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EC number: 218-827-2 | CAS number: 2244-16-8
Toxicity to reproduction
Administrative data
- Endpoint:
- two-generation reproductive toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- secondary literature
- Justification for type of information:
- Data from secondary source
Data source
Referenceopen allclose all
- Reference Type:
- secondary source
- Title:
- Scientific Opinion on the safety assessment of carvone, considering all sources of exposure
- Author:
- EFSA Scientific Committee / European Food Safety Authority (EFSA)
- Year:
- 2 014
- Bibliographic source:
- EFSA Journal 2014;12(7):3806
- Reference Type:
- secondary source
- Title:
- Proposal for Harmonised Classification and Labelling – Carvone
- Author:
- European Chemicals Agency (ECHA)
- Year:
- 2 012
- Bibliographic source:
- Proposal for Harmonised Classification and Labelling – Carvone,2012
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
- Principles of method if other than guideline:
- 2-Generation reproduction study of d-carvone was performed on Wistar Crl : (WI) BR rats.
- GLP compliance:
- not specified
- Limit test:
- no
Test material
- Reference substance name:
- (5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one
- Cas Number:
- 2244-16-8
- Molecular formula:
- C10H14O
- IUPAC Name:
- (5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one
- Details on test material:
- - Name of test material: D-Carvone- IUPAC name: (5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one- Molecular formula: C10H14O- Molecular weight: 150.22 g/mol- Substance type: Organic
Constituent 1
- Specific details on test material used for the study:
- - Name of test material: d-Carvone- IUPAC name: (5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one- Molecular formula: C10H14O- Molecular weight: 150.22 g/mole- Smiles :C1[C@H](CC=C(C1=O)C)C(C)=C- Inchl: 1S/C10H14O/c1-7(2)9-5-4-8(3)10(11)6-9/h4,9H,1,5-6H2,2-3H3/t9-/m0/s1- Substance type: Organic- Physical state: Colorless to pale-yellow liquid
Test animals
- Species:
- rat
- Strain:
- other: Wistar Crl : (WI) BR rats
- Sex:
- male/female
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Details on exposure:
- Details on exposurePREPARATION OF DOSING SOLUTIONS:The test material diluted with corn oil.DIET PREPARATION- Rate of preparation of diet (frequency):No data available- Mixing appropriate amounts with (Type of food )- Storage temperature of food: No data availableVEHICLE- Justification for use and choice of vehicle (if other than water): test material mixed with corn oil as it is not soluble in water - Concentration in vehicle: f0: 0, 3, 10 or 30 mg/kg/dayF1: 0, 10, 30 and 90 mg/kg/day.- Amount of vehicle (if gavage): No data available- Lot/batch no. (if required): No data available- Purity: No data available
- Details on mating procedure:
- - M/F ratio per cage:1:1 - Length of cohabitation: - Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy - After ... days of unsuccessful pairing replacement of first male by another male with proven fertility. - Further matings after two unsuccessful attempts: [no / yes (explain)] - After successful mating each pregnant female was caged (how): - Any other deviations from standard protocol:
- Analytical verification of doses or concentrations:
- not specified
- Duration of treatment / exposure:
- 90 days
- Frequency of treatment:
- daily
- Details on study schedule:
- - F1 parental animals not mated until [...] weeks after selected from the F1 litters.for 10 weeks - Selection of parents from F1 generation when pups were [...] days of age. - Age at mating of the mated animals in the study: [...] weeks
Doses / concentrations
- Remarks:
- F0: 0, 3, 10 or 30 mg/kg/dayF1: 0, 10, 30 and 90 mg/kg/day.
- No. of animals per sex per dose:
- Total:2000 mg/kg bw/day:25 male and 25 female 3 mg/kg bw/day:25 male and 25 female10 mg/kg bw/day:25 male and 25 female30 mg/kg bw/day:25 male and 25 female
- Control animals:
- yes, concurrent vehicle
Examinations
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes - Time schedule: daily - Cage side observations checked in table [No.?] were included. DETAILED CLINICAL OBSERVATIONS: Yes - Time schedule: daily BODY WEIGHT: Yes - Time schedule for examinations:weekly FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): weekly - Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data - Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data - Time schedule for examinations: OTHER:
- Sperm parameters (parental animals):
- Parameters examined in [all/P/F1/F2] male parental generations:sperm was assessed for motility and morphology plus one testis and epididymis was frozen for enumeration of homogenisation-resistant spermatids and cauda epididymal sperm reserves.
