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EC number: 701-246-8 | CAS number: -
- 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 cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study performed under GLP
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 995
- Report date:
- 1995
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
Test material
- Reference substance name:
- Nu-Film-17
- IUPAC Name:
- Nu-Film-17
- Reference substance name:
- Oligomerisation products of beta-pinene
- EC Number:
- 701-246-8
- Molecular formula:
- Variable (dimer = C20-H34)
- IUPAC Name:
- Oligomerisation products of beta-pinene
- Details on test material:
- - Batch number: 154293
- Pinene oligomers content 96 wt%
Constituent 1
Constituent 2
Method
- Target gene:
- Endpoint investigated = chromosome damage
Species / strain
- Species / strain / cell type:
- lymphocytes: Human
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254 induced rat liver S9 mix
- Test concentrations with justification for top dose:
- First test concentrations: 2.0, 3.9, 7.8, 15.6, 31.3, 62.5, 125, 250, 500 and 1000 ug/ml.
Second test concentrations:
- Without S9 mix, 18 hour harvest: 62.5, 125, 250, 375, 500, 750 and 1000 ug/ml
- With S9 mix, 18 hour harvest: 125, 250, 500 and 1000 ug/ml
- Without S9 mix, 32 hour harvest: 15.6, 31.3, 62.5, 125, 250, 375, 500, 750 and 1000 ug/ml
- With S9 mix, 32 hour harvest: 125, 250, 500 and 1000 ug/ml - Vehicle / solvent:
- Nu-Film-17 (Pinolene) was immiscible in DMSO and so was prepared directly in the tissue culture medium.
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- no
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: ethyl methanesulphonate, cyclophosphamide
- Details on test system and experimental conditions:
- First test: Human lymphocyte cultures (0.9E06 cells/ml) that had been incubated for 48 hours were centrifuged at 200g. The cell pellets were resuspended in appropriate dosing solutions (5ml) to give the desired final concentrations of Nu-Film-17 (Pinolene) (2.0, 3.9, 7.8, 15.6, 31.3, 62.5, 125, 250. 500 and 1000 ug/ml). Duplicate cultures in the absence of S9 mix were prepared for each concentration of the test substance and four cultures were left untreated to serve as the negative control group. For the positive control group, ethyl methansulphonate was used at concentrations of 500 and 750 ul/ml in duplicate cultures. The cultures without S9 mix were incubated for 18 hours.
The test was also performed in duplicate cultures in the presence of S9 mix. For the positive control group with S9 mix, cyclophosphamide was added to duplicate cultures at concentrations of 10 and 15 ug/ml. Three hours after dosing cultures with the S9 mix, these were centrifuged and cell pellets resuspended in fresh medium and incubated for an additional 15 hours.
Second test: Human lymphocyte cultures (1E06 cells/ml) that had been incubated for 48 hours were centrifuged at 200g. The cell pellets were resuspended in appropriate dosing solutions (5ml) to give the desired final concentrations of Nu-Film-17 (Pinolene):
- without S9 mix, 18 hour harvest: 62.5, 125, 250, 375, 500, 750 and 1000 ug/ml
- with S9 mix, 18 hour harvest: 125, 250, 500 and 1000 ug/ml
- without S9 mix, 32 hour harvest: 15.6, 31.3, 62.5, 125, 250, 375, 500, 750 and 1000 ug/ml
- with S9 mix, 32 hour harvest: 125, 250, 500 and 1000 ug/ml
Duplicate cultures were prepared and used for each concentration of the test substance and four cultures were left untreated. For the 32 hour harvest ethyl methansulphonate was used at concentrations of 250 and 500 ul/ml in duplicate cultures (without S9 mix) and cyclophosphamide was used at concentrations of 2.5 and 5 ug/ml (with S9 mix). Three hours after dosing cultures with the S9 mix, these were centrifuged and cell pellets resuspended in fresh medium and incubated for an additional 15 or 29 hours. Cultures that were treated without S9 mix were incubated for 18 or 32 hours.
