Cyclododeca-1,5,9-triene

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Robust summary for 1,5,9-cyclododecatriene (revised).

Author:

DuPont Safety, Health & Environmental Excellence Center, Wilmington (Del., USA)

Year:

2003

Bibliographic source:

U.S. EPA, 46 pp

Reference 2

Reference Type:

publication

Title:

Unnamed

Year:

2002

Reference 3

Reference Type:

study report

Title:

Unnamed

Year:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Cyclododeca-1,5,9-triene

EC Number:

225-533-8

EC Name:

Cyclododeca-1,5,9-triene

Cas Number:

4904-61-4

Molecular formula:

C12H18

IUPAC Name:

cyclododeca-1,5,9-triene

Constituent 2

Reference substance name:

1,5,9-cyclododecatriene

IUPAC Name:

1,5,9-cyclododecatriene

Details on test material:

purity 99.86%

Test animals

Species:

rat

Strain:

other: Crl:CD (SD)IGS BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ORGANISMS
- Age: males 59 days, females 52 days (approximately)
- Number of animals: 10 per sex and per dose group
- Further information: see references

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

ADMINISTRATION / EXPOSURE
- Vehicle: corn oil
- Concentration in vehicle: 6, 20, 60 mg/ml
- Total volume applied: 5 ml/kg bw/dose

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analysis of the test substance indicated that the test substance was homogeneously distributed at all levels and that the concentrations were at the
targed level. Stability results indicated that the test substance was stable at all concentrations.

Duration of treatment / exposure:

Females were dosed during a premating period of approximately 4 weeks, a mating period of approximately 1 to 2 weeks, a gestation period of
approximately 3 weeks, and a lactation period of approximately 4 days.
Males were dosed through test day 55.

Frequency of treatment:

daily

Doses / concentrations

Remarks:

Doses / Concentrations:
30, 100, 300 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

Following the 4-week premating period, each female was paired with a male of the same dosage group during a 1-2 week mating period.

Positive control:

no

Examinations

Observations and examinations performed and frequency:

PARAMETERS ASSESSED DURING STUDY P: 
- Clinical signs: at least once daily
- Mortality: at least twice daily
- Body weight: weekly, for females also on lactation days 0 and 4
- Food consumption: weekly
- Ophthalmoscopic examination: Both eyes of all rats prior to study and  on test day 24, prior to clinical pathology evaluation (surviving rats).
- Clinical pathology evaluation on all rats after 4 weeks of dosing, and  on male rats at the time of scheduled sacrifice:   
- hematology / coagulation: erythrocyte count, total leukocyte count,  platelet count, hemoglobin concentration, hematocrit, differential  
leukocyte count, mean corpuscular volume, mean corpuscular hemoglobin,  mean corpuscular hemoglobin concentration, absolute reticulocyte  counts,  red cell distribution width, microscopic blood examination, activated  partial thromoboplastin time, prothrombin time.   
- clinical chemistry: alkaline phosphatase, alanine aminotransferase,  aspartate aminotransferase, sorbitol dehydrogenase, glucose, urea  nitrogen,  calcium, inorganic phosphorus, total bilirubin, cholesterol,  triglyceride, creatinine, total protein, albumin, globulin, sodium,  potassium, chloride.- urine: volume, specific gravity, urobilinogen, blood, glucose,  protein, appearance (quality, transparency, color), pH, bilirubin,  ketones, sediment  microscopy.
- Other: Neurobehavioral Functional Observational Battery (FOB) on all  study rats prior to exposure and following approximately 4 weeks of test  
substance administration.
OFFSPRING: gestation length, mating index, gestation index, fecundity  index, implantation site numbers, implantation efficiency, sex ratio,  pups 
born alive, viability index

Sacrifice and pathology:

ORGANS EXAMINED AT NECROPSY (MACROSCOPIC AND MICROSCOPIC):
- Time: Sacrifice and necropsy of males on day 56 and of females on days  56-64.
- Organ weights P: liver, kidneys, adrenal glands, thymus, brain, spleen,  heart, testes, epididymides. Calculation of ratios to final body and  brain
 weights.
- Histopathology P:    
all high dose and control rats: testes, epididymides, ovaries, gross  lesions;   
5 high dose and 5 control rats per sex: additional 37 tissues   5 females from other groups with >= 1 offspring: liver

Other examinations:

no other examinations

Statistics:

STATISTICAL METHODS: 
- All weight parameters (P, not F), gestation length, clinical pathology,  grip strength, foot splay: One-way analysis of variance followed with  
Dunnett's test or Kruskal-Wallis test; followed with Dunn's test.
- Incidence of clinical and FOB observations; mating, gestation,  fecundity indexes: Sequential application of Cochran-Armitage test for  trend
- Implantation site number and efficiency, sex ratio, pups born alive,  viability index: Jonckheere's test - Mean pup weights: Linear contrast of the least  square means
- Motor activity: Levene's and Shapiro-Wilk tests followed by repeated  measures analysis of variance followed by contrasts of Jonckheere's trend  test

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

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):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Details on results:

ACTUAL DOSE RECEIVED BY DOSE LEVEL BY SEX: At the targeted level
- Number of deaths at each dose:    
One high-dose male was sacrificed in extremis due to a dosing-related  injury.   
One high-dose female was found dead on day 57 from dystocia.   
No other deaths occured during the study.
TOXIC RESPONSE/EFFECTS BY DOSE LEVEL: 
- Body weight:   Decreased body weight gain was observed and considered to be  compound-related and of biological significance:   
300 mg/kg males (-19 % for days 1-56, not statistically significant;  weight -7%);   
100 and 300 mg/kg females (only during gestation: gains -13% and -20%,  weights -7% and -12%, respectively, statistically significant).   
Body weights of pups in the 300 mg/kg group were significantly  decreased (-17% on lactation day 4).
- Food/water consumption: Increased food consumption was observed and  considered to be compound-related and of biological significance:   
300 mg/kg males (+19%)   
100 mg/kg (+7%, not statistically significant) and 300 mg/kg females  (+13%, statistically significant) during gestation.   
As a consequence, food efficiency was significantly reduced:   
300 mg/kg males (-33%)   100 and 300 mg/kg females during gestation (+14 and +29%, respectively,  statistically significant).
- Ophthalmoscopic examination: no test substance-related effects
- Description, severity, time of onset and duration of clinical signs:  
There were no test substance-related effects on clinical observations as  well as in neurobehavioral parameters or motor activity.
- Fertility index: no test substance-related effects
- Duration of gestation: no test substance-related effects
- Gestation index: no test substance-related effects
- Hematological findings incidence and severity: no test  substance-related effects
- Clinical biochemistry findings incidence and severity: no test  substance-related effects
- Gross pathology incidence and severity: no test substance-related  effects
- Number of implantations: no test substance-related effects
- Organ weight changes: no test substance-related adverse effects.   
- Liver weights were statistically significantly increased in several  dosed groups:   
Males: 30 mg/kg +10% relative; 100 mg/kg +14 % absolute, +21% relative;  300 mg/kg +34 % absolute, +45% relative.   
Females: 100 mg/kg +15% relative; 300 mg/kg +36% absolute, +42%  relative.   
In females it was associated with microscopic centrilobular  hepatocellular hypertrophy, which is indicative of a physiological  response-induction 
of enzymes associated with metabolism, and is not  considered biologically adverse.   
In males, minimal diffuse hypertrophy may have occurred, which is hard  to identify histologically.   
- Kidney weights were statistically significantly increased in several  dosed groups:   
Males: 100 mg/kg +16% absolute, +21% relative; 300 mg/kg +22%, +37%  relative   
Females: 100 mg/kg +11% relative; 300 mg/kg +15% absolute, +17% relative   
These findings were associated with some evidence of diuresis in  high-dose males and females, but there were no compound-related changes  in 
any other kidney parameter including histopathology. These weight  changes may be an adaptive response to the physiological changes that  
occur following administration of large doses of a test material. They  were not considered to be biologically adverse.
- Histopathology incidence and severity: no test substance-related  adverse effects
- Sex and sex ratios: no test substance-related effects
- Viability index: no test substance-related effects
- Other observations   
Mating index: no test substance-related effects   
Implantation efficiency: no test substance-related effects   
Pups born alive: no test substance-related effects

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

30 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Decreased of body weight gain was osberved at 100 and 300 mg/kg bw/d.

2

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Decreased of body weight gain was osberved at 300 mg/kg bw/d.

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

no further remarks

Applicant's summary and conclusion

Conclusions:

Based on the decreased of bobyweight gain, the NOAEL was 30 mg/kg bw/d for female rat, and 100 mg/kg bw/d for male rats, after oral administration of 1,5,9-cyclododecatriene.

Executive summary:

Male and female rats were administered an oral daily dose of 0, 30, 100, or 300 mg/kg/day 1,5,9-cyclododecatriene by gavage. Females were dosed during a premating period of approximately 4 weeks, a mating period of approximately 1 to 2 weeks, a gestation period of approximately 3 weeks, and a lactation period of approximately 4 days. Females that were not gestating were

dosed through the day prior to sacrifice. Males were dosed through test day 55.

A test-substance related, biologically significant decrease in body weight gains occurred in male rats administered 300 mg/kg/day. Decreased body weight gain in the 300 mg/kg/day males was accompanied by increased food consumption and decreased food efficiency. Females administered 100 or 300 mg/kg/day had test substance-related, significantly decreased body weight and

body weight gain during gestation that was accompanied by a significant increase in food consumption (300 mg/kg/day

only) and significantly decreased food efficiency in 100 and 300 mg/kg/day females.

There were no test substance-related effects on clinical observations in males and females during the premating phase or in females during gestation or lactation. There were no test substance-related effects on reproductive parameters. There were no test substance-related changes in neurobehavioral parameters or motor activity. There were no toxicologically significant changes in

hematology, coagulation, clinical chemistry, or urinalysis parameters in males or females. There were no test substance-related, biologically adverse changes in organ weights or tissue morphology in males or females.

Based on the decreased of bobyweight gain, the NOAEL was 30 mg/kg bw/d for female rat, and 100 mg/kg bw/d for male rats, after oral administration of 1,5,9-cyclododecatriene.