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EC number: 235-697-2 | CAS number: 12542-30-2
- Life Cycle description
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- Endpoint summary
- Appearance / physical state / colour
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- Density
- Particle size distribution (Granulometry)
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- Additional physico-chemical information
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- Endpoint summary
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- 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
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- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
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- Acute Toxicity
- Irritation / corrosion
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- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 12 Oct - 5 Dec 1994
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: OECD guideline study conducted in compliance with GLP regulations
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- testing lab.
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- HPRT locus
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- rat liver S-9 fraction
- Test concentrations with justification for top dose:
- 0; 0.0001; 0.0005; 0.001; 0.005; 0.01 mg/mL in experiment 1 without metabolic activation and 0; 0.005; 0.01; 0.05; 0.1; 0.5 mg/mL in experiment 1 with metabolic activation.
0; 0.0005; 0.001; 0.005; 0.0075; 0.01 mg/mL in experiment 2 without metabolic activation and 0; 0.0005; 0.005; 0.05; 0.1; 0.5 mg/mL in experiment 2 with metabolic activation. - Vehicle / solvent:
- DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: without S-9 mix: Ethylmethanesulfonate; with S-9 mix: 3-Methylcholanthrene
- Details on test system and experimental conditions:
DURATION
- Attachment period: 20-24 h
- Exposure duration: 4 h
- Expression time (cells in growth medium): 1 week
- Selection time (if incubation with a selection agent): 1 week
- Fixation time (start of exposure up to fixation or harvest of cells): 2 weeks
SELECTION AGENT (mutation assays): 6-Thioguanine
DETERMINATION OF CYTOTOXICITY
- Method:
cloning efficiency (Cytotoxicity 1): 17 - 24 hours after termination of the exposure period approx. 200 cells of each replicate (pooled from 2 flasks) were taken in duplicates, seeded into Ham's F12 medium (25 cm2 flasks) and allowed for colony formation (incubator; 1 week). After the end of the incubation period the colonies were fixed, stained and counted.
cloning efficiency (Cytotoxicity 2):
-Same procedure as "Cytotoxicity 1".
-Timepoint: parallel to selection (7 days after exposure)
Other: examination of precipitation- Evaluation criteria:
- The criteria for a positive response are:
- Increases of the corrected mutation frequencies above the concurrent negative control values and above 15 mutants per 10E +06 clonable cells and/or the evidence of a dose-response relationship in the increase in mutant frequencies.
- Evidence of reproducibility of any increase in mutant frequencies. - Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: Reduced cell densities/cloning efficiencies were found at concentrations > 0.005 mg/mL in the experiments without metabolic activation. In the presence of S-9 mix cytotoxicity was observed at 0.5 mg/mL.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of test substance after addition to test medium was observed at concentrations >=0.1 mg/mL
RANGE-FINDING/SCREENING STUDIES:
-Treatment with Dihydrodicyclopentadienylacrylat at concentrations ranging from 0.005 to 5 mg/mL (with and without S-9 mix). Cytotoxicity was observed at concentrations >= 0.005 mg/mL in the absence of S9-mix, and at concentrations >=0.5 mg/mL in the presence of metabolic activation. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
Reference
Table 1: Mutant frequencies-experiment 1 without metabolic activation.
Concentration (mg/ml) |
No. of coloniesa |
Mean mutant frequency (per 106cells) |
|||||||
Not corrected |
Correctedb |
||||||||
0 |
A |
0 |
0 |
0 |
0 |
0 |
0 |
2.22 |
2.73 |
|
B |
0 |
1 |
1 |
2 |
2 |
2 |
||
1% DMSO |
A |
0 |
0 |
0 |
0 |
1 |
1 |
1.95 |
2.15 |
|
B |
0 |
0 |
1 |
1 |
1 |
2 |
||
0.0001 |
A |
0 |
0 |
0 |
0 |
0 |
0 |
0.56 |
0.68 |
|
B |
0 |
0 |
0 |
0 |
1 |
1 |
||
0.0005 |
A |
0 |
0 |
0 |
0 |
0 |
0 |
0.56 |
0.78 |
|
B |
0 |
0 |
0 |
0 |
1 |
1 |
||
0.001 |
A |
0 |
1 |
2 |
3 |
3 |
3 |
5.84 |
7.13 |
|
B |
0 |
1 |
1 |
2 |
2 |
3 |
||
0.005 |
A |
0 |
0 |
0 |
0 |
0 |
0 |
3.06 |
4.01 |
|
B |
1 |
1 |
1 |
2 |
2 |
4 |
||
0.01 |
A |
0 |
0 |
0 |
0 |
0 |
0 |
0.00 |
0.00 |
|
B |
0 |
0 |
0 |
0 |
0 |
0 |
||
EMS 0.3 |
A |
22 |
27 |
27 |
29 |
30 |
40 |
98.06 |
116.56 |
|
B |
20 |
25 |
29 |
30 |
32 |
42 |
||
|
|
|
|
|
|
|
|
|
|
a: No of colonies 7 days after seeding, about 300000 cells/flask into selection medium (6 flasks/replicate) |
|||||||||
b: corrected on the basis of the absolute cloning efficiency at the end of the expression period (cytotoxicity 2). |
Table 2: Mutant frequencies-experiment 1 with metabolic activation.
