3,3'-oxybis(ethyleneoxy)bis(propylamine)

Endpoint summary

Description of key information

OECD 422, rat, gavage (GLP, reliability 1, 2013):
NOAEL systemic parental toxicity: 600 mg/kg bw/d
NOAEL locally irritating effect (stomach): 100 mg/kg bw/d (females)
NOAEL locally irritating effect (stomach): < 100 mg/kg bw/d (males)

OECD 408, GLP, oral, rat: Animal study planned

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Jan - Dec 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP compliant

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:

1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA, Health Effects Test Guidelines; OPPTS 870.3650: Combined Repeated Dose Toxicity Study With the Reproduction/Developmental Toxicity Screening Test

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht; 22.10.2009

Limit test:

no

Specific details on test material used for the study:

- Name of test substance: 3,3'-Oxybis(ethyleneoxy)bis(propylamine)
- Test substance No.: 11/0401-1
- Batch identification: O 2895
- Purity: 99.7% (project No: 11L00203)
- Homogeneity: given (visually, project No: 11L00203)
- Stability: expiry date: 15 Jun 2013
- Date of production: 15 Jun 2011
- Physical state/ appearance: liquid / colorless, clear
- Storage conditions: room temperature
- Other: The stability of the test substance under storage conditions over the test period was guaranteed by the sponsor.

Species:

rat

Strain:

Wistar

Details on species / strain selection:

The rat is the preferred animal species for reproduction studies according to the various test guidelines and the Wistar strain was selected.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at supplied: 10-11 weeks
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sulzfeld, Germany
- Weight at study initiation: Males: 321 - 322 g, Females 205 - 209 g
- Fasting period before study: no
- Housing: individually in Makrolon type M III cages;
- Exceptions:
- During overnight matings, male and female mating partners were housed together in Makrolon type M III cages
- Pregnant animals and their litters were housed together until PND 4 (end of lactation).
- For motor activity (MA) measurements the animals were housed individually in polycarbonate cages (floor area of about 800 cm2) and
small amounts of bedding material
- The cages with the test animals were arranged on the racks in such a way that uniform experimental conditions (ventilation and light) were ensured.
- Diet, ad libitum: Ground Kliba maintenance diet mouse-rat “GLP” meal, supplied by Provimi Kliba SA, Kaiseraugst, Switzerland
- Water, ad libitum: drinking water
- Acclimation period: On the day of arrival the animals were subjected to an appropriate acclimatization period.

ENVIRONMENTAL CONDITIONS
- Temperature: 20-24°C
- Humidity: 30-70%
- Air changes (per hr): 15
- Photoperiod: 12-hour light/12-hour dark cycle

IN-LIFE DATES: From: 2012-01-09 To: 2013-03-12

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The appropriate amount of test substance was weighed out depending on the desired concentration. Then, drinking water was filled up to the desired volume, subsequently released with a magnetic stirrer. During administration of the test substance, preparations were kept homogeneous by stirring with a magnetic stirrer. The test substance preparations were produced at least once a week and were stored at room temperature.

VEHICLE
- Justification for use and choice of vehicle: solubility
- Concentration in vehicle: 1.00 mg/100 mL (100 mg/kg bw/d)*, 3.00 mg/100 mL (300 mg/kg bw/d)*, 10.00 and 6.00 mg/100 mL (1000 and 600 mg/kg bw/d)*
*) The dose refers to the body weight of the individual rats determined most recently.
- Administration volume:: 10 mL/kg

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The stability of the test substance in drinking water for a period of 7 days at room temperature was proven during the study. Homogeneity and concentration control analyses of the test-substance preparations were performed in all concentrations at the start of the administration period. Additionally, samples from all concentrations as reverse samples for concentration control analysis were taken at the end of the study. The various analyses confirmed the stability over 7 days and homogeneity and concentration in the vehicle.
The determined mean and single concentrations of the test substance in drinking water were found to be in the range of 90 % – 110 % of the nominal
concentrations. These results demonstrated the correctness of the concentrations.

Duration of treatment / exposure:

Males were exposed for 59 days (prior to mating, during mating, and up to termination) and females 62 days (prior to mating, during mating, during post-coitum, and at least 4 days of lactation).

