3,7-dimethylocta-2,6-dienenitrile

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from study report by Commission of the European Communities

Data source

Reference

Reference 1

Reference Type:

publication

Title:

COMPARATIVE METABOLISM OF GERANYL NITRILE AND CITRONELLYL NITRILE IN MOUSE, RAT, AND HUMAN HEPATOCYTES

Author:

Raymond A. Kemper, Diane L. Nabb, Shawn A. Gannon, Timothy A. Snow, and Anne Marie Api

Year:

2006

Bibliographic source:

DRUG METABOLISM AND DISPOSITION, 2006 vol.34:pp1019-1029

Reference 2

Reference Type:

other:

Title:

Commission of the European Communities DG XI

Author:

Prepared by-Germany

Year:

2007

Bibliographic source:

CLASSIFICATION AND LABELLING OF DANGEROUS SUBSTANCES Recommended form to be used for the proposed classification and labelling of a dangerous substance under Directive 67/548/EEC, January 2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Principles of method if other than guideline:

Repeated dose 28 day oral toxiicty study was performed to determine the toxic nature of the test compound geranyl nitrile

GLP compliance:

not specified

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3,7-dimethylocta-2,6-dienenitrile

EC Number:

225-918-0

EC Name:

3,7-dimethylocta-2,6-dienenitrile

Cas Number:

5146-66-7

Molecular formula:

C10H15N

IUPAC Name:

3,7-dimethylocta-2,6-dienenitrile

Test material form:

other: Liquid

Details on test material:

- Name of test material: Geranyl nitrile
- Molecular formula: C10H15N
- Molecular weight: 149.236 g/mol
- Smiles notation: N#C\C=C(\CC\C=C(\C)C)C
- InChl: 1S/C10H15N/c1-9(2)5-4-6-10(3)7-8-11/h5,7H,4,6H2,1-3H3/b10-7+
- Substance type: Organic
- Physical state: Liquid

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

No details available

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

No data
PREPARATION OF DOSING SOLUTIONS: The test chemical was dissolved in olive oil at dose levels of 0, 50, 150 or 450 mg/Kg bw/day

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): Olive oil
- Concentration in vehicle: 0, 50, 150 or 450 mg/Kg bw/day
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

28 days + 14 days recovery

Frequency of treatment:

No data

Doses / concentrations

Remarks:

Doses / Concentrations:
0 (vehicle control), 50, 150 and 450 mg/kg bw/day
Basis:
no data

No. of animals per sex per dose:

Total: 60
0 mg/Kg bw/day: 10 males and 10 females
50 mg/Kg bw/day: 5 males and 5 females
150 mg/Kg bw/day: 5 males and 5 females
450 mfg/Kg bw/day: 10 males and 10 females

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:No data
- Rationale for animal assignment (if not random):No data
- Rationale for selecting satellite groups: After 4 weeks of treatment 5 animals per
sex of all dose groups were sacrificed (main group). The remaining 5 animals per sex of
control and high dose group were maintained for another 14 days without administration of
the test substance (recovery groups).
- Post-exposure recovery period in satellite groups: 14 days
- Section schedule rationale (if not random): No data

Positive control:

No data

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once a day
- Parameters noted: Mortality, signs of toxicity such as salivation and piloerection

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: prior to the start of the administration and weekly thereafter

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, weekly
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes (Food consumption)
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY: Yes
- 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): Yes
- Time schedule for examinations:weekly

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: end of the administration period
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. :No

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: end of the administration period
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined.: Total bilirubin, alanine transferase, urea, transitional epithelial cells, epithelial cell casts and granular casts, alkaline phosphatase, inorganic phosphate, glucose levels, triglycerides and protein storage (α2u-globulin)

URINALYSIS: Yes
- Time schedule for collection of urine: end of the administration period and recovery period
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined.: No data

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: after 4 weeks of treatment of administration and at the end of recovery period
- Dose groups that were examined:No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: Functional observational battery (FOB) as well as measurement of motor activity (MA)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, all animals were assessed by gross pathology
HISTOPATHOLOGY: Yes, all animals were assessed by histopathological examinations

Other examinations:

No data

Statistics:

No data

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

The salivation observed throughout all dose groups in both sexes is more likely caused by the physico-chemical properties (intensive smell and taste) of the test compound than reflecting an adverse effect.

Mortality:

mortality observed, treatment-related

Description (incidence):

The salivation observed throughout all dose groups in both sexes is more likely caused by the physico-chemical properties (intensive smell and taste) of the test compound than reflecting an adverse effect.

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

effects observed, treatment-related

Water consumption and compound intake (if drinking water study):

effects observed, treatment-related

Ophthalmological findings:

not specified

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Changes in clinicochemical parameters in the serum are assessed as treatment-related and are indicative of mild impairment of liver function.

Urinalysis findings:

effects observed, treatment-related

Behaviour (functional findings):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

No detailed data available

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Histopathology did not reveal any morphological correlate in the liver of any of the treated groups

Histopathological findings: neoplastic:

not specified

Details on results:

-Clinical signs and mortality:
In 450 mg/kg bw: Piloerection was noetd in 4 males nad 5 females, anogenital region was smeared with urine, salivation of different severity in all
animals of both sexes on several days of the study, predominantly after treatment was noted

Recovery period: Piloerection was noted in 2 females until study day 35

In 150 mg/Kg bw/day: anogenital region was smeared with urine in 1 male (during FOB) as well as 1 female (during clinical examinations) and salivation of different severity in all animals of both sexes on several days of the study, predominantly after treatment was noted

In 50 mg/Kg bw: Slight salivation in 3 males and 2 females on several days of the study, predominantly after treatment was noted

-Body weight and weight gain:
In 450 mg/kg bw day group:
Main dose group: Reduced body weight was noted in both sexes, up to -22% in males and -12% in females on day 28. Impaired body weight change in both sexes was observed, with a maximum of -58% in males and -73% in females, on day 7.

