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: 209-013-8 | CAS number: 552-45-4
- 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:
- weight of evidence
- Study period:
- 22 May 2018 - 06 June 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- according to GLP
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- adopted July 21, 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- dated May 31, 2008
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Version / remarks:
- August 1998
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- α-chloro-o-xylene
- EC Number:
- 209-013-8
- EC Name:
- α-chloro-o-xylene
- Cas Number:
- 552-45-4
- Molecular formula:
- C8H9Cl
- IUPAC Name:
- 1-(chloromethyl)-2-methylbenzene
- Test material form:
- liquid
Constituent 1
- Specific details on test material used for the study:
- purity: 98.3 g / 100 g (substance characterization available)
physical state / appearance: colorless to yellowish, clear, liquid
storage conditions: +2 to +8°C
Method
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: Tinova Biochem GmbH (Gießen, Germany)
- Suitability of cells: as published by Ames et al.
Regular checking of the properties of the Salmonella typhimurium and Escherichia coli strains regarding the membrane permeability, ampicillin resistance; UV sensitivity, and amino acid requirement as well as normal spontaneous mutation rates is performed in Envigo CRS GmbH according to Ames et al. (1977) and Maron and Ames (1983). Thus, it is ensured that the experimental conditions set down by Ames are fulfilled.
- Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital/β-naphthoflavone induced rat liver S9
- Test concentrations with justification for top dose:
- According to the current OECD Guideline No. 471 the maximum concentration should be 5000 μg/plate, unless limited by toxicity or solubility of the test item.
In the pre-experiment the concentration range of the test item was 3 – 5000 μg/plate. The pre-experiment met the acceptance criteria (cf. 4.8.3), thus, it is reported as experiment I. The concentration range included two logarithmic decades.
The following concentrations were tested in experiment II:
Strain TA 100: 1, 3, 10; 33; 100; 333; 1000; and 2500 μg/plate
The remaining strains: 3; 10; 33; 100; 333; 1000; 2500; and 5000 μg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solvent was chosen because of its solubility properties and its relative nontoxicity to the bacteria (Maron et al.; 1981)
Controls
- Untreated negative controls:
- yes
- Remarks:
- untreated
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Remarks:
- sterility controls
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- methylmethanesulfonate
- other: 4-nitro-o-phenylene-diamine (4-NOPD); 2-Aminoanthracene (2-AA)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation); preincubation
For each strain and dose level, including the controls, three plates were used.
Experiment I (Plate Incorporation)
The following materials were mixed in a test tube and poured onto the selective agar plates:
100 μL Test solution at each dose level (solvent or reference mutagen solution (positive control)),
500 μL S9 mix (for test with metabolic activation) or S9 mix substitution buffer (for test without metabolic activation),
100 μL Bacteria suspension (cf. test system, pre-culture of the strains),
2000 μL Overlay agar
Experiment II (Pre-Incubation)
The following materials were mixed in a test tube and incubated at 37°C for 60 minutes:
100 μL Test solution at each dose level (solvent or reference mutagen solution (positive control)),
500 μL S9 mix (for test with metabolic activation) or S9 mix substitution buffer (for test without metabolic activation),
100 μL Bacteria suspension (cf. test system, pre-culture of the strains)
After pre-incubation 2000 μL overlay agar (approx. 45°C) was added to each tube. Then, the mixture was poured on selective agar plates.
After solidification the plates were incubated upside down for at least 48 hours at 37°C in the dark.
In parallel to each test a sterile control of the test item was performed and documented in the raw data. Therefore, 100 μL of the stock solution, 500 μl S9 mix / S9 mix substitution buffer were mixed with 2.0 mL overlay agar and poured on minimal agar plates. - Evaluation criteria:
- A test item is considered as a mutagen if a biologically relevant increase in the number of revertants of two-fold or above (strains TA 98, TA 100, and WP2 uvrA) or of three-fold or above (strains TA 1535 and TA 1537) the spontaneous mutation rate of the corresponding solvent control is observed.
A dose dependent increase is considered biologically relevant if the threshold is reached or exceeded at more than one concentration.