Concentration (mg/ml) |
No. of coloniesa |
Mean mutant frequency (per 106cells) |
|||||||
Not corrected |
Correctedb |
||||||||
0 |
A |
0 |
0 |
1 |
1 |
2 |
2 |
6.39 |
8.23 |
|
B |
1 |
3 |
3 |
3 |
3 |
4 |
||
1% DMSO |
A |
0 |
0 |
0 |
0 |
1 |
2 |
0.84 |
1.12 |
|
B |
0 |
0 |
0 |
0 |
0 |
0 |
||
0.005 |
A |
0 |
0 |
0 |
0 |
0 |
0 |
0.00 |
0.00 |
|
B |
0 |
0 |
0 |
0 |
0 |
0 |
||
0.01 |
A |
0 |
1 |
1 |
2 |
2 |
2 |
3.33 |
3.95 |
|
B |
0 |
0 |
1 |
1 |
1 |
1 |
||
0.05 |
A |
0 |
0 |
0 |
0 |
0 |
1 |
0.56 |
0.69 |
|
B |
0 |
0 |
0 |
0 |
0 |
1 |
||
0.1 |
A |
0 |
0 |
1 |
1 |
2 |
3 |
3.61 |
4.69 |
|
B |
0 |
0 |
1 |
1 |
2 |
2 |
||
0.5 |
A |
0 |
0 |
0 |
0 |
0 |
0 |
3.34 |
4.51 |
|
B |
1 |
1 |
2 |
2 |
2 |
4 |
||
MCA 0.01 |
A |
37 |
40 |
45 |
49 |
50 |
51 |
129.17 |
188.84 |
|
B |
27 |
29 |
30 |
33 |
34 |
40 |
||
|
|
|
|
|
|
|
|
|
|
a: No of colonies 7 days after seeding, about 300000 cells/flask into selection medium (6 flasks/replicate) |
|||||||||
b: corrected on the basis of the absolute cloning efficiency at the end of the expression period (cytotoxicity 2). |
Table 3: Mutant frequencies-experiment 2 without metabolic activation.
Concentration (mg/ml) |
No. of coloniesa |
Mean mutant frequency (per 106cells) |
|||||||
Not corrected |
Correctedb |
||||||||
0 |
A |
0 |
0 |
0 |
1 |
1 |
1 |
2.50 |
3.64 |
|
B |
0 |
0 |
0 |
1 |
2 |
3 |
||
1% DMSO |
A |
0 |
0 |
0 |
0 |
0 |
0 |
1.67 |
2.30 |
|
B |
0 |
0 |
1 |
1 |
1 |
3 |
||
0.0005 |
A |
0 |
0 |
0 |
0 |
0 |
0 |
0.00 |
0.00 |
|
B |
0 |
0 |
0 |
0 |
0 |
0 |
||
0.001 |
A |
0 |
0 |
0 |
0 |
0 |
1 |
3.06 |
4.72 |
|
B |
1 |
1 |
2 |
2 |
2 |
2 |
||
0.005 |
A |
0 |
0 |
0 |
0 |
1 |
1 |
1.67 |
2.5 |
|
B |
0 |
0 |
0 |
1 |
1 |
2 |
||
0.0075 |
A |
0 |
0 |
0 |
0 |
0 |
0 |
2.78 |
4.26 |
|
B |
0 |
1 |
1 |
2 |
2 |
4 |
||
0.01 |
A |
0 |
0 |
0 |
c |
c |
c |
0.00 |
0.00 |
|
B |
0 |
0 |
c |
c |
c |
C |
||
EMS 0.3 |
A |
34 |
37 |
41 |
43 |
50 |
50 |
149.45 |
262.33 |
|
B |
26 |
41 |
44 |
50 |
58 |
64 |
||
|
|
|
|
|
|
|
|
|
|
a: No of colonies 7 days after seeding, about 300000 cells/flask into selection medium (6 flasks/replicate) |
|||||||||
b: corrected on the basis of the absolute cloning efficiency at the end of the expression period (cytotoxicity 2). |
|||||||||
c: not seeded because not enough cells (due to toxicity) |
Table 4: Mutant frequencies-experiment 2 with metabolic activation.