Harvesting and fixation: Colchicine was added to each culture to stop mitotic activity 2 hours before harvesting. Cell suspensions were centrifuged and cell pellets treated with a hypotonic solution (20% Hanks balanced salt solution). After treatment with the hypotonic solution, cell suspensions were again centrifuged and cell pellets fixed with freshly prepared fixative (3 parts methanol:1 part glacial acetic acid). These pellets were allowed to fixed for 2 hours upon which were centrifuged and resuspended in fresh fixative. These cell suspensions were placed onto microscope slides and allowed to air-dry and then stained with Giemsa and mounted in DPX.
Results and discussion
Test results
- Species / strain:
- lymphocytes: human
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Only at highest concentration - not used for metaphase analysis
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Toxicity:
- First test: The mitotic indices of Nu-Film-17 (Pinolene) treated human lymphocytes can be found in table 1. A reduction in mitotic index was observed at the 500 ug/ml dose group in the absence of S9 mix. The higher dose group (1000 ug/ml) was too toxic for analysis, thus the concentrations selected for metaphase analysis were 62.5, 250 and 500 ug/ml. In the presence of S9 mix, no significant reduction in mitotic index was observed when compared to the control value, thus the doses selected for metaphase analysis were 125, 500 and 1000 ug/ml.
- Second test, 18 hour harvest: The mitotic indices of Nu-Film-17 (Pinolene) treated human lymphocytes can be found in table 1. A reduction in mitotic index was observed at the 500 ug/ml dose group in the absence of S9 mix. The higher dose group (1000 ug/ml) was too toxic for analysis, thus the concentrations selected for metaphase analysis were 62.5, 250 and 500 ug/ml. In the presence of S9 mix, no significant reduction in mitotic index was observed when compared to the control value, thus the doses selected for metaphase analysis were 125, 500 and 1000 ug/ml.
- Second test, 32 hour harvest: The mitotic indices of Nu-Film-17 (Pinolene) treated human lymphocytes can be found in table 1. A reduction in mitotic index was observed at the 250 and 375 ug/ml dose groups in the absence of S9 mix, thus the concentrations selected for metaphase analysis were 31.3, 125 and 250 ug/ml. In the presence of S9 mix, a slight reduction in mitotic index was observed in the 1000 ug/ml dose group when compared to the control value. The doses selected for metaphase analysis were 125, 500 and 1000 ug/ml.
Metaphase analysis:
- First test: See table 2 for metaphasis analysis data. No statistically significant increases of chromosome aberrations were observed at any concentration either in the presence or absence of S9 mix. The positive control groups (with ethyl methanesulphonate or cyclophosphamide) caused statistically significant increases of aberrant cells.
- Second test, 18 hour harvest: See table 2 for metaphasis analysis data. No statistically significant increases of chromosome aberrations were observed at any concentration either in the presence or absence of S9 mix. The positive control groups (with ethyl methanesulphonate or cyclophosphamide) caused statistically significant increases of aberrant cells.
