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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 948-032-2 | 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
Endpoint summary
Administrative data
Description of key information
Several studies and assessment reports from competent authorities are available for the skin sensitising properties of alpha-pinene, beta-pinene and limonene.
As a result from these studies all of these substance were identified as inducing skin sensitisation.
Even if the DPRA study of Terpene Dimers shows negative results, this information is not sufficient enought to dissipate the doubts. That´s why we follow the worts-case approach and classify the substance as skin senstitiser.
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2006
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Remarks:
- publication
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- data from publication
- Species:
- guinea pig
- Strain:
- Hartley
- Sex:
- female
- Details on test animals and environmental conditions:
- weight: 300-350 g
temperature: 25 oC
humidity: 50-70% - Route:
- intradermal
- Vehicle:
- DMSO
- Concentration / amount:
- 4%
- Day(s)/duration:
- 7 days
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- DMSO
- Concentration / amount:
- 4%
- Day(s)/duration:
- 14 days
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- DMSO
- Concentration / amount:
- 0.8%
- Day(s)/duration:
- 1 day
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- 6
- Details on study design:
- For induction, 6 intradermal injections of 0.1 ml of compound and Freund's adjuvant were administered in the shaved scapular region. After 7 d, an occluded patch of 0.15 ml of compound was placed on the injection site for 48 h. In the control guinea pigs, the compound was replaced by the solvent. After 14 d, all the guinea pigs (including controls) were exposed to a challenge dose on the shaved flank for 24 h. Skin reactions were observed and scored.
- Positive control substance(s):
- no
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 0
- No. with + reactions:
- 0
- Total no. in group:
- 6
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 0.8%
- No. with + reactions:
- 0
- Total no. in group:
- 6
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- In a Guinea-Pig Maximisation Test alpha-pinene was tested for its skin sensitising properties in female Hartley guinea pigs. None of the tested animals showed a positive response after challenge exposure. Therefore it can be concluded that alpha-pinene was not identified as a skin sensitizer in the Guinea-Pig Maximisation Test .
- Executive summary:
In a Guinea-Pig Maximisation Test alpha-pinene was tested in female Hartley guinea pigs. None of the tested animals showed a positive response after challenge exposure. Therefore it can be concluded that alpha-pinene was not identified as a skin sensitizer in the Guinea-Pig Maximisation Test .
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Remarks:
- publication
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- CBA:J
- Sex:
- female
- Details on test animals and environmental conditions:
- age: 8-12 weeks
temperature: 25oC
humidity: 50-70% - Vehicle:
- acetone/olive oil (4:1 v/v)
- Concentration:
- 0,1, 25, 100%
- No. of animals per dose:
- 4
- Details on study design:
- Groups of mice (4 for each group) received topical administration of 25 µl of various concentrations of the test chemicals on the dorsum of both ears, or the same volume of the relevant vehicle alone, daily for 3 consecutive days. Five days after initiation of exposure, all mice were injected via the tail vein with 250 µl phosphate-buffered saline (PBS) containing 20 µCi [3H] methyl thymidine. Five hours later, the mice were sacrificed and the draining auricular lymph nodes excised and pooled for each experimental group.
- Positive control substance(s):
- hexyl cinnamic aldehyde (CAS No 101-86-0)
- Positive control results:
- HCA: EC3: 10.86%
- Key result
- Parameter:
- SI
- Value:
- 1
- Test group / Remarks:
- control
- Key result
- Parameter:
- SI
- Value:
- 1.03
- Test group / Remarks:
- 1%
- Key result
- Parameter:
- SI
- Value:
- 1.65
- Test group / Remarks:
- 25%
- Key result
- Parameter:
- SI
- Value:
- 2.59
- Test group / Remarks:
- 100%
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- α-pinene was assessed in the local lymph note assay (LLNA) for its skin sensitising properties in CBA/J female mice. The substance failed in all tested concentrations to induce a positive response. Even at the higest test concentration of 100 % a SI of 2.59 was estimated. Therefore it can be concluded that α-pinene did not induce an allergic reaction in the described LLNA-test.
- Executive summary:
α-pinene was assessed in the local lymph note assay (LLNA) for its skin sensitising properties in CBA/J female mice. The substance failed in all tested concentrations to induce a positive response. Even at the higest test concentration of 100 % a SI of 2.59 was estimated. Therefore it can be concluded that α-pinene did not induce an allergic reaction in the described LLNA-test.
