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EC number: 402-860-6 | CAS number: 110553-27-0 CG 25-1320; IRGANOX 1520; TK 12229/1
- 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 cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 991
- Report date:
- 1991
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
Test material
- Reference substance name:
- 4,6-bis(octylthiomethyl)-o-cresol
- EC Number:
- 402-860-6
- EC Name:
- 4,6-bis(octylthiomethyl)-o-cresol
- Cas Number:
- 110553-27-0
- Molecular formula:
- C25 H44 O S2
- IUPAC Name:
- 2-methyl-4,6-bis[(octylsulfanyl)methyl]phenol
- Details on test material:
- - Physical state: yellow liquid
- Storage condition of test material: room temperature
Constituent 1
Method
Species / strain
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- CELLS USED
- Type and source of cells: CHO cells
- Suitability of cells: proven
- Normal cell cycle time (negative control): 16 - 20 h in stock cultures
For cell lines:
- Absence of Mycoplasma contamination: yes
- Number of passages if applicable: twice per week
- Methods for maintenance in cell culture: The cell cultures were incubated at 37°C and 15.0 % carbon dioxide atmosphere
- Modal number of chromosomes: 20
- Periodically checked for karyotype stability: yes
MEDIA USED
- Type and composition of media, CO2 concentration, humidity level, temperature, if applicable: DMEM/F12 (Dulbecco's modified eagle medium; mixture 1:1) supplemented with 10 % fetal calf serum (FCS)
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix from rat liver induced with Aroclor 1254
- Test concentrations with justification for top dose:
- without and with S9 mix:
8 h: 10.0; 20.0; 30.0; 40.0; 50.0 µg/mL
24 h: 1.0; 5.0; 10.0; 20.0; 30.0; 40.0; 50.0 µg/mL
30 h: 10.0; 20.0; 30.0; 40.0; 50.0 µg/mL - Vehicle / solvent:
- DMSO
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Without metabolic activation: EMS; EthyImethanesulfonate; With metabolic activation: CPA; Cyclophosphamide
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in suspension
DURATION
- Preincubation period: 48 h
- Exposure duration: 5, 21, 27 h
- Fixation time (start of exposure up to fixation or harvest of cells): 8, 24, 30 h
The treatment interval was 4 h.
SPINDLE INHIBITOR (cytogenetic assays): colcemid (3 h)
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 100 per slide = 200 per test group
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index - Evaluation criteria:
- A test article is classified as mutagenic if it induces either a significant dose-related increase in the number of structural chromosomal aberrations or a significant positive response for at least one of the test points.
A test article producing neither a significant dose-related increase in the number of structural chromosomal aberrations nor a significant positive response at any one of the test points is considered non-mutagenic in this system.
However, both biological and statistical significance should be considered together. - Statistics:
- Statistical significance at the five per cent level (p < 0.05) was evaluated by means of the chi-square test. Evaluation was performed only for cells carrying aberrations exclusive gaps.
Results and discussion
Test results
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- the mitotic index was slightly reduced after treatment with the highest dose level
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- RANGE-FINDING/SCREENING STUDIES (if applicable):
In the pre-experiment on toxicity (colony forming ability) in the absence and presence of 89 mix even after treatment with the highest concentration (50.0 ng/ml) the colony forming ability was only slightly reduced. Higher concentrations precipitated strongly in the culture medium.
STUDY RESULTS
Both, in the absence and presence of S9 mix the test article did not increase the frequency of cells with aberrations at any dose level either to a biologically or to statistically relevant extend. The aberration rates after treatment with the test article (0.5 % - 5.0 %) are in our historical control data range (0.0 % - 5.0 %) or near to the range of the actual control values: 0.5 % - 2.0 %. In this experiment, there was a clonal effect in the CHO cell line revealing dicentric marker chromosomes distributed randomly in the treatment and control groups. Therefore, dicentric chromsomes were recorded but not included in the calculation of the aberration rates. Due to the random distribution this measure did not affect the validation of the study.
Any other information on results incl. tables
PRE-EXPERIEMNT FOR TOXICITY
In the pre-experiment the toxicity of the test article was examined with the plating efficiency (colony forming ability). The results are given below:
Table 1: Plating Efficiency Assay (PE) without metabolic activation Per flask 496 single cells were seeded.
colonies counted | ||||
conc. per ml | flask I | flask II | mean | relative Plating Efficiency % |
negative control | 550 | 439 | 494.5 | |
solvent control | 383 | 360 | 371.5 | 100.0 |
0.01 | 457 | 494 | 475.5 | 128.0 |
0.10 | 490 | 464 | 477.0 | 128.4 |
0.30 | 428 | 405 | 416.5 | 112.1 |
1.00 | 389 | 433 | 411.0 | 110.6 |
3.00 | 332 | 350 | 341.0 | 91.8 |
10.00 | 314 | 339 | 326.0 | 87.9 |
25.00 | 253 | 278 | 265.0 | 71.5 |
50.00 | 286 | 314 | 300.0 | 80.8 |
Table 2: Plating Efficiency Assay (PE) with metabolic activation Per flask 496 single cells were seeded.
