N-[3-(dimethoxymethylsilyl)propyl]ethylenediamine

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 Oct 2021 (study initiation) - 17 Jan 2023

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2023

Report date:

2023

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

adopted 25 June 2018

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N-[3-(dimethoxymethylsilyl)propyl]ethylenediamine

EC Number:

221-336-6

EC Name:

N-[3-(dimethoxymethylsilyl)propyl]ethylenediamine

Cas Number:

3069-29-2

Molecular formula:

C8H22N2O2Si

IUPAC Name:

N-[3-[dimethoxy(methyl)silyl]propyl]ethane-1,2-diamine

Test material form:

liquid

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Crl: WI(Han)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Age at study initiation: 16-17 weeks (males at start pairing), 11-12 weeks (females, at arrival)
- Weight at study initiation: 375 - 472 g (males), 197 - 247 g (females), before initiation of pairing
- Fasting period before study: No
- Housing: Type III H, polysulphone cages, housed individually except during mating
- Diet: Altromin 1324 maintenance diet for rats and mice, ad libitum
- Water: Tap water, sulphur acidified to a pH of approximately 2.8, ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 55 +/- 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 5 Nov 2020 To: 18 Feb 2021

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Remarks:

dried and de-acidified

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test item was dissolved in dried and de-acidified corn oil. Corn oil was filtered through a mixture of activated silica gel 60 and aluminium oxide (1:1, volume/volume), which had been filled into a glass chromatography column to three quarters of its height. For filtering, a vacuum of 75 mbar was applied. The dried and de-acidified vehicle was overlaid with argon and stored until usage.

The test item was weighed into a tared glass vial on a suitable precision balance and the vehicle was added to give the appropriate final concentration of the test item. The formulation was vortexed and/or stirred until visual homogeneity was achieved. After homogenization, the formulation was overlaid with argon to prevent instability caused by repeated contact of the test item formulation with air.

Based on the results of stability testing (separate study), the test item formulations were prepared fresh on each day of administration, and used within 2 hours. The prepared formulation was stored protected from light and at room temperature.

The test item formulation and vehicle were administered as a single daily dose to the animals by oral gavage. The application volume for all groups was 4 ml/kg bw. For each animal, the individual dosing volume was calculated on the basis of the body weight most recently measured.

A specified procedure was required for cleaning of the gavage apparatus (needle, flexible cannula or tube) after drawing up the test item and before administering the item by gavage as follows: Following completion of filling of the syringe attached to the gavage apparatus with the required amount of test item, the outside of the gavage tube was wiped clean of all test item residue. The gavage tube was then lightly tapped onto a clean towel to remove the remaining droplet from the tip of the tubing. Additionally, the canula was dipped into 5% aqueous glucose solution immediately prior to application to further reduce the risk of irritation during administration.

VEHICLE
- Justification for use and choice of vehicle: Suggested by sponsor based on the test item’s characteristics
- Concentration in vehicle: 0, 25, 150, 187.50, 250 mg/ml
- Amount of vehicle: 4 ml/kg, total dosing volume

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

In a separate study, test item formulations (15, 25, 250 mg/ml) were homgenous and stable for 2 hr at room temperature and for 2 days at -15 to -35C.

Details on mating procedure:

Mating was performed using a ratio of 1:2 (male to female). Females were paired for cohabitation in batches in order to control the number of animals for terminal sacrifice on a particular day. On the subsequent mornings, the vaginal smear of the females was checked to confirm evidence of mating. The day on which sperm is observed in the vaginal smear was considered as gestation day (GD) 0. After getting 92 sperm-positive females, the remaining females and males were discarded without any observation. However, in the case of the high dose (HD) group, an additional 3 females were mated due to mortality at 1000 mg/kg bw/day and assigned to the new 750 mg/kg bw/day HD group in order to obtain at least 20 pregnant females per group.