Frequency of treatment:

Daily

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

analytical verified concentration

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

analytical verified concentration

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

reduced to 600 mg/kg bw/day on study day 7; analytical verified concentration

No. of animals per sex per dose:

10 animals

Control animals:

yes, concurrent vehicle

Details on study design:

The oral route was selected since this was proven to be suitable for the detection of a toxicological hazard.
Fasting period before blood sampling for clinical biochemistry: 16 hours

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
A cageside examination was conducted at least once daily for any signs of morbidity, pertinent behavioral changes and signs of overt toxicity.
The littering and lactation behavior of the dams was generally evaluated in the mornings in combination with the daily clinical inspection of the dams.
On weekdays (except public holidays) the parturition behavior of the dams was inspected in the afternoons in addition to the evaluations in the mornings.

DETAILED CLINICAL OBSERVATIONS: Yes
Detailed clinical observations (DCO) were performed in all animals prior to the administration period and thereafter at weekly intervals. The animals were transferred to a standard arena (50 × 37.5 cm with sides of 25 cm high).
The following parameters were examined: abnormal behavior when handled, fur, skin, posture, salivation, respiration, activity/arousal level, tremors, convulsions, abnormal movements, impairment of gait, lacrimation, palpebral closure, exophthalmus, feces (appearance/consistency), urine, pupil size

BODY WEIGHT: Yes
Body weight was determined before the start of the administration period in order to randomize the animals. During the administration period body weight was determined on study day 0 (start of the administration period) and thereafter once a week at the same time of the day (in the morning).
The body weight change of the animals was calculated from these results.
The following exceptions are notable for the female animals:
- During the mating period the parental females were weighed on the day of positive evidence of sperm (GD 0) and on GD 7, 14 and 20.
- Females with litter were weighed on the day of parturition (PND 0) and on PND 4.
- Females without a litter and without positive evidence of sperm in the vaginal smear were weighed weekly. These body weight data were solely used for the calculations of the dose volume.

FOOD CONSUMPTION:
Generally, food consumption was determined once a week for male and female parental animals, with the following exceptions:
- Food consumption was not determined during the mating period (male and female F0 animals)
- Food consumption of the F0 females with evidence of sperm was determined on GD 0-7, 7-14, 14-20.
- Food consumption of F0 females, which gave birth to a litter, was determined for PND 1-4.
Food consumption was not determined in females without positive evidence of sperm (during the mating period of dams used in parallel) and females without litter (during the lactation period of dams used in parallel).

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
Drinking water consumption was monitored by daily visual inspection of the water bottles for any changes in volume.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: In the morning towards the end of the administrative period
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: first 5 surviving parental males and the first 5 surviving females with litter (in order of delivery) per group
- Parameters examined: see Table 1 in 'Any other information on materials and methods incl. tables'.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: In the morning towards the end of the administrative period
- Animals fasted: Yes
- How many animals: first 5 surviving parental males and the first 5 surviving females with litter (in order of delivery) per group
- Parameters examined: see Table 2 in 'Any other information on materials and methods incl. tables'.

URINALYSIS: Yes
- Time schedule for collection of urine: overnight towards the end of the administrative period
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters examined: see Table 3 in 'Any other information on materials and methods incl. tables'.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Functional observational battery (FOB) was performed in all animals towards the end of the administration period
- Dose groups that were examined: all animals

- Battery of functions tested:
Home cage observations:
Attention was paid to: Posture, Tremors, Convulsions, Abnormal movements, Gait abnormalities.
Open field observations:
Behavior when removed from cage, fur, skin, salivation, nose discharge, lacrimation, eyes/pupil size, posture, palpebral closure, respiration, tremors, convulsions, abnormal movements, impairment of gait, activity/arousal level, feces (number of fecal pellets/appearance/consistency) within two minutes, urine (appearance/quantity) within two minutes, number of rearings within two minutes.
Sensorimotor tests/reflexes:
Approach response, touch response, vision ("visual placing response"), pupillary reflex, pinna reflex, audition ("startle response"), coordination of movements ("righting response"), behavior during "handling", vocalization, pain perception ("tail pinch"), grip strength of forelimbs, grip strength of hindlimbs, landing foot-splay test, other findings.