- Litter observations:
- STANDARDISATION OF LITTERS- Performed on day 4 postpartum: yes- If yes, maximum of [...] pups/litter ([...]/sex/litter as nearly as possible); excess pups were killed and discarded. PARAMETERS EXAMINEDThe following parameters were examined in [F1 ] offspring:Each litter was examined for number of live and dead pups as soon as possible after birth. The individual weights of all live pups were recorded at day 1, 4, 7, 14 and 21. The sex of all pups was recorded, as was the number with any physical or behavioural abnormality. For the F1 weanlings selected for mating the day of vaginal opening or balanopreputial separation was recordedGROSS EXAMINATION OF DEAD PUPS:[no / yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead.]ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:
- Postmortem examinations (parental animals):
- SACRIFICE - Male animals:. F0 males were killed as soon as possible after successful mating while F1 males were only killed after successful delivery of the dams - Maternal animals: Females from both generations were killed at day 21 post partum. GROSS NECROPSY: yes All parental animals were subject to a necropsy and some tissues weighed (adrenals, brain, epididymides, kidneys, liver, ovaries, pituitary, prostate, seminal vesicles, spleen, testes, thyroid, uterus with oviducts and cervix) also a range of tissues (adrenals, brain, coagulating glands, epididymides, identification marks, kidneys, liver, ovaries, pituitary, prostate, seminal vesicles, spleen, testes, thyroid, uterus with oviducts and cervix, vagina) preserved for histological examination. HISTOPATHOLOGY / ORGAN WEIGHTS The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
- Postmortem examinations (offspring):
- SACRIFICEAll pups not selected for mating were taken for necropsy at day 21 post partum or shortly thereafter; organ weights were recorded for brain, spleen and thymus. -
- Reproductive indices:
- Reproductive parameters recorded were percentage mating, fertility index, conception rate, gestation index, duration of gestation
- Offspring viability indices:
- percentage live offspring by sex, percentage of post-natal loss, percentage of breeding loss, percentage live at weaning by sex, viability index, weaning index.
Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- No treatment-related clinical signs were observed
- Dermal irritation (if dermal study):
- not specified
- Mortality:
- no mortality observed
- Description (incidence):
- Animals were found dead or killed in extremis due to bad health or delivery difficulties. The deaths are not considered to be treatment-related.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- The reduction in body weight and body weight gain in females of the F0 generation treated with 30 mg/kg bw/day is considered not oftoxicological significance
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not specified
- Haematological findings:
- not specified
- Clinical biochemistry findings:
- not specified
- Urinalysis findings:
- not specified
- Behaviour (functional findings):
- not specified
- Immunological findings:
- not specified
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Histopathology of the male kidneys showed changes typical of 2u -globulin nephropathy at all doses. No other histopathological changes were reported and no similar changes were observed in females
- Histopathological findings: neoplastic:
- not specified
- Other effects:
- not specified
Reproductive function / performance (P0)
- Reproductive function: oestrous cycle:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Reproductive function: sperm measures:
- no effects observed
- Description (incidence and severity):
- There were no differences between treated and control animals in the results of the sperm morphology and motility measurements
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- There were no differences between treated and control animals in any of the indices of reproductive performance
Effect levels (P0)
open allclose all
- Dose descriptor:
- NOAEL
- Effect level:
- 30 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- clinical signs
- mortality
- body weight and weight gain
- organ weights and organ / body weight ratios
- gross pathology
- histopathology: non-neoplastic
- Remarks on result:
- other: No effects on systemic toxicity
- Dose descriptor:
- NOAEL
- Effect level:
- 90 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- reproductive function (oestrous cycle)
- reproductive function (sperm measures)
- reproductive performance
- Remarks on result:
- other: overall reproductive performance
Target system / organ toxicity (P0)
- Critical effects observed:
- not specified
- System:
- other: not specified
- Organ:
- not specified
- Treatment related:
- not specified
- Dose response relationship:
- not specified
- Relevant for humans:
- not specified
Results: P1 (second parental generation)
General toxicity (P1)
- Clinical signs:
- no effects observed
- Dermal irritation (if dermal study):
- not specified
- Mortality:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- Females of the 90 mg/kg bw/day group had in increased body weight and body weight gain. The study authors considers this finding not toxicologically relevant
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not specified
- Ophthalmological findings:
- not specified
- Haematological findings:
- not specified
- Clinical biochemistry findings:
- not specified
- Urinalysis findings:
- not specified
- Behaviour (functional findings):
- not specified
- Immunological findings:
- not specified
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- Males of the F1 generation given 30 and 90 mg/kg bw/day had increased mean relative liver and kidney weights.