Duration of treatment / exposure:

GDs 5-19

Frequency of treatment:

Once daily

Duration of test:

GDs 0-20

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

600 mg/kg bw/day (nominal)

Doses / concentrations 4

Dose / conc.:

750 mg/kg bw/day (nominal)

Remarks:

Due to severe clinical signs and subsequent morbidity, the HD level was reduced from 1000 mg/kg bw/day to 750 mg/kg bw/day on 2 Dec 2020. The respective gestation days of the HD animals on this date are provided in Table 1 under other information on method and materials.

Doses / concentrations 5

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

HD level from study initiation through 1 Dec 2020

No. of animals per sex per dose:

At least 20 pregnant females per group

Control animals:

yes

Details on study design:

- Dose selection rationale: The highest dose level was chosen with the aim of inducing toxic effects, but not death or severe suffering. Thereafter, a descending sequence of dose levels was selected with a view to demonstrate any dose-related response and a NOAEL. Due to severe clinical signs and subsequent morbidity, the HD level was reduced from 1000 mg/kg bw/day to 750 mg/kg bw/day on 2 Dec 2020. The respective gestation days of the HD animals on this date are provided in Table 1 under other information on method and materials.
- Rationale for animal assignment: Random
- Fasting period before blood sampling for (rat) dam thyroid hormones: Not specified
- Time of day for (rat) dam blood sampling: At termination, time of day not specified

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
Twice daily, all animals were observed for morbidity and mortality except on weekends and public holidays when observations were made once daily.

DETAILED CLINICAL OBSERVATIONS: Yes
General clinical observations were made at least once a day, preferably at the same time each day.

BODY WEIGHT: Yes
All animals were weighed once before initiation of pairing to ensure that the body weights were within 20% variation. Sperm-positive females were weighed on GDs 0, 5, 8, 11, 14, 17 and 20, with body weights presented for these intervals: GDs 0-5, 5-8, 8-11, 11-14, 14-17 and 17-20. Males were not weighed in this study except once before initiation of pairing.

FOOD CONSUMPTION: Yes
Food consumption of sperm-positive females was determined for GDs 0-5, 5-8, 8-11, 11-14, 14-17 and 17-20. Food consumption was not measured for males during the entire study or for either sex during the mating period.

WATER CONSUMPTION: No

SACRIFICE:
On GD 20, sperm-positive (presumed pregnant) females were sacrificed using anaesthesia (ketamine/xylazin) and subjected to caesarean section.

Immediately after the termination or as soon as possible after death, the uteri were removed, the pregnancy status of the dams was confirmed, and the uterine content of pregnant females examined.

ORGAN WEIGHT: Yes
Each gravid uterus with cervix was weighed. However, the gravid uterus obtained from dead animals was not weighed.

The weight of thyroid/parathyroid glands was measured after 24 hours fixation in 4% neutral-buffered formaldehyde. Relative weights of thyroid/parathyroid glands were calculated in relation to the body weight (measured at necropsy) and were presented as percentage.

GROSS PATHOLOGY: Yes
At the time of termination or death during the study, each dam (presumed pregnant female) was examined macroscopically for any structural abnormalities or pathological changes which may have influenced the pregnancy. Any macroscopic findings were preserved in 4% neutral-buffered formaldehyde.

HISTOPATHOLOGY: Yes
A histopathological evaluation was carried out on the thyroid/parathyroid glands from all dams sacrificed at the end of treatment.

Ovaries and uterine content:

The uterine content of pregnant females was examined after termination including:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Position and number of foetuses in each uterine horn: Yes
- Number of live and dead foetuses: Yes

Blood sampling:

At termination, dam thyroid hormone levels (T3, T4, TSH) in abdominal aorta serum were assessed at the end of treatment using ELISA.

Fetal examinations:

SACRIFICE:
On GD20 via dam sacrifice with anaesthesia (ketamine/xylazin)

BODY WEIGHT: Yes
All foetuses were weighed on GD 20.

