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Administrative data

Description of key information

Carcinogenicity by oral route

Under the conditions of these 2-year gavage studies, d-carvone (2244-16-8) was considered to be not carcinogenic as there was no evidence of carcinogenic activity was observed, When male or female B6C3F1 mice administered d-carvone (2244-16-8)at 375 or 750 mg/ kg, 5 days per week for 2 years.

Key value for chemical safety assessment

Carcinogenicity: via oral route

Link to relevant study records
Reference
Endpoint:
carcinogenicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data from experimental study report
Reference:
Composition 0
Composition 0
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 451 (Carcinogenicity Studies)
Principles of method if other than guideline:
The carcinogenicity study of d-carvone was performed in B6C3F1 mice.
GLP compliance:
not specified
Test material information:
Composition 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
Species:
mouse
Strain:
B6C3F1
Details on species / strain selection:
no data available
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: Frederick Cancer Research Facility(Frederick, MDl)- Age at study initiation: 8weeks ,6days - Weight at study initiation: no data available- Fasting period before study: no data available - Housing: Animals were housed five per cage,Polycarbonate (Hazleton Systems, Inc., Aberdeen, MD , Bedding: Beta Chips (Northeastern Products, Inc., Warrensburg, NY) Cage Filters Reemaye spun-bonded polyester filters (Snow Filtration, Cincinnati, om)- Diet NIH 07 Rat and Mouse Ration (ZeiglerBros., Inc., Gardners, PA); availablead libitum- Water (e.g. ad libitum): Automatic watering system (Edstrom Industries, Waterford, WI); available ad libitum - Acclimation period: 13 days ENVIRONMENTAL CONDITIONS- Temperature (°C): 15-28°C- Humidity (%):23% 90%- Air changes (per hr): 6-12room air Changes/h- Photoperiod (hrs dark / hrs light): 12h/dIN-LIFE DATES: From: To:
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:Test material soluble in corn oil DIET PREPARATION- Rate of preparation of diet (frequency): no data available- Mixing appropriate amounts with (Type of food): no data - Storage temperature of food: no data availableVEHICLE- Justification for use and choice of vehicle (if other than water): Because the feed blends of d-carvone were found to be unstable under the feed blending and simulated animal exposure conditions and because d carvoneis insoluble in water, - Concentration in vehicle: 0,375, or 750 mglkg- Amount of vehicle (if gavage):10ml/kg - Lot/batch no. (if required):no data - Purity:
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The corn oil solutions were extracted with methanol and analysed by high-performance liquid chromatography with a Brownlee RP-18 column and ultraviolet detection at 229 nm
Duration of treatment / exposure:
103 wk (some mice received 1or 2 doses during wk 104)
Frequency of treatment:
5 days /week
Post exposure period:
no data available
Remarks:
0,375, or 750 mg/kg
No. of animals per sex per dose:
Total:3000mg/kg:50 male and 50 female 375mg/kg:50 male and 50 female750mg/kg:50 male and 50 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Because 1,500 mg/kg was lethal to both male and female mice in the 13-week studies, doses of d-carvone selected for mice for the 2-year studies were 375 and 750 mg/ kg, administered in corn oil by gavage, 5 days per week.
Positive control:
no data available
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes - Time schedule: twice day - Cage side observations checked in table [No.?] were included. DETAILED CLINICAL OBSERVATIONS: Yes - Time schedule: DERMAL IRRITATION (if dermal study): No data - Time schedule for examinations: BODY WEIGHT: Yes - Time schedule for examinations:weighed initially, 1 X wk for 13 wk, and then at least 1 X month FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data - Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data FOOD EFFICIENCY: - Body weight gain in kg/food consumption in kg per unit time X 100 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: OPHTHALMOSCOPIC EXAMINATION: No data - Time schedule for examinations: - Dose groups that were examined: HAEMATOLOGY: No data - Time schedule for collection of blood: - Anaesthetic used for blood collection: Yes (identity) / No / No data - Animals fasted: Yes / No / No data - How many animals: - Parameters checked in table [No.?] were examined. CLINICAL CHEMISTRY: No data - Time schedule for collection of blood: - Animals fasted: Yes / No / No data - How many animals: - Parameters checked in table [No.?] were examined. URINALYSIS:No data - Time schedule for collection of urine: - Metabolism cages used for collection of urine: Yes / No / No data - Animals fasted: Yes / No / No data - Parameters checked in table [No.?] were examined. NEUROBEHAVIOURAL EXAMINATION: No data - Time schedule for examinations: - Dose groups that were examined: - Battery of functions tested: sensory activity / grip strength / motor activity / other: OTHER:
