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EC number: 289-612-9 | CAS number: 89957-91-5 Extractives and their physically modified derivatives such as tinctures, concretes, absolutes, essential oils, oleoresins, terpenes, terpene-free fractions, distillates, residues, etc., obtained from Citrus bergamia risso, Rutaceae.
- 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
Developmental toxicity / teratogenicity
Administrative data
- Endpoint:
- developmental toxicity
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- supporting study
- Study period:
- 1975
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- other: Original reference in Japanese language
Data source
Reference
- Reference Type:
- publication
- Title:
- Unnamed
- Year:
- 1 975
Materials and methods
- Principles of method if other than guideline:
- Prenatal developmental toxicity study: Groups of pregnant Wistar rats (20 animals/dose: 15 animals for teratogenicity study, 5 animals for postnatal development) were administered with d-limonene at dose levels of 0, 591 and 2869 mg/kg bw/day suspended with 1% gum-arabic solution (via oral route) for 7 days during gestation day 9 to 15. Maternal toxicity and developmental toxicity were evaluated.
- 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:
- (4R)-1-methyl-4-(prop-1-en-2-yl)cyclohexene
- Details on test material:
- - Name of test material (as cited in study report): d-limonene; d-p-mentha-1,8-diene
- Boiling point: 176-177 °C
- Specific gravity (at 25 °C): 0.8340-0.8420
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Wistar
- Details on test animals or test system and environmental conditions:
- No data
Administration / exposure
- Route of administration:
- oral: unspecified
- Vehicle:
- other: 1% gum-arabic solution
- Details on exposure:
- Volume administered: 5 mL/kg bw for all doses
- Analytical verification of doses or concentrations:
- not specified
- Details on analytical verification of doses or concentrations:
- None
- Details on mating procedure:
- no data
- Duration of treatment / exposure:
- 7 days (gestation Day 9-15)
- Frequency of treatment:
- Once daily
- Duration of test:
- Gestation Day 0 to postnatal week 7
Doses / concentrations
- Remarks:
- Doses / Concentrations:
0, 591 and 2869 mg/kg bw/day
Basis:
actual ingested
- No. of animals per sex per dose:
- 20 pregnant rats
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- No data
Examinations
- Maternal examinations:
- See result tables
- Ovaries and uterine content:
- See result tables
- Fetal examinations:
- See result tables
- 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:
Maternal deaths (40%) and decreased bodyweight gain at 2869 mg/kg bw/day
Effect levels (maternal animals)
open allclose all
- Dose descriptor:
- NOAEL
- Effect level:
- 591 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: maternal toxicity
- Dose descriptor:
- NOAEL
- Effect level:
- 591 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: developmental toxicity
Results (fetuses)
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:yes
Details on embryotoxic / teratogenic effects:
Significantly delayed ossification of metacarpal bones and proximal phalanxes in fetuses, significantly decreased bodyweight gain (male offspring) and organ weights at 2869 mg/kg bw/day
Fetal abnormalities
- Abnormalities:
- not specified
Overall developmental toxicity
- Developmental effects observed:
- not specified
Any other information on results incl. tables
Maternal examinations:
- At 2869 mg/kg bw/day, maternal bodyweight decreased (significant at GD 16) and several mothers (40%) died during the treatment period. At 591 mg/kg bw/day no maternal effects were observed.
Fetal examinations:
- Significantly delayed ossification of fetal metacarpal bone and proximal phalanx was found in the 2869 mg/kg bw/day dose group, compared to the control group. This effect was restored to normal within several weeks after birth.
- A decreased tendency of bodyweight was noted in postnatal male offspring of dams administered with 2869 mg/kg bw/day, compared to the control group.
- Relative weights of thymus, spleen and ovaries were significantly decreased in offspring of dams administered with 2869 mg/kg bw/day.