EXTERNAL: Yes
Each foetus was examined for external anomalies and sexed using the measured AGD. External sex was compared to internal sex. Relative AGD was calculated as AGD / Cube root of foetus weight. Incomplete testicular descent / cryptorchidism was noted in male foetuses.

VISCERAL: Yes
One half of each litter (20 litters per dose) was examined for soft tissue anomalies by a microdissection technique. Particular attention was paid to the reproductive tract which was examined for signs of altered development.

HEAD: Yes
The same half of each litter as for visceral (20 litters per dose) was subjected to craniofacial examination (eyes, brain, nasal passage and tongue) by razor blade serial sectioning technique.

SKELETAL: Yes
The remaining half of foetuses from each litter (20 litters per dose) was processed by Alizarin red staining and were examined for skeletal alterations.

Statistics:

All results were reported in tabular form (summarised in mean or summary tables and/or listed in individual data tables). A statistical assessment of the results of the body weight and food consumption was performed by comparing test item-treated groups with control using a one-way ANOVA and a post-hoc Dunnett Test. Results of absolute and relative organ weights, thyroid hormones and foetal evaluation parameters like external, visceral, craniofacial and skeletal parameters were statistically analysed by comparing values of treated animals with control animals using either a parametric one-way ANOVA and a post-hoc Dunnett Test or a non-parametric Kruskal-Wallis Test and a post-hoc Dunn’s Test, based on the results of homogeneity and normality tests. The statistics were performed with GraphPad Prism V.6.01 software or Ascentos 1.3.4 software (p<0.05 was considered as statistically significant).

Indices:

Not specified

Historical control data:

Historical control data for the strain of rat used (Wistar Han) mentioned for the prenatal data, foetal weights, visceral findings, and craniofacial findings.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Test item-related adverse clinical signs were observed in all animals at 600 mg/kg bw/day (MD) and 750/1000 mg/kg bw/day (HD), with the severity of signs occurring in a dose-dependent manner. No test item-related clincal signs were seen at 100 mg/kg bw/day (LD).

Predominant clinical signs in the MD and HD groups (with control and LD also shown) included:
- increased salivation (slight, moderate/severe; control-1/23, LD-1/23, MD-10/23, HD-20/26)
- moving the bedding (control-0/23, LD-1/23, MD-21/23, HD-25/26)
- abnormal breathing (control-0/23, LD-0/23, MD-6/23, HD-11/26)
- piloerection (control-0/23, LD-1/23, MD-3/23, HD-7/26)
- spontaneous reduced activity (slight/moderate/severe; control-0/23, LD-0/23, MD-2/23, HD-2/26).

A few incidences of both eyelids closed, dehydration, nasal discharge and coughing/sneezing were also observed in the MD and HD groups.

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, treatment-related

Description (incidence):

Test item-related mortality was observed in 600 mg/kg bw/day (MD) and 1000 mg/kg bw/day (initial HD). No mortality was observed at 100 mg/kg bw/day (LD), at 750 mg/kg bw/day (HD as of 2 Dec 2020), or in the control group.

Prior to reducing the HD level, one MD animal (no. 47) and four HD animals (nos. 70, 75, 76 and 77) were moribund sacrificed. After reducing the HD from 1000 to 750 mg/kg bw/day, no mortality was observed during the study and all other animals survived until the end of the study.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

A test item-related adverse effect on body weight gain was observed at 750/1000 mg/kg bw/day (HD) when compared with the control group. No test item-related effect on this endpoint was seen at 100 mg/kg bw/day (LD) or 600 mg/kg bw/day (MD). For mean body weight, no test item-related effect was identified at any dose.

For body weight gain over GDs 5-20, there was a slight (not significant) decrease in mean body weight gain observed in the HD group (9.52% below control) when compared to the control group. This effect was considered test item-related and adverse.