colonies counted | ||||
conc. per ml | flask I | flask II | mean | relative Plating Efficiency % |
negative control | 356 | 328 | 342.0 | |
solvent control | 316 | 331 | 323.5 | 100.0 |
0.01 | 337 | 358 | 247.5 | 107.4 |
0.10 | 320 | 319 | 319.5 | 98.8 |
0.30 | 319 | 375 | 347.0 | 107.3 |
1.00 | 322 | 355 | 338.5 | 104.6 |
3.00 | 345 | 379 | 362.0 | 111.9 |
10.00 | 386 | 353 | 369.5 | 114.2 |
25.00 | 347 | 389 | 368.0 | 113.8 |
50.00 | 305 | 190 | 247.5 | 76.5 |
fixation interval: 8 h |
|||||
Concentration (µg/mL) | Metabolic activation | Metaphases | Aberrations incl. Gaps (%) | Aberrations excl. Gaps (%) | Exchanges (%) |
Vehicle Ctrl. | without | 200 | 4.5 | 2.0 | 0.0 |
with | 200 | 3.0 | 2.0 | 0.0 | |
50.0 | without | 200 | 1.0 | 0.5 | 0.0 |
with | 200 | 7.0 | 5.0 | 0.0 | |
fixation interval: 24 h | |||||
Concentration (µg/mL) | Metabolic activation | Metaphases | Aberrations incl. Gaps (%) | Aberrations excl. Gaps (%) | Exchanges (%) |
Negative Ctrl. | without | 200 | 4.0 | 2.0 | 0.0 |
with | 200 | 2.5 | 2.0 | 1.0 | |
Vehicle Ctrl. | without | 200 | 5.5 | 2.0 | 0.0 |
with | 200 | 1.0 | 0.5 | 0.0 | |
5.0 | without | 200 | 2.5 | 2.5 | 0.0 |
with | 200 | 5.0 | 3.0 | 0.0 | |
20.0 | without | 200 | 2.0 | 2.0 | 0.0 |
with | 200 | 3.5 | 2.0 | 0.0 | |
50.0 | without | 200 | 8.0 | 5.0 | 0.0 |
with | 200 | 2.0 | 1.0 | 0.0 | |
Positive Ctrl. | without | 100 | 28.5 | 26.0 | 13.5 |
with | 100 | 15.0 | 12.0 | 5.0 | |
fixation interval: 30 h | |||||
Concentration (µg/mL) | Metabolic activation | Metaphases | Aberrations incl. Gaps (%) | Aberrations excl. Gaps (%) | Exchanges (%) |
Vehicle Ctrl. | without | 200 | 2.0 | 1.0 | 0.0 |
with | 200 | 2.0 | 0.5 | 0.0 | |
50.0 | without | 200 | 2.0 | 0.5 | 0.0 |
with | 200 | 2.0 | 1.5 | 0.0 |
Applicant's summary and conclusion
- Conclusions:
- Under the experimental conditions reported, the test article did not induce structural chromosome aberrations in the CHO cell line.
- Executive summary:
The test article was assessed for its potential to induce structural chromosome aberrations in CHO cells in vitro. Preparation of chromosomes was done 8 h (high dose), 24 h (low, medium and high dose) and 30 h (high dose) after start of treatment with the test article. The treatment interval was 4 h. In each experimental group two parallel cultures were used. Per culture 100 metaphases were scored for structural chromosomal aberrations. The following dose levels were evaluated both without S9 mix and with S9 mix:
8 h: 50.0 µ/ml
24 h: 5.0; 20.0; 50.0 µg/ml
30 h: 50.0 µg/ml
The concentration range of the test article applied had been determined in a pre-experiment using the plating efficiency assay as indicator for toxicity response. Treatment of the cells even with the highest dose level
(50.0 µg/ml) reduced only slightly the plating efficiency. Higher concentrations than 50.0 µg/ml precipitated strongly in the culture medium. Also, the mitotic index was slightly reduced with the highest concentration at each fixation interval in the absence and at interval 8 h in the presence of S9 mix. There was no relevant increase in cells with structural aberrations after treatment with the test article at any fixation interval either without or with metabolic activation by S9 mix. Appropriate reference mutagens were used as positive controls and showed distinct increases of cells with structural chromosome aberrations.
In conclusion, it can be stated that in the study described and under the experimental conditions reported, the test article did not induce structural chromosome aberrations as determined by the chromosomal aberration test in the CHO Chinese Hamster cell line. Therefore, the test item is considered to be non-mutagenic in this chromosomal aberration test.
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