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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Description of key information

Oral

2000: According to OECD 422; GLP; 10 Sprague Dawley rats; 30, 100, 300 mg/kg bw/day; no specific target organ toxicity; no mortality; NOAEL males and females: 100 and 30 mg/kg bw/day, resp. (K1)

Inhalation

1999: No guideline stated; GLP not specified; 10 Sprague Dawley rats; 5, 50, 260 ppm; no specific target organ toxicity; no mortality; NOAEC 33.4 mg/m³ (K2)

Dermal

No study available

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
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:
guideline study
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
1986
GLP compliance:
yes
Limit test:
no
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
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:
males through test day 55; females 4 weeks premating through 4-day lactation period
Frequency of treatment:
daily
Dose / conc.:
30 other: mg/kg bw/day
Dose / conc.:
100 other: mg/kg bw/day
Dose / conc.:
300 other: mg/kg bw/day
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
Post-exposure period: none
MATING PROCEDURES: mating period approximately 1-2 weeks
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
Clinical signs:
no effects observed
Description (incidence and severity):
There were no test substance-related effects on clinical observations
Mortality:
mortality observed, non-treatment-related
Description (incidence):
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.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
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).
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
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.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
300 mg/kg males (-33%)
100 and 300 mg/kg females during gestation (+14 and +29%, respectively, statistically significant).
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No test substance-related effects
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No test substance-related effects in neurobehavioral parameters and motor activity
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
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.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No test substance-related effects
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
- Fertility index: no test substance-related effects
- Duration of gestation: no test substance-related effects
- Gestation index: no test substance-related effects
- Number of implantations: no test substance-related effects
- Mating index: no test substance-related effects
- Implantation efficiency: no test substance-related effects
- Pups born alive: no test substance-related effects
Details on results:
TOXIC RESPONSE/EFFECTS BY DOSE LEVEL:
- Body weights of pups in the 300 mg/kg group were significantly decreased (-17% on lactation day 4).
- Sex and sex ratios: no test substance-related effects
- Viability index: no test substance-related effects
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
body weight and weight gain
Remarks on result:
other: associated with increased FC and decreased food efficiency
Dose descriptor:
NOAEL
Effect level:
30 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
body weight and weight gain
Critical effects observed:
no
Conclusions:
According to the outcome of the study it is concluded that the no-observed -adverse-effect level (NOAEL) was 100 mg/kg for male rats and 30 mg/kg/day for female rats.
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.

According to the outcome of the study it is concluded that the no-observed -adverse-effect level (NOAEL) was 100 mg/kg for male rats and 30 mg/kg/day for female rats.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
30 mg/kg bw/day
Study duration:
subacute
Species:
rat

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Principles of method if other than guideline:
Method: as described in the reference (Bamberger et al)
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl:CD(R)(SD)BR
Sex:
male
Details on test animals or test system and environmental conditions:
Strain: Crl:CD(R)(SD)BR
- Source: Charles River Breeding Laboratories, Raleigh (USA)
- Age: ca. 8 weeks
- Weight at study initiation: ca. 245 g (mean)
further information : see reference (Bamberger et al)
Route of administration:
inhalation
Type of inhalation exposure:
nose only
Vehicle:
air
Remarks on MMAD:
MMAD / GSD: Particle size: vapour at 5 and 50 ppm;
 aerosol at 260 ppm;   particle size: MMAD 3.5 or 3.7 um,    35 or 36% of particles < 3 um;   98 or 99% of particles < 10 um
Details on inhalation exposure:
see reference
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
-analysed by gas chromatography and gravimetric analysis in 60 min intervalls
Duration of treatment / exposure:
2 weeks
Frequency of treatment:
6 hours/day, 5 days/week, total 9 exposures
Dose / conc.:
5 ppm
Remarks:
33.4 mg/m3; 5 +/- 0 ppm (analytical concentration)
Dose / conc.:
50 ppm
Remarks:
334 mg/m3; 51 +/- 1.0 ppm (analytical concentration)
Dose / conc.:
260 ppm
Remarks:
1740 mg/m3; 260 +/- 5.7 ppm (analytical concentration)
No. of animals per sex per dose:
10 per group
Control animals:
yes, concurrent no treatment
Details on study design:
- Number of animals: 10 animals per group
-  2 groups per concentration:     
1 for standard toxicological evaluations      
1 for neurotoxicity testing
-Post-exposure period: 2 weeks (half of the animals)
- SATELLITE GROUPS AND REASONS THEY WERE ADDED: neurotoxicity
Observations and examinations performed and frequency:
- Clinical signs: groupwise during exposure, individually after each  exposure
- Mortality: before / during / after exposure
- Body weight: before each exposure; daily except weekends during  recovery period
- Haematology: end of exposure period
- Biochemistry: end of exposure period
- Urinalysis: end of exposure period
Sacrifice and pathology:
ORGANS EXAMINED AT NECROPSY (MACROSCOPIC AND MICROSCOPIC): 

