Oligomerisation products of 2,2'- oxydiethanol with ethylene oxide and propylene oxide

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

NOAEL for male (general toxicity) 1000 mg/kg bw/day

NOAEL for female (general toxicity): 300 mg/kg bw/day

NOAEL (reproduction/developmental toxicity) >= 1000 mg/kg bw/day

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

February – March, 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Strain: Hsd Cpb:WU
- Source: Harlan-Winkelmann GmbH, Borchen Germany
- Age at start of treatment: males: 11-12 weeks, females: 12-13 weeks
- Weight at study initiation: males: 321-348g; females: 181-206g
- Housing: individual
- Diet (e.g. ad libitum): ad libitum(Provimi Kliba maintenance Diet)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1°C
- Humidity (%): 50
- Air changes (per hr): ≥ 10 passages/hour
- Photoperiod (hrs dark / hrs light): 12 hours rhythm

IN-LIFE DATES: From: January 31st to May 10, 2005

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

Amount of vehicle (if gavage): 10 ml/kg bw

Details on mating procedure:

MATING PROCEDURES: Pairing was performed overnight by placing one F0 female animal together with one F0 male rat. If sperm was detected in the vaginal smear taken on the morning following mating, this day was regarded as day 0 of gestation. Animals were paired daily during the 2-week mating and one week remating period. Females in which insemination had not been detected by the end of the 2-week mating period, were mated for another week with another male of the respective dose group which had successfully inseminated a female paired with it. F0 females found sperm-positive after the first matings but where body weight gain did not indicate pregnancy by the end of the 2-week mating period were paired again for 7 days during the remating period.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Liquid chromatography.

Frequency of treatment:

Daily

Remarks:

Doses / Concentrations:
0
Basis:
nominal in water

Remarks:

Doses / Concentrations:
100
Basis:
nominal in water

Remarks:

Doses / Concentrations:
300
Basis:
nominal in water

Remarks:

Doses / Concentrations:
1000
Basis:
nominal in water

No. of animals per sex per dose:

12 animals per sex per dose

Control animals:

yes, concurrent vehicle

Details on study design:

Doses were selected according to a preceding subacute rat study were the same doses were applied over 4 weeks by gavage.

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: The experimental animals were inspected twice a day for morbidity and mortality (once on weekends and public holidays). General clinical examinations (in the home cage) were made daily (especially findings occurring during littering e.g. prolonged parturition) some time (about 30 to 60 minutes) after the administration and recorded, if any.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: F0-Males: Prior to the treatment and then weekly up to necropsy.
F0-Females: Prior to the treatment and then weekly during premating and mating
period. Additionally, during pregnancy and lactation period daily.

BODY WEIGHT: Yes
- Time schedule for examinations: The individual body weights of all parental animals were determined just prior before the first treatment of animals and then daily thereafter.

FOOD CONSUMPTION: Yes
- Time schedule for examinations: The food intake of F0 males was measured weekly during the premating period on a seven day basis.
In F0 females food intake was measured in the same way during the premating period. During gestation period determination of food intake was done on post-coital days 0-7, 7-14 and 14-20. During lactation period determination of food intake was done on day 0-4 p.p.

WATER CONSUMPTION : No

Litter observations:

-The numbers of live and dead pups as well as the sex of the pups (including those of dead pups, if possible) were determined shortly after birth (on postpartum day 0) and on day 4 p.p. At these time points individual body weights and clinical signs were recorded as well. Note was taken of any apparent malformations.

Postmortem examinations (parental animals):

Gross pathological examination:
- Gross pathological examination was performed for all females and males at necrosy. In F0 females the number of implantation sites was counted and the number of corpora lutea was determined.
HISTOPATHOLOGY / ORGAN WEIGHTS
- Organ weights: testes, epididymides,
- Abnormalities and the following organs were preserved: testes, epididymides, prostate, seminal vesicles with coagulating glands, uterus with vagina, ovaries with oviducts.
Histopathological examination of reproductive organs (testes, epididymides, ovaries) from control and 1000 mg/kg (highest dose) group.

Statistics:

Analysis of Variance (ANOVA) and in case of significant results Dunnett test as post hoc test 2*N CHi2 test; in case of significant differences Fisher's exact test with Bonferroni correction CHi2 test and Fisher’s Exact test.

Reproductive indices:

Reproductive indices:
- mating performance
- insemination index
- fertility index
- gestation index

Clinical signs:

effects observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Clinical signs: Transient salivation in in both sexes at 1000 mg/kg. Body weight: Slight body weight loss occurred in females of the 1000 mg/kg dose group during lactation. (Marginal body weight gains during premating period all doses).
Gross pathology: Findings in one animal found in moribund condition an immediately sacrificed - not considered test material related.

Dose descriptor:

NOEL

Effect level:

300 mg/kg bw/day (nominal)

Sex:

male

Basis for effect level:

other: based on salivation - as concluded in the report.

Dose descriptor:

NOEL

Effect level:

100 mg/kg bw/day (nominal)

Sex:

female

Basis for effect level:

other: based on salivation - as concluded in the report.

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw (total dose)

Sex:

female

Basis for effect level:

other: NOAEL as concluded in the report based on general toxicity (loss of body weight during lactation) as a potential relation to dosing could not be ruled out by the author.

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Sex:

male

Basis for effect level:

other: NOAEL as concluded in the report. No adverse effects observed with males in the Study.

