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EC number: 287-484-9 | CAS number: 85536-04-5
- 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 mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 08 Mar - 29 Jun 1994
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 994
- Report date:
- 1994
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Version / remarks:
- adopted April 1984
- Deviations:
- no
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Version / remarks:
- 2016
- Deviations:
- yes
- Remarks:
- limited documentation, less then 2 million cells cultured during expression period
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OTS 798.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Hessisches Ministerium für Umwelt, Energie und Bundesangelegenheiten, Wiesbaden, Germany
- Type of assay:
- in vitro mammalian cell gene mutation test using the Hprt and xprt genes
Test material
- Reference substance name:
- 3687-45-4 (purity: 55-70%)
- IUPAC Name:
- 3687-45-4 (purity: 55-70%)
Constituent 1
Method
- Target gene:
- HPRT locus
Species / strain
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: minimal essential medium (MEM) supplemented with 10 % fetal calf serum (FCS)
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes, by treatment with HAT-medium
- Doubling time 12 - 16 h in stock cultures
- Metabolic activation:
- with and without
- Metabolic activation system:
- Co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with 500 mg/kg bw Aroclor 1254 i.p. 5 days prior to sacrifice.
- Test concentrations with justification for top dose:
- 10, 30, 60 and 100 µg/mL, with and without metabolic activation
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol. The final concentration of ethanol in the culture medium did not exceed 1 % v/v.
- Justification for choice of solvent/vehicle: The solvent was chosen due to its solubility properties and its non-toxicity to the cells.
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- ethanol
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: ethylmethanesulphonate (600 µg/mL in MEM without FCS, -S9) and 7,12-dimethylbenz(a)anthracene (3.85 µg/mL in DMSO, +S9)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 7 days
- Selection time (if incubation with a selection agent):
Cloning efficiency: 7 days
Mutant selection: 9 and 12 days (experiments 2 and 1, respectively)
- Fixation time (start of exposure up to fixation or harvest of cells):
Cloning efficiency: 7 and 14 days
Mutant selection: 16 and 19 days (experiments 2 and 1, respectively)
Counting of colonies: the colonies were stained with 10 % methylene blue in 0.01 % KOH solution. Stained colonies with more than 50 cells were counted. In doubt the colony size was checked with a preparation microscope.
SELECTION AGENT (mutation assays): thioguanine, 11 µg/mL medium (Sigma, Deisenhofen, Germany)
NUMBER OF REPLICATIONS: 1 replication in 2 independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: determination of concentration-related cloning efficiency on day 7 after start of exposure; determination of cell survival on day 14 after start of exposure (end of selection time) - Evaluation criteria:
- The test substance is considered to be positive if it induces either a concentration-related increase of the mutant frequency or a reproducible positive response for one of the test points.
A test material producing neither a concentration-related increase in the mutant frequency nor a reproducible positive response in any of the test points is considered non-mutagenic in this system.
A significant response is:
1) test substance induces reproducibly with one of the concentrations a mutation frequency that is 3 times higher than the spontaneous mutation frequency in the experiment
2) test substance induces reproducible concentration-related increase of the mutation frequency. May also be considered in case a 3-fold increase of the mutant frequency is not observed.
In a case-by-case evaluation the decision depends on the level of the corresponding negative control data.
Results and discussion
Test results
- Key result
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Remarks:
- precipitation at limit of water solubility at 100 µg/mL observed.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: Due to limited solubility of the test substance higher concentrations than 100 µg/mL for the cytogenetic evaluation were not feasible.
- Precipitation: observed at concentrations higher than 100 µg/mL
RANGE-FINDING/SCREENING STUDIES
Data on the pre-test were obtained from the chromosomal aberration study (IUCLID section 7.6.1: key, Emery, 1994, ChrAb, RL1)
A pre-test was performed to determine the toxicity of the test article. The test substance was dissolved in ethanol due to its solubility properties. The highest attainable concentration was 100 µg/mL, at which a slight precipitation was observed. No toxic effects were observed up to and including the highest dose level.
