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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
![](https://poisoncentres.echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Developmental toxicity / teratogenicity
Administrative data
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 1977
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- documentation insufficient for assessment
- Remarks:
- Original reference in Japanese language
Data source
Reference
- Reference Type:
- publication
- Title:
- Unnamed
- Year:
- 1 977
Materials and methods
- Principles of method if other than guideline:
- Prenatal developmental toxicity study: Groups of pregnant Japanese white rabbits were administered orally with d-limonene at dose levels of 250, 500 and 1000 mg/kg bw/day for 13 days from Day 6 to 18 of gestation and evaluated for teratogenicity.
- GLP compliance:
- not specified
- Limit test:
- no
Test material
- Reference substance name:
- (R)-p-mentha-1,8-diene
- EC Number:
- 227-813-5
- EC Name:
- (R)-p-mentha-1,8-diene
- Cas Number:
- 5989-27-5
- Molecular formula:
- C10H16
- IUPAC Name:
- 4-isopropenyl-1-methylcyclohexene
- Details on test material:
- - Name of test material (as cited in study report): d-limonene; d-p-mentha-1,8-diene
Constituent 1
Test animals
- Species:
- rabbit
- Strain:
- other: Japanese white
- Details on test animals or test system and environmental conditions:
- no data
Administration / exposure
- Route of administration:
- oral: unspecified
- Vehicle:
- not specified
- Details on exposure:
- no data
- Analytical verification of doses or concentrations:
- not specified
- Details on analytical verification of doses or concentrations:
- no data
- Details on mating procedure:
- No data
- Duration of treatment / exposure:
- 13 days (gestation Day 6-18)
- Frequency of treatment:
- Once daily
- Duration of test:
- Gestatation Day 0 to postnatal Day 49
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day
- Dose / conc.:
- 250 mg/kg bw/day
- Dose / conc.:
- 500 mg/kg bw/day
- Dose / conc.:
- 1 000 mg/kg bw/day
- No. of animals per sex per dose:
- 10 (in 0-500 mg/kg bw/day groups) or 18 (in 1000 mg/kg bw/day group) pregnant females
- Control animals:
- yes
- Details on study design:
- no data
Examinations
- Maternal examinations:
- CAGE SIDE OBSERVATIONS: No data
General behaviour observed, but no data regarding the frequency of observation
BODY WEIGHT: Yes
- Time schedule for examinations: daily
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
mean daily food consumption by treatment group is reported
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
POST-MORTEM EXAMINATIONS: No data
OTHER: - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: No data
Examinations included: Number of implantations, number of resorptions and foetus bodyweight and placental weight - Fetal examinations:
- - External examinations: Yes: about 90% per litter
- Visceral examinations: Yes: about 90% per litter
- Skeletal examinations: Yes: about 90% per litter - Statistics:
- statistical significance difference of effects from controls were calculated at 5% level.
- Indices:
- no data
- Historical control data:
- no data
Results and discussion
Results: maternal animals
Maternal developmental toxicity
- Details on maternal toxic effects:
- Maternal toxic effects:yes
Details on maternal toxic effects:
Treatment with the highest dose level (1000 mg/kg) of d-limonene resulted in death of dams with less than 40% mortality. The significant decrease of body- weight gain and food consumption were temporarily observed in dams given 500 and 1000 mg/kg of d-limonene, but no anomalies were observed in the general behaviour of dams given 250 and 500 mg/kg of d-limonene during the gestation.
Effect levels (maternal animals)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 250 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: maternal toxicity
Maternal abnormalities
- Key result
- Abnormalities:
- no effects observed
Results (fetuses)
- External malformations:
- no effects observed
- Skeletal malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- - Visceral and skeletal examinations revealed some anormalies such as incomplete lobulation of the lungs, enlargement of the foramen ovale and retarded ossification of the middle phalanx of fore limbs in addition to the 5th sternebrae. These did not appear to be dose-dependent and restored to normal during the postnatal development.
- Visceral malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- - Visceral and skeletal examinations revealed some anormalies such as incomplete lobulation of the lungs, enlargement of the foramen ovale and retarded ossification of the middle phalanx of fore limbs in addition to the 5th sternebrae. These did not appear to be dose-dependent and restored to normal during the postnatal development.