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

Clinical signs: 4 pups with filiformed tip at 1000 mg/kg compared to 3 pups in control. In line with occurrences of filiformed tip in historical control.
Sex ratio in F1: not affected by treatment.
Not considered test material related. Other singular findings all not considered treatment related.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Reproductive effects observed:

not specified

Conclusions:

The registrant considers the mild bodyweight loss observed with females at the highest dose group (1000 mg/kg bw/day) as non adverse treatment related effect as it follows a statistically significant increased body weight gain, as compared to control, in the premating phase.
The No Adverse Effect Level is therefore concluded to be >=1000 mg/kg bw/day.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Adequate testing has been undertaken on a sufficient number of the core substances and repeating units. None of the tested core substances and none of the repeating units is classifiable as a reproductive toxin. Hence it would be anticipated that the NLP polyols, as a category, would also not be reproductive toxins. Three of the NLP polyols, nitrilotriethanol, propoxylated, diaminotoluene, propoxylated and ethylenediamine, ethoxylated and propoxylated, were tested to fulfil the requirements of Annex VIII (>10 tonnes/y).

Toxicity to reproduction of Nitrilotriethanol, propoxylated was investigated in an OECD 421 screening study. Treatment with 2,2',2''-Nitrilotriethanol, propoxylated resulted in increased incidence of salivation at the 1000 mg/kg dose in both genders and possibly transiently in females at 300 mg/kg dose group. Females of the 1000 mg/kg dose group showed body weight loss during lactation for which treatment relationship could not be completely excluded.

The reported NOAEL for male (general toxicity) 1000 mg/kg bw/day

The reported NOAEL for female (general toxicity): 300 mg/kg bw/day

The registrant considers the mild bodyweight loss observed with females at the highest dose group (1000 mg/kg bw/day) as non adverse treatment related effect as it follows a statistically significant increased body weight gain, as compared to control, in the premating phase. Possible minor effects on bodyweight do not constitute 'serious damage', the requirement relevant to classification.

The No Adverse Effect Level is therefore concluded to be >=1000 mg/kg bw/day.

There were no detectable effects in offspring. Thus the reported NOAEL (reproduction/developmental toxicity) >= 1000 mg/kg bw/day.

The results from these tests confirmed the pre-existing information: The NLP polyols are not reproductive or developmental toxins. Furthermore, a range of studies has been conducted on core substances and repeating units and screening tests have been conducted on most substances in the NLP polyol categories. It is possible to ‘read across’ the results from all of these sources to all substances in these categories. Sufficient data exist to permit robust conclusions that the substances are not reproductive or developmental toxins and that no further testing is required.

Short description of key information:

NOAEL (reproduction/developmental toxicity, OECD 421) >=1000 mg/kg bw/day.

Effects on developmental toxicity

Description of key information

NOAEL (maternal tox) 1000 mg/kg bw/day

NOAEL (developmental toxicity/ teratogenicity, OECD 414) >=1000 mg/kg bw/day

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and Guideline study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Qualifier:

according to guideline

Guideline:

EU Method B.31 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Crl:CD(SD)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River
- Age at study initiation: sexually mature adult
- Weight at study initiation: 200-250 g
- Housing: 1/cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 (range of 20-26)
- Humidity (%): 50 (range of 30-70)
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Dosing solutions were prepared by mixing the test material in ultrapure water at concentrations of 0, 25, 75 or 250 mg/ml and administered at a dose volume of 4 ml/kg body weight. Dose volumes were adjusted daily based on individual body weights. The control rats were dosed with ultrapure water at 4 ml/kg body weight. Dose solutions were prepared periodically throughout the study based on the established stability.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Dose confirmation of dose solutions from the first mix was performed pre-exposure. The homogeneity of the low- and high-dose test solutions was determined concurrent with dose confirmation. The method used for analyzing the test material in ultrapure water was liquid chromatography-mass spectrometry (LC/MS).

Details on mating procedure:

- Impregnation procedure: purchased timed pregnant

Duration of treatment / exposure:

GD 6-20

Frequency of treatment:

once daily, 7 days/week

Duration of test:

up to GD 21

No. of animals per sex per dose:

24/dose level

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose levels for this study were selected on the basis of the developmental toxicity probe study. Oral administration of Ethylenediamine, ethoxylated and propoxylated (>1 – <8.5 mol of EO and PO) to time-mated Crl:CD(SD) rats up to and including the limit dose of 1000 mg/kg/day resulted in no maternal toxicity and no indication of embryo/fetal lethality; therefore, no observable toxicity was expected. The high-dose of 1000 mg/kg/day represents a limit dose as defined in the Health Effects Test Guideline of the United States Environmental Protection Agency (OPPTS 870.3700 Prenatal Developmental Toxicity Study). The lower dose levels were selected to provide dose response data for any toxicity that may have been observed among the high-dose group rats and to establish a no-observed-effect level (NOEL).
- Rationale for animal assignment: Animals were stratified by GD 0 body weight and then randomly assigned to treatment groups using a computer program designed to increase the probability of uniform mean group weights and standard deviations at the start of the study.

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily
- All rats were observed in their cages for significant clinical abnormalities clearly visible upon a limited examination and to monitor the general health of the animals.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least once daily; approximately 1 hour after dosing.

