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EC number: 210-258-8 | CAS number: 611-19-8
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- Ecotoxicological Summary
- Aquatic toxicity
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- Short-term toxicity to fish
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- Long-term toxicity to aquatic invertebrates
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Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- September 1983
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 983
- Report date:
- 1983
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- no follow-up experiments were done for confirmation of negative results
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- α,2-dichlorotoluene
- EC Number:
- 210-258-8
- EC Name:
- α,2-dichlorotoluene
- Cas Number:
- 611-19-8
- Molecular formula:
- C7H6Cl2
- IUPAC Name:
- 1-chloro-2-(chloromethyl)benzene
- Details on test material:
- - Source: HOECHST AG
Constituent 1
Method
- Target gene:
- In the Salmonella typhimurium strains (TA 1535, TA 100, TA 1537, TA 1538, TA 98) the amino acid histidine locus is the target gene.
In the Escherichia coli strain (WP2 uvrA) the amino acid tryptophan locus is the target gene.
Species / strain
- Species / strain / cell type:
- other: Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA1538, Escherichia coli WP2 uvrA
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat liver S-9 mix from Arochlor 1254 induced rats
- Test concentrations with justification for top dose:
- 0.8, 4, 20, 100, 500, 1,500 µg/plate
- Vehicle / solvent:
- - Vehicle/solvent used: DMSO
- Justification for choice of solvent/vehicle: solubility of test item
Controlsopen allclose all
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- yes
- Remarks:
- 2-aminoanthracene (-S9: 0.5 µg/plate; TA98, TA100, TA1538)
- Positive controls:
- yes
- Remarks:
- TA98, TA100, TA1538
- Positive control substance:
- other: -S9: methylhydrazone derivative (5 µg/plate), +S9: 2-aminoanthracene (0.5 µg/plate)
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- yes
- Remarks:
- 2-aminoanthracene (-S9: 1 µg/plate (TA1535, TA 1537), 10 µg/plate (WP2 uvr A))
- Positive controls:
- yes
- Remarks:
- TA 1557, TA 1535, WP2 uvr A
- Positive control substance:
- other: -S9: streptocotocine (5 µg/plate (TA 1535)), 9-aminoacridine (100 µg/plate (TA 1537)), N-ethyl-N-nitro-N-nitrosoguanidine (2 µg/plate, WP2 uvr A); +S9: 2-amioanthracene (1 µg/plate (TA 1535, TA1537), 10 µg/plate (WP2 uvr A))
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: Overlay (soft) agar (plate incorporation method)
DURATION
- Preincubation period: No preincubation was performed. After mixing of all components, soft agar was distributed directly on the plates
- Exposure duration/ incubation time : Plates with microorganisms / test item situated in the Topagar and Controls were incubated upside down for 48 hours at 37 °C.
SELECTION AGENT (mutation assays): The selection is done by using a minimal agar that contains no histidine for the Salmonella typhimurium strains and a minimal agar that contains no tryptophane for the Escherichia coli strains.
NUMBER OF REPLICATIONS: 4 - Evaluation criteria:
- The test substance is considered positive in this assay if the following criteria is met:
- Doubling of the revertant colonies compared to controls
Results and discussion
Test results
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: Without metabolic activation: >= 500 µg/plate, with metabolic activation: >= 1500 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- see below
- Remarks on result:
- other: other: Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA1538, Escherichia coli WP2 uvrA
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
The number of revertant colonies did not increase compare
with the solvent control in S. typhimurium , and E. coli WP2 uvrA,
both with and without metabolic activation.
Table 1
========================================================
With Test Mean number of
or substance revertant colonies/plate
without dose --------------------------------
S9mix (ug/plate) Base-pair Frameshift
substitution type
type
--------------------------------
TA TA WP2 TA TA TA
100 1535 urvA 98 1537 1538
---------------------------------------------------------
0 154 12 30 25 12 10
0.8 169 15 26 22 12 12
without 4 159 13 26 22 14 11
(-) 20 152 12 29 19 13 8
100 186 13 28 14 0.5 9
500 * * 13 * * *
1,500 ** ** * ** ** **
---------------------------------------------------------
0 163 16 49 30 15 19
0.8 158 11 52 29 15 16
with 4 151 15 44 32 15 19
(+) 20 175 14 44 32 14 16
100 188 11 44 23 13 15
500 123 6 27 18 6 12
1,500 * ** 19 7 * *
---------------------------------------------------------
[Positive control without S9mix]
Chemical MD SC ENNG MD 9AA MD
Dose(ug/plate) 5 5 2 5 100 5
Mean number of
Colonies/plate 3405 >5000 633 3170 >5000 3035
---------------------------------------------------------
[Positive control with S9mix]
Chemical 2AA 2AA 2AA 2AA 2AA 2AA
Dose(ug/plate) 0.5 1 10 0.5 1 0.5
Mean number of
Colonies/plate 715 156 2330 750 136 740
========================================================
MD: Methylhydrazone Derivative, SC: Streptocotocine,
ENNG: N-Ethyl-N-nitro-N-nitrosoguanidine,
9AA: 9-Aminoacridine, 2AA: 2-Aminoanthracene
*: no colony growth, **: no bacterial growth
These results have led to the conclusion that OCBC was not
mutagenic under the conditions of this study.
Applicant's summary and conclusion
- Executive summary:
In a reverse gene mutation assay in bacteria Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, TA1538 and Escherichia coli strain WP2 uvrA were exposed to o-Chlorobenzylchloride at concentrations of 0, 0.8, 4, 20, 100, 500, 1,500 µg/plate in the presence and absence of mammalian metabolic activation.No preincubation was performed. There was no evidence of induced mutant colonies over background up to a concentration of 1.5 mg o-Chlorobenzylchloride/plate. The positive controls induced the appropriate responses in the corresponding strains. In conclusion, o-Chlorobenzylchloride did not show any mutagenic activity in any tester strains regardless of metabolic activation.
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