An increase of revertant colonies equal or above the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- starting at 2500 µg/plate, except for Experiment II without S9 mix (precipitation was observed)
- 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:
- cytotoxicity
- Remarks:
- starting at 5000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- starting at 2500 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:
- cytotoxicity
- Remarks:
- starting at 1000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- starting at 2500 µg/plate, except for experiment I without S9 mix (precipitation was observed)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: test item precipitated in the overlay agar in the test tubes from 2500 μg/plate up to the highest investigated dose in all strains with S9 mix. Precipitation of the test item in the overlay agar on the incubated agar plates was observed from 2500 μg/plate up to the highest investigated dose in all strains with and without S9 mix. The undissolved particles had no influence on the data recording.
Any other information on results incl. tables
Summary of Experiment I
Metabolic Activation |
Test Group |
Dose Level (per plate) |
Revertant Colony Counts (Mean ±SD) |
||||
|
|
|
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
WP2 uvrA |
Without Activation |
DMSO |
|
11 ± 2 |
10 ± 5 |
23 ± 7 |
208 ± 10 |
44 ± 5 |
Untreated |
|
10 ± 5 |
11 ± 3 |
33 ± 8 |
198 ± 13 |
42 ± 10 |
|
2-Methylbenzyl chloride |
3 μg |
11 ± 3 |
10 ± 3 |
25 ± 5 |
201 ± 25 |
39 ± 10 |
|
10 μg |
11 ± 0 |
11 ± 4 |
28 ± 4 |
211 ± 12 |
30 ± 4 |
||
33 μg |
15 ± 2 |
8 ± 2 |
27 ± 5 |
214 ± 6 |
35 ± 6 |
||
100 μg |
15 ± 1 |
12 ± 3 |
24 ± 6 |
216 ± 14 |
51 ± 7 |
||
333 μg |
9 ± 4 |
9 ± 4 |
20 ± 1 |
168 ± 31 |
40 ± 3 |
||
1000 μg |
11 ± 1 |
11 ± 3 |
21 ± 5 |
52 ± 12 |
24 ± 4 |
||
2500 μg |
4 ± 1PM |
5 ± 1PM |
11 ± 3PM |
48 ± 8PM |
23 ± 3P |
||
5000 μg |
4 ± 1PM |
3 ± 1PM |
4 ± 1PM |
33 ± 7PM |
23 ± 8P |
||
NaN3 |
10 μg |
1331 ± 117 |
|
|
1790 ± 101 |
|
|
4-NOPD |
10 μg |
|
|
539 ± 32 |
|
|
|
4-NOPD |
50 μg |
|
75 ± 10 |
|
|
|
|
MMS |
2.0 μL |
|
|
|
|
842 ± 11 |
|
|
|
|
|
|
|
|
|
With Activation |
DMSO |
|
15 ± 2 |
8 ± 2 |
43 ± 3 |
207 ± 6 |
46 ± 13 |
Untreated |
|
15 ± 4 |
12 ± 2 |
44 ± 13 |
213 ± 5 |
55 ± 8 |
|
2-Methylbenzyl chloride |
3 μg |
13 ± 2 |
11 ± 1 |
45 ± 9 |
194 ± 21 |
55 ± 4 |
|
10 μg |
16 ± 6 |
9 ± 3 |
41 ± 5 |
179 ± 18 |
43 ± 14 |
||
33 μg |
18 ± 4 |
10 ± 4 |
42 ± 7 |
202 ± 22 |
57 ± 6 |
||
100 μg |
21 ± 2 |
13 ± 4 |
40 ± 11 |
213 ± 8 |
60 ± 10 |
||
333 μg |
17 ± 5 |
13 ± 1 |