BODY WEIGHT: Yes
- Time schedule for examinations: Recorded on GD 0 by the supplier and daily from GD 6-21. Statistical analysis of body weights was performed using data collected on GD 0, 6, 9, 12, 15, 18, and 21. Statistical analysis of body weight gains was conducted for the following intervals: GD 0-6, 6-9, 9-12, 12-15, 15-18, 18-21, 6-21, and 0-21.

FOOD CONSUMPTION: Yes, recorded every 3 days from GD 3-21.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 21
- Organs examined: The maternal necropsy included an examination of the external tissues and all orifices. The skin was reflected from the carcass, the thoracic and abdominal cavities were opened and the viscera were examined. The stomach, liver, and kidneys were dissected from the carcass and were incised. Any obvious gross pathologic alterations were recorded.

OTHER: The weight of the liver, kidneys, and gravid uterus were recorded. The ratios of liver and kidney weights to terminal body weight were calculated. Representative portions of liver, kidneys, and gross lesions were preserved in neutral, phosphate-buffered 10% formalin. Miccoscopic examination of the liver, kidneys, and gross lesions was not conducted.

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: Yes
- Number of late resorptions: Yes

OTHER: The uteri of females lacking visible implantations was stained with a 10% aqueous solution of sodium sulfide based on (Kopf et al., 1964) and examined for evidence of early resorptions in order to verify pregnancy status.

Fetal examinations:

- External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [ half per litter]
- Skeletal examinations: Yes: [ half per litter]
- Head examinations: Yes: [half per litter]

Statistics:

Maternal body weights, maternal body weight gains, maternal organ weights, gravid uterine weights, fetal body weights, feed consumption: Bartlett's test, parametric or non-parametric ANOVA, Dunnett's test or Wilcoxon Rank-Sum test with Bonferroni's correction
Frequency of pre- and post-implantation loss and fetal alterations: censored Wilcoxon test with Bonferroni's correction
Number of corpora lutea, implantations, and litter size: non-parametric ANOVA, Wilcoxon Rank-Sum test with Bonferroni's correction
Pregnancy rates: Fisher exact probability test with Bonferroni's correction
Fetal sex ratios: binomial distribution test
Statistical outliers were identified using a sequential method and, if excluded, were excluded for sound scientific reasons.

Indices:

Pre/post-implantation loss

Historical control data:

Yes

Clinical signs:

no effects observed

Description (incidence and severity):

Clinical and cage-side examinations performed on all animals revealed no treatment-related findings. Observations were sporadic and transient and were therefore considered unrelated to treatment. See 'Overall remarks, attachments' below for data.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

There were no deaths. See 'Overall remarks, attachments' below for data.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no treatment-related differences in the body weights or body weight gains in any of the treated groups when compared to controls. See 'Overall remarks, attachments' below for data.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no treatment-related differences in the amount of feed consumed in any of the treated groups when compared to controls. See 'Overall remarks, attachments' below for data.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

There were no treatment-related differences in any of the measured parameters in any of the treated groups when compared to controls. See 'Overall remarks, attachments' below for data.

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were no treatment-related gross pathologic observations.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Number of abortions:

no effects observed

Description (incidence and severity):

See 'Overall remarks, attachments' below for data.

Pre- and post-implantation loss:

effects observed, non-treatment-related

Description (incidence and severity):

There were no treatment-related effects on numbers of implantations, percent pre-implantation loss, percent postimplantation loss, in any of the treated groups compared to controls. Mean percent post-implantation loss was statistically identified as higher in the 1000 mg/kg/day group when compared to the control group; however, this value was well within the range of historical control data and, therefore, was not considered treatment related. See 'any other information on results incl. tables' and 'Overall remarks, attachments' below for data.

Total litter losses by resorption:

no effects observed

Description (incidence and severity):

There were no treatment-related effects on resorption rates. See 'Overall remarks, attachments' below for data.

Early or late resorptions:

no effects observed

Description (incidence and severity):

See 'Overall remarks, attachments' below for data.

Dead fetuses:

no effects observed

Description (incidence and severity):

See 'Overall remarks, attachments' below for data.

Changes in pregnancy duration:

no effects observed

Description (incidence and severity):

See 'Overall remarks, attachments' below for data.
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.DescriptionIncidenceAndSeverityEffectsOnPregnancyDuration): See 'Overall remarks, attachments' below for data.

Changes in number of pregnant:

no effects observed

Description (incidence and severity):

See 'Overall remarks, attachments' below for data.

Other effects:

not specified

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
Clinical and cage-side examinations performed on all animals revealed no treatment-related findings. Observations were sporadic and transient and were therefore considered unrelated to treatment.
There were no treatment-related differences in the body weights or body weight gains in any of the treated groups when compared to controls.
There were no treatment-related differences in teh amount of feed consumed in any of the treated groups when compared to controls.
There were no treatment-related differences in terminal body, liver, or kidney weights in any of the treated groups when compared to controls.
There were no treatment-related gross pathological observations.
There were no treatment-related effects on pregnancy rates, resorption rates, litter size, numbers of corpora lutea or implantations, percent pre-implantation loss, percent post-implantation loss, fetal sex ratios, fetal body weights or gravid uterine weights in any of the treated groups compared to controls. Mean percent post-implantation loss was statistically identified as higher in the 1000 mg/kg/day group when compared to the control group; however, this value was well within the range of historical control data and, therefore, was not considered treatment related. See 'any other information on results incl. tables' and 'Overall remarks, attachments' below for data.