Table 1: Body weight changes in pregnant rats treated orally with d-limonene
Dose (mg/kg bw) |
Gestational days |
Gain |
||||
0 |
9 |
12 |
16 |
20 |
||
Control |
214.80 ± 32.55 |
248.70 ± 28.92 |
265.60 ± 30.53 |
289.85 ± 35.01 |
325.30 ± 44.01 |
105.50 ± 29.67 |
591 |
221.25 ± 39.58 |
254.65 ± 41.16 |
264.80 ± 39.94 |
290.10 ± 38.37 |
325.60 ± 52.75 |
103.95 ± 19.38 |
2869 |
214.75 ± 4.57 |
258.75 ± 32.76
|
248.92 ± 26.27 |
263.17 ± 22.96 * |
305.00 ± 27.07 |
90.25 ± 22.38 |
* Significantly different from the control, P <0.05
Table 2: Effects of d-limonene on rat fetuses
Dose (mg/kg bw) |
Control |
591 |
2869 |
No. of mothers |
15 |
15 |
15 |
Mortality of mothers (%) |
0 |
0 |
40 |
No. of total implants |
12.73 ± 2.96 |
12.18 ± 3.65 |
10.44 ± 3.71 |
No. of dead fetuses |
0 |
0 |
0 |
No. of resorbed fetuses |
1.00 ± 1.10 |
1.47 ± 2.42 |
0.89 ± 0.73 |
No. of live fetuses |
176 |
162 |
87 |
Sex ratio (Male/Female) |
0.69 |
1.22 |
0.85 |
Fetuses body weight (g) Male |
3.71 ± 0.45 |
3.53 ± 0.35 |
3.73 ± 0.52 |
Female |
3.46 ± 0.44 |
3.38 ± 0.45 |
3.63 ± 0.40 |
Placental weight(g) Male |
0.49 ± 0.07 |
0.49 ± 0.10 |
0.48 ± 0.06 |
Female |
0.47 ± 0.07 |
0.46 ± 0.06 |
0.44 ± 0.05 |
Malformation External |
0 |
0 |
0 |
Visceral |
1 |
0 |
0 |
Table 3: Effects of d-limonene on skeletal development of rat fetuses
Dose (mg/kg bw) |
Control |
591 |
2869 |
No. of examined fetuses |
83 |
84 |
42 |
Variation Shortness of 13th rid |
1 |
0 |
0 |
Lumbar rid |
0 |
1 |
2 |
Asymmetry of sternebrae |
0 |
0 |
1 |
Ossification Delayed ossification of parietal bone |
2 |
0 |
0 |
Non-ossification of occipital bone
|
0 |
4 |
1 |
Non-ossification of parietal bone |
0 |
3 |
0 |
No. of ossified metacarpal bone |
7.69 ± 0.72 |
7.49 ± 0.84 |
6.97 ± 0.96 * |
No. of ossified proximal phalanx (Forelimb) |
2.48 ± 1.71 |
2.25 ± 1.81 |
0.55 ± 1.28 * |
No. of ossified metatarsal bone
|
7.98 ± 0.56 |
8.01 ± 011 |
8.00 ± 0 |
No. of ossified sternebraea |
5.47 ± 0.98 |
5.60 ± 0.71 |
5.52 ± 0.73 |
No. of ossified caudal vertebrae |
3.76 ± 0.64 |
3.80 ± 0.57 |
3.95 ± 0.68 |
* Significantly different from the control, P <0.05
Table 4: Body weight changes of postnatal rat offsprings born to mothers treated orally with d-limonene
Postnatal weeks |
Males |
Females |
||||
Dose (mg/kg bw) |
Dose (mg/kg bw) |
|||||
Control |
591 |
2869 |
Control |
591 |
2869 |
|
0 |
5.19 ± 0.55 |
5.46 ± 0.52 |
4.79 ± 0.46 |
4.93 ± 0.62 |
5.09 ± 0.68 |
4.89 ± 0.66 |
1 |
13.06 ± 1.50 |
12.49 ± 0.99 |
10.62 ± 1.54 * |
12.82 ± 1.59 |
12.22 ± 1.07 |
10.66 ± 1.84 |
2 |
26.06 ± 3.12 |
24.86 ± 2.74 |
22.67 ± 5.05 * |
25.88 ± 3.35 |
24.31 ± 2.42 |
22.72 ±.3.92 |
3 |
41.91 ± 5.89 |
39.55 ± 5.14 |
40.77 ± 5.16 |
41.08 ± 5.59 |
38.56 ± 4.37 |
38.39 ± 4.96 |
4 |
73.12 ± 9.89 |
71.33 ± 8.87 |
67.67 ± 8.02 |
69.66 ± 9.43 |
67.38 ± 6.71 |
66.01 ± 9.29 |
5 |
122.32 ± 12.25 |
116.47 ± 12.78 |
112.77 ± 12.65 * |
109.57 ± 10.61 |
107.58 ± 7.95 |
107.10 ± 13.34 |
6 |
176.04 ± 15.80 |
164.68 ± 16.69 |
163.11 ± 17 .85 * |
141.39 ± 10.54 |