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Assessment report from a highly reputed government agency.
- Qualifier:
- no guideline followed
- GLP compliance:
- no
- Type of study:
- other: review of data
- Species:
- mouse
- Strain:
- CBA:J
- Sex:
- not specified
- Vehicle:
- acetone/olive oil (4:1 v/v)
- Concentration:
- 1,25, 100%
- No. of animals per dose:
- 1
- Key result
- Parameter:
- SI
- Value:
- 1.89
- Test group / Remarks:
- 1%
- Key result
- Parameter:
- SI
- Value:
- 1.82
- Test group / Remarks:
- 25%
- Key result
- Parameter:
- SI
- Value:
- 2.55
- Test group / Remarks:
- 100%
- Interpretation of results:
- Category 1 (skin sensitising) based on GHS criteria
- Executive summary:
Citation from assessment report:
"Based on the available human data, the chemicals are considered to be skin sensitisers warranting hazard classification.
A popliteal lymph node (PLN) assay was conducted to assess the sensitisation potential of (–)-alpha-pinene. Female Wistar rats were treated by subcutaneous injection with 50 µL of (–)-alpha-pinene (0.5, 2.5 or 5 mg) into the right hind footpad while the contralateral footpad was injected with the vehicle (DMSO) alone. Weight and cellularity indices (WI and CI) for draining PLNs were determined 7 days after treatment. The chemical was positive (WI>2 and CI≤5) under these test conditions. In a secondary PLN assay (T-cell priming test), animals whose immune systems had been primed to the chemical (5 mg), were re-administered the chemical via subcutaneous injection (0.5 mg). Assessment of WI and CI were conducted 5 and 7 days following the second injection. The chemical was negative in the second test. The chemical induced a clear immuno-stimulatory response due to its irritant properties but it was not considered to be a sensitising agent (Friedrich et al., 2007).
The chemical alpha-pinene was assessed for its potential to produce skin sensitisation in a guinea pig maximisation test (GPMT). Female Hartley guinea pigs (6/dose group) were induced with 0.1 mL of the chemical at 4 % (in acetone: olive oil (v/v 4:1)). After 7 days, an occluded patch was applied to the shaved skin with 0.15 mL of the chemical. After 14 days, animals were exposed to a challenge dose (0.8 %) in the same vehicle as the induction dose. The chemical was negative for sensitisation under these test conditions (Wei et al., 2006).
A study was conducted to assess the sensitisation potential of the components of the Myoga (Zingiber Myoga Roscoe) plant, one of which is alpha-pinene. In a Local Lymph Node Assay (LLNA), alpha-pinene was topically-administered to the dorsum of both ears of 4 CBN/J mice. The chemical was applied at concentrations of 0, 1, 25 and 100 %. No EC3 value (the estimated concentration needed to produce a 3-fold increase in lymphocyte proliferation) could be determined up to 100 %, indicating that it was not a skin sensitiser under these test conditions (Wei et al., 2010).
The Quantitative Structure–Activity Relationship (QSAR) modelling for skin sensitisation using the OECD QSAR Toolbox (version 3.4) indicated that there were protein binding alerts for the predicted auto-oxidation metabolic products of alpha-pinene.
The chemical alpha-pinene is predicted to be a skin sensitiser using the VEGA skin sensitisation model (CAESAR). The chemical falls within the applicability domain of the model.
The chemical (–)- alpha-pinene is predicted to be a moderate skin sensitiser (predicted LLNA EC3%: 9.6) using Derek Nexus v5.0.2. The prediction is based on the triggered structured alert for terpenoids. The prediction strength is 'equivocal' (REACHb).
In the EU, the ECHA Guidance on the Application of the CLP Criteria (2013), the decision logic was used to determine the skin sensitising potential of alpha-pinene multiconstituent.
The chemicals (–)-beta-pinene and alpha-pinene are structurally-similar (both are bicyclic monounsaturated terpenes and are positional isomers). They also share very similar physico-chemical properties. (-)-beta-pinene has been classified as a Category1B skin sensitiser (based on an EC3 value of 29 %, obtained in a Local Lymph Node Assay) and is present in alpha-pinene multiconstituent above the generic concentration limit of 1 % for classification as skin sensitiser (REACHa).
