1,2-Ethanediamine, N-{3-(trimethoxysilyl)propyl}-,N-{(ethenylphenyl)methyl}derivate,hydrochlorides

Endpoint summary

Description of key information

Screening study for reproductive/developmental toxicity (OECD 422, oral, rat): NOAEL systemic toxicity = 100 mg/kg bw/day

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

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-04-11 to 2018-03-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

July 2016

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Crl:WI(Han) (outbred, SPF-Quality)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: P 10 wks for males (supplementary males in groups 1 and 3: 12 weeks) 12 wks for females (supplementary females in groups 1 and 3: 14 weeks)
- Weight at study initiation: not reported
- Housing: Macrolon plastic cages
- Diet: pelleted rodent diet provided ad libitum
- Water: tap-water provided ad libitum
- Acclimation period: at least 5 days prior to start of pretest (females) or treatment (males)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 22
- Humidity (%): 43 - 69
- Air changes (per hr): at least 10 room air changes/hour
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test item was dosed undiluted (i.e. without the use of a vehicle). Correction was made for specific gravity (factor: 0.9). No correction was made for purity/composition. On each morning of dosing, the required amount of test item was transferred into a container, flushed with nitrogen and stored at room temperature in the tightly closed container until use. This was done for Groups 2, 3 and 4 separately to minimize the time that the test item was exposed to oxygen in the air during dosing.
For dosing of control Group 1, water (Elix, Millipore S.A.S., Molsheim, France) was used.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

No chemical analyses were conducted since the test substance was dosed undiluted (i.e. without the use of a vehicle).

Duration of treatment / exposure:

For Groups 0, 50, 100 mg/kg bw/day, males were treated for 29 days, i.e. 2 weeks prior to mating, during mating, and up to the day prior to scheduled necropsy. Females that delivered were treated for 50-55 days, i.e. during 2 weeks prior to mating (with the objective of covering at least two complete estrous cycles), the variable time to conception, the duration of the pregnancy and at 13-15 days after delivery up to and including the day before scheduled necropsy. Females which failed to deliver healthy offspring were treated for 41 or 54 days.
For the highest dose group (200 mg/kg bw/day), treatment at the high dose was terminated after 10 days of dosing due to unacceptable high toxicity.

Frequency of treatment:

Once daily for 7 days per week, approximately the same time each day with a maximum of 6 hours difference between the earliest and latest dose.

Dose / conc.:

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

Dose / conc.:

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

Dose / conc.:

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

No. of animals per sex per dose:

Group 1 (control group): 10 males and 10 females
Group 2 (50 mg/kg bw/day): 10 males and 10 females
Group 3 (100 mg/kg bw/day): 10 males and 12 females
Group 4 (200 mg/kg bw/day): 19 males and 19 females

Control animals:

yes

Details on study design:

- Dose selection rationale: Based on the results of a 14-day dose range finder in which dose limiting effects were noted from 300 mg/kg bw/day onward, the dose levels for this combined 28-day oral gavage study with reproduction/developmental toxicity screening test were selected to be 50, 100 and 200 mg/kg bw/day.

- Rationale for animal assignment: Before initiation of pretest (females) or treatment (males), by computer-generated random algorithm according to body weight, with all animals within ± 20% of the sex mean.

Positive control:

No

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once prior to start of treatment and at weekly intervals during the treatment period this was also performed outside the home cage in a standard arena

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least twice daily from start of treatment onwards up to the day prior to necro psy, detailed clinical observations were made for all animals, immediately after treatment and 1 hour
thereafter (i.e. on the peak period of anticipated effects after treatment).

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of treatment (prior to first dosing) and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on PND 1, 4, 7 and 13.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as gfood/kg body weight/day: Yes; weekly, except for males and females which were housed together for mating and for females without evidence of mating. Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on PND 1, 4, 7 and 13.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Day of scheduled necropsy between 7.00 and 10.30 a.m.
- Anaesthetic used for blood collection: Yes; isoflurane
- Animals fasted: Yes
- How many animals: 5 animals/sex from Groups 1, 2 and 3 (note: Group 4 was terminated preterm)
- Parameters checked in tables 1 and 2 were examined.