- Gross pathological findings:
- no effects observed
- Neuropathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Histopathology of the male kidneys showed changes typical of 2u -globulin nephropathy at all doses. No differences were seen in females and no other histopathological changes were reported. .
- Histopathological findings: neoplastic:
- not specified
- Other effects:
- not specified
Reproductive function / performance (P1)
- Reproductive function: oestrous cycle:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Reproductive function: sperm measures:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Reproductive performance:
- no effects observed
Effect levels (P1)
- Dose descriptor:
- NOAEL
- Effect level:
- 90 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- clinical signs
- mortality
- body weight and weight gain
- food consumption and compound intake
- organ weights and organ / body weight ratios
- gross pathology
- histopathology: non-neoplastic
- reproductive function (oestrous cycle)
- reproductive function (sperm measures)
- reproductive performance
- Remarks on result:
- other: overall no reproductive toxic effects was observed
Target system / organ toxicity (P1)
- Critical effects observed:
- not specified
- System:
- other: not specified
- Organ:
- not specified
- Treatment related:
- not specified
- Dose response relationship:
- not specified
- Relevant for humans:
- not specified
Results: F1 generation
General toxicity (F1)
- Clinical signs:
- not specified
- Dermal irritation (if dermal study):
- not specified
- Mortality / viability:
- no mortality observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Food consumption and compound intake (if feeding study):
- not specified
- Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not specified
- Haematological findings:
- not specified
- Clinical biochemistry findings:
- not specified
- Urinalysis findings:
- not specified
- Sexual maturation:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Histopathological findings:
- not examined
- Other effects:
- not specified
Developmental neurotoxicity (F1)
- Behaviour (functional findings):
- not specified
Developmental immunotoxicity (F1)
- Developmental immunotoxicity:
- not specified
Effect levels (F1)
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 90 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- viability
- sexual maturation
- mortality
- body weight and weight gain
- organ weights and organ / body weight ratios
- gross pathology
- Remarks on result:
- other: No overall developmental toxic effects was observed
Target system / organ toxicity (F1)
- Critical effects observed:
- not specified
- System:
- other: not specified
- Organ:
- not specified
- Treatment related:
- not specified
- Dose response relationship:
- not specified
- Relevant for humans:
- not specified
Results: F2 generation
General toxicity (F2)
- Clinical signs:
- not specified
- Dermal irritation (if dermal study):
- not specified
- Mortality / viability:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Food consumption and compound intake (if feeding study):
- not specified
- Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not specified
- Haematological findings:
- not specified
- Clinical biochemistry findings:
- not specified
- Urinalysis findings:
- not specified
- Sexual maturation:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- no toxicologically relevant effect
- Histopathological findings:
- not examined
- Other effects:
- not specified
Developmental neurotoxicity (F2)
- Behaviour (functional findings):
- not specified
Developmental immunotoxicity (F2)
- Developmental immunotoxicity:
- not specified
Effect levels (F2)
- Dose descriptor:
- NOAEL
- Generation:
- F2
- Effect level:
- 90 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- not specified