-5 animals per group sacrificed 1 day after last exposure; other animals sacrificed after (additional) 14 days of recovery

- Macroscopic:    weights of liver, kidneys, lungs, testes, brain;   examination of liver, kidneys, lungs, duodenum, heart, spleen, brain  (cerebrum, midbrain, cerebellum, medulla / pons), spinal cord, stomach,  jejunum, ileum, pancreas, cecum, colon, rectum, mesenteric lymph node,  thymus, adrenal glands, sciatic nerve, thyroid gland, parathyroid glands,  trachea, esophagus, pharynx / larynx, eyes, prostate, seminal vesicles,  urinary bladder, testes epididymides, sternum (with bone marrow), nose;  samples of all plus of gross lesions were saved

- Microscopic:    control and high dose animals sacrificed without recovery period: all  organs listed above (macroscopic examination);   animals from other dose groups sacrificed without recovery and control  and high dose animals sacrificed after recovery: nose, pharynx / larynx,  lungs, liver, kidneys
Other examinations:
OTHER EXAMINATIONS: 
Neurotoxicity groups   Functional observational battery (FOB) of tests   + motor activity (MA) evaluations   immediately after 4th and 9th exposures;   Neuropathology evaluation in 6/10 control and high dose animals after  9th exposure
Statistics:
- Mean, standard deviation, standard error;
- Incidences: Cochran-Armitage test for trend;
- Body & organ weights: one-way analysis of variance;
- Pairwise comparison with controls: Dunnett's test
- Homogeneity of variances: Bartlett's test
- Clinical pathology: one-way analysis of variance, Bartlett's test;  Dunnett's test for comparison of means to control; Kruskal-Wallis test  and Mann-Whitney U test when Bartlett's test showed significance  (p<0.005) 
- FOB: Cochran-Armitage test for trend, Fisher's exact test for  significance of deviations from control 
- Other test in neurobehavioral evaluations: Bartlett's test for  homogeneity (p<0.005), univariate analysis of variance, Shapiro-Wilk's  test, Levene's test, Kruskal-Wallis followed by Dunn's test, block (all  p<0.05)
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Irregular respiration in the high dose group. Other clinical observations occurred sporadically in all groups including controls and were considered incidental.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
significantly reduced at >= 50 ppm, reversible
Haematological findings:
no effects observed
Description (incidence and severity):
Compound-related or biologically adverse changes did not occur during this study.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Compound-related or biologically adverse changes did not occur during this study.
Urinalysis findings:
no effects observed
Description (incidence and severity):
Statistically significant or biologically adverse changes did not occur during this study.
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
A diminished or absent response to an alerting stimulus was observed in the high dose group. Mean foot splay was significantly decreased on days 4 and 9 in the high dose group. The incidence of defecation was decreased in the 50 and 260 ppm groups during the motor activity assessments. Beyond this, there were no statistically significant or toxicologically remarkable neurobehavioral differences between the control and treatment groups.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment-related changes were observed. (Absolute but not relative lung weights were reduced in the high dose group.)
Gross pathological findings:
no effects observed
Description (incidence and severity):
No treatment-related changes were observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Effects only occurred in the nasal tissue of the high dose group. Minimal degeneration / necrosis of olfactory epithelium occurred in the anterior portion of the nose of 4/5 rats exposed to 260 ppm. This lesion was observed neither in the 260 ppm recovery group, which suggests reversibility, nor in the lower dosed groups.
Dose descriptor:
NOAEC
Effect level:
33.4 mg/m³ air
Sex:
male
Basis for effect level:
other: no adverse effects observed
Dose descriptor:
LOAEC
Effect level:
334 mg/m³ air
Based on:
test mat.
Sex:
male
Basis for effect level:
body weight and weight gain
Critical effects observed:
no
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEC
33.4 mg/m³
Study duration:
subacute
Experimental exposure time per week (hours/week):
30
Species:
rat

