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Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD 422 Combined repeated dose oral toxicity study with the reproduction/developmental toxicity screening test
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name: Fatty acid, tall oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetramine
Structural Formula: UVCB substance
Colour: dark
Appearance: concentrated suspension
Declared purity: 97%
Molecular weight:>700

Test animals

Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
Young healthy male and nulliparous, non pregnant female rats [strain: Wistar Crl:WI] (Full-Barrier), were used in this study. The animals were derived from a controlled full barrier maintained breeding system (SPF) (Source: Charles River, 97633 Sulzfeld, Germany).

At the beginning of the study, the age of the animals was 8-9 weeks. The range of the body weight was:
Females: 154 to 183 g, (mean: 167.93 g, ± 20%= 33.59 g)
Males: 213 to 233 g, (mean: 222.30 g, ± 20%= 44.46 g)

After an adequate acclimatisation period (at least five days) the animals were barrier maintained (full barrier) in air-conditioned rooms under the following conditions:
- Temperature: 22 3 °C
- Relative humidity: 55 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (lot no.: xx).
- Free access to tap water, sulphur acidified to a pH of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- Housed individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
VEHICLE
Name: Corn oil
Justification for use and choice of vehicle (if other than water):The vehicle was chosen as suggested by sponsor and the test item‟s solubility.
Batch No.: 11KBC6753
Storage conditions: at room temperature (RT),
Safety precautions: Routine hygienic procedures were sufficient to assure personnel health and safety
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
Animals were paired in the ratio of 1:1 (male to female). The subsequent morning and the next morning there onwards the vaginal smear of female were checked to confirm the evidence of mating. The day of vaginal plug and/or sperm was considered as day 0 of gestation. Cages were arranged in such a way that possible effects due to cage placement was minimised.
Duration of treatment / exposure:
The animals will be dosed with the test item on 7 days per week for a period of approximately 54 days. The test substance will be administered daily during 14 days pre mating and 14 days mating in both male and in female, during gestation period and up to post natal day 3 in females. Males will be dosed after the mating period until the minimum total dosing period of 28 days has been completed.
Frequency of treatment:
daily, the animals were dosed with the test item on 7 days per week basis
Duration of test:
approximately 54 days
Doses / concentrationsopen allclose all
Dose / conc.:
300 mg/kg bw/day
Dose / conc.:
600 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Remarks:
vehicle control
No. of animals per sex per dose:
10 females and 10 males/ group
4 groups: control group, 300 mg/kg bw/day group, 600 mg/kg bw/day group, 1000 mg/kg bw/day group
Control animals:
yes, concurrent vehicle

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
DETAILED CLINICAL OBSERVATIONS: Yes
BODY WEIGHT: Yes
FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on post natal day 4 for both females and pups. Males were sacrificed at the end of the mating period (total dosing: 28 - 29 days)

Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: No data
- Number of late resorptions: No data
Fetal examinations:
- External examinations: Yes
- Soft tissue examinations: Yes
- Skeletal examinations: No data
- Head examinations: no data
Statistics:
Parameters like body weight gain and food consumption were calculated for each animal as the difference in weight measured from one week to the next. The relative organ weights were calculated in relation to the body weight (measured at necropsy) and presented as percentage.
For statistical analysis one-way analysis of variance (ANOVA) followed by Dunnett‟s multiple comparison test was carried out to reveal any differences between control- and test groups. These statistics were performed with GraphPad Prism V.5 software (p<0.05 was considered as statistical significant).
In the evaluation of laboratory parameters all values within a range of the mean value the two fold standard deviation (x 2s) are considered to be „normal“ values within a „normal“ population.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
In males, the clinical findings observed were, control-nibbling of fur (1/10 animals); LD group - nasal discharge (1/10 animals); MD group - piloerection (1/10 animals), vocalization (1/10 animals), nibbling of fur (1/10 animals) and HD group - salivation (1/10 animals), regurgitation (2/10 animals), nibbling of fur (3/10 animals) and pushing the bedding (1/10 animals).
In females, the clinical findings observed were, control - nibbling of fur (1/10 animals), salivation (1/10 animals); LD group - salivation (1/10 animals), nibbling of fur (1/10 animals); MD group - piloerection (1/10 animals).
Mortality:
no mortality observed
Description (incidence):
No test item related mortalities were observed in male and female animals during the entire study period.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
The body weight and food consumption in both males and females remained unaffected due to treatment when compared to the corresponding controls. However, statistical significant deviation was observed for body weight change in females during premating days 7 to 14 in MD group.
Haematological findings:
no effects observed
Description (incidence and severity):
In male and female, statistically no significant effect was observed for any of the hematological and clinical biochemistry parameters when compared to their corresponding control. However, there were some individual deviations observed from the historical control range for the respective parameters.
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Urinalysis in five randomly selected males from each group revealed no test item related effect.
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No test item related effect on functional and behavioural endpoints were observed in male and female animals during the entire study period.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
In males, statistically no significant differences in the absolute and relative organ weights of the treatment groups was observed when compared with the controls.
In females, statistically significant difference was observed for relative brain weight in LD and HD group compared to controls.
In the female and male reproductive organs, no histopathological lesions considered to be test item-related were noted. Three females were recorded non pregnant during the study, but in histopathology showed normal cycling activity.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Necropsy revealed macroscopic changes associated with liver, lung and epididymides in most of the animals of all groups, but microscopically only in lungs, the histopathological changes were observed in the animals of LD, MD and HD groups.
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Test item-related histopathological lung changes like multifocal subacute bronchopneumonia, characterized by peribronchial foci of prominent fibrosis, with re-epithelialization, infiltration with mononuclear cells, histiocytes and occasional multinucleated cells, was observed in a small proportion of treated males and females of all dose groups, without dose relationship. In addition, a mild amount of intrahistiocytic black material was seen in the lung of each one male in LD and HD group.