- Other effects:
- effects observed, non-treatment-related
- Description (incidence and severity):
- - Other non specific anormalies involved the lumber ribs in fetuses and offsprings, formation of the accessory ossification center of the 5th sternebrae in offsprings and the atrial septal defect detected in only 2 fetuses of a litter from dams treated with 250 mg/kg bw/day of d-limonene.
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:no effects
Effect levels (fetuses)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- > 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: fetotoxicity
Fetal abnormalities
- Key result
- Abnormalities:
- no effects observed
Overall developmental toxicity
- Key result
- Developmental effects observed:
- no
Any other information on results incl. tables
Table 1: Effect of d-limonene on prenatal development of rabbit fetuses
Dose (mg/kg bw) |
Control |
250 |
500 |
1000 |
No. of pregnant animals |
10 |
10 |
10 |
18 |
No. of dead clams |
0 |
0 |
0 |
6 |
(%) |
|
|
|
33 |
No. of examined clams |
10 |
10 |
10 |
10 |
No. of implantations |
96 |
94 |
85 |
91 |
(mean ± S.E.) |
9.50 ± 0.25 |
9.40 ± 0.21 |
8.50 ± 0.33 |
9.10 ± 0.25 |
No. of resorbed fetuses |
5 |
4 |
4 |
8 |
No. of dead fetuses |
3 |
5 |
0 |
3 |
No. of live fetuses |
88 |
85 |
81 |
80 |
Sex ratio (Male/Female) |
0.73 (37/51) |
1.13 (45/50) |
0.62 (31/50) |
1.11 (38/42) |
Fetus body weight (g) |
|
|
|
|
Male (mean ± S.E.) |
44.39 ± 1.33 |
48.09 ± 1.07 * |
44.76 ± 1.51 |
43.22 ± 0.96 |
Female (mean ± S.E.) |
45.64 ± 1.00 |
47.45 ± 1.08 |
46.14 ± 1.21 |
45.13 ± 1.10 |
Placental weight (g) |
|
|
|
|
Male (mean ± S.E.) |
5.76 ± 0.17 |
5.84 ± 0.17 |
5.95 ± 0.29 |
5.77 ± 0.19 |
Female (mean ± S.E.) |
5.87 ± 0.19 |
5.70 ± 0.15 |
6.16 ± 0.18 |
5.87 ± 0.23 |
* Significantly different from the control at 5% level
Table 2: Prenatal examinations of rabbit fetuses
Dose (mg/kg bw) |
Control |
250 |
500 |
1000 |
External examination |
|
|
|
|
No. of examined fetuses |
91 |
90 |
81 |
83 |
No. of malforrned fetuses |
0 |
0 |
0 |
0 |
Visceral examination |
|
|
|
|
No. of examined fetuses |
88 |
85 |
81 |
80 |
No. of malformed fetuses |
|
|
|
|
Atrial septal defect (%) |
0 |
2 (2.4) |
0 |
0 |
No. of minor abnormality |
|
|
|
|
Incomplete lobulation of lungs (%) |
11 (12.5) |
16 (18.8) |
19 (23.5) |
19 (23.8) |
Enlargement of foramen ovale (%) |
2 (2.3) |
2 (2.4) |
5 (6.2) |
4 (5.0) |
Skeletal examination |
|
|
|
|
No. of examined fetuses |
86 |
87 |
81 |
80 |
No. of malformed fetuses |
0 |
0 |
0 |
0 |
No. of variation |
|
|
|
|
Left lumbar rib (%) |
18 (20.9) |
26 (29.9) |
14 (17.3) |
25 (31.3) |
Right lumbar rib (%) |
16 (18.6) |
22 (25.3) |
14 (17.3) |
22( 27.5) |
Ossification pattern |
|
|
|
|
Retarded ossification of 5th sternebrae (%) |
11 (12.8) |
14 (16.1) |