Repeated dose toxicity: inhalation - local effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Principles of method if other than guideline:
Method: as described in the reference (Bamberger et al)
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl:CD(R)(SD)BR
Sex:
male
Details on test animals or test system and environmental conditions:
Strain: Crl:CD(R)(SD)BR
- Source: Charles River Breeding Laboratories, Raleigh (USA)
- Age: ca. 8 weeks
- Weight at study initiation: ca. 245 g (mean)
further information : see reference (Bamberger et al)
Route of administration:
inhalation
Type of inhalation exposure:
nose only
Vehicle:
air
Remarks on MMAD:
MMAD / GSD: Particle size: vapour at 5 and 50 ppm;
 aerosol at 260 ppm;   particle size: MMAD 3.5 or 3.7 um,    35 or 36% of particles < 3 um;   98 or 99% of particles < 10 um
Details on inhalation exposure:
see reference
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
-analysed by gas chromatography and gravimetric analysis in 60 min intervalls
Duration of treatment / exposure:
2 weeks
Frequency of treatment:
6 hours/day, 5 days/week, total 9 exposures
Dose / conc.:
5 ppm
Remarks:
33.4 mg/m3; 5 +/- 0 ppm (analytical concentration)
Dose / conc.:
50 ppm
Remarks:
334 mg/m3; 51 +/- 1.0 ppm (analytical concentration)
Dose / conc.:
260 ppm
Remarks:
1740 mg/m3; 260 +/- 5.7 ppm (analytical concentration)
No. of animals per sex per dose:
10 per group
Control animals:
yes, concurrent no treatment
Details on study design:
- Number of animals: 10 animals per group
-  2 groups per concentration:     
1 for standard toxicological evaluations      
1 for neurotoxicity testing
-Post-exposure period: 2 weeks (half of the animals)
- SATELLITE GROUPS AND REASONS THEY WERE ADDED: neurotoxicity
Observations and examinations performed and frequency:
- Clinical signs: groupwise during exposure, individually after each  exposure
- Mortality: before / during / after exposure
- Body weight: before each exposure; daily except weekends during  recovery period
- Haematology: end of exposure period
- Biochemistry: end of exposure period
- Urinalysis: end of exposure period
Sacrifice and pathology:
ORGANS EXAMINED AT NECROPSY (MACROSCOPIC AND MICROSCOPIC): 

-5 animals per group sacrificed 1 day after last exposure; other animals sacrificed after (additional) 14 days of recovery

- Macroscopic:    weights of liver, kidneys, lungs, testes, brain;   examination of liver, kidneys, lungs, duodenum, heart, spleen, brain  (cerebrum, midbrain, cerebellum, medulla / pons), spinal cord, stomach,  jejunum, ileum, pancreas, cecum, colon, rectum, mesenteric lymph node,  thymus, adrenal glands, sciatic nerve, thyroid gland, parathyroid glands,  trachea, esophagus, pharynx / larynx, eyes, prostate, seminal vesicles,  urinary bladder, testes epididymides, sternum (with bone marrow), nose;  samples of all plus of gross lesions were saved