- Second test, 32 hour harvest: See table 2 for metaphasis analysis data. There were no statistically significant increases in aberrant cells in any dose group in the presence of S9 mix. However, in the absence of S9 mix there was a statistically significant increase in chromosomal damage in the 250 ug/ml dose group, but this was associated with high cytotoxicity and was not considered to be of biological significance. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
Table 1. Mitotic indices
Test substance concentration (ug/ml) | S9 mix | Mean mitotic index (%) | Relative mitotic index (%) | |
First test, 18 hour harvest | Untreated | - | 3.6 | 100 |
2 | - | 4.0 | 111 | |
3.9 | - | 4.5 | 125 | |
7.8 | - | 4.0 | 111 | |
15.6 | - | 4.3 | 119 | |
31.3 | - | 4.3 | 119 | |
62.5 | - | 3.3 | 92 | |
125 | - | 4.4 | 122 | |
250 | - | 4.3 | 119 | |
500 | - | 1.8 | 50 | |
1000 | - | 0.3 | 8 | |
Untreated | + | 3.8 | 100 | |
2 | + | 3.6 | 95 | |
3.9 | + | 3.0 | 79 | |
7.8 | + | 3.0 | 79 | |
15.6 | + | 4.5 | 118 | |
31.3 | + | 4.0 | 105 | |
62.5 | + | 4.2 | 111 | |
125 | + | 4.7 | 124 | |
250 | + | 4.0 | 105 | |
500 | + | 3.7 | 97 | |
1000 | + | 4.9 | 129 | |
Second test, 18 hour harvest | Untreated | - | 4.0 | 100 |
62.5 | - | 3.3 | 83 | |
125 | - | 2.9 | 73 | |
250 | - | 2.7 | 68 | |
375 | - | 2.0 | 50 | |
500 | - | 1.8 | 45 | |
750 | - | 0.3 | 8 | |
1000 | - | 0.0 | 0 | |
Untreated | + | 2.6 | 100 | |
125 | + | 2.5 | 96 | |
250 | + | 2.2 | 85 | |
500 | + | 2.3 | 88 | |
1000 | + | 2.2 | 85 | |
Second test, 32 hour harvest | Untreated | - | 4.8 | 100 |
15.6 | - | 4.2 | 88 | |
31.3 | - | 3.0 | 63 | |
62.5 | - | 3.3 | 69 | |
125 | - | 2.8 | 58 | |
250 | - | 1.6 | 33 | |
375 | - | 1.6 | 33 | |
500 | - | 0.5 | 10 | |
750 | - | 0.1 | 2 | |
1000 | - | 0.0 | 0 | |
Untreated | + | 3.6 | 100 | |
125 | + | 3.9 | 108 | |
250 | + | 3.1 | 86 | |
500 | + | 3.5 | 97 | |
1000 | + | 2.2 | 61 |
+ = Presence of S9 mix
- = Absence of S9 mix
Table 2. Metaphase analysis
Test | Test material | Concentration | S9 mix | Number of aberrant cells | |
Mean % (Excluding gaps) | Mean % (Including gaps) | ||||
First test | Untreated | - | - | 1.25 | 1.25 |
Nu-Film-17 (Pinolene) | 62.5 | - | 1.00 | 1.00 | |
250 | - | 1.00 | 1.00 | ||
500 | - | 2.50 | 2.50 | ||
Ethyl methanesulphonate | 500 | - | 19.00* | 19.5* | |
Untreated | - | + | 1.25 | 1.25 | |
Nu-Film-17 (Pinolene) | 125 | + | 3.50 | 3.50 | |
500 | + | 3.00 | 3.50 | ||
1000 | + | 3.00 | 3.50 | ||
Cyclophosphamide | 15 | + | 16.5* | 16.5* | |
Second test, 18 hour harvest | Untreated | - | 2.50 | 2.50 | |
Nu-Film-17 (Pinolene) | 32.5 | - | 1.50 | 1.50 | |
250 | - | 1.00 | 1.00 | ||
500 | - | 4.50 | 4.50 | ||
Ethyl methanesulphonate | 500 | - | 18.5* | 18.5* | |
Untreated | - | + | 0.50 | 0.50 | |
Nu-Film-17 (Pinolene) | 125 | + | 0.00 | 0.00 | |
500 | + | 0.00 | 0.50 | ||
1000 | + | 0.50 | 1.00 | ||
Cyclophosphamide | 10 | + | 13.0* | 13.0* | |
Second test, 32 hour harvest | Untreated | - | - | 1.50 | 1.50 |
Nu-Film-17 (Pinolene) | 31.3 | - | 0.50 | 0.50 | |
125 | - | 4.50 | 4.50 | ||
250 | - | 5.5* | 5.5* | ||
Ethyl methanesulphonate | 500 | - | 32.5* | 32.5* | |
Untreated | + | 1.50 | 1.50 | ||
Nu-Film-17 (Pinolene) | 125 | + | 2.00 | 2.00 | |
500 | + | 1.00 | 1.00 | ||
1000 | + | 1.50 | 1.50 | ||
Cyclophosphamide | 5 | + | 24.7* | 25.2* |
* Statistically significant
+ = Presence of S9 mix
- = Absence of S9 mix
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
Under the conditions of this study, no evidence of clastogenic activity was detected.
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