8 (9.9) |
18 (22.5) |
Retarded ossification of middle phalanx of fore limbs (%) |
2 (2.3) |
3 (3.4) |
0 |
6 (7.4) |
Table 3: Absolute organ weights of rabbit offsprings
|
Male |
Female |
||||||
|
Control |
250 |
500 |
1000 |
Control |
250 |
500 |
1000 |
No. of offsprings |
13 |
12 |
8 |
13 |
10 |
13 |
16 |
9 |
Final body weight (g) |
893.0 ± 45.3 |
1021.2 ± 45.4 ** |
931.9 ± 55.5 |
957.3 ± 52.4 |
1005.5 ± 53.7 |
1093.1 ± 46.6 |
860.0 ± 31.6 * |
1071.7 ± 58.1 |
Liver (g) |
36.49 ± 2.52 |
45.08 ± 1.52 * |
38.67 ± 3.10 |
34.91 ± 2.76 |
41.79 ± 3.39 |
13.95 ± 3.18 |
34.68 ± 1.90 |
48.30 ± 6.14 |
Lungs (g) |
5.53 ± 0.35 |
6.25 ± 0.26 |
5.98 ± 0.36 |
5.46 ± 0.18 |
5.91 ± 0.25 |
5.94 ± 0.28 |
5.72 ± 0.18 |
5.93 ± 0.45 |
Heart (g) |
2.63 ± 0.17 |
3.50 ± 0.16 ** |
2.86 ± 0.17 |
3.04 ± 0.19 |
3.19 ± 0.17 |
3.38 ± 0.21 |
2.72 ± 0.12 * |
3.34 ± 0.20 |
Spleen (g) |
0.71 ± 0.05 |
0.77 ± 0.04 |
0.71 ± 0.08 |
0.81 ± 0.04 |
0.64 ± 0.06 |
0.74 ± 0.05 |
0.74 ± 0.04 |
0.78 ± 0.07 |
Thymus (g) |
2.31 ± 0.20 |
2.51 ± 0.23 |
2.11 ± 0.38 |
1.96 ± 0.11 |
2.40 ± 0.30 |
2.77 ± 0.19 |
1.671.14 * |
2.36 ± 0.31 |
Kidneys (g) |
7.67 ± 0.42 |
9.80 ± 0.46 ** |
8.29 ± 0.29 |
8.58 ± 0.52 |
8.82 ± 0.46 |
8.40 ± 0.30 |
7.86 ± 0.37 |
9.56 ± 0.55 |
Thyroids (mg) |
75.89 ± 8.35 |
102.68 ± 4.18* |
86.74 ± 10.97 |
80.93 ± 7.41 |
82.78 ± 8.00 |
89.90 ± 4.11 |
75.75 ± 5.43 |
96.30 ± 9.67 |
Adrenals (mg) |
69.65 ± 6.02 |
9.1.93 ± 1.06 ** |
78.79 ± 5.89 |
71.36 ± 6.00 |
82.23 ± 4.37 |
94.24 ± 5.07 |
87.64 ± 4.18 |
105.39 ± 15.11 |
Testes or Ovaries (mg) |
180.42 ± 17.15 |
272.86 ± 16.46 ** |
185.62 ± 23.78 |
162.84 ± 20.59 |
46.31 ± 7.90 |
46.10 ± 2.80 |
43.53 ± 2.69 |
47.77 ± 3.59 |
* Significantly different from the control at 5% level
** Significantly different from the control at 1% level
Table 4: Relative organ weights per 100 g of rabbit offsprings
|
Male |
Female |
||||||
|
Control |
250 |
500 |
1000 |
Control |
250 |
500 |
1000 |
No. of offsprings |
13 |
12 |
8 |
13 |
10 |
13 |
16 |
9 |
Final body weight (g) |
893.0 ± 45.3 |
1021.2 ± 45.4 ** |
931.9 ± 55.5 |
957.3 ± 52.4 |
1005.5 ± 53.7 |
1093.1 ± 46.6 |
860.0 ± 31.6 * |
1071.7 ± 58.1 |
Liver (g/100 g) |
4.08 ± 0.25 |
3.99 ± 0.22 * |
4.16 ± 0.26 |
3.52 ± 0.10 |
4.25 ± 0.17 |
4.03 ± 0.21 |
4.02 ± 0.16 |
4.04 ± 0.32 |
Lungs (g/100 g) |
0.61 ± 0.03 |
0.55 ± 0.03 |
0.65 ± 0.04 |
0.58 ± 0.03 |
0.62 ± 0.02 |
0.55 ± 0.02 * |
0.67 ± 0.03 |
0.51 ± 0.02 ** |
Heart (g/100 g) |
0.29 ± 0.01 |
0.31 ± 0.01 |
0.31 ± 0.01 |
0.31 ± 0.02 |
0.33 ± 0.01 |
0.31 ± 0.01 |
0.32 ± 0.01 |
0.29 ± 0.01 * |
Spleen (g/100 g) |
0.08 ± 0.01 |
0.07 ± 0.01 |
0.08 ± 0.01 |
0.08 ± 0 |
0.07 ± 0.01 |
0.07 ± 0.01 |
0.09 ± 0 |
0.07 ± 0.01 |
Thymus (g/100 g) |
0.25 ± 0.02 |
0.22 ± 0.02 |
0.25 ± 0.03 |
0.20 ± 0.01 * |
0.24 ± 0.03 |
0.25 ± 0.01 |
0.195 ± 0.01 |
0.20 ± 0.02 |
