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EC number: 227-815-6 | CAS number: 5989-54-8
- 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
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Carcinogenicity
Administrative data
Description of key information
No studies were located on l-limonene. D‑limonene has been studied in carcinogenicity studies in F344/N rats and B6C3F1 mice (2-year study by gavage). Increased incidences of renal tubular cell hyperplasia, adenomas, and adenocarcinomas of the kidney have been observed in male rats. No increased incidences of tumors have been reported in female rats and in mice.
The mechnaism of nephrocarcinogenicity has been proven as being male-rat specific and not relevant for humans.
Key value for chemical safety assessment
Carcinogenicity: via oral route
Link to relevant study records
- Endpoint:
- carcinogenicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Read-across from an analogue substance for which there is available information (Klimish =2).
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
See cross-reference to justification of read-across. - Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across: supporting information
- Clinical signs:
- effects observed, treatment-related
- Mortality:
- mortality observed, treatment-related
- Body weight and weight changes:
- effects observed, treatment-related
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not specified
- Haematological findings:
- not specified
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- no effects observed
- Histopathological findings: non-neoplastic:
- no effects observed
- Histopathological findings: neoplastic:
- no effects observed
- Relevance of carcinogenic effects / potential:
- Not relevant as no evidence of carcinogenic activity of d-limonene for male and female B6C3F1 mice.
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 250 - <= 500 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male
- Basis for effect level:
- other: no evidence of carcinogenic activity
- Remarks on result:
- not determinable
- Remarks:
- no NOAEL identified. Effect type:carcinogenicity
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 500 - <= 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- female
- Basis for effect level:
- other: no evidence of carcinogenic activity
- Remarks on result:
- not determinable
- Remarks:
- no NOAEL identified. Effect type:carcinogenicity
- Conclusions:
- Based on the read-across from the analogue substance d-limonene, l-limonene is not expected to have any carcinogenic activity in male and female B6C3F1 mice.
- Executive summary:
A 2 -year carcinogenicity study was performed similarly to OECD Guideline 451 and in compliance with GLP with the analogue substance d-limonene which was administered through gavage to groups of 50 B6C3F1 mice/sex/dose mixed in corn oil at dose levels of 0, 250 and 500 mg/kg bw/day (in males) or 0, 500 and 1000 mg/kg bw/day (in females) for 103 weeks (5 days/week).
Survival of the low dose group of male mice was significantly lower than that of the vehicle controls at the end of the study. Survival at week 104 was 33/50 male and 43/50 female in vehicle control group; 24/50 males and 44/50 females in low dose group and 39/50 males and 43/50 females in high dose group. No treatment-related clinical signs were observed during the study. Mean bodyweights of high dose female mice were 5-15% lower than those of the vehicle controls after week 28. Mean bodyweights of dosed and vehicle control male mice were similar throughout the studies. No treatment-related statistically or biologically significant histologic effects were observed during the study.
Under the test conditions, there was no evidence of carcinogenic activity of d-limonene for male and female B6C3F1 mice.
Based on the read-across from the analogue substance d-limonene, l-limonene is not expected to have any carcinogenic activity in male and female B6C3F1 mice.
- Endpoint:
- carcinogenicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Read-across from an analogue substance for which there is available information (Klimish=4).
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
See cross-reference to justification of read-across. - Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across: supporting information
- Clinical signs:
- effects observed, treatment-related
- Mortality:
- mortality observed, treatment-related
- Body weight and weight changes:
- effects observed, treatment-related
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- no effects observed
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- effects observed, treatment-related
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Histopathological findings: neoplastic:
- effects observed, treatment-related
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 75 - <= 150 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male
- Basis for effect level:
- other: clear evidence of carcinogenic activity based on the increased incidences of tubular cell hyperplasia, adenomas and adenocarcinomas of the kidney
- Remarks on result:
- not determinable
- Remarks:
- no NOAEL identified. Effect type:carcinogenicity (migrated information)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 300 - <= 600 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- female
- Basis for effect level:
- other: no evidence of carcinogenic activity
- Remarks on result:
- not determinable
- Remarks:
- no NOAEL identified
- Key result
- Critical effects observed:
- yes
- Lowest effective dose / conc.:
- 75 mg/kg bw/day (nominal)
- System:
- urinary
- Organ:
- kidney
- Treatment related:
- yes
- Dose response relationship:
- yes
- Relevant for humans:
- no
- Conclusions:
- There was clear evidence of carcinogenic activity of d-limonene for male F344/N rats, as shown by increased incidences of tubular cell hyperplasia, adenomas, and adenocarcinomas of the kidney. There was no evidence of carcinogenic activity of d-limonene for female F344/N rats. This effect is well known to be specifics of male rats and irrelevant to humans.