It is expected that alpha-pinene multiconstituent will have the same weak potential for skin sensitisation. This, coupled with the available data, suggest classification of alpha-pinene for sensitisation is warranted.
Turpentine oil (CAS No. 8006-64-2), of which alpha-pinene is the primary constituent, is classified as hazardous with hazard category ‘Skin sensitisation – category 1’ and hazard statement ‘May cause an allergic skin reaction' (H317) in the Hazardous Chemical Information System (HCIS) (Safe Work Australia). Data for the mixture (including a positive Guinea Pig Maximisation test) support this classification (NICNASa)."
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2006
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Remarks:
- publication
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- data from publication
- Species:
- guinea pig
- Strain:
- Hartley
- Sex:
- female
- Details on test animals and environmental conditions:
- weight: 300-350 g
temperature: 25 oC
humidity: 50-70% - Route:
- intradermal
- Vehicle:
- DMSO
- Concentration / amount:
- 4%
- Day(s)/duration:
- 7 days
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- DMSO
- Concentration / amount:
- 4%
- Day(s)/duration:
- 14 days
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- DMSO
- Concentration / amount:
- 0.8%
- Day(s)/duration:
- 1 day
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- 6
- Details on study design:
- For induction, 6 intradermal injections of 0.1 ml of compound and Freund's adjuvant were administered in the shaved scapular region. After 7 d, an occluded patch of 0.15 ml of compound was placed on the injection site for 48 h. In the control guinea pigs, the compound was replaced by the solvent. After 14 d, all the guinea pigs (including controls) were exposed to a challenge dose on the shaved flank for 24 h. Skin reactions were observed and scored.
- Positive control substance(s):
- no
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 0
- No. with + reactions:
- 0
- Total no. in group:
- 6
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 0.8%
- No. with + reactions:
- 0
- Total no. in group:
- 6
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- In a Guinea-Pig Maximisation Test beta-pinene was tested for its skin sensitising properties in female Hartley guinea pigs. None of the tested animals showed a positive response after challenge exposure. Therefore it can be concluded that beta-pinene was not identified as a skin sensitizer in the Guinea-Pig Maximisation Test .
- Executive summary:
In a Guinea-Pig Maximisation Test beta-pinene was tested in female Hartley guinea pigs. None of the tested animals showed a positive response after challenge exposure. Therefore it can be concluded that beta-pinene was not identified as a skin sensitizer in the Guinea-Pig Maximisation Test .
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Remarks:
- publication
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- CBA:J
- Sex:
- female
- Details on test animals and environmental conditions:
- age: 8-12 weeks
temperature: 25oC
humidity: 50-70% - Vehicle:
- acetone/olive oil (4:1 v/v)
- Concentration:
- 0,1, 25, 100%
- No. of animals per dose:
- 4
- Details on study design:
- Groups of mice (4 for each group) received topical administration of 25 µl of various concentrations of the test chemicals on the dorsum of both ears, or the same volume of the relevant vehicle alone, daily for 3 consecutive days. Five days after initiation of exposure, all mice were injected via the tail vein with 250 µl phosphate-buffered saline (PBS) containing 20 µCi [3H] methyl thymidine. Five hours later, the mice were sacrificed and the draining auricular lymph nodes excised and pooled for each experimental group.
- Positive control substance(s):
- hexyl cinnamic aldehyde (CAS No 101-86-0)
- Positive control results:
- HCA: EC3: 10.86%
- Key result
- Parameter:
- SI
- Value:
- 1
- Test group / Remarks:
- control
- Key result
- Parameter:
- SI
- Value:
- 1.89
- Test group / Remarks:
- 1%
- Key result
- Parameter:
- SI
- Value:
- 1.82
- Test group / Remarks:
- 25%
- Key result
- Parameter:
- SI
- Value:
- 2.55
- Test group / Remarks:
- 100%
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- β-pinene was assessed in the local lymph note assay (LLNA) for its skin sensitising properties in CBA/J female mice. The substance failed in all tested concentrations to induce a positive response. Even at the higest test concentration of 100 % a SI of 2.55 was estimated. Therefore it can be concluded that β-pinene did not induce an allergic reaction in the described LLNA-test.
