than the reversion rate of the solvent control.
According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.
A test item producing neither a dose related increase in the number of revertants nor a reproducible biologically relevant positive response at any of the dose groups is considered to be non-mutagenic in this system.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Remarks:
- 5000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Remarks:
- 5000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Remarks:
- 5000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Remarks:
- 5000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Remarks:
- 5000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation observed at 2500 µg/plate and higher in all tester strains used in Experiment I and II
RANGE-FINDING/SCREENING STUDIES:The toxicity of the test item was determined with tester strains TA 98 and TA 100 in a pre-experiment.
Eight concentrations were tested for toxicity and induction of mutations with three plates each. The experimental conditions in this pre-experiment were the same as described below for the main experiment I (plate incorporation test).
Toxicity may be detected by a clearing or rather diminution of the background lawn or a reduction in the number of revertants down to a mutation factor of approximately ≤ 0.5 in relation to the solvent control.
The test item was tested in the pre-experiment with the following concentrations:
3.16, 10.0, 31.6, 100, 316, 1000, 2500 and 5000 µg/plate.
No toxicity was observed.
HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data (2013-2015):
Without S9:
TA 98: 4-NOPD 10µg: min 141 max 2213 mean 444.5 SD 180.0
TA 100: NaN3 10 µg: min 132 max 1498 mean 627.6 SD 2255.3
TA 1535: NaN3 10 µg: min 34 max 1854 mean 795.5 SD 323.7
TA 1537: 4-NOPD 40µg: min 32 max 273 mean 92.0 SD 24.8
TA 102: MMS 1µL: min 162 max 3321 mean 1801.0 SD 483.5
With S9:
TA 98: 2-AA 2.5 µg: min 113 max 3606 mean 2126.7 SD 679.0
TA 100: 2-AA 2.5 µg: min 169 max 3132 mean 1786.6 SD 494.6
TA 1535: 2-AA 2.5 µg: min 20 max 1384 mean 103.4 SD 64.9
TA 1537: 2-AA 2.5 µg: min 26 max 489 mean 221.8 SD 92.2
TA 102: 2-AA 10 µg: min 137 max 1520 mean 748.8 SD 176.3
- Negative (solvent/vehicle) historical control data (2013-2015):
Without S9:
TA 98: min 13 max 54 mean 23.1 SD 6.1
TA 100: min 49 max 139 mean 89.2 SD 13.2
TA 1535: min 4 max 39 mean 12.0 SD 5.8
TA 1537: min 2 max 35 mean 7.4 SD 2.6
TA 102: min 141 max 472 mean 251.4 SD 55.6
With S9:
TA 98: min 13 max 61 mean 29.9 SD 7.0
TA 100: min 67 max 162 mean 98.3 SD 14.2
TA 1535: min 4 max 32 mean 9.4 SD 3.7
TA 1537: min 3 max 36 mean 7.4 SD 2.8
TA 102: min 91 max 586 mean 317.7 SD 76.6
- Measurement of cytotoxicity used: [complete, e.g. CBPI or RI in the case of the cytokinesis-block method; RICC, RPD or PI when cytokinesis block is not used]
- Other observations when applicable: [complete, e.g. confluency, apoptosis, necrosis, metaphase counting, frequency of binucleated cells] - Remarks on result:
- other: Precipitation observed at 2500 µg/plate and higher
Applicant's summary and conclusion
- Conclusions:
- During the described mutagenicity test and under the experimental conditions reported, 2,6-Di-tert-butyl-4-nonylphenol did not cause gene mutations by
base pair changes or frameshifts in the genome of the tester strains used.
Therefore, 2,6-Di-tert-butyl-4-nonylphenol is considered to be non-mutagenic in th is bacterial reverse mutation assay. - Executive summary:
In order to investigate the potential of 2,6-Di-tert-butyl-4-nonylphenol for its ability to induce gene mutations the plate incorporation test (experiment I) and the pre-incubation test (experiment II) were performed with the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102.
In two independent experiments several concentrations of the test item were used. Each assay was conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicate. The following concentrations of the test item were prepared and used in the experiments: 31.6, 100, 316, 1000, 2500 and 5000 µg/plate
Precipitation was observed in all tester strains used in experiment I and II (with and without metabolic activation).
No toxic effects of the test item were noted in any of the five tester strains used up to the highest dose group evaluated (with and without metabolic activation) in experiment I and II.
No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with 2,6-Di-tert-butyl-4-nonylphenol at any concentration level, neither in the presence nor absence of metabolic activation in experiment I and II.
All criteria of validity were met.
In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, 2,6-Di-tert-butyl-4-nonylphenol did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used.
Therefore, 2,6-Di-tert-butyl-4-nonylphenol is considered to be non-mutagenic in this bacterial reverse mutation assay.
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.
