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EC number: 226-009-1 | CAS number: 5216-25-1
- 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:
- 1986-11-5 to 1986-11-13
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 986
- Report date:
- 1986
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- α,α,α,4-tetrachlorotoluene
- EC Number:
- 226-009-1
- EC Name:
- α,α,α,4-tetrachlorotoluene
- Cas Number:
- 5216-25-1
- Molecular formula:
- C7H4Cl4
- IUPAC Name:
- 1-chloro-4-(trichloromethyl)benzene
Constituent 1
Method
- Target gene:
- Histidine: Salmonella typhimurium strains
One of the genes for tryptophan biosynthesis: Escherichia coli strain
No more data
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- other: deficient in the complete structure of the lipopolysaccharide layer and in DNA excision repair system
- Species / strain / cell type:
- S. typhimurium TA 1538
- Additional strain / cell type characteristics:
- other: deficient in the complete structure of the lipopolysaccharide layer and in DNA excision repair system
- Species / strain / cell type:
- E. coli WP2 uvr A
- Additional strain / cell type characteristics:
- other: deficient in the uvrA system of DNA repair
- Metabolic activation:
- with and without
- Metabolic activation system:
- rat liver homogenate (S-9 mix)
- Test concentrations with justification for top dose:
- Dose range: 0.16 to 500 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol
No further data available
Controlsopen allclose all
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- executed for strain S. typhimurium TA 100 without metabolic activation
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- TA 100, TA 1535; without metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- TA 1537; without metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- Remarks:
- TA 98, TA 1538; without metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)
- Remarks:
- WP2uvrA; without metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- executed for strain S. typhimurium TA 100 with metabolic activation
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- TA 98, TA 100, TA 1535, TA 1537, TA 1538, WP2uvrA; with metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- executed for strain S. typhimurium TA 100 with metabolic activation
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- TA 98, TA 100, TA 1535, TA 1537, TA 1538, WP2uvrA; with metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 to 72h
NUMBER OF REPLICATIONS: 3
No further data available - Evaluation criteria:
- Amino-acid requiring strains of bacteria are used to detect reverse gene mutations. Based on the number of revertants one can assess the mutagenicity of the test compound.
- Statistics:
- No data
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Remarks:
- see table 1 and 2 in overall remarks sections
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- thinning of bacterial lawn and reduction in number of colonies at 20 or 100 µg/plate depending on strain
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- see table 1 and 2 in overall remarks sections
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- thinning of bacterial lawn and reduction in number of colonies at 100 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- see table 1 and 2 in overall remarks sections
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- thinning of bacterial lawn and reduction in number of colonies at 100 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: visible precipitation of the test compound on the plates at 2500 and 10000 µg/plate for all strains with and without metabolic activiation
RANGE-FINDING/SCREENING STUDIES:
An experiment was performed with all tester strains (3 replicates per dose) to obtain information on mutagenicity and toxicity for calculation of an appropriate dose range. As indicators for toxicity a reduced rate of spontaneously occuring colonies as well as visible thinning of the bacterial lawn (controlled microscopically) were used.
Based on this test 500 µg/plate was chosen as highest dose for mutagenicity testing.
No further data available - Remarks on result:
- other: all strains/cell types tested
Any other information on results incl. tables
Table 1: Result for the mutagenicity experiment with 4 -chloro-benzotrichloride without metabolic activation for all tested strains, including controls. The values presented are the mean value obtained from the three replicates. (ibl: incomplete bacterial lawn; nbl: no bacterial lawn)
S. typhimurium | S. typhimurium | S. typhimurium | S. typhimurium | S. typhimurium | E. coli | ||
TA-100 | TA-1535 | TA-1537 | TA-1538 | TA-98 | WP2uvrA | ||
Positive control* | |||||||
Test compound | 0 µg/plate | 168 | 19 | 8 | 17 | 26 | 46 |
0.16 µg/plate | 7 | 28 | |||||