- Executive summary:
β-pinene was assessed in the local lymph note assay (LLNA) for its skin sensitising properties in CBA/J female mice. The substance failed in all tested concentrations to induce a positive response. Even at the higest test concentration of 100 % a SI of 2.55 was estimated. Therefore it can be concluded that β-pinene did not induce an allergic reaction in the described LLNA-test.
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Assessment report from a highly reputed government agency.
- Qualifier:
- no guideline followed
- GLP compliance:
- no
- Type of study:
- other: review of data
- Species:
- mouse
- Strain:
- CBA:J
- Sex:
- female
- Vehicle:
- acetone/olive oil (4:1 v/v)
- Concentration:
- 5, 10, 25, 50, 100%
- No. of animals per dose:
- 5
- Key result
- Parameter:
- SI
- Value:
- 2.29
- Test group / Remarks:
- 5%
- Key result
- Parameter:
- SI
- Value:
- 1.16
- Test group / Remarks:
- 10%
- Key result
- Parameter:
- SI
- Value:
- 2.23
- Test group / Remarks:
- 25%
- Key result
- Parameter:
- SI
- Value:
- 7.17
- Test group / Remarks:
- 50%
- Key result
- Parameter:
- SI
- Value:
- 14.7
- Test group / Remarks:
- 100%
- Key result
- Parameter:
- EC3
- Value:
- 29
- Interpretation of results:
- Category 1 (skin sensitising) based on GHS criteria
- Executive summary:
Citation from assessment report:
"Based on the weight of evidence from the available animal, human and in silico data, the chemicals are skin sensitisers and warrant hazard classification.
Hydroperoxide species originating from auto-oxidation are thought to be the major contributors.
In a local lymph node assay (LLNA) performed in accordance with OECD TG 429, female CBA/J mice (5/dose) received topical applications of 5.0, 10, 25, 50 or 100 % (v/v) beta-pinene in acetone/olive oil on three consecutive days. The reported stimulation indices (SI) were 2.29, 1.16, 2.23, 7.17, 6.47 and 14.7 for concentrations of 5.0, 10, 25, 50 and 100 % respectively. The reported concentration producing a three-fold increase in lymphocyte proliferation (EC3) was 29 %, indicating weak sensitisation potential (REACH).
In a non-guideline LLNA, CBN/J (1/dose) mice received topical applications of 1, 25 and 100 % of the beta-pinene in acetone/olive oil on three consecutive days. The reported stimulation indices (SI) were 1.89, 1.82 and 2.55 for the concentrations 1, 25 and 100 %, respectively. As all SI values were below 3, an EC3 value could not be determined and the chemical was; therefore; considered a non-sensitiser (Wei et al., 2010).
No structural alerts for skin sensitisation were present for beta-pinenes (OECD QSAR Toolbox v3.4). However, when auto-oxidation was simulated, mechanistic alerts, including alerts for protein binding via nucleophilic addition and free radical formation were present for the metabolites.
In domain skin sensitisation predictions for beta-pinenes using OASIS–TIMES 2.27.19 were negative for the unmetabolised chemicals. Several auto-oxidised metabolites of beta-pinene were predicted to be weak sensitisers which were supported by mechanistic alerts for hydroperoxide free radical decomposition and Michael type addition on conjugated systems with electron withdrawing groups."
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2006
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Remarks:
- publication
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- data from publication
- Species:
- guinea pig
- Strain:
- Hartley
- Sex:
- female
- Details on test animals and environmental conditions:
- weight: 300-350 g
temperature: 25 oC
humidity: 50-70% - Route:
- intradermal
- Vehicle:
- DMSO
- Concentration / amount:
- 4%
- Day(s)/duration:
- 7 days
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- DMSO
- Concentration / amount:
- 4%
- Day(s)/duration:
- 14 days
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- DMSO
- Concentration / amount:
- 0.8%
- Day(s)/duration:
- 1 day
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- 6
- Details on study design:
- For induction, 6 intradermal injections of 0.1 ml of compound and Freund's adjuvant were administered in the shaved scapular region. After 7 d, an occluded patch of 0.15 ml of compound was placed on the injection site for 48 h. In the control guinea pigs, the compound was replaced by the solvent. After 14 d, all the guinea pigs (including controls) were exposed to a challenge dose on the shaved flank for 24 h. Skin reactions were observed and scored.