COMPARISON WITH HISTORICAL CONTROL DATA: yes (data not presented)
ADDITIONAL INFORMATION ON CYTOTOXICITY:
In parallel to the mutagenicity testing the cloning efficiency was tested. Without metabolic activation the relative cloning efficiencies were between 64.2 - 91.8 % in treated cells compared to control cells. With metabolic activation the relative cloning efficiencies were between 92.4 - 109.7 % in treated cells compared to control cells.
No statistically significant increase in gene mutations was observed (see Table 1 and 2). The positive controls were shown to be valid.
Any other information on results incl. tables
Table 1: mutagenicity data, experiment 1
|
Concentration (µg/mL) |
Metabolic activation |
No. of mutant colonies after plating in TG medium* (mean of 5 flasks) |
% cell survival |
No. of mutant colonies per 106 surviving cells |
% cloning efficiency |
Negative control |
- |
- |
2.2 |
54 |
9.4 |
100.0 |
Vehicle control |
- |
- |
1.0 |
66 |
3.9 |
100.0 |
EMS |
600 |
- |
89.6 |
50 |
416.4 |
73.8 |
Test substance |
10 |
- |
1.0 |
72 |
3.3 |
91.8 |
Test substance |
30 |
- |
2.0 |
72 |
6.8 |
72.0 |
Test substance |
60 |
- |
0.2 |
81 |
0.6 |
68.1 |
Test substance |
100 |
- |
2.8 |
65 |
10.2 |
64.2 |
Negative control |
- |
+ |
2.0 |
69 |
7.3 |
100.0 |
Vehicle control |
- |
+ |
3.2 |
68 |
11.4 |
100.0 |
DMBA |
3.85 |
+ |
112.0 |
57 |
496.2 |
53.1 |
Test substance |
10 |
+ |
5.4 |
78 |
16.1 |
92.4 |
Test substance |
30 |
+ |
0.2 |
65 |
0.8 |
95.6 |
Test substance |
60 |
+ |
1.4 |
67 |
5.4 |
97.3 |
Test substance |
100 |
+ |
0.2 |
74 |
0.7 |
109.7 |
*only colonies with more than 50 cells 8 days after seeding in TG selection medium were scored
** ration of mean number of cells found after 7 days in normal growth medium / cell number seeded per flask
TG = Thioguanine
EMS = Ethylmethanesulphonate
DMBA = 7,12-dimethylbenz(a)anthracene
Table 2: mutagenicity data, experiment 2
|
Concentration (µg/mL) |
Metabolic activation |
No. of mutant colonies after plating in TG medium* (mean of 5 flasks) |
% cell survival |
No. of mutant colonies per 106 surviving cells |
% cloning efficiency |
Negative control |
- |
- |
2.0 |
69 |
6.6 |
100.0 |
Vehicle control |
- |
- |
1.8 |
64 |
6.4 |
100.0 |
EMS |
600 |
- |
145.8 |
60 |
496.9 |
61.5 |
Test substance |
10 |
- |
4.2 |
62 |
18.4 |
93.6 |
Test substance |
30 |
- |
1.0 |
63 |
4.1 |
107.8 |
Test substance |
60 |
- |
3.2 |
60 |
13.5 |
56.4 |
Test substance |
100 |
- |
0.8 |
59 |
3.5 |
61.4 |
Negative control |
- |
+ |
2.4 |
60 |
10.3 |
100.0 |
Vehicle control |
- |
+ |
2.2 |
61 |
9.0 |
100.0 |
DMBA |
3.85 |
+ |
141.6 |
64 |
550.4 |
50.3 |
Test substance |
10 |
+ |
1.4 |
61 |
5.7 |
104.8 |
Test substance |
30 |
+ |
2.2 |
72 |
7.5 |
106.1 |
Test substance |
60 |
+ |
0.6 |
69 |
2.1 |
102.8 |
Test substance |
100 |
+ |
1.4 |
79 |
4.4 |
107.7 |
*only colonies with more than 50 cells 8 days after seeding in TG selection medium were scored
** ratio of mean number of cells found after 7 days in normal growth medium / cell number seeded per flask
TG = Thioguanine
EMS = Ethylmethanesulphonate
DMBA = 7,12-dimethylbenz(a)anthracene
Applicant's summary and conclusion
- Conclusions:
- CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.
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