140.11 ± 9.40 |
139.45 ± 14.22 |
7 |
235.52 ± 17.72 |
222.43 ± 18.57 |
213.64 ± 20.10 * |
173.06 ± 8.89 |
169.65 ± 11.13 |
167.84 ± 15.86 |
* Significantly different from the control, P <0.05
Table 5: Effects of d-limonene on postnatal development of the rats
Dose (mg/ kg bw) |
Days of postnatal development |
|||||
Opening of the ear-shell |
Coating with |
Odontiasis |
Opening of the eyelid |
Descending of the testis |
Opening of the vaginal orifice |
|
Control |
2.55 ± 0.76 |
5.51 ± 0.91 |
10.1 ± 0.96 |
14.83 ± 0.55 |
22.5 ± 1.30 |
35.6 ± 2.50 |
591 |
2.09 ± 0.82 |
6.00 ± 0 |
10.4 ± 0.71 |
15.00 ± 0.76 |
21.6 ± 1.39 |
35.5 ± 1.75 |
2869 |
2.41 ± 0.49 |
8.50 ±0.50 |
10.4 ± 1.85 |
15.14 ± 0.75 |
21.27 ± 0.57 |
35.93 ± 2.20 |
Table 6: Effects of d-limonene on development of rat offsprings
Dose (mg/kg) |
Control |
591 |
2869 |
No. of mothers |
5 |
5 |
5 |
Mortality of mothers |
0 |
0 |
40 |
No. of offspring from birth to the 7th week 0 |
61 |
65 |
33 |
1 |
53 |
63 |
30 |
2 |
53 |
63 |
30 |
3 |
53 |
63 |
28 |
4 |
53 |
63 |
28 |
5 |
53 |
63 |
28 |
6 |
53 |
63 |
28 |
7 |
53 |
63 |
28 |
External abnormality |
0 |
0 |
0 |
No. of total implants |
13.6 ± 3.1 |
14.8 ± 1.7 |
13.7 ± 1.7 |
No. of dead fetuses at birth |
4 |
4 |
5 |
Parturition rate |
95 |
92 |
93 |
Weaning rate |
89 |
97 |
85 |
Table 7: Absolute organ weights of postnatal rat offsprings born to mothers treated orally with d-limonene
Sex |
Dose (mg/kg bw) |
No. of offsprings. |
Final BW (g) |
Pituitary (mg) |
Thyroids (mg) |
Thymus (g) |
Lungs (g) |
Heart (g) |
Spleen (g) |
Kidneys (g) |
Liver (g) |
Adrenals (g) |
Testes (g) or Ovaries (mg) |
Male |
Control |
27 |
235.5 ± 17.7 |
10.04 ± 1.95 |
14.06 ± 2.52 |
0.77 ± 0.08 |
1.15 ± 0.10 |
0.82 ± 0.08 |
0.75 ± 0.11 |
2.18 ± 0.36 |
11.55 ± 1.14 |
38.85 ± 7.37 |
2.15 ± 0.17 |
591 |
29 |
222.4 ± 18.6 |
9.58 ± 4.84 |
13.12 ± 2.77 |
0.72 ± 0.09 |
1.10 ± 0.11 |
0.81 ± 0.08 |
0.69 ± 0.08 |
2.10 ± 0.20 |
11.24 ± 1 .49 |
36.77 ± 6.93 |
2.13 ± 0.28 |
|
2869 |
11 |
213.6 ± 20.1 |
9.69 ± 0.65 |
14.41 ± 3.60 |
0.66 ± 0.08 * |
1.19 ± 0.17 |
0.80 ± 0.07 |
0.63 ± 0.07 * |
2.23 ± 0.38 |
11.64 ± 1.64 |
43.23 ± 10.91 |
2.18 ± 0.14 |
|
Female |
Control |
25 |
173.1 ± 8.9 |
11.18 ± 3.15 |
12.26 ± 1.32 |
0.59 ± 0.08 |
0.97 ± 0.11 |
0.66 ± 0.07 |
0.52 ± 0.06 |
1.72 ± 0.22 |
8.88 ± 0.85 |
45.51 ± 8.01 |
77.24 ± 22.01 |
591 |
34 |
169.7 ± 11.7 |
10.05 ± 3.34 |
11.57 ± 1.62 |
0.54 ± 0.07 |
0.96 ± 0.07 |
0.67 ± 0.05 |
0.50 ± 0.06 |
1.60 ± 0.15 * |
8.16 ± 0.85 |
46.56 ± 8.45 |
85.84 ± 42.52 * |
|
2869 |
17 |
167.8 ± 15.9 |
9.95 ± 1.87 |
12.31 ± 1.80 |
0.51 ± 0.07 * |
0.94 ± 0.10 |
0.62 ± 0.06 |
0.44 ± 0.04 * |
1.62 ± 0.17 * |
8.50 ± 0.58 |
47.15 ± 6.63 |
63.15 ± 7.99 |
* Significantly different from the control, P <0.05
Table 8: Relative organ weights per 100 g body weights of postnatal rat offsprings born to mothers treated orally with d-limonene
Sex |
Dose (mg/kg bw) |
No. of offsprings. |
Final BW (g) |
Pituitary (mg/100 g) |
Thyroids (mg/100 g) |
Thymus (mg/100 g) |