Dose descriptor:

NOEL

Effect level:

1 000 mg/kg bw/day

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOEL

Effect level:

1 000 mg/kg bw/day

Basis for effect level:

other: developmental toxicity

Abnormalities:

not specified

Fetal body weight changes:

no effects observed

Description (incidence and severity):

There were no treatment related effects on fetal body weights. See 'Overall remarks, attachments' below for data.

                                              
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): See 'Overall remarks, attachments' below for data.

Reduction in number of live offspring:

no effects observed

Description (incidence and severity):

See 'Overall remarks, attachments' below for data.

Changes in sex ratio:

no effects observed

Description (incidence and severity):

See 'Overall remarks, attachments' below for data.

Changes in litter size and weights:

no effects observed

Description (incidence and severity):

See 'Overall remarks, attachments' below for data.

Changes in postnatal survival:

no effects observed

Description (incidence and severity):

See 'Overall remarks, attachments' below for data.

External malformations:

no effects observed

Description (incidence and severity):

There were no treatment-related external alterations in any dose group. Incidental findings bearing no relationship to treatment included the malformations of agnathia and astomia in a single control fetus. See 'any other information on results incl. tables' and 'Overall remarks, attachments' below for data.

Skeletal malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were no statically significant or treatment-related skeletal alterations in any dose group. Incidental findings bearing no relationship to treatment included the malformations sternoschisis and fused ribs and the variations delayed ossification (DO) interparietal, supernumerary skull bone, DO cervical centra, extra site of ossification sternebrae, irregular pattern of ossification sternebrae, extra 1st lumbar rib, DO thoracic centra, and DO pubis. Given that these observations occurred in the control group, at low frequencies, and/or lacked a dose response, these observations were considered spurious and unrelated to treatment. See 'any other information on results incl. tables' and 'Overall remarks, attachments' below for data.

Visceral malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were no treatment-related visceral alterations in any dose group. Incidental findings bearing no relationship to treatment included the malformations situs inversus, retroesophageal aortic arch, and hydronephosis and the variations fused lung, hemorrhage adrenal, pale liver, and supernumerary hepatic liver lobule. Given that these observations occurred in the control group, at low frequencies, and/or lacked a dose response, these observations were considered spurious and unrelated to treatment. See 'any other information on results incl. tables' and 'Overall remarks, attachments' below for data.

Other effects:

no effects observed

Description (incidence and severity):

Craniofacial Examination
There were no treatment-related craniofacial alterations in any dose group. Incidental findings bearing no relationship to treatment included the malformation
microphthalmia in a single control fetus.

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
There were no statisticaly signficant differences in the incidence of any fetal alteration in any of the treatment groups compared to controls. The small number of alterations observed in fetuses from dams administered Ethylenediamine, ethoxylated and propoxylated either occurred at low frequences, were within recent historical control values, and/or were not dose related.

Dose descriptor:

NOEL

Effect level:

1 000 mg/kg bw/day

Basis for effect level:

other: embryofetal development

Abnormalities:

no effects observed

Developmental effects observed:

no

Post-implantation Loss:

Dose Level (mg/kg/day)	0	100	300	1000
Mean % Post-implantation Loss	3.3 +/- 3.7	3.5 +/- 7.7	2.2 +/- 3.9	6.2* +/- 5.1

*Statistically different from control mean by Consored Wilcoxon's test alpha =0.05

Historal Control Post-implantation Loss:

(D=Dietary, G=Gavage)

Study Route Year

1 D 2010

2 D 2010

3 D 2010

4 D 2012

5 D 2012

6 D 2014

7 G 2014

8 D 2014

9 G 2014

10 G 2015

11 G 2015

12 D 2015

13 G 2015

Mean % Post-implantation Loss

1.9 +/- 4.3

1.4 +/-2.8

2.2 +/-3.4

4.1 +/-9.8

2.0 +/-3.5

4.0 +/-6.5

5.1 +/-4.0

4.2 +/-6.0

3.9 +/-11.1

3.4 +/-4.9

8.8 +/-16.8

2.3 +/-5.3

3.7 +/-5.9

D=Dietary, G=Gavage

Bold type indicates the highest observed value for the endpoint.

Incidences of External Malformations

Dose (mg/kg/day)		0	100	300	1000
Agnathia	F L	1/303^a  1/22	0/326 0/24	0/303 0/22	0/303 0/22
Astomia	F L	1/303^a  1/22	0/326 0/24	0/303 0/22	0/303 0/23

F = fetuses; L = litters

^aMalformations denoted with the same superscript were noted in a single fetus.

Incidences of External Malformations:

Dose (mg/kg/day)		0	100	300	1000
Microphthalmia	F L	1/153 1/22	0/160 0/24	0/150 0/22	0/151 0/23

F=fetuses; L= litters

Incidences of Visceral Malformations:

Dose (mg/kg/day)		0	100	300	1000
Situs Inversus	F L	0/153 0/22	0/160 0/24	1/150 1/22	0/151 0/23
Retroesophageal Aortic Arch	F L	0/153 0/22	0/160 0/24	0/150 0/22	1/151 1/23
Hydronephrosis	F L	1/153 0/22	1/160 1/24	1/150 0/22	0/151 0/23

F=fetuses; L=litters

Incidences of Accessory Blood Vessel Kidney:

Dose (mg/kg/day)		0	100	300	1000
Accessory Blood Vessel Kidney	F L	1/153 (0.7%) 1/22 (4.5%)	1/160 (0.6%) 1/24(4.2%)	4/150 (2.7%) 2/22 (9.1%)	3/151 (2.0%) 3/23 (13.0%)

