Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 201-291-9 | CAS number: 80-56-8
- 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
![](https://echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- September 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Cross-referenceopen allclose all
- Reason / purpose for cross-reference:
- reference to same study
- Reason / purpose for cross-reference:
- reference to other study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
- Report date:
- 2017
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- (+)-pin-2(3)-ene
- EC Number:
- 232-087-8
- EC Name:
- (+)-pin-2(3)-ene
- Cas Number:
- 7785-70-8
- Molecular formula:
- C10H16
- IUPAC Name:
- (1R,5R)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene
- Reference substance name:
- (-)-pin-2(3)-ene
- EC Number:
- 232-077-3
- EC Name:
- (-)-pin-2(3)-ene
- Cas Number:
- 7785-26-4
- Molecular formula:
- C10H16
- IUPAC Name:
- (1S,5S)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene
- Reference substance name:
- (1R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
- EC Number:
- 227-336-2
- EC Name:
- (1R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
- Cas Number:
- 5794-03-6
- Molecular formula:
- C10H16
- IUPAC Name:
- (1R,4S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
- Reference substance name:
- (1S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
- EC Number:
- 227-337-8
- EC Name:
- (1S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
- Cas Number:
- 5794-04-7
- Molecular formula:
- C10H16
- IUPAC Name:
- (1S,4R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
- Reference substance name:
- (-)-pin-2(10)-ene
- EC Number:
- 242-060-2
- EC Name:
- (-)-pin-2(10)-ene
- Cas Number:
- 18172-67-3
- Molecular formula:
- C10H16
- IUPAC Name:
- (1S,5S)-6,6-dimethyl-2-methylenebicyclo[3.1.1]heptane
- Reference substance name:
- 1,7,7-trimethyltricyclo[2.2.1.02,6]heptane
- EC Number:
- 208-083-7
- EC Name:
- 1,7,7-trimethyltricyclo[2.2.1.02,6]heptane
- Cas Number:
- 508-32-7
- Molecular formula:
- C10H16
- IUPAC Name:
- 1,7,7-trimethyltricyclo[2.2.1.0~2,6~]heptane
- Reference substance name:
- 7,7-dimethyl-2-methylene-(1R,4S)-bicyclo[2.2.1]heptane
- Cas Number:
- 116724-26-6
- Molecular formula:
- C10H16
- IUPAC Name:
- 7,7-dimethyl-2-methylene-(1R,4S)-bicyclo[2.2.1]heptane
- Reference substance name:
- 7,7-dimethyl-2-methylene-, (1S,4R)-bicyclo[2.2.1]heptane
- Cas Number:
- 7378-37-2
- Molecular formula:
- C10H16
- IUPAC Name:
- 7,7-dimethyl-2-methylene-, (1S,4R)-bicyclo[2.2.1]heptane
- Reference substance name:
- (1R,5R)-6,6-dimethyl-2-methylenebicyclo[3.1.1]heptane
- Cas Number:
- 19902-08-0
- Molecular formula:
- C10H16
- IUPAC Name:
- (1R,5R)-6,6-dimethyl-2-methylenebicyclo[3.1.1]heptane
- Test material form:
- liquid
- Details on test material:
- Batch No. : FAB-29062016
Purity : 96.2% (sum of the two main constituents)
Name of test material (as cited in study report): alpha-pinene multiconstituent
Physical state: colourless liquid
Storage Conditions: +2°C to +8°C, under nitrogen and protected from light
Expiry Date: 28 June 2017
Constituent 1
Constituent 2
impurity 1
impurity 2
impurity 3
impurity 4
impurity 5
impurity 6
impurity 7
Method
- Target gene:
- His+ for S. typhimurium; trp+ for E. coli
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Metabolic activation system:
- 10 % (v/v) S9 mix; S9 fraction prepared from liver homogenates of rats induced with Phenobarbital sodium/5,6-benzoflavone.