Kidneys (g/100 g) |
0.85 ± 0.04 |
0.86 ± 0.04 |
0.90 ± 0.03 |
0.88 ± 0.02 |
0.91 ± 0.02 |
0.80 ± 0.01 |
0.91 ± 0.01 |
0.83 ± 0.03 * |
Thyroids (mg/100 g) |
8.21 ± 0.69 |
9.10 ± 0.51 |
9.21 ± 0.87 |
8.18 ± 0.42 |
8.40 ± 0.56 |
8.41 ± 0.45 |
8.81 ± 0.57 |
8.13 ± 0.41 |
Adrenals (mg/100 g) |
7.82 ± 0.60 |
8.37 ± 0.40 |
8.58 ± 0.77 |
7.18 ± 0.28 |
8.61 ± 0.51 |
8.79 ± 0.57 |
9.15 ± 0.42 |
9.04 ± 0.14 |
Testes or Ovaries (mg/100 g) |
23.68 ± 1.31 |
19.68 ± 0.94 * |
19.48 ± 1.69 |
16.35 ± 1.56 |
5.15 ± 1.19 |
4.33 ± 0.32 |
5.19 ± 0.43 |
3.84 ± 0.32 |
* Significantly different from the control at 5% level
** Significantly different from the control at 1% level
Table 5: Effects of d-limonene on gross differentiations of rabbit offsprings
|
Control |
250 |
500 |
1000 |
No. of examined offsprings |
23 |
25 |
24 |
22 |
Days of gross differentiation after birth Opening of the ear-shell |
|
|
|
|
6th day (%) |
0 |
0 |
1 (4.2) |
0 |
7th day (%) |
23 (100) |
25 (100) |
23 (95.8) |
22 (100) |
Coating with the hair |
|
|||
2nd day (%) |
7 (30.4) |
0 |
0 |
0 |
3rd day (%) |
16 (69.6) |
25 (100) |
24 (100) |
22 (100) |
Odontiasis |
|
|||
At birth (%) |
23 (100) |
25 (100) |
24 (100) |
22 (100) |
Opening of the eyelids |
|
|||
9th day (%) |
0 |
0 |
0 |
3 (13.6) |
10th day (%) |
11 (47.8) |
4 (16.0) |
13 (54.2) |
5 (22.7) |
11th day (%) |
4 (17.4) |
15 (60.0) |
10 (41.7) |
12 (54.5) |
12th day (%) |
3 (13.0) |
5 (20.0) |
1 (4.2) |
2 (9.1) |
13th day (%) |
5 (21.7) |
1 (4.0) |
0 |
0 |
Table 6: Effects of d-limonene on postnatal development of rabbit offsprings
|
Control |
250 |
500 |
1000 |
No of dams |
3 |
3 |
3 |
3 |
No. of still-birth (Male/Female) |
1 (1/0) |
0 |
0 |
1 (0/1) |
No. of offsprings (Male/Female) At birth |
28 (15/13) |
27 (14/13) |
26 (8/18) |
27 (15/12) |
1st week |
28 (15/13) |
27 (14/13) |
25 (8/17) |
26 (14/12) |
2nd week |
26 (14/12) |
27 (14/13) |
25 (8/17) |
26 (14/12) |
3rd week |
24 (14/10) |
27 (14/13) |
25 (8/17) |
25 (14/11) |
4th week |
23 (13/10) |
26 (13/13) |
25 (8/17) |
22 (13/ 9) |
5th week |
23 (13/10) |
25 (12/13) |
25 (8/17) |
22 (13/ 9) |
6th week |
23 (13/10) |
25 (12/13) |
25 (8/17) |
22 (13/ 9) |
7th week |
23 (13/10) |
25 (12/13) |
24 (8/16) |
22 (13/ 9) |
Weanling rate (%) |
79.3 (81.2/76.9) |
92.6 (85.7/100) |
92.3 (100/88.9) |
78.6 (86.7/69.2) |
Table 7: Postnatal examinations of rabbit offsprings
|
Control |
250 |
500 |
1000 |
No. of dams |
3 |
3 |
3 |
3 |
No. of examined offsprings |
23 |
25 |
24 |
22 |
Sensory function |
Normal |
Normal |
Normal |
Normal |
External examination No. of malformed offsprings |
0 |
0 |
0 |
0 |
Visceral examination No. of malformed offsprings |
0 |
0 |
0 |
0 |
No. of minor abnormality Incomplete lobulation of lungs (%) |
2 (8.7) |
1 (4.0) |
0 |
0 |
Accessory spleen (%) |
2 (8.7) |
0 |
0 |
0 |