For two GD subintervals, the mean body weight gain was found to be decreased: on GDs 5-8 in all treated groups (LD-28.24%, MD-11.01% and HD-35.92% below control), and on GDs 11-14 in the MD (9.53% below control) and HD (29.47% below control) groups without statistical significance. The MD and HD decreases in mean body weight gain during these intervals improved at later gestation days, and were considered not related to the test item.

Mean body weight increased with the progress of the study in all the treated groups and the control group, and no test item-related effect was identified for this endpoint.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

A test item-related adverse effect on food consumption was observed at 750/1000 mg/kg bw/day (HD). No test item-related effect on this endpoint was seen at 100 mg/kg bw/day (LD) or 600 mg/kg bw/day (MD).

For GDs 5-20, mean food consumption observed in the LD, MD and HD groups were comparable to the control group. However, a slight (statistically significant) decrease in food consumption was observed in the HD group (7.77% below control) when compared to the control group. This effect was considered test item-related and adverse.

For a few GD subintervals, there was a slight decrease in food consumption observed in the HD group (8.99% on GDs 5-8, 6.61% on GDs 11-14, and 11.06% on GDs 14-17), but this effect was considered not related to the test item.

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

Description (incidence and severity):

The results for thyroid hormones in serum are presented under endocrine findings.

Endocrine findings:

effects observed, non-treatment-related

Description (incidence and severity):

No test item-related effects on thyroid hormone levels in serum were observed up to 750/1000 mg/kg bw/day (HD).

With one exception, the mean T3, T4 and TSH hormone levels in aortic serum were comparable between the treatment groups and the control group. At 600 mg/kg/kg bw/day (MD), there was a statistically significant increase in mean TSH. In the absence of dose dependency, this finding was not considered to be test item-related.

See also organ weight and histopathology findings.

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):

No statistically significant differences were observed for uterine weight (gravid with cervix) up to 750/1000 mg/kg bw/day (HD) when compared to the control group, and all values were within the historical control range for this strain of rat.

No statistically significant differences were observed for absolute and relative (to body weight) thyroid/parathyroid weights up to the HD when compared to the control group.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Test item-related adverse macroscopic lesions were observed at 600 mg/kg bw/day (MD) and 750/1000 mg/kg bw/day (HD), but not at 100 mg/kg bw/day (LD).

For the MD and HD groups, test item-related adverse macroscopic lesions were observed in moribund sacrificed animals. Animal no. 47 (MD group) had a gas-filled stomach, duodenum, jejunum, ileum and caecum. Animal no. 70 (HD group) had a gas filled stomach, duodenum, jejunum, ileum and caecum. Animal no. 75 (HD group) had many red foci in the glandular mucosa of the stomach and the stomach, and the jejunum, caecum and colon were gas filled. Female no. 76 (HD group) had a gas filled stomach, an enlarged thymus and enlarged hindlimb and female no. 77 (HD group) had a small thymus. The macroscopic findings observed in the gastrointestinal tract were most likely due to the irritating properties of the test item and, hence, considered to be test item-related and adverse.

No test item-related macroscopic lesions were observed in the LD group. Animal no. 34 of the LD group had a hard, brown mass in the liver facing the thoracic cavity near the diaphragmatic surface, which was considered incidental.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

No test item-related effects on post-fixed thyroid/parathyroid histopathology were observed up to 750/1000 mg/kg bw/day (HD) when compared to the control group.

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Maternal developmental toxicity

Number of abortions:

no effects observed

Description (incidence and severity):

None of the females showed signs of abortion or premature delivery prior to the scheduled sacrifice up to 750/1000 mg/kg bw/day (HD).

Pre- and post-implantation loss:

no effects observed

Description (incidence and severity):

No statistical significance was observed for pre- or post-implantation up to 750/1000 mg/kg bw/day (HD) when compared to the control group, and all values were within the historical control range for this strain of rat.

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Description (incidence and severity):

No statistical significance was observed for early or late resorptions up to 750/1000 mg/kg bw/day (HD) when compared to the control group, and all values were within the historical control range for this strain of rat.