- Test concentrations with justification for top dose:
- Experiment 1 (plate incorporation method): 5, 15, 50, 150, 500, 1500 and 5000 µg/plate with and without S9 mix in all strains
Experiment 2 (pre-incubation method): 0,05, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate with and without S9 mix in all strains - Vehicle / solvent:
- DMSO
Controlsopen allclose all
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- benzo(a)pyrene
- other: 2-Aminoanthracene
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- Details on test system and experimental conditions:
- SOURCE OF TEST SYSTEM: Strains of S. typhimurium and E. coli were obtained from
METHOD OF APPLICATION:
Experiment 1: In agar (direct plate incorporation)
Experiment 2: preincubation method
DURATION
- Preincubation period: Exp. 2, 30 minutes at 37 °C
- Incubation period: Approximately 48-72 h at 37 °C for both direct plate incorporation and preincubation methods
NUMBER OF REPLICATIONS:
-3 plates/dose for treatment, vehicle and positive controls
DETERMINATION OF CYTOTOXICITY
- Method: Any toxic effects of the test substance may be detected by a substantial reduction in mean revertant colony counts, by a sparse or absent background bacterial lawn, or both.
OTHER: After 72 h of incubation at 37 °C, the appearance of the background bacterial lawn was examined and revertant colonies counted using an automated colony counter (Perceptive Instruments Sorcerer). - Evaluation criteria:
- - If exposure to a test substance produces a reproducible increase in revertant colony numbers of at least twice (three times in the case of strains TA1535 and TA1537) that of the concurrent vehicle controls, with some evidence of a positive concentration-response relationship, it is considered to exhibit mutagenic activity in this test system.
- If exposure to a test substance does not produce a reproducible increase in revertant colony numbers, it is considered to show no evidence of mutagenic activity in this test system. No statistical analysis is performed.
- If the results obtained fail to satisfy the criteria for a clear “positive” or “negative” response, even after additional testing, the test data may be subjected to analysis to determine the statistical significance of any increases in revertant colony numbers. The statistical procedures used are those described by Mahon et al (1989) and are usually Dunnett’s test followed, if appropriate, by trend analysis. Biological importance will be considered along with statistical significance. In general, treatment-associated increases in revertant colony numbers below two or three times those of the vehicle controls (as described above) are not considered biologically important. It should be noted that it is acceptable to conclude an equivocal response if no clear results can be obtained.
- Occasionally, these criteria may not be appropriate to the test data and, in such cases, the Study Director would use his/her scientific judgement. - Statistics:
- - Statistical procedures used are those described by Mahon et al (1989) and are usually Dunnett’s test followed, if appropriate, by trend analysis.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium TA 1535
- Remarks:
- Plate incorporation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Remarks:
- Plate incorporation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Remarks:
- Plate incorporation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- hinning of background lawn observed at 5000 µg per plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Remarks:
- Plate incorporation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A pKM 101
- Remarks:
- Plate incorporation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Remarks:
- : Pre-incubation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Slight thinning of background lawn observed at 15 µg per plate, without metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Remarks:
- Pre-incubation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Slight thinning of background lawn observed at 15 µg per plate without metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Remarks:
- Pre-incubation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Slight thinning of background lawn observed at 15µg per plate without metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- not valid
- Species / strain:
- S. typhimurium TA 100
- Remarks:
- Pre-incubation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Slight thinning of background lawn observed at 15 µg per plate without metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A pKM 101
- Remarks:
- Pre-incubation assay
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
Any other information on results incl. tables
Table 1: Positive control results (Plate incorporation assay, without metabolic activation) :
Strain |
Addition |
Concentration per plate |
Mean revertants per plate |
Standard Deviation |
Fold increase relative to vehicle |
Individual revertant |
TA98 |
2NF |
2 µg |
238.3 |
57.3 |
11.7 |
174, 284, 257 |
TA100 |
NaN3 |
2 µg |
554.0 |
19.9 |
3.1 |
531, 565, 566 |
TA1535 |
NaN3 |
2 µg |
635.7 |
38.3 |
38.1 |
647, 593, 667 |
TA1537 |
AAC |
50 µg |
214.7 |
186.2 |
16.5 |
71, 148, 425 |
WP2 uvrA (pKM101) |
NQO |
2 µg |
2692.7 |
110.4 |
16.2 |
2682, 2808, 2588 |
Table 2: Positive control results (Plate incorporation assay, with metabolic activation) :
Strain |
Addition |
Concentration per plate |
Mean revertants per plate |
Standard Deviation |
Fold increase relative to vehicle |
Individual revertant |
TA98 |
B[a]P |
5 µg |
173.7 |
17.2 |
5.3 |
177, 155, 189 |
TA100 |
AAN |
5 µg |
3118.0 |
192.7 |
16.1 |
3269, 2901, 3184 |
TA1535 |
AAN |
5 µg |
210.3 |
24.4 |
16.6 |
205, 237, 189 |
TA1537 |
B[a]P |
5 µg |
167.7 |
34.1 |
14.4 |
147, 207, 149 |
WP2 uvrA (pKM101) |
AAN |
10 µg |
1432.0 |
66.6 |
7.0 |
1435, 1497, 1364 |
Table 3: Positive control results (Pre-incubation assay, without metabolic activation):
Strain |
Addition |
Concentration per plate |
Mean revertants per plate |
Standard Deviation |
Fold increase relative to vehicle |
Individual revertant |
TA98 |
2NF |
2 µg |
295.7 |
40.0 |
12.5 |
308, 328, 251 |
TA100 |
NaN3 |
2 µg |
624.3 |
9.3 |
4.1 |
614, 627, 632 |
TA1535 |
NaN3 |
2 µg |
632.3 |
49.2 |
41.2 |
577, 649, 671 |
TA1537 |
AAC |
50 µg |
226.3 |
66.3 |
33.9 |
282, 244, 153 |
WP2 uvrA (pKM101) |
NQO |
2 µg |
2622.3 |
12.9 |
16.7 |
2613, 2617, 2637 |
Table 4: Positive control results (Pre-incubation assay, with metabolic activation):
Strain |
Addition |
Concentration per plate |
Mean revertants per plate |
Standard Deviation |
Fold increase relative to vehicle |
Individual revertant |
TA98 |
B[a]P |
5 µg |
236.0 |
22.1 |
8.6 |
244, 253, 211 |
TA100 |
AAN |
5 µg |
912.0 |
327.7 |
5.9 |
1286, 775, 675 |
TA1535 |
AAN |
5 µg |
314.3 |
22.1 |
24.8 |
335, 291, 317 |
TA1537 |
B[a]P |
5 µg |
157.3 |
3.2 |
9.4 |
155, 156, 161 |
WP2 uvrA (pKM101) |
AAN |
10 µg |
1661.0 |
48.1 |
9.1 |
1716, 1627, 1640 |
Applicant's summary and conclusion
- Conclusions:
- alpha-Pinene multiconstituent is not considered as mutagenic in S. typhimurium (TA 1535, TA 1537, TA 98 and TA 100) and E. coli (WP2uvrA) strains.
- Executive summary:
In a reverse gene mutation assay in bacteria, performed according to the OECD 471 Guideline and in compliance with GLP, strains of Salmonella typhimurium (TA 1535, TA 1537, TA 98 and TA 100) and Escherichia coli (WP2uvrA) were exposed to alpha-pinene multiconstituent at the following concentrations:
Experiment 1 (plate incorporation method) 5, 15, 50, 150, 500, 1500 and 5000 µg/plate with and without S9 mix in all strains;
Experiment 2 (pre-incubation method) 0.05, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate with and without S9 mix in all strains.
Metabolic activation system used in this test was10 % (v/v) S9 mix;S9 fraction prepared from liver homogenates of rats induced with Phenobarbital sodium/5,6-benzoflavone.Vehicle and positive control groups were also included in mutagenicity tests.
No signs of toxicity towards the tester strains were observed in the first experiment following exposure to test item. Toxicity, observed as a reduction in revertant colony numbers, ans slight to severe thinning of the background lawn, was obtained in all strains in the second experiment following exposure to test item at 15 µg/plate in the absence of S9 mix. No substantial increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to alpha-pinene multiconstituent, at any concentration up to and including 5000 µg/plate in either the presence or absence of S9 mix. The positive and vehicle controls induced the appropriate responses in the corresponding strains indicating the validity of the study.
Therefore, alpha-pinene multiconsituent is not considered as mutagenic in these bacterial systems
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
![ECHA](/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/echa_logo.png)