- Basis for effect level:
- viability
- sexual maturation
- body weight and weight gain
- organ weights and organ / body weight ratios
- gross pathology
- Remarks on result:
- other: no overall developmental toxic effects was observed
Target system / organ toxicity (F2)
- Critical effects observed:
- not specified
- System:
- other: not specified
- Organ:
- not specified
- Treatment related:
- not specified
- Dose response relationship:
- not specified
- Relevant for humans:
- not specified
Overall reproductive toxicity
- Reproductive effects observed:
- not specified
- Treatment related:
- not specified
- Relation to other toxic effects:
- not specified
- Dose response relationship:
- not specified
- Relevant for humans:
- not specified
Any other information on results incl. tables
Summary table of the 2 generation study
| Dose (mg/kg bw/day) | 0 |
| 3 |
| 10 |
| 30 |
| 90 |
| dr |
| Sex | m | f | m | f | m | f | m | f | m | f |
|
F0 | Mortality A | 2 | 1 |
| 1 |
| 1 |
| 1 |
|
|
|
| Clinical signs | no toxicologically relevant effect |
|
|
|
| ||||||
| Body weight (gain)B |
|
|
|
|
|
|
| ds |
|
|
|
| Food consumptionC |
|
|
|
|
|
|
| ds |
|
|
|
| Oestrus cycle | no toxicologically relevant effect |
|
|
|
| ||||||
| Sperm parameters | no toxicologically relevant effect |
|
|
|
| ||||||
| Organ weights -kidney -thyroids |
|
|
|
|
Isr |
Isr |
|
|
|
|
|
| Pathology | no toxicologically relevant effect |
|
|
| |||||||
| - macroscopy | no toxicologically relevant effect |
|
|
| |||||||
| - microscopyD | no toxicologically relevant effect |
|
|
| |||||||
F1 pups | Litter size | no toxicologically relevant effect |
|
|
| |||||||
| Survival index | no toxicologically relevant effect |
|
|
| |||||||
| Sex ratio | no toxicologically relevant effect |
|
|
| |||||||
| Body weight | no toxicologically relevant effect |
|
|
| |||||||
| Organ weight | no toxicologically relevant effect |
|
|
| |||||||
| Pathology |
|
|
|
| |||||||
| - macroscopy | no toxicologically relevant effect |
|
|
| |||||||
| - microscopy (weanlings) | not performed |
|
|
| |||||||
F1 animals | MortalityA
|
| 1 |
|
|
| 1 |
| 2 |
| 1 |
|
| Clinical signs |
|
|
|
| no toxicologically relevant effect |
| |||||
| Body weightE |
|
|
|
| no toxicologically relevant effect |
| |||||
| Food consumption |
|
|
|
| no toxicologically relevant effect |
| |||||
| Mating, fertility, gestation |
|
|
|
| no toxicologically relevant effect |
| |||||
| Oestrus cycle |
|
|
|
| no toxicologically relevant effect |
| |||||
| Sperm parameters |
|
|
|
| no toxicologically relevant effect |
| |||||
| Organ weights -liver -kidney |
|
|
|
|
|
| isr isar |
| isr isar |
| dr dr |
| Pathology |
|
|
|
|
|
|
| ||||
| - macroscopy |
|
|
|
| no toxicologically relevant effect |
| |||||
| - microscopyD |
|
|
|
| no toxicologically relevant effect |
| |||||
F2 pups | Litter size |
|
|
|
| no toxicologically relevant effect |
| |||||
| Survival index |
|
|
|
| no toxicologically relevant effect |
| |||||
| Sex ratio |
|
|
|
| no toxicologically relevant effect |
| |||||
| Body weight |
|
|
|
| no toxicologically relevant effect |
| |||||
| Organ weight |
|
|
|
| no toxicologically relevant effect |
| |||||
| Pathology |
|
|
|
| no toxicologically relevant effect |
| |||||
| - macroscopy |
|
|
|
| no toxicologically relevant effect |
| |||||
| - microscopy (weanlings) |
|
|
|
| not performed |
| |||||
dr = dose related; i = increased; d = decreased; is = increased significantly; ds = decreased significantly, a= absolute, r=relative
A Animals were found dead or killed in extremis due to bad health or delivery difficulties. The deaths are not considered to be treatment-related.
B Females of the 30 mg/kg bw/day group showed small but significant decreases in body weight gain during lactation on days
4, 7 and 14, and body weight loss on day 4 during lactation.