Dead fetuses:

no effects observed

Description (incidence and severity):

No dead foetuses were noted up to 750/1000 mg/kg bw/day (HD).

Changes in pregnancy duration:

not examined

Description (incidence and severity):

Changes in pregnancy duration were not observed (no signs of abortion or premature delivery) prior to the scheduled sacrifice up to 750/1000 mg/kg bw/day (HD).

Changes in number of pregnant:

no effects observed

Description (incidence and severity):

Successful mating resulted in 21/23 pregnancies at 100 mg/kg bw/day (LD), 22/22 at 600 mg/kg bw/day (MD) and 20/22 at 750/1000 mg/kg bw/day (HD) compared to 20/23 pregnancies in the control group.

Other effects:

not examined

Effect levels (maternal animals)

Key result

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

gross pathology

mortality

Remarks on result:

other: Based on maternal systemic effects at 600 and 750/1000 mg/kg bw/day

Maternal abnormalities

Key result

Abnormalities:

no effects observed

Description (incidence and severity):

other: No maternal abnormalities related to developmental toxicity observed up to 750/1000 mg/kg bw/day

Results (fetuses)

Fetal body weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

No test item-related effects on foetal body weight were observed up to 750/1000 mg/kg bw/day (HD).

On an individual basis (sum of weight of all foetuses in the group divided by the total number of foetuses in respective group), the mean male and female foetal weights observed in the LD (100 mg/kg bw/day), MD (600 mg/kg bw/day) and HD groups were comparable to the control group. However, a statistically significant decrease was observed in the MD group in both male (4.90% below control) and female (4.52% below control) foetal weights on a per foetal basis when compared to the control group. This finding was considered incidental since there was no dose dependency and the values were within the historical control range for this strain of rat.

Reduction in number of live offspring:

no effects observed

Description (incidence and severity):

No statistical significance was observed for the number of live foetuses up to 750/1000 mg/kg bw/day (HD) when compared to the control group, and all values were within the historical control range for this strain of rat.

Changes in sex ratio:

no effects observed

Description (incidence and severity):

No statistical significance was observed for sex ratio up to 750/1000 mg/kg bw/day (HD) when compared to the control group, and all values were within the historical control range for this strain of rat.

Changes in litter size and weights:

no effects observed

Description (incidence and severity):

On a per litter basis (group mean of individual litter mean), no statistically significant effects on mean foetal weight, male and female foetal weight were observed up to 750/1000 mg/kg bw/day (HD) when compared to the control group.

Anogenital distance of all rodent fetuses:

effects observed, non-treatment-related

Description (incidence and severity):

ANOGENITAL DISTANCE:
No test item-related effects on AGD were observed up to 750/1000 mg/kg bw/day (HD).

The absolute and relative AGD in treatment groups remained unaffected when compared to the control group, except for a statistically significant increase in relative AGD of male foetuses at the MD (600 mg/kg bw/day). However, this finding occurred without any dose dependency and was not considered to be test item-related.

TESTICULAR DESCENT:
No effects observed. Testicular descent (abdominal) was complete in all male foetuses up to 750/1000 mg/kg bw/day (HD).

Changes in postnatal survival:

not examined

External malformations:

no effects observed

Description (incidence and severity):

There were no external abnormalities observed in any of the foetuses up to 750/1000 mg/kg bw/day (HD) or in the control group.

Skeletal malformations:

effects observed, non-treatment-related

Description (incidence and severity):

No test item-related skeletal findings were observed up to 750/1000 mg/kg bw/day (HD) when compared to the control group and historical control data.