Protrusion of gall bladder (%) |
1 (4.3) |
1 (4.0) |
0 |
0 |
Skeletal examination No. of malformed offsprings |
0 |
0 |
0 |
0 |
No. of variation Left lumbar rib (%) |
4 (17.4) |
4 (16.0) |
4 (16.7) |
4 (18.2) |
Right lumbar rib (%) |
2 (8.7) |
6 (24.0) |
6 (25.0) |
4 (18.2) |
Translocation of caudal vertebrae (%) |
1 (4.3) |
0 |
1 (4.2) |
0 |
Ossification pattern Retarded ossification of 5th sternebrae (%) |
0 |
2 (8.0) |
0 |
1 (4.5) |
Accessory ossification center of 5th sternebrae (%) |
1 (4.3) |
2 (8.0) |
0 |
3 (13.6) |
Applicant's summary and conclusion
- Conclusions:
- Under the test conditions, d-limonene was not teratogenic in rabbit fetuses and the NOAEL for fetal toxicity was considered to be greater than 1000 mg/kg bw/day. The NOAEL for maternal toxicity was considered to be 250 mg/kg bw/day based on the decreased bodyweight gain.
- Executive summary:
In a prenatal developmental toxicity study, d-limonene was administered orally to groups of pregnant Japanese white rabbits at dose levels of 250, 500 and 1000 mg/kg bw/day for 13 days from Day 6 to 18 of gestation. Food consumption and bodyweights of pregnant rabbits were recorded during organogenesis. Caesarean sections were performed and the number of dead, live or resorbed fetuses, sex ratio and number of implantation sites were recorded. Fetuses were weighed and examined for external, visceral and skeletal malformations.
Treatment with the highest dose level (1000 mg/kg bw/day) of d-limonene resulted in death of 6/18 dams (33% mortality). The significant decrease of bodyweight gain and food consumption were temporarily observed in dams given 500 and 1000 mg/kg bw/day of d-limonene, but no anormalies were observed in the general behavior of dams given 250 and 500 mg/kg bw/day of d-limonene during the gestation. External examination of fetuses showed no anormalies. Visceral and skeletal examinations revealed some anormalies such as incomplete lobulation of the lungs, enlargement of the foramen ovale and retarded ossification of the middle phalanx of fore limbs in addition to the 5th sternebrae. These did not appear to be dose-dependent and restored to normal during the postnatal development. Other non specific anormalies involved the lumber ribs in fetuses and offsprings, formation of the accessory ossification center of the 5th sternebrae in offsprings and the atrial septal defect detected in only 2 fetuses of a litter from dams treated with 250 mg/kg bw/day of d-limonene.
Under the test conditions, d-limonene was not teratogenic in rabbit fetuses and the NOAEL for fetal toxicity was considered to be greater than 1000 mg/kg bw/day. The NOAEL for maternal toxicity was considered to be 250 mg/kg bw/day based on the decreased bodyweight gain.
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