- Microscopic:    control and high dose animals sacrificed without recovery period: all  organs listed above (macroscopic examination);   animals from other dose groups sacrificed without recovery and control  and high dose animals sacrificed after recovery: nose, pharynx / larynx,  lungs, liver, kidneys
Other examinations:
OTHER EXAMINATIONS: 
Neurotoxicity groups   Functional observational battery (FOB) of tests   + motor activity (MA) evaluations   immediately after 4th and 9th exposures;   Neuropathology evaluation in 6/10 control and high dose animals after  9th exposure
Statistics:
- Mean, standard deviation, standard error;
- Incidences: Cochran-Armitage test for trend;
- Body & organ weights: one-way analysis of variance;
- Pairwise comparison with controls: Dunnett's test
- Homogeneity of variances: Bartlett's test
- Clinical pathology: one-way analysis of variance, Bartlett's test;  Dunnett's test for comparison of means to control; Kruskal-Wallis test  and Mann-Whitney U test when Bartlett's test showed significance  (p<0.005) 
- FOB: Cochran-Armitage test for trend, Fisher's exact test for  significance of deviations from control 
- Other test in neurobehavioral evaluations: Bartlett's test for  homogeneity (p<0.005), univariate analysis of variance, Shapiro-Wilk's  test, Levene's test, Kruskal-Wallis followed by Dunn's test, block (all  p<0.05)
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Irregular respiration in the high dose group. Other clinical observations occurred sporadically in all groups including controls and were considered incidental.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
significantly reduced at >= 50 ppm, reversible
Haematological findings:
no effects observed
Description (incidence and severity):
Compound-related or biologically adverse changes did not occur during this study.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Compound-related or biologically adverse changes did not occur during this study.
Urinalysis findings:
no effects observed
Description (incidence and severity):
Statistically significant or biologically adverse changes did not occur during this study.
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
A diminished or absent response to an alerting stimulus was observed in the high dose group. Mean foot splay was significantly decreased on days 4 and 9 in the high dose group. The incidence of defecation was decreased in the 50 and 260 ppm groups during the motor activity assessments. Beyond this, there were no statistically significant or toxicologically remarkable neurobehavioral differences between the control and treatment groups.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment-related changes were observed. (Absolute but not relative lung weights were reduced in the high dose group.)
Gross pathological findings:
no effects observed
Description (incidence and severity):
No treatment-related changes were observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Effects only occurred in the nasal tissue of the high dose group. Minimal degeneration / necrosis of olfactory epithelium occurred in the anterior portion of the nose of 4/5 rats exposed to 260 ppm. This lesion was observed neither in the 260 ppm recovery group, which suggests reversibility, nor in the lower dosed groups.
Dose descriptor:
NOAEC
Effect level:
33.4 mg/m³ air
Sex:
male
Basis for effect level:
other: no adverse effects observed
Dose descriptor:
LOAEC
Effect level:
334 mg/m³ air
Based on:
test mat.
Sex:
male
Basis for effect level:
body weight and weight gain
Critical effects observed:
no
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEC
334 mg/m³
Study duration:
subacute
Species:
rat

