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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 vivo
Administrative data
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- 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:
- 1 996
- Report date:
- 1996
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)
- Principles of method if other than guideline:
- Mouse bone marrow micronucleus test
- GLP compliance:
- yes
- Type of assay:
- micronucleus assay
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 oilgomers content 96 wt%
Constituent 1
Constituent 2
Test animals
- Species:
- mouse
- Strain:
- CD-1
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Breeding Laboratories (UK), Margate, Kent, England.
- Age: 4 - 5 weeks old on arrival
- Weight: 14.9 - 21.6 g on arrival
- Assigned to test groups randomly: yes, using computer generated random numbers
- Fasting period before study:
- Housing: Animals were housed in polypropylene cages with stainless steel tops.
- Diet: Laboratory animal diet RM1 (E)SQC (Special Diet Services Ltd., Witham, Essex, England) ad libitum
- Water: Drinkning water from the public supply, ad libitum
- Acclimation period: At least 4 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 23
- Humidity (%): 45 - 60
- Air changes (per hr): 15
- Photoperiod: 12 hour light / 12 hour dark
Administration / exposure
- Route of administration:
- intraperitoneal
- Vehicle:
- - Vehicle: sterile water
- Details on exposure:
- The test material formed a doseable emulsion when mixed with sterile water (purified by reverse osmosis) at a maximum concentration of 200 mg/ml. Dosing formulations were prepared with sterile water on the day of dosing. The test material or dosing vehicle (sterile water) was administered once in a dose volume of 10 ml/kg via intraperitoneal injection to groups of male and female mice.
- Duration of treatment / exposure:
- Preliminary study: 48 hours
Main study: 24 and 48 hours - Frequency of treatment:
- Single dose (both preliminary and main study)
- Post exposure period:
- 24 hours after treatment, 5 test animals of each sex from each dose group were killed and smears were prepared from bone marrow cells of the femur. 48 hours after treatment the remaining 5 animals (per sex) from the 2000 mg/kg dose group and vehicle control group were killed and smears prepared from the bone marrow cells of the femur.
Doses / concentrationsopen allclose all
- Remarks:
- Doses / Concentrations:
Main study: 500, 1000 and 2000 mg/kg
Basis:
nominal in water
intraperitoneal injection
- Remarks:
- Doses / Concentrations:
Preliminary study: 500, 1000, 1500 and 2000 mg/kg
Basis:
nominal in water
intraperitoneal injection
- No. of animals per sex per dose:
- Preliminary test: 2 animals/sex/dose
Main test: For the 500 and 1000 mg/kg test substance dose groups used 5 animals of each sex. For the 2000 mg/kg test substance dose group and vehicle control group, 10 animals of each sex were used. - Control animals:
- yes
- Positive control(s):
- Chlorambucil (reference substance) was administered by intraperitoneal injection at a dosage of 30 mg/kg in aqueous 10% ethanol.
Examinations
- Tissues and cell types examined:
- Smears were prepared from the bone marrow cells of the femur.
- Details of tissue and slide preparation:
- Following carbon dioxide inhalation, animals were killed by cervical dislocation. The femurs of each animal were dissected out and cleaned of adherent tissue. The epiphyses were cut off to gain access to the marrow canal. Marrow cells were washed out with 2.5 ml foetal calf serum using a syringe and needle. These cells were centrifuged and the majority of the supernantant fluid was discarded, the remaining fluid was used to resuspend the cell pellet. Single drops of the cell suspension were placed onto clean dry slides, three smears (two smears for the preliminary study) were prepared and the slides left to dry. Cells were then fixed with methanol and stained using 5% Giemsa stain. After staining, slide were washed with buffer, air-dried and cleared in xylene, coverslips were applied using a DPX mountant.
- Evaluation criteria:
- The test substance can be considered to have caused clastogenic or other damage leading to micronucleus production if the following criteria are met:
- A statistically significant increase in micronucleated polychromatic erythrocyte are observed at least one dose levels in either or both sex.
- the increases are reproducible in animals of the same sex and same treatment group(s)
- the increases exceed the lab's historical range of the control group
- Evidence of a dose-response relationship increases in both sexes or increases at more than one sacrifice time will be considered to support the conclusion.
Results and discussion
Test results
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Remarks:
- weight loss seen in all mice at 1000 and 2000 mg/kg was of uncertain significance
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Preliminary study: All animals survived to the scheduled termination. Treatment related effect were observed at all test concentrations and included: underactivity (14 mice), piloerection (4 mice) and flat posture (1 mouse). One male of the 2000 mg/kg dose group was unclean at the base of the tail (observed from day 2 until scheduled termination).
Weight loss was apparent in 16 mice 24 hours after treatment and in 14 mice after 48 hours.