- Executive summary:
In a 2 year carcinogenicity study performed similarly to OECD Guideline 451 and in compliance with GLP, d-limonene was administered through gavage to groups of 50 F344/N rats/sex/dose mixed in corn oil at dose levels of 0, 75 and 150 mg/kg bw/day (in males) or 0, 300 and 600 mg/kg bw/day (in females) for 103 weeks (5 days/week). Animals were observed twice daily for clinical signs of toxicity and bodyweights were recorded once per week for 12 weeks and once per month thereafter. Necropsy performed on all animals and microscopic examination of specified tissues was performed for all animals dying during the studies, all vehicle controls, all low dose female rats and all high dose animals at the end of the treatment period.
Survival of the high dose female rats after week 39 and of the vehicle control male rats after week 81 was significantly reduced. Survival at week 104 was 29/50 male and 42/50 female in vehicle control group; 33/50 males and 40/50 females in low dose group and 40/50 males and 20/50 females in high dose group. Mean body weights of high dose rats were generally 4-7% lower than those of the vehicle controls from week 2 (in males) or 28 (in females) to the end of the studies. No treatment-related clinical signs were observed during the study. Kidney was confirmed as the primary target organ for chemically related lesions. No lesions were observed in female rats. For males, the nonneoplastic lesions included exacerbation of the age-related nephropathy, linear deposits of mineral in the renal medulla and papilla, and focal hyperplasia of the transitional epithelium overlying the renal papilla. Uncommon tubular cell adenomas and adenocarcinomas of the kidney also occurred in dosed male rats and this effect was supported by a dose-related increased incidence of tubular cell hyperplasia.
Under the test conditions, there was clear evidence of carcinogenic activity of d-limonene for male F344/N rats, as shown by increased incidences of tubular cell hyperplasia, adenomas, and adenocarcinomas of the kidney. There was no evidence of carcinogenic activity of d-limonene for female F344/N rats. This mechanism of nephrocarcinogenicity has been proven as being male-rat specific and not relevant for humans.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- LOAEL
- 75 mg/kg bw/day
- Study duration:
- chronic
- Species:
- rat
- System:
- urinary
- Organ:
- kidney
Carcinogenicity: via inhalation route
Endpoint conclusion
- Endpoint conclusion:
- no study available
Carcinogenicity: via dermal route
Endpoint conclusion
- Endpoint conclusion:
- no study available
Mode of Action Analysis / Human Relevance Framework
No studies were located on l-limonene.For further information on read-across justification, see section 13: point "read-across approach"
D‑limonene has been studied in carcinogenicity studies in F344/N rats and B6C3F1 mice (2-year study by gavage). Increased incidences of renal tubular cell hyperplasia, adenomas, and adenocarcinomas of the kidney have been observed in male rats. No increased incidences of tumors have been reported in female rats and in mice.The mechnaism of nephrocarcinogenicity has been proven as being male-rat specific and not relevant for humans.
Mechanisms and specificity of toxicity of d-limonene on kidney of male rats and its non relevance for human are well known.
d-limonene produces hyaline droplets, tubular necrosis, regeneration, hyperplasia adenomas and carcinomas in the kidneys of male rats. These effects are not observed in mice. The key events are:
- Metabolism to d-limonene epoxide,
- Binding of the metabolite to serum alpha2u -globulin, which appears to be the rate-limiting step in the development of the nephropathy and tumours,
- Accumulation of alpha2u-globulin in the form of hyaline droplets in the renal tubules,
- Chronic renal cellular protein overload,
- Renal cell necrosis and compensatory cell proliferation, which is the critical event and must be sustained for tumours to develop,
- Renal tubule adenomas and carcinomas.
The key events can be demonstrated by measurement of d-limonene epoxide in plasma, demonstration of alpha2u-globulin in renal tubules and histology of kidney tumours. Alpha2u-globulin is a male rat specific protein with no equivalent in mice or humans capable of binding d-limonene epoxide. Thus, the conclusion of the assessment is that the proven mechanism of action is not relevant for humans. (Crome S. et al., 2008, Developments in Lifesciences Vol B No.2)
A study was also available, which showed that there was no clear evidence of skin tumour promoting activity of d-limonene for female CD-1 mice.
Justification for classification or non-classification
Harmonized classification:
No harmonized classification is available according to the Regulation (EC) No 1272/2008.
Self classification:
CLP Annex I, 3.9.2.8.1. (e) states that d-limonene induces a male-rat specific carcinogenic effect on kidney, which is not deemed relevant for classification of d-limonene as a carcinogenic substance. Therefore, l-limonene does not need to be classified for carcinogenicity.
Additional information
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.
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