Sacrifice and pathology:
GROSS PATHOLOGY: Yes HISTOPATHOLOGY: Yes (all organs and tissues were examined for grossly visible lesions. Tissues were preserved in 10% neutral buffered formalin, embeddedin paraffin, sectioned, and stained with hematoxylin and eosin.)
Other examinations:
no data available
Statistics:
The logistic regression, alternativemethods of statistical analysis were used, andThese include the life table test (Cox, 1972; Tarone, 1975), appropriate for rapidly lethal tumors, and the Fisher exact test and the Cochran-Armitage trend test (Armitage,1971; Gart et aI., 1979), procedures based on the overall proportion of tumor-bearing animals.Tests of significance include pairwise comparisons of each dosed group with vehicle controls and a test for an overall dose-response trend. Continuity-corrected tests were used in the analysis of tumor incidence, and reported P values are one-sided. The procedures described above also were used to evaluate selected non neoplastic lesions
Clinical signs:
no effects observed
Description (incidence and severity):
No compound-related clinical signs were observed.
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, non-treatment-related
Description (incidence):
The survival of both the 375mg/kg dose (after week 101) and 750 mg/kg dose (after week 92) groups of female mice were significantly greater than that of the vehicle controls. No significant differences were observed between any groups of male mice
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Mean body weights of dosed and vehicle control male mice were similar throughout most of the studies; mean body weights of dosed female mice were within 7% of those of vehicle controls throughout most of the studies.
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
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Atrophy of the olfactory epithelium and hyperplasia of the underlying Bowman's glands occurred together. These may have been related to reflux of d-carvone into the nose after the gavage needle was withdrawn, because inflammatory exudate and "foreign material" were often found in the nasal passage of dosed animals. Fibromas, sarcomas, fibrosarcomas, or neurofibrosarcomas (combined) were observed with a negative trend in male mice, and the reduced incidence was significant in 375mg/kg dose male mice (vehicle control, 9/50; 375mg/kg dose, 1/50; 750mg/kg dose, 3/50). Three haemangioma or haemangiosarcomas were observed in vehicle control male mice, but none was seen in dosed males; the difference was not significant. Abscesses of the ovary and the uterus occurred at a high incidence in vehicle control female mice and at much lower incidences in dosed female mice (ovary: vehicle control, 26/50; 375mg/kg dose, 9/48; 750mg/kg dose, 1/48; uterus: 10/50; 3/50; 0/50). The lesions were similar to those observed in other studies associated with Krebsiella sp. infections and are believed to be the cause of reduced survival in vehicle control female mice relative to that of dosed female mice.
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
No increases in tumor incidences were observed in mice administered d-carvone.
Other effects:
not specified
Relevance of carcinogenic effects / potential:
Under the conditions of these 2-year gavage studies, d-carvone (2244-16-8) was considered to be not carcinogenic as no increases in tumor incidences were observed in mice administered d-carvone. Despite the absence of clearly chemically related toxic effects in the 2-year studies, the doses used were considered adequate for assessment of potential carcinogenicity because higher doses caused deaths in the 13-week studies.
Dose descriptor:
NOAEL
Effect level:
750 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
histopathology: non-neoplastic
histopathology: neoplastic
Remarks on result:
other: No increases in tumor incidences were observed in mice administered d-carvone.
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