Motor activity (MA) measured on the same day as the FOB was performed.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
All parental animals were sacrificed by decapitation under isoflurane anesthesia. The exsanguinated animals were necropsied and assessed by gross pathology.
Organ weights: Weight assessment was carried out on all animals.
- The following weights were determined: Anesthetized animals, Epididymides, Testes.
- The following weights were determined in 5 animals/sex and test group (females with litters, same animals as used for clinical pathology examinations): Adrenal glands, Brain, Heart, Kidneys, Liver, Spleen, Thymus

HISTOPATHOLOGY: Yes
The following organs/ tissues were preserved in neutral-buffered 4% formaldehyde or in modified Davidson’s solution: Adrenal glands, All gross lesions, Aorta, Bone marrow (femur), Brain, Cecum, Cervix, Coagulating glands, Colon, Duodenum, Eyes with optic nerve, Esophagus, Extraorbital lacrimal gland, Epididymides (modified Davidson’s solution), Femur with knee joint, Heart, Ileum, Jejunum (with Peyer’s patches), Kidneys, Larynx, Liver, Lungs, Lymph nodes (axillary and mesenteric), Mammary gland (male and female), Nose (nasal cavity), Ovaries (modified Davidson’s solution), Oviducts, Pancreas, Parathyroid glands, Pharynx, Pituitary gland, Prostate gland, Rectum, Salivary glands, (mandibular and sublingual), Sciatic nerve, Seminal vesicles, Skeletal muscle, Spinal cord (cervical, thoracic and lumbar cord), Spleen, Sternum with marrow, Stomach (forestomach and glandular stomach), Target organs,Testes (modified Davidson’s solution), Thymus, Thyroid glands, Trachea, Urinary bladder, Uterus, Vagina.
For further evaluation proceeding, see Table 4 in 'Any other information on materials and methods incl. tables'.

Other examinations:

The supplier assayed the food used in the study for chemical and microbiological contaminants.
The drinking water is regularly assayed for chemical contaminants.
The bedding and the enrichment are regularly assayed for contaminants (chlorinated hydrocarbons and heavy metals).

Statistics:

Please refer to 'Any other information on materials and methods incl. tables'.

Clinical signs:

no effects observed

Description (incidence and severity):

No test substance-related, adverse findings were noted.
A temporary, short appearance of salivation immediately after dosing was concluded to be induced by a bad taste of the test substance or local affection of the upper digestive tract.

Mortality:

no mortality observed

Description (incidence):

No rat died prematurely in the present study.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

On study day 7 (premating) mean body weight of male animals in test group 3 (1000 and 600 mg/kg bw/d) was significantly decreased (-6%). Although not significantly altered, mean body weight loss in female animals was observed after 7 days of treatment.
Mean body weight change values during premating were significantly decreased in male animals of test group (1000 and 600 mg/kg bw/d) between study days 0-7 (-84%) and 0-13 (-48%). Although not significantly altered mean body weight change value in female animals was observed between study days 0-7 days. All mentioned findings were assessed as being related to treatment.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

In test group 3 (1000 and 600 mg/kg bw/d) food consumption during premating was significantly decreased in male animals between study days 0-7 (-17%) and 0-13 (-11%) as well as in female animals between study days 0-7 (-19%). These findings were assessed as being related to treatment with the test substance by irritating the upper digestive tract.
No other findings were observed for male and female animals in test group 1 and 2 (100 and 300 mg/kg bw/d) nor for male and female animals of test group 3 (1000 and 600 mg/kg bw/d) after reducing the dose level.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

no effects observed

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

Description (incidence and severity):

Home cage observations: No test substance-related effects were observed.
Open field observations: No test substance-related effects were observed.
Sensorimotor tests/reflexes: No test substance-related effects were observed.
Quantitative Parameters: No test substance-related effects were observed.
Motor activity measurement: There were no significant deviations concerning the overall motor activity (summation of all intervals) and regarding the single intervals in male and female animals of all test groups in comparison to the concurrent control group.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