C Females of the 30 mg/kg bw/day group showed a significant decrease in food consumption from day 1-4 during lactation.
D Males of all treated groups showed histopathological changes in the kidney consistent with accumulation ofα2u-globulin.
E Females of the 90 mg/kg bw/day group had in increased body weight and body weight gain. The study authors consider this finding not toxicologically relevant. The present reviewers endorse this view
Applicant's summary and conclusion
- Conclusions:
- The NOAEL for systemic toxicity is 30 mg/kg bw/day. No effects on reproductive parameters were observed. Based on this the NOAEL for reproductive effects for treatment of two generations of rats is 90 mg/kg bw/day. When male and female Wistar Crl : (WI) BR rats were treated with d-carvone (2244-16-8) orally.
- Executive summary:
2-Generation reproduction study of d-carvone(2244-16-8) was performed on male and female Wistar Crl : (WI) BR rat according to GLP and to OECD Guideline 416 (OECD, 2001). The test material soluble in corn oil in dose concentration 0, 3, 10 or 30 mg/kg/day and administered by oral gavage for 10 weeks prior to mating until termination.Groups of 25 male and 25 female per dose group were used in F0 generation.For the F1 generation the dose levels were revised to 0, 10, 30 and 90 mg/kg/day. Females were mated individually and remained with the litters during weaning. Pups were not treated during the weaning period. On day 4 after birth all litters of more than 8 pups were culled to approximately 4 males and 4 females. After weaning of the F1 generation, animals were selected and again treated for 10 weeks prior to mating and up to termination. F0 males were killed as soon as possible after successful mating while F1 males were only killed after successful delivery of the dams. Females from both generations were killed at day 21 post partum. All animals were checked at least once daily for clinical signs and condition. Body weight and food consumption measurements were recorded weekly. All parental animals were subject to a necropsy and some tissues weighed (adrenals, brain, epididymides, kidneys, liver, ovaries, pituitary, prostate, seminal vesicles, spleen, testes, thyroid, uterus with oviducts and cervix) also a range of tissues (adrenals, brain, coagulating glands, epididymides, identification marks, kidneys, liver, ovaries, pituitary, prostate, seminal vesicles, spleen, testes, thyroid, uterus with oviducts and cervix, vagina) preserved for histological examination. All pups not selected for mating were taken for necropsy at day 21 post partum or shortly thereafter; organ weights were recorded for brain, spleen and thymus.
For males surviving to scheduled necropsy, sperm was assessed for motility and morphology plus one testis and epididymis was frozen for enumeration of homogenisation-resistant spermatids and cauda epididymal sperm reserves. For females, the ovaries and uterus were examined prior to fixation and the number of corpora lutea and implantation sites recorded. Each litter was examined for number of live and dead pups as soon as possible after birth. The individual weights of all live pups were recorded at day 1, 4, 7, 14 and 21. The sex of all pups was recorded, as was the number with any physical or behavioural abnormality. For the F1 weanlings selected for mating the day of vaginal opening or balanopreputial separation was recorded. Reproductive parameters recorded were percentage mating, fertility index, conception rate, gestation index, duration of gestation, percentage live offspring by sex, percentage of post-natal loss, percentage of breeding loss, percentage live at weaning by sex, viability index, weaning index.
There were no differences between treated and control animals in any of the indices of reproductive performance or in the results of the sperm morphology and motility measurements. Males of the F0 generation given 30 mg/kg bw/day had increased relative kidney weights. Histopathology of the male kidneys showed changes typical of 2u -globulin nephropathy at all doses. No other histopathological changes were reported and no similar changes were observed in females. Males of the F1 generation given 30 and 90 mg/kg bw/day had increased mean relative liver and kidney weights. Histopathology of the male kidneys showed changes typical of 2u -globulin nephropathy at all doses. No differences were seen in females and no other histopathological changes were reported.Hence The NOAEL for systemic toxicity is 30 mg/kg bw/day. No effects on reproductive parameters were observed. Based on this the NOAEL for reproductive effects for treatment of two generations of rats is 90 mg/kg bw/day.When male and femaleWistar Crl : (WI) BR ratswere treated with d-carvone(2244-16-8)orally.
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