During recovery periods: Reduced body weight change in both sexes, -30% in males and -29% in females at the end of the recovery period. Decreased mean terminal body weight was observed in male rats (-16%)

In 150 mg/Kg bw/day: Reduction in body weight change was noted in male animals upto -9% on day 28. ALso impaired body weight change was noted in male rats on day 28

50 mg/kg bw/day: Impaired body weight change was noted in males on study day 28 (-11%)

-Food consumption and compound intake:
450 mg/kg bw/day: Reduced food consumption in both sexes during the whole study period was observed, up to -25% on day 7 in males and -26% on day 7 in females.
Recovery group: Reduction in food consumption was noted (12%) in males on day 35

150 mg/kg bw/day: Reduction in food consumption was noted in males on day 7 (-11%) and day 14 (-8%)

-Food efficiency:
450 mg/kg bw/day:Reduced food efficiency was noted in both sexes

-Water consumption and compound intake No data available

-Opthalmoscopic examination No data available

-Haematology: No detailed data available

-Clinical chemistry:
450 mg/kg bw/day: Increased total bilirubin (48% and 36% in males and females) and urinary specific gravity was noted in both sexes (+2.8% and 2.1% in males and females). Increased alanine aminotransferase (+54%), urea (+19%), urinary urobilinogen, transitional epithelial cells, epithelial cell casts and granular casts in the males as well as alkaline phosphatase (+48%) and inorganic phosphate (+15%) in the females. Decreased glucose (-17%) and
triglycerides (-50%) in the males and increased protien storage (α2u-globulin) was noted in the kidenys of male rats

Recovery group: Increased alanine aminotransferase in males (+20%) and alkaline phosphatase in females (+32%) was noted at the end of the recovery period

150 mg/kg bw/day group: Increased urea (+20%), urinary urobilinogen, transitional epithelial cells, epithelial cell casts and granular casts in the males. Increased alkaline phosphatase (+44%), total bilirubin (+21%) in the females was observed.

50 mg/Kg bw/day: an increase in the protein storage (α2u-globulin) in the kidneys of male rats was noted

-Urinanalysis:
450 mg/kg bw/day:a decrase in the urinary volume in both sexes (-28% and -31% in males and females) was noted

150 mg/kg bw/day group: A decrease in the urinary volume was observed in the females (-28%)

-Neurobehaviour:
450 mg/kg bw/day: Reduced rearing in females (-48%), impaired foot splay test in females (-12%) and reduced overall motor activity in male animals was observed

-Organ weights: No data available

-Gross pathology:No data available

-Histopathology: Histopathology did not reveal any morphological correlate in the liver of any of the treated groups

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

50 other: mg/kg/day

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No significant changes were noted at the dose mentioned and changes in the liver enzyme levels were noted in the 150 mg/Kg bw/day female rats

2

Dose descriptor:

NOAEL

Effect level:

< 50 other: mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Slightly decreased body weight change was noted in male rats at the end of treatment period

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

The slightly impaired body weight change in the high dose females and down to the low dose males was assessed as an adverse compound-related effect. There were no indication of any other treatment-related adverse effects of the investigated organs as above-mentioned. Moreover, no adverse findings were obtained within the central or peripheral nervous system, the reproductive system, and the immune system by the test article.

Applicant's summary and conclusion

Conclusions:

The No observed adverse effect level (NOAEL) was found to be 50 mg/kg bw/day in female rats and less than 50 mg/kg bw/day in male rats when exposed by Dimethylocta-2,6-dienenitrile to rats by gavage route.

Executive summary:

The 28-days subacute repeated dose study was performed according to OECD TG 407 on Male and female wistar rats by gavage route with the dose concentration of 0 (vehicle control), 50, 150 and 450 mg/kg bw/day in olive oil as vechicle. Control and high dose group (450 mg/Kg bw/day) consisted of each 10 animals per sex whereas low (50 mg/Kg bw/day) and mid (150 mg/Kg bw/day) dose group consisted of each 5 animals per sex. After 4 weeks of treatment 5 animals per sex of all dose groups were sacrificed (main group). The remaining 5 animals per sex of control and high dose group were maintained for another 14 days without administration of the test substance (recovery groups). The salivation observed throughout all dose groups in both sexes is more likely caused by the physico-chemical properties (intensive smell and taste) of the test compound than reflecting an adverse effect. Changes in clinicochemical parameters in the serum are assessed as treatment-related and are indicative of mild impairment of liver function. However, histopathology did not reveal any morphological correlate in the liver of any of the treated groups.There were no indication of any other treatment-related adverse effects of the investigated organs as above-mentioned. Hence, the No Observed Adverse Effect Level (NOAEL) was 50 mg/kg bw/day in female rats due to the changed liver enzyme levels in mid dose animals (150 mg/kg bw/day) and less than 50 mg/kg bw/day in male rats due to the slight decreased body weight change at the end of the treatment period.