The mean absolute weights of the kidney were significantly increased in females of the highest dose (9% increased compared to control). The increase in absolute kidney weights of females of test group 3 (1000 and 600 mg/kg bw/d) was not regarded to be treatment-related due to a missing histopathological correlate and a missing dose-response relationship.
The increase of relative liver weights in females of test group 2 (300 mg/kg bw/d) by 6% was also not regarded to be treatment-related because the livers examined histopathologically in
animals of test group 3 (1000 and 600 mg/kg bw/d) did not show any finding which could explain the weight increase.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Glandular stomach (discoloration, thickening of wall), duodenum (thickening of wall)
The obove mentioned gross lesions in males in the glandular stomach and duodenum were regarded to be treatment related. All other gross lesions noted were single observations and they were regarded to have developed spontaneously and unrelated to compound and treatment.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Treatment of male and female Wistar rats with up to 1000 and 600 mg/kg bw/d of the test substance led to treatment-related findings in the upper digestive tract (starting at 100 mg/kg bw/d in males and 300 mg/kg bw/d in females). The hyperplasia, hydropic degeneration of squamous cells and inflammatory cell infiltrates, occasionally with multinucleated giant cells in the forestomach as well as the hyperemia, eosinophilic cytoplasmic change, increased mitotic figures, submucosal edema and inflammatory cell infiltrates in the glandular stomach were regarded to be signs of a slight irritating effect of the test substance. These findings were almost exclusively observed around or at the margo plicatus and in the glandular stomach in the fundic area. They were regarded to be adverse. The increase in thickness in the duodenum (only males >= 300 mg/kg bw/d) was also regarded to be treatment-related and as the specific mechanism could not be determined, it was also judged to be adverse in nature. All these adverse findings described above are regarded to be local irritating effects but no systemic toxicity.

Histopathological findings: neoplastic:

no effects observed

Details on results:

Findings which occurred either individually or were biologically equally distributed over control and treatment groups, were considered to be incidental or spontaneous in origin and without any relation to treatment.

Dose descriptor:

NOAEL

Remarks:

parental systemic toxicity

Effect level:

600 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effect up to and including the highest tested dose (starting dose: 1000 mg/kg bw/d, reduced to 600 mg/kg bw/d on study day 7)

Dose descriptor:

NOAEL

Remarks:

local

Effect level:

100 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

histopathology: non-neoplastic

Dose descriptor:

LOAEL

Remarks:

local

Effect level:

100 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

histopathology: non-neoplastic

Dose descriptor:

NOAEL

Remarks:

reproductive toxicity

Effect level:

600 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effect up to and including the highest tested dose (starting dose: 1000 mg/kg bw/d, reduced to 600 mg/kg bw/d on study day 7)

Dose descriptor:

NOAEL

Remarks:

developmental toxicity

Effect level:

600 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Starting dose: 1000 mg/kg bw/d, reduced to 600 mg/kg bw/d on study day 7

Critical effects observed:

not specified

Table 5: Histopathology findings (10 animals in each dose group)

	Males				Females
Test group (mg/kg bw/d)	0	100	300	1000 (600)	0		100	300	1000 (600)
Forestomach
• Hyperplasia squamous, focal			2	4				1
• Degeneration, hydropic, (m)focal		1	2	2		1		3	5
•Inflammatory cell infiltrates, (m)focal			1	6					6
Glandular stomach
• Hyperemia, mucosal		1	1	2
• Eosinophilic cytoplasmic change		1	5	4				1	8
• Mitotic figures increased			4	3				1	3
• Edema, submucosal		2	3	2				1	2
• Inflammatory cell infiltrates, (m)focal		3	5	7				1	6
Duodenum
•Thickness, increased			1	5

Conclusions:

The no observed adverse effect level (NOAEL) for general systemic toxicity was 600 mg/kg bw/d in males and in females, the NOAEL for the locally
irritating effect in the stomach was 100 mg/kg bw/d in females and below that in males.