0.8 µg/plate | 175 | 16 | 10 | 15 | 29 | 50 | |
4 µg/plate | 216 | 22 | 10 | 17 | 37 | 58 | |
20 µg/plate | 318 (ibl) | 20 | 17 | 16 | 67 | 63 | |
60 µg/plate | 113 (ibl) | 15 (ibl) | 22 (ibl) | 15 (ibl) | 36 (ibl) | 67 | |
100 µg/plate | 85 (nbl) | 2 (ibl) | 10 (ibl) | 7 (ibl) | 20 (ibl) | 22 (ibl) | |
500 µg/plate | nbl | nbl | nbl | nbl | nbl | ibl |
* | TA-100 | Sodium-azide | 1 µg/plate | 602 |
TA-1535 | Sodium-azide | 1 µg/plate | 510 | |
TA-1537 | 9-aminoacridine | 50 µg/plate | 242 | |
TA-1538 | 2-nitrofluorene | 2.5 µg/plate | 603 | |
TA-98 | 2-nitrofluorene | 2.5 µg/plate | 366 | |
WP2uvrA | MNNG | 2.5 µg/plate | 330 | |
p-chlorbenzotrichlorid TTR | 0.8 µg/plate | 0 |
Table 2: Result for the mutagenicity experiment with 4 -chloro-benzotrichloride with metabolic activation for all tested strains, including controls. The values presented are the mean value obtained from the three replicates. (ibl: incomplete bacterial lawn; nbl: no bacterial lawn)
Metabolic activation (rat liver) | S. typhimurium | S. typhimurium | S. typhimurium | S. typhimurium | S. typhimurium | E. coli | |
TA-100 | TA-1535 | TA-1537 | TA-1538 | TA-98 | WP2uvrA | ||
Positive control** | |||||||
Test compound | 0 µg/plate | 182 | 23 | 10 | 20 | 34 | 54 |
0.16 µg/plate | 6 | 43 |
|||||
0.8 µg/plate | 216 | 23 | 9 | 18 | 39 | 54 | |
4 µg/plate | 274 | 26 | 13 | 19 | 43 | 55 | |
20 µg/plate | 392 (ibl) | 23 | 24 | 23 | 75 | 65 | |
60 µg/plate | 173 (ibl) | 13 (ibl) | 14 (ibl) | 18 (ibl) | 66 (ibl) | 71 | |
100 µg/plate | 97 (ibl) | 13 (ibl) | 9 (ibl) | 11 (ibl) | 25 (ibl) | 34 (ibl) | |
500 µg/plate | nbl | nbl | nbl | nbl | 20 (nbl) | nbl |
** | TA-100 | 2-aminoanthracen | 0.5 µg/plate | 718 |
TA-100 | Benzo(a)pyrene | 10 µg/plate | 619 | |
TA-1535 | 2-aminoanthracen | 1 µg/plate | 160 | |
TA-1535 | Benzo(a)pyrene | 10 µg/plate | 25 | |
TA-1537 | 2-aminoanthracen | 1 µg/plate | 74 | |
TA-1537 | Benzo(a)pyrene | 10 µg/plate | 112 | |
TA-1538 | 2-aminoanthracen | 0.5 µg/plate | 470 | |
TA-1538 | Benzo(a)pyrene | 10 µg/plate | 162 | |
TA-98 | 2-aminoanthracen | 0.5 µg/plate | 444 | |
TA-98 | Benzo(a)pyrene | 10 µg/plate | 652 | |
WP2uvrA | 2-aminoanthracen | 10 µg/plate | 292 | |
WP2uvrA | Benzo(a)pyrene | 10 µg/plate | 88 | |
S-9 mix | 500 µl | 0 | ||
p-chlorbenzotrichlorid TTR | 0.8 µg/plate | 0 |
Table 3: Surviving fraction (i.e. ratio of number of colonies from solvent control plates and from plates with test compound) obtained using the solvent control with the Salmonella strain TA 100
Surviving fraction |
|||
Without metabolic activation | With metabolic activation | ||
Test compound | 0 µg/plate | 1.0 | 1.0 |
0.8 µg/plate | 1.0 |
1.0 |
|
4 µg/plate | 1.0 | 1.0 | |
20 µg/plate | 0.9 | 1.0 | |
60 µg/plate | 0.00 | 0.8 | |
100 µg/plate | 0.00 | 0.7 | |
500 µg/plate | 0.00 | 0.00 |
Applicant's summary and conclusion
- Conclusions:
- Mutagenic
- Executive summary:
The authors tested the mutagenicity of 4-chloro-benzotrichloride (CAS n° 5216 -25 -1) in the absence and presence of a metabolizing system with a methodology similar to the OECD guideline 471. The Salmonella typhimurium strains TA 100, TA 1535, TA 1537, TA 1538 and TA 98 and the Escherichia coli strain WP2uvrA were used and the metabolizing system was obtained from rat liver homogenates. The number of revertants per plate was estimated and was a basis for comparison with solvent controls.
Based on the range-finding/screening test, 500 µg/plate was chosen as top dose level for the mutagenicity test and the test compound proved to be toxic to most of the bacterial strains at 20 or 100 µg/plate according to observed diminuation or even complete absence of the bacterial lawn.
In the mutagenicity test, seven different doses ranging from 0.16 to 500 µg/plate were used (3 replicates per dose) and the strains were exposed for 48 to 72 h. Control plates without mutagen showed the described number of spontaneous revertant colonies reported in literature and all the positive control compounds gave the expected increase in the number of revertant colonies.
Furthermore the mutagenicity test showed a dose dependant increase in the number of revertant colonies with the S. typhimurium strain TA 98 in absence of the metabolic activation system (S-9 mix) after exposure to the test compound. In the presence of the metabolic activation system, treatment with the test substance resulted in a relevant increase in the number of revertant colonies in the S. typhimurium strain TA 98. Also slightly increased numbers of revertant colonies were obtained in the absence and presence of the metabolic activation system for S. typhimurium strains TA 100 and TA 1537. Considering the overall data, thus, 4 -chloro-benzotrichloride can be considered mutagenic with and without metabolic activation system for the tested strains.
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