Maternal developmental toxicity

Number of abortions:
no effects observed
Description (incidence and severity):
All pregnancesies resulted in normel births. Successful mating resulted in 9/10 pregnancies in the control and LD group, 8/9 in MD groups and 10/10 pregnancies in HD.
Pre- and post-implantation loss:
effects observed, non-treatment-related
Description (incidence and severity):
The deviation observed for pre implantation loss in MD and HD groups may not be considered treatment related as the loss were very low compared to LD group and controls.
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Details on maternal toxic effects:
The group mean number of corpora lutea, number of pre-impantation sites, number of live pups (born on PND 0 and 4), percent post implantation lossremained unaffected due to treatment when compared to controls.

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
clinical signs
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
no systemic toxicity was seen, but only local, non-systemic effects

Maternal abnormalities

Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
not examined
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
Survival of the pups from PND 0 to PND 4 remained unaffected due to treatment in all treatment groups.
No test item related changes were observed in pups at necropsy or death during the study.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
Statistical analysis of reproduction and litter data revealed no treatment related effect on group litter sex ratio
Changes in litter size and weights:
effects observed, non-treatment-related
Description (incidence and severity):
Except for group mean litter weight on PND 4 (MD and HD group), the statistical analysis of litter weight data revealed no deviation for total litter weight, male and female litter weight on PND 0 and PND 4 when compared with corresponding controls.
The deviation in group mean litter weight on PND 4 in MD and HD group cannot be attributed to treatment due to lack of consistent findings.
Changes in postnatal survival:
no effects observed
Description (incidence and severity):
Survival of the pups from PND 0 to PND 4 remained unaffected due to treatment in all treatment groups.
External malformations:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
Gross necropsy of dead pups revealed no treatment related findings.

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects

Fetal abnormalities

Abnormalities:
no effects observed

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
In conclusion, the repeated dose administration of fatty acids, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine in corn oil to the male (28 - 29 days) and female (maximum 54 days) Wistar rats at dosages of 300, 600 and 1000 mg/kg body weight revealed test item-related histopathological lung changes in males and females of all treatment group.
Based on the data generated from this combined repeated dose toxicity and reproduction/developmental toxicity screening test with Fatty acids, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine, no effects were reported on reproductive/developmental toxicity parameters measured in this study. There were also no effects reported on general toxicity parameters except for the reported macroscopic/microscopic lung changes, that was considered a local indirect effect due to suspected chemical pneumonitis. It is known that low viscosity chemicals can generate residues in the esophagus that can be aspired in the trachea and deposited in lungs creating such local inflammation as seen in the study. Dark material (the test material) seen in some animals in lung histiocytes do support this conclusion.
For reproductive/developmental toxicity, the NOAEL could be set at 1000 mg/kg bw, due to absence of developmental and reproductive effects.
Executive summary:

The test substance fatty acids, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine, was assessed for reproductive toxicity within an OECD 422 combined repeated dose, reproductive toxicity screening study.

The repeated dose administration of fatty acids, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine in corn oil to the male (28 - 29 days) and female (maximum 54 days) Wistar rats at dosages of 300, 600 and 1000 mg/kg body weight revealed test item-related histopathological lung changes in parental males and females of all treatment group.

Based on the data generated from this combined repeated dose toxicity and reproduction/developmental toxicity screening study with fatty acids, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine, no effects were reported on reproductive/developmental toxicity parameters measured in this study. There were also no effects reported on general toxicity parameters except for the reported macroscopic/microscopic lung changes. This local effect was seen in lungs but considered an indirect effect due to suspected chemical pneumonitis. It is known that low viscosity chemicals can generate residues in the esophagus that can be aspired in the trachea and deposited in lungs creating such local inflammation as seen in the study. Dark material (the test material) seen in some animals in lung histiocytes do support such a conclusion. Due to the lack of clear dose-response relationship (solely restricted to histopathological lung changes) observed in this study, the suitable NOAEL (No observed adverse effect level) for general toxicity could not be determined, but for systemic toxicity is set to 1000 mg/kg bw/d. Furthermore, for reproductive/ developmental toxicity, in absence of any adverse findings, the NOAEL could be set at 1000 mg/kg bw.