- Positive control substance(s):
- no
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 0
- No. with + reactions:
- 0
- Total no. in group:
- 6
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 0.8%
- No. with + reactions:
- 2
- Total no. in group:
- 6
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- Category 1 (skin sensitising) based on GHS criteria
- Conclusions:
- In a Guinea-Pig Maximisation Test R-(+)-limonene was tested for its skin sensitising properties in female Hartley guinea pigs. 2 of the 6 tested animals showed a positive response after challenge exposure. Therefore it can be concluded that R-(+)-limonene was identified as a skin sensitizer in the Guinea-Pig Maximisation Test .
- Executive summary:
In a Guinea-Pig Maximisation Test R-(+)-limonene was tested in female Hartley guinea pigs. Two of the six tested animals showed a positive response after challenge exposure. Therefore it can be concluded that R-(+)-limonene was identified as a skin sensitizer in the Guinea-Pig Maximisation Test .
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Remarks:
- publication
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- CBA:J
- Sex:
- female
- Details on test animals and environmental conditions:
- age: 8-12 weeks
temperature: 25oC
humidity: 50-70% - Vehicle:
- acetone/olive oil (4:1 v/v)
- Concentration:
- 0,1, 25, 100%
- No. of animals per dose:
- 4
- Details on study design:
- Groups of mice (4 for each group) received topical administration of 25 µl of various concentrations of the test chemicals on the dorsum of both ears, or the same volume of the relevant vehicle alone, daily for 3 consecutive days. Five days after initiation of exposure, all mice were injected via the tail vein with 250 µl phosphate-buffered saline (PBS) containing 20 µCi [3H] methyl thymidine. Five hours later, the mice were sacrificed and the draining auricular lymph nodes excised and pooled for each experimental group.
- Positive control substance(s):
- hexyl cinnamic aldehyde (CAS No 101-86-0)
- Positive control results:
- HCA: EC3: 10.86%
- Key result
- Parameter:
- SI
- Value:
- 1
- Test group / Remarks:
- control
- Key result
- Parameter:
- SI
- Value:
- 1.6
- Test group / Remarks:
- 1%
- Key result
- Parameter:
- SI
- Value:
- 2.31
- Test group / Remarks:
- 25%
- Key result
- Parameter:
- SI
- Value:
- 7.09
- Test group / Remarks:
- 100%
- Key result
- Parameter:
- EC3
- Value:
- 35.8
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- Limonene was assessed in the local lymph note assay (LLNA) for its skin sensitising properties in CBA/J female mice. The substance induced a positive response and the EC3 value was 35.8%.
Therefore it can be concluded that limonen induced an allergic reaction in the described LLNA-test. - Executive summary:
Limonene was assessed in the local lymph note assay (LLNA) for its skin sensitising properties in CBA/J female mice. The substance induced a positive response and the EC3 value was 35.8%.
Therefore it can be concluded that limonen induced an allergic reaction in the described LLNA-test.
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Assessment report from a highly reputed government agency.
- Qualifier:
- no guideline followed
- GLP compliance:
- no
- Key result
- Parameter:
- SI
- Remarks on result:
- other: Assessment report from competent authority, results are only summarised.
- Interpretation of results:
- GHS criteria not met
- Executive summary:
Citation from assessment report:
"Although d‑limonene was once considered the main allergen in citrus fruits, data from more recent studies in animals have revealed air‑oxidizedd‑limonene,rather than unoxidized d‑limonene,to be the sensitising agent. When limonene (unspecified form and unknown purity of the testmaterial) was tested in four different sensitisation assays with guinea‑pigs (Open Epicutaneous Test,Maximization Test, Draize's Test, and a test with Freund's Complete Adjuvant),it was sensitising in all but Draize's Test*(Klecak et al., 1977). In another study in mice,d‑limonene did not induce sensitisation*(Maisey & Miller, 1986). Hydroperoxides and other oxidation products of d-limonene formed on exposure to the air have proved to be potent contact allergens when tested with Freund's Complete Adjuvant in guinea‑pigs, whereas unoxidized d-limonene did not cause any sensitisation(Karlberg et al., 1991;Karlberg et al., 1992).
It has been suggested that limonene has a quenching effect (inhibition of the sensitising capacity of another substance) on the induction of skin sensitisation to citral. No effect was found in trials on guinea pigs or in the murine local lymph node assay (LLNA)(European Union, 2000)."