The polychromatic:mature erythrocyte ratios are given in table 1 (below). Bone marrow toxicity was suggested by a reduction in the ratio of polychromatic to mature erythrocytes to 0.7 and 0.6 in mice of the 1500 and 2000 mg/kg dose groups respectively. In view of the small sample of animals used in this test and absence of concurrent control, this data should be treated with caution. From this data the highest dose selected for the main test was 2000 mg/kg.
Main test: although changes in bodyweight were observed in the majority of test animals in this study, these were not considered to be significant changes due to the short timespan between weighings. In the positive control group (chlorambucil) weight loss was observed in all mice during the 24 hour period between dosing and sacrifice. Clinical signs were observed at all concentrations tested, these included: underactivity (29 mice), piloerection (8 mice) and hunched postures (5 mice). Mice of the 2000 mg/kg dose group showed signs that included flat posture (2 mice) and unclean fur at the base of the tail (6 mice).
No statistically significant differences in the frequencies of micronucleated polychromatic cells were observed between sexes in any dose group (see table 2). The positive control group (chlorambucil) produced a statistically significant increase in the frequency of micronucleated polychromatic erythrocytes and this increase confirms the sensitivity of the test system.
The polychromatic to mature erythrocyte ratio for each dose group was similar to corresponding vehicle control group values at each sacrifice time.
Any other information on results incl. tables
Table 1. Preliminary study: polychromatic: mature erythrocyte ratios
Group | Treatment (mg/kg) | Animal number | Polychromatic cells (P) | P:M ratio | |||
Mature cells (M) | By animal | By sex | By group | ||||
1 | 500 | 1M | 1184 | 100 | 1.2 | 1 | 0.9 |
2M | 1001 | 1110 | 0.9 | ||||
9F | 1013 | 1184 | 0.9 | 0.8 | |||
10F | 1042 | 1249 | 0.8 | ||||
2 | 1000 | 3M | 1023 | 1092 | 0.9 | 1 | 0.9 |
4M | 1017 | 1050 | 1 | ||||
11F | 1006 | 1066 | 0.9 | 0.8 | |||
12F | 1030 | 1365 | 0.8 | ||||
3 | 1500 | 5M | 1000 | 1838 | 0.5 | 0.5 | 0.7 |
6M | 954 | 2054 | 0.5 | ||||
13F | 1013 | 1090 | 0.9 | 0.8 | |||
14F | 1026 | 1419 | 0.7 | ||||
4 | 2000 | 7M | 1005 | 1702 | 0.6 | 0.6 | 0.6 |
8M | 1004 | 1827 | 0.5 | ||||
15F | 1001 | 2022 | 0.5 | 0.6 | |||
16F | 1019 | 1593 | 0.6 |
P:M ratio = Ratio of polychromatic to mature erythrocytes
Table 2. Main study. Polychromatic: mature erythrocyte ratio
Group | Treatment (mg/kg) | Frequency of micronucleated polychromatic erythrocytes | P:M ratio | |
Mean +/- SD | Range | |||
24 hour sacrifice time | ||||
1 | Sterile water | 0.6 +/-0.8 | 0.0 - 1.8 | 1 |
2 | Nu-Film-17 (500 mg/kg) | 0.9 +/- 0.7 | 0.0 - 2.0 | 0.9 |
3 | Nu-Film-17 (1000 mg/kg) | 0.6 +/- 0.6 | 0.0 - 1.5 | 0.9 |
4 | Nu-Film-17 (2000 mg/kg) | 0.9 +/- 1.0 | 0.0 - 2.8 | 0.8 |
5 | Chlorambucil (30 mg/kg) | 45.6 +/- 18.9* | 20.7 - 87.1 | 1 |
48 hour sacrifice time | ||||
1 | Sterile water | 1.0 +/- 0.8 | 0.0 - 2.0 | 1 |
4 | Nu-Film-17 (2000 mg/kg) | 0.4 +/- 0.6 | 0.0 - 1.8 | 0.9 |
* Highly significant (P<0.01)
P:M ratio: ratio of polychromatic to mature erythrocytes
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information): negative
Under the conditions of this study, no evidence of chromosomal or other damage leading to micronucleus formation in polychromatic erythrocytes was obtained following intraperitoneal administration at high dosage. - Executive summary:
In this mouse bone marrow micronucleus assay, 5 animals/sex/dose (the highest dose group and negative control group used 10 animal/sex) were treated intraperitoneally at doses of 0, 500, 1000 and 2000 mg/kg bw. Bone marrow cells were harvested at 24 h (for all dose groups) and 48 hours (high dose group only) post-treatment. The vehicle was sterile water.
There were signs of toxicity during the study which included underactivity (29 mice), piloerection (8 mice) and hunched postures (5 mice). Mice of the 2000 mg/kg dose group showed signs that included flat posture (2 mice) and unclean at the base of the tail (6 mice). The positive control induced the appropriate response. There was no significant increase in the frequency of micronucleated polychromatic erythrocytes in bone marrow in test group animals.
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