F=fetuses; L=litters

Historical Control Data for Accessory Blood Vessel Kidney:

Study    Route Year

1 D 2010

2 D 2010

3 D 2010

4 D 2012

5 D 2012

6 D 2014

7 G 2014

8 D 2014

9 G 2014

10 G 2015

11 G 2015

12 D 2015

13 G 2015

Accessory Blood Vessel Kidney

F L

0/138 (0.0%) 0/22 (0.0%)

0/168 (0.0%) 0/24 (0.0%)

0/179 (0.0%) 0/26 (0.0)

0/123 (0.0%) 0/22 (0.0%)

0/150 (0.0%) 0/24 (0.0%)

0/137 (0.0%)  0/23 (0.0%)

0/136 (0.0%) 0/23 (0.0%)

0/133 (0.0%) 0/24 (0.0%)

0/140 (0.0%) 0/24 (0.0%)

0/148 (0.0%)   0/24 (0.0%)

0/124 (0.0%) 0/21 (0.0%)

0/153 (0.0%) 0/23 (0.0%)

3/146 (2.1%) 3/24 (12.5%)

D=Dietary, G=Gavage

F = fetuses; L = litters

Bold type indicates the highest observed value for the endpoint.

Incidences of Skeletal Malformations

Dose (mg/kg/day)		0	100	300	1000
Sternoschisis	F L	1/150 1/22	0/166 0/24	0/153  0/22	0/152 0/23
Fused Ribs	F L	0/150 0/22	1/166 1/24	0/153 0/22	0/152 0/23

F=fetuses; L=litters

Incidences of Selected Skeletal Variations:

Dose (mg/kg/day)		0	100	300	1000
DO Sternebrae	F L	0/150 (0.0%) 0/22 (0.0%)	1/166 (0.6%) 1/24 (4.2%)	3/153 (2.0%) 1/22 (4.5%)	3/152 (2.0%) 3/23 (13.0%)
Class I Wavy Ribs	F L	0/150 (0.0%) 0/22 (0.0%)	1/166 (0.6%) 1/24 (0.0%)	0/153 (0.0%) 0/22 (0.0%)	2/152 (1.3%) 2/23 (8.7%)
Class II Wavy Ribs	F L	0/150 (0.0%) 0/22 (0.0.0%)	1/166 (0.6%) 1/24 (4.2%)	0/153 (0.0%) 0/22 (0.0%)	1/152 (0.7%) 1/23 (4.3%)
Calloused Ribs	F L	1/150 (0.7%) 1/22 (4.5%)	2/166 (1.2%) 1/24 (4.2%)	1/153 (0.7%) 1/24 (4.5%)	2/152 (1.3%) 2/23 (8.7%)

F=fetuses; L=litters

Historical Control Data for Selected Skeletal Variations:

Study Route Year

1 D 2010

2 D 2010

3 D 2010

4 D 2012

5 D 2012

6 D 2014

7 G 2014

8 D 2014

9 G 2014

10 G 2015

11 G 2015

12 D 2015

13 G 2015

DO Sternebrae

F L

1/138 (0.7%)  1/22 (4.5%)

2/152 (1.3%) 2/24 (8.3%)

3/180 (1.7%) 2/26 (7.7%)

2/131 (1.5%) 2/22 (9.1%)

2/151 (1.3%) 2/24 (8.3%)

1/139 (0.7%) 1/23 (4.3%)

0/135 (0.0%) 0/23 (0.0%)

1/134 (0.7%) 1/24 (4.2%)

0/145 (0.0%) 0/24 (0.0%)

2/151 (1.3%) 2/24 (8.3%)

0/125 (0.0%) 0/21 (0.0%)

0/152 (0.0%) 0/23 (0.0%)

2/147 (1.4%) 2/24 (8.3%)

Class I Wavy Ribs

F L

0/138 (0.0%) 0/22 (0.0%)

0/152 (0.0%) 0/24 (0.0%)

2/180 (1.1%) 2/26 (7.7%)

1/131 (0.8%) 1/22 (4.5%)

2/151 (1.3%) 2/24 (8.3%)

0/139 (0.0%) 0/23 (0.0%)

1/135 (0.7%) 1/23 (4.3%)

4/134 (30%) 1/24 (4.2%)

0/145 (0.0%) 0/24 (0.0%)

0/151 (0.0%) 0/24 (0.0%)

0/125 (0.0%) 0/21 (0.0%)

1/152 (0.7%) 1/23 (4.3%)

2/147 (1.4%) 2/24 (8.3%)

Class II Wavy Ribs

F L

0/138 (0.0%) 0/22 (0.0%)

0/152 (0.0%) 0/24 (0.0%)

0/180 (0.0%)  0/26 (0.0%)

0/131 (0.8%) 0/22 (0.0%)

0/151 (0.0%) 0/24 (0.0%)

0/139 (0.0%) 0/23 (0.0%)

0/135 (0.0%) 0/23 (0.0%)

1/134 (0.7%) 1/24 (4.2%)

0/145 (0.0%) 0/24 (0.0%)

0/151 (0.0%) 0/24 (0.0%)

0/125 (0.0%) 0/21 (0.0%)

0/152 (0.0%) 0/23 (0.0%)

2/147 (1.4%) 2/24 (8.3%)

Calloused Ribs

F L

0/138 (0.0%) 0/22 (0.0%)

0/152 (0.0%) 0/24 (0.0%)

1/180 (0.6%) 1/26 (3.8%)

0/131 (0.8%) 0/22 (0.0%)

1/151 (0.7%) 1/24 (4.2%)

0/139 (0.0%) 0/23 (0.0%)

3/135 (2.2%) 3/23 (13.0%)

4/134 (30%) 1/24 (4.2%)

0/145 (0.0%) 0/24 (0.0%)

0/151 (0.0%) 0/24 (0.0%)

0/125 (0.0%) 0/21 (0.0%)

0/152 (0.0%) 0/23 (0.0%)

2/147 (1.4%) 2/24 (8.3%)

D=Dietary, G=Gavage

F = fetuses; L = litters

Bold type indicates the highest observed value for the endpoint.