38 ± 6 |
224 ± 2 |
52 ± 13 |
||
1000 μg |
16 ± 5 |
15 ± 1 |
33 ± 7 |
106 ± 16 |
47 ± 11 |
||
2500 μg |
17 ± 4PM |
11 ± 2PM |
17 ± 3P |
61 ± 16PM |
44 ± 8P |
||
5000 μg |
4 ± 1PM |
0 ± 1PM |
1 ± 1PM |
8 ± 3PM |
16 ± 2PM |
||
2-AA |
2.5 μg |
417 ± 34 |
195 ± 29 |
3686 ± 348 |
4705 ± 78 |
|
|
2-AA |
10.0 μg |
|
|
|
|
465 ± 6 |
Key to Positive Controls |
Key to Plate Postfix Codes |
||
NaN3 2-AA 4-NOPD MMS |
sodium azide 2-aminoanthracene 4-nitro-o-phenylene-diamine methyl methane sulfonate |
P M |
Precipitate Manual count |
Summary of Experiment II
Metabolic Activation |
Test Group |
Dose Level (per plate) |
Revertant Colony Counts (Mean ±SD) |
||||
|
|
|
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
WP2 uvrA |
Without Activation
|
DMSO |
|
10 ± 1 |
9 ± 3 |
19 ± 4 |
176 ± 8 |
36 ± 8 |
Untreated |
|
12 ± 2 |
6 ± 3 |
23 ± 3 |
207 ± 5 |
41 ± 3 |
|
2-Methylbenzyl chloride
|
1 μg |
|
|
|
174 ± 9 |
|
|
3 μg |
8 ± 2 |
9 ± 3 |
24 ± 7 |
175 ± 10 |
29 ± 3 |
||
10μg |
12 ± 4 |
9 ± 2 |
26 ± 7 |
198 ± 15 |
30 ± 9 |
||
33μg |
10 ± 1 |
8 ± 3 |
29 ± 5 |
203 ± 10 |
36 ± 9 |
||
100μg |
13 ± 1 |
10 ± 1 |
26 ± 6 |
213 ± 14 |
40 ± 11 |
||
333μg |
12 ± 2 |
10 ± 1 |
19 ± 6 |
109 ± 14 |
25 ± 4 |
||
1000μg |
6 ± 1 |
6 ± 1 |
23 ± 3 |
77 ± 26 |
24 ± 8 |
||
2500μg |
8 ± 3P |
1 ± 1P |
16 ± 3P |
2 ± 2P |
35 ± 8P |
||
5000μg |
6 ± 1P |
0 ± 0P |
0 ± 1P |
|
1 ± 1P |
||
NaN3 |
10 μg |
1220 ± 4 |
|
|
1886 ± 47 |
|
|
4-NOPD |
10 μg |
|
|
470 ± 28 |
|
|
|
4-NOPD |
50 μg |
|
84 ± 11 |
|
|
|
|
MMS |
2.0 μL |
|
|
|
|
346 ± 45 |
|
|
|
|
|
|
|
|
|
With Activation
|
DMSO |
|
14 ± 5 |
14 ± 2 |
34 ± 5 |
152 ± 10 |
39 ± 3 |
Untreated |
|
13 ± 4 |
13 ± 3 |
41 ± 3 |
214 ± 5 |
48 ± 7 |
|
2-Methylbenzyl chloride
|
1 μg |
|
|
|
124 ± 23 |
|
|
3 μg |
10 ± 2 |
14 ± 2 |
31 ± 4 |
150 ± 12 |
45 ± 15 |
||
10μg |
12 ± 2 |
14 ± 3 |
38 ± 2 |
122 ± 10 |
45 ± 2 |
||
33μg |
14 ± 4 |
14 ± 4 |
35 ± 9 |
139 ± 19 |
51 ± 3 |
||
100μg |
15 ± 5 |
12 ± 3 |
35 ± 9 |
173 ± 26 |
62 ± 11 |
||
333μg |
11 ± 4 |
18 ± 5 |
39 ± 8 |
72 ± 20 |
52 ± 5 |
||
1000μg |
7 ± 2 |
15 ± 5 |
33 ± 6 |
55 ± 15 |
31 ± 4 |
||
2500μg |
1 ± 1P |
11 ± 5P |
6 ± 1P |
2 ± 1P |
2 ± 1P |
||
5000μg |
0 ± 0P |
1 ± 1P |
0 ± 1P |
|
1 ± 1P |
||
2-AA |
2.5 μg |
341 ± 20 |
209 ± 13 |
2915 ± 654 |
2773 ± 347 |
|
|
2-AA |
10.0 μg |
|
|
|
|
404 ± 13 |
Key to Positive Controls |
Key to Plate Postfix Codes |
||
NaN3 2-AA 4-NOPD MMS |
sodium azide 2-aminoanthracene 4-nitro-o-phenylene-diamine methyl methane sulfonate |
P |
Precipitate |
Applicant's summary and conclusion
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.