Concentration (mg/ml) |
No. of coloniesa |
Mean mutant frequency (per 106cells) |
|||||||
Not corrected |
Correctedb |
||||||||
0 |
A |
0 |
0 |
0 |
0 |
0 |
1 |
2.78 |
4.22 |
|
B |
0 |
1 |
1 |
1 |
2 |
4 |
||
1% DMSO |
A |
0 |
0 |
0 |
0 |
0 |
0 |
0.00 |
0.00 |
|
B |
0 |
0 |
0 |
0 |
0 |
0 |
||
0.0005 |
A |
1 |
2 |
2 |
3 |
3 |
5 |
5.56 |
9.25 |
|
B |
0 |
0 |
0 |
1 |
1 |
2 |
||
0.005 |
A |
0 |
0 |
0 |
0 |
0 |
1 |
1.12 |
1.74 |
|
B |
0 |
0 |
0 |
1 |
1 |
1 |
||
0.05 |
A |
0 |
0 |
0 |
0 |
0 |
1 |
1.12 |
1.70 |
|
B |
0 |
0 |
0 |
1 |
1 |
1 |
||
0.1 |
A |
1 |
1 |
1 |
3 |
3 |
5 |
5.28 |
8.01 |
|
B |
0 |
0 |
1 |
1 |
1 |
2 |
||
0.5 |
A |
0 |
c |
c |
c |
c |
c |
0.00 |
0.00 |
|
B |
0 |
c |
c |
c |
c |
c |
||
MCA 0.01 |
A |
2 |
5 |
6 |
7 |
9 |
13 |
27.78 |
43.53 |
|
B |
7 |
9 |
9 |
9 |
11 |
13 |
||
|
|
|
|
|
|
|
|
|
|
a: No of colonies 7 days after seeding, about 300000 cells/flask into selection medium (6 flasks/replicate) |
|||||||||
b: corrected on the basis of the absolute cloning efficiency at the end of the expression period (cytotoxicity 2). |
|||||||||
c: not seeded because not enough cells (due to toxicity) |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
In-vitro studies: Bacterial systems
One bacterial gene mutation assay according to OECD guideline 471 is available with dihydrodicyclopentadienyl acrylate (BASF SE, 1994). The assay was negative at doses up to 5000 μg/plate (standard plate test) and 120 µg/plate (preincubation test), respectively with and without S-9 mix in Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537. The selected Salmonella strains are not capable to detect oxidizing or cross-linking damage, thus leading to a limited conclusion. Since the experiment was negative in the strains tested, with and without metabolic activation, there is no indication for a mutagenic potential of the test substance in bacterial cells, not taking in account oxidizing or cross-linking damage as the underlying mechanism.
In-vitro studies: Mammalian cell gene mutation test
One mammalian cell gene mutation test (HPRT test, according to OECD TG No. 476) is available for dihydrodicyclopentadienyl acrylate (BASF AG, 1994).
Doses of up to 0.01 mg/mL (without S-9 mix) or 0.5 mg/mL (with S-9 mix from Aroclor-induced rat livers) were tested in a 4-hour treatment. Reduced cell densities/cloning efficiencies were found at concentrations >0.005 mg/mL in the experiments without metabolic activation. In the presence of S-9 mix cytotoxicity was observed at 0.5 mg/mL. No significant increase of mutant frequencies nor a dose-response relationship were observed, indicating that the substance has no mutagenic potential in mammalian CHO cells.
In-vitro studies: chromosomal aberration
For analysis of chromosomal aberrations, V79 hamster lung fibroblasts were treated with 1.25 -15 µg/mL (without S9-mix) and 25 – 200 µg/mL (with S9-Mix), respectively. Cells were treated for 18 and 28 h (without S9-Mix), and were sampled afterwards. Treatment with S9-mix was 4 h with subsequent sampling 18 and 28 h after beginning of treatment. 1000 cells were examined for mitotic index and 100 mitoses per dose were analyzed for chromosomal damage. No increase in aberration frequency was observed in treated cultures, thus there is no evidence for clastogenicity of dihydrodicyclopentadienyl acrylate.
Justification for selection of genetic toxicity endpoint
The key study was selected
Justification for classification or non-classification
Dangerous Substance Directive (67/548/EEC)
No classification required for genetic toxicity.
Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008 (last amended by EC/286/2008 (2011-03-10)):
No classification required for genetic toxicity.
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