Conclusions:

Gavage administration of Ethylenediamine, ethoxylated and propoxylated (>1-<8.5 mol of EO and PO) up to, and including the limit dose of 1000 mg/kg/day, resulted in no treatment-related maternal toxicity and no indication of embryo/fetal toxicity or teratogenicity. Therefore, under the conditions of this study, the no-observed-effect level (NOEL) for maternal toxicity and developmental toxicity was the limit dose of 1000 mg/kg/day.

Executive summary:

The purpose of this study was to evaluate the maternal and developmental toxicity of Ethylenediamine, ethoxylated and propoxylated (>1-<8.5 mol of EO and PO) in Crl:CD(SD) rats following repeated gavage administration.

Groups of 24 time-mated female rats were administered Ethylenediamine, ethoxylated and propoxylated (>1-<8.5 mol of EO and PO) in ultrapure water by gavage at dose levels of 0, 100, 300, or 1000 mg/kg/day on gestation day (GD) 6 through 20. In-life maternal study parameters included clinical observations, body weight, body weight gain and feed consumption. On GD 21, all rats were euthanized and examined for gross pathologic alterations. Liver, kidneys and gravid uterine weights were recorded, along with the number of corpora lutea, uterine implantations, resorptions and live/dead fetuses. All fetuses were weighed, sexed and examined for external alterations. Approximately one half of the fetuses were examined for visceral and craniofacial alterations while skeletal examinations were conducted on the remaining fetuses.

Gavage administration of Ethylenediamine, ethoxylated and propoxylated (>1-<8.5 mol of EO and PO) up to, and including the limit dose of 1000 mg/kg/day, resulted in no treatment-related maternal toxicity and no indication of embryo/fetal toxicity or teratogenicity.

Therefore, under the conditions of this study, the no-observed-effect level (NOEL) for maternal toxicity and developmental toxicity was the limit dose of 1000 mg/kg/day.

Reference 2

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

(2001)

Qualifier:

according to guideline

Guideline:

EU Method B.31 (Prenatal Developmental Toxicity Study)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)

Qualifier:

according to guideline

Guideline:

other: Japanese MAFF guidelines: Guideline on the Compiling of Test Results on Toxicity “Teratology Study”, 12-Nousan No. 8147 of November 24, 2000, amended June 26, 2001

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Strain: Hsd Cpb:WU (SPF-bred)
- Source: Harlan-Nederlands, 5960 AD Horst, The Netherlands
- Age at study initiation: 12-17 weeks
- Weight at study initiation: 220 - 265 g (females); 428 - 536 g (males)
- Housing: Starting from gestation day 0 individually in Type IIIh Makrolon cages on low-dust wood shavings.
- Diet and water: ad libitum
- Acclimation period: at least 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): approximately 55 %
- Air changes (per hr): > 10 per hour
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

other: demineralized water

Details on exposure:

Administration volume: 10 mL/kg bw

PREPARATION OF DOSING SOLUTIONS: The formulations were prepared as needed taking into account the analytically determined stability.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The test substance was administered as a solution in the vehicle. The formulations were stored at room temperature.
Before the start of treatment, the suitability of the formulation was confirmed by the analysis of concentration and stability of dosage forms prepared in the same way as it was done in the study (0.5 and 300 mg/mL, covering the concentration range used after 8 days of storage). Analyses were carried out before the start of the study. Content checks of the active ingredient in samples with concentrations of 10, 30, and 100 mg/mL were carried out twice during the study.

Details on mating procedure:

The animals were mated by placing two females overnight into a Type IIIh cage together with one male rat. If sperm was detected in the vaginal smear taken on the morning following mating, this day was regarded as day 0 of gestation.

Duration of treatment / exposure:

Day 6 - 20 p. c.

Frequency of treatment:

once daily (between 06:00 and 12:00 CET)

Duration of test:

from days 0 to 21 p.c.

Remarks:

Doses / Concentrations:
100, 300, 1000 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

25 females

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose levels were selected based on the results of a previous dose toleration study in rats, which revealed no treatment related findings after 1000 mg/kg.

Maternal examinations:

CLINICAL EXAMINATIONS: Yes
- Time schedule: The females were inspected from days 0 to 21 p.c. twice daily (once daily only on weekends, on public holidays, and on day 21 p.c.), and all findings were recorded. Attention was paid to disturbances in the general condition of the rats (appearance, behavior), and any alterations concerning their excretory products.

BODY WEIGHT: Yes
- Time schedule for examinations: The body weight of the females was determined on day 0 p.c. and daily from day 6 to day 21 p.c. Corrected body weight gain was determined by subtracting the uterus weight on day 21 p.c. from the body weight gain from days 0 to 21 p.c.