At GD 20, incomplete ossification was seen in several bones of litters in all treated groups and the control group. Mostly, bones of the skull, sternum, paws, limbs, and vertebra were affected by variations in the status of expected ossification, described in terms of incomplete ossification, irregular ossification, unossified or increased ossification throughout all experimental groups. However, no dose dependency and no trend towards a treatment-related ossification effects of bones were observed. Statistical analysis revealed only a significantly lower foetal incidence of skull frontal, incomplete ossifications in the 100 mg/kg bw/day (LD), 600 mg/kg bw/day (MD) and HD groups (0%) when compared to the control group (2.14%). This finding was considered incidental in nature and not a test item-related effect. Overall, observed ossification-related findings were observed at lower or higher incidences, without dose-dependency, and almost entirely without statistical significance and thus considered not related to the test item.

Other skeletal findings in the treated groups also were considered not test-item related, occurring at slightly higher or lower litter incidences than the control group and without any statistical significance or dose dependency.

Visceral malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were no visceral findings related to treatment observed up to 750/1000 mg/kg bw/day (HD) when compared to the control group and historical control data.

No statistical significance was observed in any of the treated groups when compared to the control group. Higher litter incidences for umbilical artery, malpositioned were observed at 100 mg/kg bw/day (LD; 76.2% compared to 63.2% in the control group). Higher litter incidences of abdomen internal haemorrhage were observed (66.7%, 68.2% and 78.9% in the LD, MD (600 mg/kg bw/day) and HD groups, respectively, compared to 63.2% in the control group).

Visceral findings observed in the treated groups were at frequencies generally comparable to or in some cases slightly higher or lower in frequency compared to the controls. As the observed findings were either minor variations and/or lacking dose dependency and consistency, no toxicological significance can be attributed and the findings were considered to be spontaneous in nature. All litter incidences from treated groups were well within the historical control data range and statistically insignificant when compared to the control group.

Other effects:

effects observed, non-treatment-related

Description (incidence and severity):

CRANIOFACIAL:
There were no test item-related craniofacial findings observed up to 750/1000 mg/kg bw/day (HD).

Craniofacial examination revealed midbrain, haematoma, subdural in the 600 mg/kg bw/day (MD) and HD groups (13.6% and 10.5%, respectively) at lower incidences when compared to the control group (21.1%). These findings were considered spontaneous in nature and not related to the treatment with the test item. Statistical analysis of the data revealed no statistical significance for any of these findings. In addition, all values were within the historical control range for this strain.

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

750 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Remarks on result:

other: Based on the absence of test item-related foetal effects up to 750/1000 mg/kg bw/day.

Fetal abnormalities

Key result

Abnormalities:

no effects observed

Description (incidence and severity):

other: No test item-related foetal abnormalities observed up to 750/1000 mg/kg bw/day.

Overall developmental toxicity

Key result

Developmental effects observed:

no

Any other information on results incl. tables

Mean and summary tables are attached in full, please see below additional information for location of specific tables:

Hormone measurements in tabular format (average results/ dose group is sufficient): Page 8

Caesarean section data in tabular format:

• Anogenital distance - Page 9
• Prenatal and litter data - Starts on Page 12
• Fetal weight - Page 15
• External fetal findings - Page 17
• Fetal Visceral findings - Page 18
• Fetal Craniofacial findings - Page 22
• Fetal skeletal findings - Page 23
• Maternal weight/fetal weight - Page 37
• Body and organ weights - Page 38

Applicant's summary and conclusion

Conclusions:

In a study conducted according to OECD Test Guideline 414 and in compliance with GLP (reliability score 1), pregnant Wistar Han rats were administered N-[3-(dimethoxymethylsilyl)propyl]ethylenediamine at 0, 100, 600, and 750/1000 mg/kg bw/day via oral gavage (dried and deacidified corn oil vehicle) during days 5 through 19 of gestation. The maternal systemic NOAEL was 100 mg/kg bw/day, based on findings considered to be test item-related and adverse (mortality, clinical signs, decreased body weight gain / food consumption, and gross pathology) at 600 and/or 750/1000 mg/kg bw/day. No effects were observed on mating or developmental parameters. The developmental NOAEL was identified as 750 mg/kg bw/day based on the absence of test item-related foetal effects up to 750/1000 mg/kg bw/day.