Summary table of the carcinogenicity study

Dose

Mg/kg bw /day

0

375

750

Dose related

 

m

f

m

f

m

f

 

Mortality

13/50

36/50

8/50

21/50

14/50

12/50

 

Mean survival (days

679

639

694

652

631

676

 

Clinical signs

no toxicologically relevant effects

 

Body weight gain

no toxicologically relevant effects

 

Histopathology

 

 

 

 

 

 

 

-forestomach

 

 

 

 

 

 

 

acanthosis, focal

2/48

5/47

3/48

2/47

5/47

7/49

 

acanthosis, multifocal

2/48

1/47

2/48

0/47

6/47

0/49

 

-Nasal cavity

 

 

 

 

 

 

 

glands hyperplasia

3/50

19/49

42/50*

45/49*

44/49*

49/50*

dr

atrophy olfact. epithelium

11/50

25/49

42/50*

46/49*

44/49*

49/50*

dr

acute multifocal

 

 

 

 

 

 

 

inflammation turbinate

0/50

5/49

3/50

22/49*

27/49*

39/50*

dr

-Kidney

 

 

 

 

 

 

 

chronic focal inflammation

2/50

1/50

5/50

2/50

7/50

4/50

 

-Rectum

 

 

 

 

 

 

 

acute focal inflammation

19/48

5/47

15/45

15/45

18/44

22/45

dr(f)

-Uterus

 

 

 

 

 

 

 

dilatation

-

5/50

-

7/50

-

14/50

 

endometrium hyperplasia

-

5/50

-

26/50

-

27/50

 

-Lymph node, mesenteric

 

 

 

 

 

 

 

multifocal lymph. hyperplas.

11/50

2/46

7/50

3/47

10/48

14/48

(f)

-Spleen

 

 

 

 

 

 

 

diffuse lymph. hyperplasia

4/50

4/50

2/50

3/49

2/48

16/50

(f)

multifocal lymph. hyperplas.

12/50

1/50

11/50

2/49

10/48

3/50

 

Tumor incidence (see also

below)

no toxicologically relevant effects

 

 

 

m/f = male/female, i/d = increased/decreased, is/ds = increased/decreased significantly, np = not performed, a/r =

absolute/relative, dr = dose-related. The probability of survival was estimated by the procedure of Kaplan and Meier. Life

Table tests, Logistic regression tests, Cochran-Armitage test and Fisher Exact test were applied to assess significance.

 

Summary of the incidence of neoplasms

Dose (mg/kg bw)

0

375

750

Liver

m

f

m

f

m

f

Hepatocellular carcinoma

50

50

50

50

49

50

Hepatocellular adenoma

5

0

3

2

3

1

Lymphoma malignant mixed

2

1

4

0

4

0

Stomach, forestomach

48

47

48

47

47

49

Papilloma squamous

1

0

1

3

0

0

Uterus

-

50

-

50

-

50

Polyp

-

0

-

0

-

2

Lymph node, mesenteric

50

46

50

47

49

48

Lymphoma malignant mixed

1

0

1

3

1

2

Spleen

50

50

50

49

48

50

Lymphoma malignant mixed

2

1

1

4

1

4

Skin

50

50

50

50

50

50

Back, subcutaneous tissue, fibroma

2

0

0

0

1

0

Subcutaneous tissue, neurofibrosarcoma

2

0

1

0

0

0

Subcutaneous tissue, sarcoma

4

0

0

1

2

0

Lung

50

50

50

50

50

50

Alveolar/bronchiolar adenoma

7

1

4

6

5

3

Harderian gland

50

50

50

50

50

50

adenoma

1

2

2

0

1

0

Kidney

50

50

50

50

50

50

Lymphoma malignant mixed

1

0

0

1

0

2

Multiple organs

50

50

50

50

50

50

Hemangioma

2

0

0

0

0

0

Lymphoma malignant mixed

4

1

1

4

1

4

Conclusions:
Under the conditions of these 2-year gavage studies, d-carvone (2244-16-8) was considered to be not carcinogenic as there was no evidence of carcinogenic activity was observed, When male or female B6C3F1 mice administered d-carvone (2244-16-8)at 375 or 750 mg/ kg, 5 days per week for 2 years.
Executive summary:

The carcinogenicity ofd-carvone (2244-16-8)was studied in male and femaleB6C3Flmicefor2years.50 males and 50 female per dose group were used. The test material in dose concentration0,375, or 750 mg/kg administered by oral gavage route in dose volume 10ml/kg for 5days /week in 103 wk (some mice received 1or 2 doses during wk 104). The feed blends of d-carvone were found to be unstable under the feed blending and simulated animal exposure conditions and becausedcarvone is insoluble in water, corn oil gavage was selected as the route of administration for these studies. The 21-day stability of d-carvone in corn oil at 0.5%(5mg/g) stored at room temperature or at 5° C was determined. The corn oil solutions were extracted with methanol and analysed by high-performance liquid chromatography with a Brownlee RP-18 column and ultraviolet detection at 229 nm. The d-carvone/corn oil solutions were found to be stable for at least 21 days when stored in the dark at room temperature or at 5° C. The corn oil solutions were also stable under simulated dosing conditions for at least 3 hours. Animals were housed five per cage. Feed and water were available ad libitum. All animals were observed two times per day. Body weights were recorded once per week for the first 13 weeks of the study and at least once per month thereafter. During necropsy, all organs and tissues were examined for grossly visible lesions. Tissues were preserved in 10% neutral buffered formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin.

 

No compound-related clinical signs were observed. The survival of both the 375mg/kg dose (after week 101) and 750 mg/kgdose(after week 92) groups of female mice were significantly greater than that of the vehicle controls. No significant differences were observed between any groups of male mice. Mean body weights of dosed and vehicle control male mice were similar throughout most of the studies; mean body weights of dosed female mice were within 7% of those of vehicle controls throughout most of the studies.

Atrophy of the olfactory epithelium and hyperplasia of the underlying Bowman's glands occurred together. These may have been related to reflux of d-carvone into the nose after the gavage needle was withdrawn, because inflammatory exudate and "foreign material" were often found in the nasal passage of dosed animals. Fibromas, sarcomas, fibrosarcomas, or neurofibrosarcomas (combined) were observed with a negative trend in male mice, and the reduced incidence was significant in 375mg/kg dose male mice (vehicle control, 9/50; 375mg/kg dose, 1/50; 750mg/kg dose, 3/50). Three haemangioma or haemangiosarcomas were observed in vehicle control male mice, but none was seen in dosed males; the difference was not significant. Abscesses of the ovary and the uterus occurred at a high incidence in vehicle control female mice and at much lower incidences in dosed female mice (ovary: vehicle control, 26/50; 375mg/kg dose, 9/48; 750mg/kg dose, 1/48; uterus: 10/50; 3/50; 0/50). The lesions were similar to those observed in other studies associated with Krebsiella sp. infections and are believed to be the cause of reduced survival in vehicle control female mice relative to that of dosed female mice.No increases in tumor incidences were observed in mice administered d-carvone. Despite the absence of clearly chemically relatedtoxic effects in the 2-year studies, the doses used were considered adequate for assessment of potential carcinogenicity because higher doses caused deaths in the 13-week studies. Hence Under the conditions of these 2-year gavage studies, d-carvone (2244-16-8) was considered to be not carcinogenic as there wasno evidence of carcinogenic activitywas observed, When male or female B6C3F1mice administered d-carvone (2244-16-8)at 375 or 750 mg/ kg, 5 days per week for 2 years.

 

 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
750 mg/kg bw/day
Study duration:
chronic
Species:
mouse
Quality of whole database:
Data from NTP report
System:
other: not specified
Organ:
not specified

Carcinogenicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Carcinogenicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Carcinogenicity by oral routeI

In given study, d-carvone (2244-16-8) has been investigated for Carcinogenicity to a greater or lesser extent. Often the studies based on in vivo experiments in rodents, i.e. most commonly in mice for d-carvone (2244-16-8) summaries below.