FOOD CONSUMPTION: Yes
- The food intake of the animals was determined from the difference in weight between the food offered and the food not consumed for the following days of gestation: Days 0 - 3, 3 - 6, 6 - 9, 9 - 12, 12 - 15, 15 - 18, and 18 -21.

WATER CONSUMPTION: Yes
- Water intake was assessed daily by visual estimation of the quantities left over and reported together with clinical findings.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Cesarean sections were performed on day 21 p.c. without knowledge of treatment groups.
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Number of dead and live fetuses: Yes
- Other: individual weight and appearance of the placentas

Fetal examinations:

- Sex of live fetuses
- Individual weights of live fetuses
- External examinations: Yes: [findings in alive and dead fetuses are included]
- Soft tissue and head examinations : Yes: [evaluation of about half of alive fetuses per litter]
- Skeletal and cartilage abnormalities : Yes: [evaluation in about half of alive fetuses per litter]

Statistics:

Differences between the control and test item treated groups were considered to be significant when p < 0.05. Significant differences from the control group are indicated with * for p < 0.05 and ** for p < 0.01. Statistical evaluation was performed on an Alpha 800 5/500 computer using the following methods:
Analysis of Variance (ANOVA); in case of significance Dunnett's test for feed intakes, body weights, body weight gains, and corrected body weight gains, uterine weights, number of corpora lutea per female, number of implantations per female, number of live fetuses per female and as percentage of implantations per female, placental weights per female, fetal weights per female.
2 by N CHI2 test; in case of significant differences Fisher's exact test with Bonferroni correction for fertility rate, gestation rate, number of implantations per group, number of preimplantation losses per group, number of postimplantation losses, early resorptions, late resorptions or dead fetuses per group, number of live fetuses per group as percentage of implantations per group, number of male or female fetuses or fetuses with indeterminable sex per group, number of placentas with findings or litters with placental findings per group, number of fetuses or litters with external, visceral or skeletal findings, with malformations or with external or visceral deviations per group.
Kruskal-Wallis test and in case of significant differences Dunn's test for number of preimplantation losses per female, number of postimplantation losses, early resorptions, late resorptions or dead fetuses per female, number of male or female fetuses or fetuses with indeterminable sex per female, number of placentas with findings per female, number of fetuses with external or visceral findings, with malformations or with external or visceral deviations per female.
CHI2 test (correction according to yates) for number of fetuses or litters with cartilaginous tissue observations.

Indices:

gestation rate

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
BODY WEIGHTS:
Statistically significantly decreased body weight gain occurred from days 6-7 p.c. at the 1000 mg/kg level, for which a treatment related adverse effect is not assumed, as only this single interval revealed a decreased body weight gain, and because absolute body weight gain during treatment and gestation periods as well as corrected body weight gain were unaffected at this dose level. Statistically significantly increased body weight gain at the 100 mg/kg and 1000 mg/kg levels from days 20-21 p.c. is caused by the incidentally lower body weight gain of the current control group within this interval.
Thus, absolute and corrected body weight gains were unaffected by treatment at dose levels up to and including 1000 mg/kg. A treatment related adverse effect is not assumed for transiently decreased body weight gain from days 6-7 p.c. at the 1000 mg/kg level.

NECROPSY:
One female of the 1000 mg group showed a slight dilation of the left renal pelvis, for which a treatment related effect is not assumed due to its single occurrence, and since this finding is known as a spontaneous finding in the rat strain used. Furthermore, one female without implantation sites at the 1000 mg/kg level revealed a uterus tightly filled with clear fluid, for which a treatment related effect is not assumed, as this finding was also evident in one female of the current control group, and because this finding is known as a spontaneous finding in the rat strain used.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
WEIGHT and APPEARANCE of PLACENTAS:
Two double placentas out of one litter occurred at the 1000 mg/kg level, for which a treatment related effect is not assumed, as the incidence of this finding lay within the range of historical control data. Furthermore, one pale placenta was shown at the 300 mg/kg level, for which a treatment related effect is not assumed due to its single occurrence, because of a lacking dose dependency, and since this finding is known as a spontaneous finding in the rat strain used.
FETAL MALFORMATIONS:
The overall incidences of fetuses with malformations showed no dose dependency at dose levels up to and including 1000 mg/kg and revealed the highest values on a fetal and a litter basis in the 100 mg/kg group. The overall incidences of affected fetuses and litters lay well within the range of historical control data of the rat strain used .

number of fetuses per group/number of fetuses with malformations/malformed fetuses per group (%)/number of litters per group/number of litters with malformations/malformed litters per group (%)
Vehicle control: 262/0/0.0/21/0/0.0
100 mg/kg b.w. and day: 261/6/2.3/20/4/20.0
300 mg/kg b.w. and day: 288/4/1.4/21/4/19.0
1000 mg/kg b.w. and day: 262/3/1.1/20/3/15.0

FETAL EXTERNAL and VISCERAL DEVIATIONS

Circumscribed hard whitish areas in different localizations of the head occurred in all groups, without showing a dose dependency. Histopathological evaluation of this finding was performed in a recently conducted prenatal developmental toxicity study with the same rat strain and revealed calcium concrements without connection to the underlying tissue in the affected localizations. Calcium might have been dissolved from the fetal bones by the Wilson fixative and precipitated so that these findings were regarded as artifacts. The finding of a small hollow structure in different localizations of the head also occurred in all groups, without showing a dose dependency, and is also considered as a fixation artifact.
The remaining findings of the dose groups revealed no dose dependency in most of the cases or were isolated findings. The incidences were comparable with the current control group and/or lay within the range of historical control data.