Executive summary:

In this reliabe OECD 422 study performed in compliance with GLP (reliability 1), the test substance was administered orally via gavage to groups of 10 male and 10 female Wistar rats (F0 animals) at dose levels of 0 mg/kg body weight/day (mg/kg bw/d; test group 0), 100 mg/kg bw/d (test group 1), 300 mg/kg bw/d (test group 2) and 1000 mg/kg bw/d (test group 3). Because of body weight loss in several animals of both sexes in test group 3 (1000 mg/kg bw/d) the animals were treated with 600 mg/kg bw/d from study day 7 onwards. Drinking water served as vehicle. The objective of the study was to detect possible effects of the test substance on the integrity and performance of male and female reproductive systems including gonadal function, mating behavior, conception, gestation and parturition. Furthermore, it was intended to obtain information about the general toxicological profile including target organs and the no observed adverse effect level (NOAEL) after repeated oral administration. The duration of treatment covered a 2-week pre-mating and mating period in both sexes, approximately 4 week post-mating in males, and the entire gestation period as well as 4 days of lactation in females followed by an additional treatment until one day before sacrifice.

Parental animals of the highest dose (1000 mg(kg bw(d) showed piloerection after treatment (1 of 10 male animal and 2 of 10 female animals) during the first study week. Food consumption during premating was significantly decreased in male animals between study days 0-7 (-17%) and 0-13 (-11%) as well as in female animals between study days 0-7 (-19%). Mean body weight of male animals was significantly decreased (-6%) on study day 7. Although not significantly altered mean body weight loss in female animals was observed after 7 days of treatment. Mean body weight change values during premating were significantly decreased in male animals between study days 0-7 (-84%) and 0-13 (-48%). Although not significantly reduced mean body weight change value in female animals was observed between study days 0-7 days. After exposure dose reduction from study day 7 (600 mg/kg bw/d) onwards, solely piloerection was observed in 1 of 10 male animals during the second study week. In the lower dose groups (100 and 300 mg/kg bw/d) no substance-related adverse findings in respect of clinical examinations and clinical pathology were observed. Local toxicity was observed after substance administration for all exposure settings. Target organ was the stomach for male and female animals and, in addition, the duodenum for males. The hyperplasia, hydropic degeneration of squamous cells and inflammatory cell infiltrates, occasionally with multinucleated giant cells in the forestomach as well as the hyperemia, eosinophilic cytoplasmic change, increased mitotic figures, submucosal edema and inflammatory cell infiltrates in the glandular stomach were regarded to be signs of a slight irritating effect of the test substance. These findings were almost exclusively observed around or at the margo plicatus and in the glandular stomach in the fundic area. They were regarded to be adverse. The increase in thickness in the duodenum (only males >= 300 mg/kg bw/d) was also regarded to be treatment-related and as the specific mechanism could not be determined, it was also judged to be adverse in nature. All these adverse findings described above are regarded to be local irritating effects but no systemic toxicity.

Thus, the no observed adverse effect level (NOAEL) for general systemic toxicity was 600 mg/kg bw/d in males and in females, the NOAEL for the locally

irritating effect in the stomach was 100 mg/kg bw/d in females and below that in males.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

600 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Guideline study conducted in accordance with GLP.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

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

In a combined repeated dose toxicity study with the reproduction/developmental toxicity screening test (BASF SE, 2013) the test substance was administered orally by gavage to groups of 10 male and 10 female Wistar rats (F0 animals) at dose levels of 0 mg/kg bw/day (test group 0), 100 mg/kg bw/d (test group 1), 300 mg/kg bw/d (test group 2) and 1000 mg/kg bw/d (test group 3). Because of body weight loss in several animals of both sexes in test group 3 (1000 mg/kg bw/d) the animals were treated with 600 mg/kg bw/d from study day 7 onwards. The animals of the control group were treated in the same way with the vehicle only (drinking water). Fourteen days after the beginning of treatment, males and females from the same test group were mated. Concerning the administration period males were exposed for 59 days (prior to mating, during mating, and up to termination) and females 62 days (prior to mating, during mating, during post-coitum, and at least 4 days of lactation).

Clinical and detailed chemical signs, food and water consumption as well as body weight development and gain were determined during the study period. Towards study end, clinicochemical and haematological examinations, urinalysis and a functional observation battery incl. motor activity measurements were performed.

