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Toxicity to reproduction

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Administrative data

Endpoint:
toxicity to reproduction
Remarks:
other: screening
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 guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Qualifier:
according to guideline
Guideline:
other: screening Test. EPA 712-C-00-368, July 2000.
GLP compliance:
yes (incl. QA statement)
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
Fatty acids, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine
EC Number:
273-601-0
EC Name:
Fatty acids, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine
Cas Number:
68990-47-6
Molecular formula:
The substance is a UVCB substance. One of the most likely and the smallest molecule arising from the reaction process is assumed to be: C18H33O. C18H31O. 2C4H12N3 . C4H2O2
IUPAC Name:
Reaction product of 2,5-Furandione with reaction products of tall-oil fatty acids, diethylenetriamine, triethylenetetramine and tetraethylenepentamine
Details on test material:
Name: Fatty acids, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine
CAS No.: 68990-47-6
Batch no.: TEE3316/22
Expiry Date: November 2011
Physical state at RT: solid
Colour: dark
Purity: Date of analysis 11 February 2010, 97% (w/w)
Storage Conditions: at room temperature, protected from light
Solubility in Water: very low
Safety precautions: Routine hygienic procedures will be sufficient to assure personnel health and safety.

Test animals

Species:
rat
Strain:
Wistar
Details on species / strain selection:
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).
According to Art. 9.2, No. 7 of the German Act on Animal Welfare the animals were bred for experimental purposes.
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)
Sex:
male/female
Details on test animals or test system 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
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.
Analytical verification of doses or concentrations:
yes
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.
Doses / concentrationsopen allclose all
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Dose / conc.:
600 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Remarks:
vehicle control
No. of animals per sex per dose:
80 animals (10 non-pregnant nulliparous females and 10 adult males /group) were included in the study.
Control animals:
yes, concurrent vehicle

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

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, non-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
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.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
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

Results: F1 generation

General toxicity (F1)

Mortality / viability:
no mortality 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.
Body weight and weight changes:
effects observed, non-treatment-related

Details on results (F1)

Statistical analysis of reproduction and litter data revealed no treatment related effect on group mean litter weight, number of males, number of females, total litter weight, male and female litter weights on PND 0 and PND 4, percent pre implantation loss, post implantation loss, total number of pups born, sex ratio, live pups, still birth and runt on PND 0, total No. of live pups and sex ratio on PND 0 and 4, precoital interval, number. of corpora lutea, number of implantation sites when compared with controls. Statistically significant deviation was observed for % pre-implantation loss and group mean litter weight (PND 4) in MD and HD groups compared to corresponding controls. This deviation was considered to be incidental.
Survival of the pups from PND 0 to PND 4 remained unaffected due to treatment in all treatment groups.

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
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

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

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.

Urinalysis in five randomly selected males from each group revealed no test item related effect.

Necropsy revealed macroscopic changes associated with liver, lung and epididymides in most of the parental animals of all groups, but microscopically only in lungs, the histopathological changes were observed in the animals of LD, MD and HD groups. No test item related changes were observed in pups at necropsy or death during the study.

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.

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.

Applicant's summary and conclusion

Conclusions:
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.
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) general toxicity could not be determined. However, for reproductive/developmental toxicity, the NOAEL could be set at 1000 mg/kg bw.
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. However, for reproductive/ developmental toxicity, in absence of any adverse findings, the NOAEL could be set at 1000 mg/kg bw.