Lungs (mg/100 g) |
Heart (mg/100 g) |
Spleen (mg/100 g) |
Kidneys (g/100 g) |
Liver (g/100 g) |
Adrenals (mg/100 g) |
Testes or Ovaries (mg/100 g) |
Male |
Control |
27 |
235.5 ± 17.7 |
4.31 ± 0.78 |
5.98 ± 0.99 |
0.33 ± 0.04 |
0.48 ± 0.04 |
0.36 ± 0.03 |
0.31 ± 0.05 |
0.93 ± 0.08 |
4.89 ± 0.36 |
16.49 ± 2.71 |
0.90 ± 0.04 |
591 |
29 |
222.4 ± 18.6 |
4.02 ± 0.50 |
5.93 ± 0.86 |
0.33 ± 0.04 |
0.49 ± 0.04 |
0.37 ± 0.04 |
0.33 ± 0.09 |
0.95 ± 0.07 |
5.10 ± 0.37 |
16.39 ± 2.53 |
0.95 ± 0.08 * |
|
2869 |
11 |
213.6 ± 20.1 |
4.23 ± 0.43 |
5.93 ± 0.66 |
0.28 ± 0.03 * |
0.51 ± 0.05 |
0.35 ± 0.02 |
0.27 ± 0.02 * |
0.96 ± 0.06 |
5.03 ± 0.27 |
17.23 ± 2.26 |
0.95 ± 0.09 |
|
Female |
Control |
25 |
173.1 ± 8.9 |
6.09 ± 1.08 |
7.08 ± 0.85 |
0.34 ± 0.05 |
0.54 ± 0.04 |
0.38 ± 0.04 |
0.30 ± 0.04 |
0.99 ± 0.12 |
5.14 ± 0.42 |
26.38 ± 4.71 |
47.94 ± 9.78 |
591 |
34 |
169.7 ± 11.7 |
5.82 ± 0.81 |
6.85 ± 0.96 |
0.32 ± 0.04 |
0.54 ± 0.12 |
0.40 ± 0.03 |
0.30 ± 0.03 |
0.95 ± 0.07 |
4.83 ± 0.37 |
27.61 ± 3.76 |
46.74 ± 10.76 |
|
2869 |
17 |
167.8 ± 15.9 |
6.27 ± 1.90 |
7.31 ± 0.90 |
0.31 ± 0.03 * |
0.57 ± 0.08 |
0.37 ± 0.03 |
0.27 ± 0.04 * |
0.94 ± 0.06 |
5.14 ± 0.33 |
26.75 ± 2.93 |
36.89 ± 4.25 * |
* Significantly different from the control, P <0.05
Applicant's summary and conclusion
- Conclusions:
- Under the test conditions, the NOAEL for maternal toxicity was considered to be 591 mg/kg bw/day based on maternal deaths (40%) and decreased bodyweight gain. The NOAEL for fetal toxicity was considered to be 591 mg/kg bw/day based on significantly delayed ossification and decreased bodyweight gain in male offspring.
- Executive summary:
In this developmental toxicity study in rats, d-limonene was administered to pregnant Wistar rats (20 animals/dose: 15 animals for teratogenicity study, 5 animals for postnatal development) at dose levels of 0, 591 and 2869 mg/kg bw/day (via oral route, suspended with 1% gum-arabic solution) for 7 days during gestation day 9-15.
At 2869 mg/kg bw/day, maternal bodyweight decreased (significant at GD 16) and several mothers (40%) died during the treatment period. At the 591 mg/kg bw/day dose level no maternal effects were observed. Significantly delayed ossification of fetal metacarpal bone and proximal phalanx was found in the 2869 mg/kg bw/day dose group, compared to the control group. This effect was restored to normal within several weeks after birth. A decreased tendency of bodyweight was noted in postnatal male offspring of dams administered with 2869 mg/kg bw/day, compared to the control group.
Under the test conditions, the NOAEL for maternal toxicity was considered to be 591 mg/kg bw/day based on maternal deaths (40%) and decreased bodyweight gain at 2869 mg/kg bw/day. The NOAEL for fetal toxicity was considered to be 591 mg/kg bw/day based on significantly delayed ossification in fetuses, significantly decreased bodyweight gain (male offspring) and organ weights at 2869 mg/kg bw/day
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