Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine

Endpoint summary

Description of key information

NOAEL systemic = 800 mg/kg bw/day (OECD 422, RTC 2013)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

The toxic effects on rats after repeated dosing with Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine, as well as any effects of the test item on male and female reproductive performance, such as gonadal function, conception, parturition and early lactation of the offspring were investigated in a study according to OECD guideline 422 and GLP (RTC, 2013). The rats were dosed at dose levels of 0, 80, 250 and 800 mg/kg bw/day.

The vehicle was corn oil. All doses were administered at a constant volume of 4 mL/kg body weight.

Males were treated for 2 weeks prior to pairing and during pairing with females until the day before necropsy, for a total of 34 days.

Females were treated for 2 weeks prior to pairing, during pairing and throughout the gestation and lactation periods until Day 3post partum.

The following investigations were performed in all groups: body weight, clinical signs (including neurotoxicity assessment, motor activity and sensory reaction to stimuli), food consumption, oestrous cycle, mating performance, clinical pathology investigations (haematology and clinical chemistry), litter data, macroscopic observations, organ weights and histopathological examination.

Clinical signs and macroscopic observations of pups were also performed.

The histopathological examination was performed only on control and high dose groups (five animals/sex/group randomly selected). The identification of the stages of the spermatogenic cycle was performed in all males of the control and high dose groups.

In addition, coagulating glands, epididymides, preputial gland, prostate gland, seminal vesicles and testes were examined histopathologically on all parental males and cervix, clitoral gland, ovaries, uterus and vagina were examined histopathologically on all parental females.

Since histopathological changes were observed in control and high dose males, the examination was then extended to the thymus of the remaining males of the control and high dose groups and in all males of Groups 2 and 3.

Mortality and fate of females

There were no compound-related effects.

One male receiving 800 mg/kg bw/day was found dead on Day 6 of the study. The cause of death of this animal is suggested to be related to complications associated with mis-dosing.

All females proved to be pregnant except one control and one receiving 800 mg/kg bw/day.

The number of females with live pups on Day 4 post partum was 9 in the control, 10 in the low dose (80 mg/kg bw/day), 10 in the mid-dose (250 mg/kg bw/day) and 9 in the high dose group (800 mg/kg bw/day).

Clinical signs and clinical observations (Functional Observation Battery Tests)

Salivation was the main clinical sign observed during the study in males at the dose levels
≥ 250 mg/kg bw/day and in females at 800 mg/kg bw/day.

Moreover the same males and females of the high dose group also showed hair loss.

Neurotoxicity assessment (removal of animals from the home cage and open arena)

Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test item.

Body weight and body weight gain

No differences of toxicological significance in body weights were recorded in animals of both sexes compared to the control group, throughout the study.

Food consumption

No effects on food consumption were observed in either males or females.

Motor activity and sensory reactivity to stimuli

No relevant differences were noted in all parameters investigated between control and treated groups.

Haematology

No changes of toxicological relevance were found in either sex at any dose.

No toxicological changes were recorded in the coagulation test.

Clinical chemistry

No adverse findings were observed by clinical chemistry examination.

Oestrous cycle, reproductive parameters, pairing combination and mating performance

Oestrous cycle, pre-coital intervals, copulatory index and fertility index did not show intergroup differences.

Implantation, pre-implantation loss data, pre-birth loss data and gestation length of females

No relevant differences were observed for these parameters between treated groups and controls.

All dams, with the exception of two not pregnant females, gave birth between Days 22 and 23 post coitum.

Litter data at birth, on Day 1 and on Day 4 post partum of females and sex ratio of pups

Litter data and sex ratio were unaffected by treatment, at birth, on Day 1 and Day 4 post partum.

Clinical signs of pups

There were no compound-related effects.

Pre-weaning clinical signs were comparable between treated and control groups.

Necropsy findings in decedent pups and in pups sacrificed on Day 4 post partum

Necropsy findings in decedent pups and in pups sacrificed on Day 4 post partum did not reveal any treatment-related effect.

Terminal body weight and organ weights

Terminal body weight was unaffected by treatment in both sexes.

Absolute and relative organ weights were, in general, comparable between treated and control groups.

Macroscopic observations

There were no compound-related effects.

Microscopic observations

No treatment-related changes were noted. In addition, seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and no alterations were noted.

Conclusions

On the basis of the results obtained in this study the NOAEL (No Observed Adverse Effect Level) for general toxicity and for reproductive and developmental toxicity was considered to be 800 mg/kg bw/day for males and females.

Based on the occurrence of salivation, which might be a local reaction to irritancy or taste of the test compound, the NOEL (No Observed Effect Level) was considered to be 80 mg/kg bw/day.

 

Justification for classification or non-classification

Based on the available data, the test substance is not classified with regard to repeated dose toxicity according to Directive 67/548/EEC (DSD) and Regulation (EC) No 1272/2008 (CLP), respectively.