After sacrifice, parent animals were examined by gross pathology. Selected organs were weighed and a histopathological examination was performed. Pups were sexed and macroscopically examined on PND0. Viability was recorded. They were weighed on PND1 and PND4. Necropsy was performed on PND4. All pups were examined macroscopically for external and visceral findings.

Obtained results:

Clinical examination: Signs of an impaired condition, i.e. reduced food consumption and impaired body weight parameters, were observed in male or female parental animals of test group 3 at a dose level of 1000 mg/kg bw/d during the first week of treatment (premating period). The animals recovered after reducing the dose level to 600 mg/kg bw/d. These findings were regarded to be related to a locally irritating effect of the test substance as described in the pathology section. Therefore, it was assumed that no systemic toxicity was observed. Salivation after treatment was seen in all animals of test groups 3 (1000 and 600 mg/kg bw/d) but not in animals of test groups 1 and 2 (100 and 300 mg/kg bw/d). From the temporary, short appearance immediately after dosing it is likely, that this finding was induced by a bad taste of the test substance or local affection of the upper digestive tract. This finding was not considered to be an adverse and toxicologically relevant effect.

Clinical chemistry, haematology and urinalysis: No treatment-related, adverse effects were observed up to a dose of the test substance of 1000 and 600 mg/kg bw/d.

Pathology: Treatment of male and female Wistar rats with up to 1000 and 600 mg/kg bw/d of the test substance led to treatment-related findings in the upper digestive tract. The hyperplasia, hydropic degeneration of squamous cells and inflammatory cell infiltrates, occasionally with multinucleated giant cells in the forestomach as well as the hyperemia, eosinophilic cytoplasmic change, increased mitotic figures, submucosal edema and inflammatory cell infiltrates in the glandular stomach were regarded to be signs of a slight irritating effect of the test substance. These findings were almost exclusively observed around or at the margo plicatus and in the glandular stomach in the fundic area. They were regarded to be adverse. The increase in thickness in the duodenum was also regarded to be treatment-related and as the specific mechanism could not be determined, it was also judged to be adverse in nature. All these adverse findings described above are regarded to be a local irritating effect but no systemic toxicity. No findings on the reproduction tract were observed. All other findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment. Furthermore, in neurobehavioural examinations no test substance-related effects were observed.

Reproduction/developmental toxicity: Fertility indices for male and female animals were not impaired by test-substance administration even at dose levels of 1000 and 600 mg/kg bw/d. In addition, live birth indices of pups in all test groups were not influenced. The viability index as indicator for pup mortality was not decreased in any test group. Clinical and macroscopical examinations of the pups did not reveal any test substance related adverse finding.

Conclusions: Under the experimental conditions of this study 3,3'-Oxybis(ethyleneoxy)bis(propylamine) revealed signs of a local irritation in the stomach at a dose level of 100 mg/kg bw/d (males only) and above (both sexes). Apart from that, no signs of systemic toxicity up to a dose level of 1000 and 600 mg/kg bw/d in animals of both sexes were observed (treatment with 600 mg/kg bw/d from study day 7 onwards; impaired body weight parameters were related to a locally irritating effect of the test substance).

Since the high dose of 1000 mg/kg bw/d was given only during the first six days of the study, the no observed adverse effect level (NOAEL) for general systemic toxicity was deemed as 600 mg/kg bw/d in males and in females.The no observed adverse effect level (NOAEL) for the locally irritating effect in the stomach was 100 mg/kg bw/d in females and below that in males.

The NOAEL for reproductive performance and fertility was set to 600 mg/kg bw/d in male and female Wistar rats. The NOAEL for developmental toxicity was deemed as 600 mg/kg bw/d. This GLP-conform study performed according to OECD TG 422 is classified as acceptable (key study).

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Only one study available. GLP-compliant guideline study

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. There were no significant systemic toxic effects at doses of 600 mg/kg bw/d upon subacute oral exposure in rats (OECD 422). As a result, the substance is not classified for repeated dose toxicity under Regulation (EC) No. 1272/2008.