In experimental study conducted by U. S. Department of Health and Human Services (National Toxicology Program, 1990, N'TP TR 381, NIH Publication No. 90-2836) The carcinogenicity of d-carvone (2244-16-8)was studied in male and femaleB6C3Flmicefor2years.50 males and 50 female per dose group were used. The test material in dose concentration0,375, or 750 mg/kg administered by oral gavage route in dose volume 10ml/kg for 5days /week in 103 wk (some mice received 1or 2 doses during wk 104). The feed blends of d-carvone were found to be unstable under the feed blending and simulated animal exposure conditions and because d carvone is insoluble in water, corn oil gavage was selected as the route of administration for these studies. The 21-day stability of d-carvone in corn oil at 0.5%(5mg/g) stored at room temperature or at 5° C was determined. The corn oil solutions were extracted with methanol and analysed by high-performance liquid chromatography with a Brownlee RP-18 column and ultraviolet detection at 229 nm. The d-carvone/corn oil solutions were found to be stable for at least 21 days when stored in the dark at room temperature or at 5° C. The corn oil solutions were also stable under simulated dosing conditions for at least 3 hours. Animals were housed five per cage. Feed and water were available ad libitum. All animals were observed two times per day. Body weights were recorded once per week for the first 13 weeks of the study and at least once per month thereafter. During necropsy, all organs and tissues were examined for grossly visible lesions. Tissues were preserved in 10% neutral buffered formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin.

 No compound-related clinical signs were observed. The survival of both the 375mg/kg dose (after week 101) and 750 mg/kg dose(after week 92) groups of female mice were significantly greater than that of the vehicle controls. No significant differences were observed between any groups of male mice. Mean body weights of dosed and vehicle control male mice were similar throughout most of the studies; mean body weights of dosed female mice were within 7% of those of vehicle controls throughout most of the studies.

Atrophy of the olfactory epithelium and hyperplasia of the underlying Bowman's glands occurred together. These may have been related to reflux of d-carvone into the nose after the gavage needle was withdrawn, because inflammatory exudate and "foreign material" were often found in the nasal passage of dosed animals. Fibromas, sarcomas, fibrosarcomas, or neurofibrosarcomas (combined) were observed with a negative trend in male mice, and the reduced incidence was significant in 375mg/kg dose male mice (vehicle control, 9/50; 375mg/kg dose, 1/50; 750mg/kg dose, 3/50). Three haemangioma or haemangiosarcomas were observed in vehicle control male mice, but none was seen in dosed males; the difference was not significant. Abscesses of the ovary and the uterus occurred at a high incidence in vehicle control female mice and at much lower incidences in dosed female mice (ovary: vehicle control, 26/50; 375mg/kg dose, 9/48; 750mg/kg dose, 1/48; uterus: 10/50; 3/50; 0/50). The lesions were similar to those observed in other studies associated with Krebsiella sp. infections and are believed to be the cause of reduced survival in vehicle control female mice relative to that of dosed female mice. No increases in tumor incidences were observed in mice administered d-carvone. Despite the absence of clearly chemically related toxic effects in the 2-year studies, the doses used were considered adequate for assessment of potential carcinogenicity because higher doses caused deaths in the 13-week studies. Hence Under the conditions of these 2-year gavage studies, d-carvone (2244-16-8) was considered to be not carcinogenic as there was no evidence of carcinogenic activity was observed, When male or female B6C3F1mice administered d-carvone (2244-16-8)at 375 or 750 mg/ kg, 5 days per week for 2 years.

Thus, comparing this value with the criteria of CLP regulation, d-carvone (2244-16-8) can be not classified for Carcinogenicity in mice by oral route exposed for 2 years.

 

 

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation, d-carvone (2244-16-8) can be not classified for Carcinogenicity in mice by oral route exposed for 2 years.