FETAL SKELETAL DEVIATIONS INCLUDING CARTIÖAGINOUS DEVIATIONS
Sporadic statistically significant skeletal findings (increased incidence of an unossified round area right at the exoccipital bone at the 100 mg/kg level, decreased incidence of asymmetrical 5th sternebrae at the 300 mg/kg level) showed no dose dependency so that a treatment related effect was not evident

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: no adverse effects with regard to the parameters examined

Abnormalities:

not specified

Developmental effects observed:

not specified

Executive summary:

A developmental toxicity study was performed with the test substance according to OECD TG 414. Twenty-five inseminated female Wistar rats each were treated daily orally by gavage with the test substance in deminaralized water from day 6 to day 20 p.c. in the following doses: 0, 100, 300, and 1000 mg/kg body weight (bw), respectively. The fetuses were delivered by cesarean section on day 21 of gestation. Investigation was performed on general tolerance of the test substance by the females as well as on its effect on intrauterine development.

Appearance, behavior, mortality, absolute and corrected body weight gains, food intake, water intake, and fecal and urinary excretions were unaffected at dose levels up to and including 1000 mg/kg bw.

No treatment related gross pathological findings occurred at dose levels up to and including 1000 mg/kg.

The gestation rate, appearance and weights of placentas, postimplantation loss and correspondingly the number of fetuses, fetal sex distribution, and fetal weights were unaffected by treatment at dose levels up to and including 1000 mg/kg.

A treatment related effect on malformations, external, visceral, and skeletal (including cartilage) deviations was not evident at dose levels up to and including 1000 mg/kg.

Summarizing and evaluating all data investigated the following no-observed-adverse-effect levels (NOAELs) were determined: for maternal toxicity 1000 mg/kg bw, for developmental toxicity 1000 mg/kg bw.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Two ‘read-across’ substances (EDA EO PO and MEG PO) are included in the evaluation of this endpoint. Both ‘read-across’ substances belong to the category of NLP polyols. By including these two substances, the structural differences as well as similarities between source and target substance are accounted for: EDA EO PO represents a polyol with an amine linkage unlike the ether-linked target substance (DEG EO PO), however it reflects the ethoxylation and propoxylation properties of the target substance. MEG PO on the other hand is an ether-linked polyol and therefore has the type of linkage in common with the target substance. Thus, including these two studies, both type of linkage and type of alkoxylation of the target substance are considered which allows a better assessment.

The first study evaluating maternal and developmental toxicity was performed with the 'read-across' substance Ethylenediamine, ethoxylated and propoxylated (>1-<8.5 mol of EO and PO) (EDA EO PO) in Crl:CD(SD) rats following repeated gavage administration according to OECD TG 414.

Groups of 24 time-mated female rats were administered Ethylenediamine, ethoxylated and propoxylated (>1-<8.5 mol of EO and PO) in ultrapure water by gavage at dose levels of 0, 100, 300, or 1000 mg/kg/day on gestation day (GD) 6 through 20. In-life maternal study parameters included clinical observations, body weight, body weight gain and feed consumption. On GD 21, all rats were euthanized and examined for gross pathologic alterations. Liver, kidneys and gravid uterine weights were recorded, along with the number of corpora lutea, uterine implantations, resorptions and live/dead fetuses. All fetuses were weighed, sexed and examined for external alterations. Approximately one half of the fetuses were examined for visceral and craniofacial alterations while skeletal examinations were conducted on the remaining fetuses.

Gavage administration of Ethylenediamine, ethoxylated and propoxylated (>1-<8.5 mol of EO and PO) up to, and including the limit dose of 1000 mg/kg/day, resulted in no treatment-related maternal toxicity and no indication of embryo/fetal toxicity or teratogenicity.

Therefore, under the conditions of this study, the no-observed-effect level (NOEL) for maternal toxicity and developmental toxicity was the limit dose of 1000 mg/kg/day.

The second developmental toxicity study was performed with the 'read-across' substance Ethane-1,2-diol, propoxylated (MEG PO) according to OECD TG 414. Twenty-five inseminated female Wistar rats each were treated daily orally by gavage with the test substance in deminaralized water from day 6 to day 20 p.c. in the following doses: 0, 100, 300, and 1000 mg/kg body weight (bw), respectively. No treatment related effects up to and including 1000 mg/kg bw were observed concerning maternal toxicity and no indication of developmental/teratogenic toxicity was observed up to levels of 1000 mg/kg bw.

Based on all evaluated parameters the following no-observed-adverse-effect levels (NOAELs) were determined:

- for maternal toxicity 1000 mg/kg bw

- for developmental toxicity 1000 mg/kg bw.

Justification for classification or non-classification

Based on the results of a valid OECD TG 421 study conducted with the 'read-across' substance 2,2',2''-Nitrilotriethanol, propoxylated a NOAEL of 1000 mg/kg bw/day in rats could be assessed. Furthermore, valid OECD TG 414 studies with the 'read-across' substances EDA EO PO and MEG PO each showed a NOAEL of 1000 mg/kg bw/day in rats.

Thus, there is no indication for classification/labelling of the the test item according to either the EU Directive 67/548/EEC or the CLP Regulation (EC) No. 1272/2008