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 1998
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Terpene dimers is primarily a mixture of terpene dimer with no more than trace quantities of unreacted monomer. The raw materials (monoterpene) react by cationic polymerisation.
Raw materials:
(R)-p-mentha-1,8-diene (D-limonene): EC: 227-813-5; ratio: 1 ~ 50
Pin-2(3)-ene (racemic mixture) (alpha-pinene); EC: 201-291-9; ratio: 15 ~ 45
(-)-pin-2(10)-ene (beta-pinene); EC: 242-060-2; ratio: 1 ~ 50
The reaction products terpene dimers are dimeric species of reactants covalently coupled twice. In other words, these species are structurally similar, i.e. they contain the same repeating reactant. It however has to be kept in mind that most of physical and chemical properties are altered significantly as a result of this increase in molecular weight going from the reactant to dimeric species. For example, the vapour pressure is very significantly reduced.
2. ANALOGUE APPROACH JUSTIFICATION
Overall, it can be assumed that the change in physical properties from starting materials to dimeric reaction products, results in a substance with significantly reduced hazardous properties.
Therefore read-across from the starting materials to terpene dimers is fully justified.
Although alpha- and beta-pinene are not classified for skin sensitisation, as a worst case assumption, the substance terpene dimers will be classified as skin sensitiser as the starting material D-limonene is classified for skin sensitisation.
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- Reliable safety assessment of the World Health Organization.
- GLP compliance:
- no
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- Published data
- Species:
- guinea pig
- Strain:
- not specified
- Sex:
- not specified
- Key result
- Reading:
- other: not stated
- Group:
- test chemical
- Dose level:
- not stated
- Interpretation of results:
- Category 1 (skin sensitising) based on GHS criteria
- Conclusions:
- When limonene (unspecified form and unknown purity of the test material) was tested in four different sensitization assays with guinea-pigs (Open
Epicutaneous Test, Maximization Test, Draize’s Test, and a test with Freund’s Complete Adjuvant), it was sensitizing in all but Draize’s Test. - Executive summary:
When limonene (unspecified form and unknown purity of the test material) was tested in four different sensitization assays with guinea-pigs (Open
Epicutaneous Test, Maximization Test, Draize’s Test, and a test with Freund’s Complete Adjuvant), it was sensitizing in all but Draize’s Test.
Overall, it can be assumed that the change in physical properties from starting materials to dimeric reaction products, results in a substance with significantly reduced hazardous properties.
Therefore read-across from the starting materials to terpene dimers is fully justified.
Although alpha- and beta-pinene are not classified for skin sensitisation, as a worst case assumption, the substance terpene dimers will be classified as skin sensitiser as the starting material D-limonene is classified for skin sensitisation.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (sensitising)
- Additional information:
Terpene dimers is primarily a mixture of terpene dimer with no more than trace quantities of unreacted monomer. The raw materials alpha-pinene, beta-pinene and limonene react by cationic polymerisation.
The raw materials are:
(R)-p-mentha-1,8-diene (D-limonene): EC: 227-813-5; ratio: 1 ~ 50
Pin-2(3)-ene (racemic mixture) (alpha-pinene); EC: 201-291-9; ratio: 15 ~ 45
(-)-pin-2(10)-ene (beta-pinene); EC: 242-060-2; ratio: 1 ~ 50
The reaction products terpene dimers are dimeric species of reactants covalently coupled twice. In other words, these species are structurally similar, i.e. they contain the same repeating reactant.
Based on the read across assumption of structural similarity the substance terpene dimers is regarded as skin sensitising substance.
As the starting materials are all classified as skin sensitising substances, the substance terpene dimers will be classified as skin sensitiser too. This can be regarded as a worst case assumption.
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
According to the CLP Regulation (EU GHS Regulation (EC) No 1272/2008) classification and labelling is required for skin sensitisation of terpene dimers, based on reliable data from the read across source substances alpha-pinene, beta-pinene and limonene.
As the starting materials for terpene dimers are all classified as skin sensitising substances, the substance terpene dimers will be classified as skin sensitiser too.
Even if the DPRA study of Terpene Dimers shows negative results, this information is not sufficient enought to dissipate the doubts. That´s why we follow the worts-case approach and classify the substance as skin senstitiser.
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.