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Oral

A combined repeated dose toxicity study with the reproduction/developmental toxicity screening test (no limit test) was performed according to OECD 422 and in compliance with GLP. Ten Sprague Dawley rats per sex per dose were treated daily orally by gavage with 30, 100, or 300 mg/kg bw/day of the test substance equivalent to 6, 20, and 60 mg/ml concentration in vehicle, respectively. The vehicle used was corn oil. The total volume applied was 5 ml/kg bw/dose. Males and females were exposed through test day 55 and from four weeks pre-mating through 4-day lactation period, respectively. Control animals were included and received concurrent vehicle treatment. No clinical signs were reported. 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 occurred during the study and the observed deaths were not considered to be treatment related. Decreased body weight gain was observed and considered to be compound-related and of biological significance. At 300 mg/kg bw/day, males experienced a decrease in body weight of -19 % for days 1-56 which was not statistically significant; weight -7%. At 100 and 300 mg/kg bw/day, females experienced a decrease only during gestation: gains -13% and -20%, weights -7% and -12%, respectively, statistically significant). Increased food consumption was observed and considered to be compound-related and of biological significance. At 300 mg/kg bw/day, males had increased food intake (+19%). At 100 and 300 mg/kg bw/day, females had increased food intake (+7%, not statistically significant and +13%, statistically significant, respectively) during gestation. Accordingly, food efficiency was also affected by treatment; at 300 mg/kg bw/day, males had decreased efficiency (-33%) and at 100 and 300 mg/kg bw/day, females during gestation had increased efficiency (+14 and +29%, respectively, statistically significant). Thus, the NOAEL for males and females was concluded to be 100 mg/kg bw/day and 30 mg/kg bw/day, respectively (2000, K1).

In addition, a combined repeated dose/reproductive developmental toxicity study in Sprague Dawley rats treated orally with test substance was only available in the original study language (Japanese) and an IUCLID dossier entry available on the NIHS Japan was used (2007, K4). Results obtained in this study are similar to those retrieved by Malley et al. (2002). However, effects on the liver were here regarded as adverse. It has to be noted that no historical ranges of the control were given and results were not properly discussed in the IUCLID dossier entry. Thus, it may be assumed that effects on the liver were adaptive and a non-adverse reaction as has been concluded for effects on the liver by Malley et al. (2002).

Inhalation

Two studies investigating the repeated exposure toxicity potential of the test substance are available.

A repeated dose toxicity (no limit test) was performed (no guideline reported) pre-GLP. Twenty male Sprague Dawley rats per dose were treated whole body by inhalation with 1.64 mg/l of the test substance. No post exposure period was included. Animals were exposed for 6 hours/day for ten days daily. No control animals were included. Peripheral vasodilation was noted throughout the study. A slight twitching only at the beginning of each exposure period through days 3 to 10 was observed. A lack of activity was observed throughout each exposure period. A LOAEC of 1.64 mg/l was concluded (1965, K2).

A repeated dose toxicity (no limit test) was performed (no guideline reported, GLP not specified). Ten male Sprague Dawley rats per dose were treated nose only by inhalation with 5, 50, and 260 ppm of the test substance. A post exposure period of two weeks was included. Animals were exposed for 6 hours/day 5 days/week for two weeks daily, at a total of 9 exposures. Control animals were included and received no treatment. Satellite groups were included investigating neurotoxicity. Irregular respiration in the high dose group. Other clinical observations occurred sporadically in all groups including controls and were considered incidental. Body weight was significantly reduced at >= 50 ppm, but was reversible. A diminished or absent response to an alerting stimulus was observed in the high dose group. Mean foot splay was significantly decreased on days 4 and 9 in the high dose group. The incidence of defecation was decreased in the 50 and 260 ppm groups during the motor activity assessments. Beyond this, there were no statistically significant or toxicologically remarkable neurobehavioral differences between the control and treatment groups. Effects only occurred in the nasal tissue of the high dose group. Minimal degeneration / necrosis of olfactory epithelium occurred in the anterior portion of the nose of 4/5 rats exposed to 260 ppm. This lesion was observed neither in the 260 ppm recovery group, which suggests reversibility, nor in the lower dosed groups. A NOAEC of 33.4 mg/m³ was concluded (1996, K2).

Although effects on body weight and certain neurobehavioral differences were observed, these effects were not considered to support classification for specific target organ toxicity following repeated exposure according to Regulation (EC) 1272/2008.

Dermal

No study investigating the dermal repeated exposure toxicity potential was available.

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result, the substance is not considered to be classified for repeated oral, inhalation or dermal dose toxicity under Regulation (EC) No. 1272/2008.