CLINICAL CHEMISTRY: Yes
- Day of scheduled necropsy between 7.00 and 10.30 a.m.
- Anaesthetic used for blood collection: Yes; isoflurane
- Animals fasted: Yes
- How many animals: 5 animals/sex from Groups 1, 2 and 3 (note: Group 4 was terminated preterm)
- Parameters checked in tables 3 were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: collected overnight (approximately 15-20 hrs during the night prior to scheduled necropsy)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes; animals were deprived of food but water was available
- Parameters checked in table 4 were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: The selected males were tested during Week 4 of treatment and the selected females were tested once during the last week of lactation (e.g. PND 6-13). These tests were performed in the morning prior to dosing.
- Dose groups that were examined: 5 animals/sex from Groups 1, 2 and 3 (note: Group 4 was euthanized preterm)
- Battery of functions tested: hearing ability, pupillary reflex, and static righting reflex; fore- and hind-limb grip strength, recorded as the mean of three measurements per animal; and locomotor activity as measured by total movements and ambulations. Ambulations represent movements characterized by a relocation of the entire body position like walking, whereas total movements represent all movements made by the animals, including ambulations but also smaller or finer movements like grooming, weaving or movements of the head.

IMMUNOLOGY: No

Sacrifice and pathology:

SACRIFICE
- Male animals: Following completion of the mating period (a minimum of 28 days of dose administration).
- Maternal animals: PND 14-16

GROSS NECROPSY
- Gross necropsy consisted of a full post mortem necropsy, with special attention being paid to the reproductive organs. Descriptions of all macroscopic abnormalities were recorded. The number of former implantation sites were recorded for all paired females. In case no macroscopically visible implantation sites were present, nongravid uteri were stained using the Salewski technique in order to detect any former implantation sites. From all animals (including animals that died spontaneously or were sacrificed in extremis), samples of the following tissues and organs shown in Table 5 were collected and fixed in 10% buffered formalin.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table 6 were prepared for microscopic examination and weighed, respectively.

Statistics:

The following statistical methods were used to analyse the data:

- If the variables could be assumed to follow a normal distribution, the Dunnett-test based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test was applied to frequency data.
- The Kruskal-Wallis nonparametric ANOVA test was applied to motor activity data to determine intergroup differences. In case intergroup differences were seen, the Wilcoxon test was applied to compare the treated groups to the control group.

All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance. Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations may have been rounded off before printing. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics values.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Rales were noted at 200 mg/kg (in most males and several females, decedents included, on several days) and, less frequently, at 100 mg/kg (in two males and two females, decedents included, on one or two days) and 50 mg/kg (in one female on one day). Additionally, a few of the 200 mg/kg animals that were terminated after 10 treatment days showed gasping, laboured respiration, hunched posture and/or piloerection, mostly at the end of their treatment period and/or on the few treatment-free days thereafter. The latter breathing abnormalities and general clinical signs of toxicity were also noted in most decedents. Piloerection was also noted in one surviving male of the 100 mg/kg group. Salivation seen after dosing among animals treated with the test item, in a dose-related manner, was considered to be a physiological response rather than a sign of systemic toxicity considering its slight severity and the time of occurrence (i.e. shortly after dosing).
No additional clinical signs of toxicity were noted during the weekly arena observations.
Any other clinical signs noted incidentally occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study and showed no dose-related trend. At the incidence observed, these were considered to be unrelated to treatment.

Mortality:

mortality observed, treatment-related

Description (incidence):

Severe toxicity was observed at the high dose, leading to the termination of this dose group. At this dose level, there were eight animals euthanized in extremis (male nos. 38, 42, 43 and 51, female nos. 111, 113, 119 and 123, sacrificed between Days 3-13 of treatment). At 100 mg/kg bw/day, one male (no. 30) was euthanized in extremis on Day 14 of treatment and one female (no. 94) was found dead on Day 11 of treatment.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Males and females treated at 200 mg/kg bw/day showed reduced body weight gain (statistically significant in males) during their two-week study period. In males this resulted in statistically significantly reduced body weights at study Days 8 and 15. No treatment-related changes in body weights or body weight gain were noted in rats treated up to 100 mg/kg bw/day.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Males and females treated at 200 mg/kg bw/day consumed less food than controls throughout their two-week study period. Their food consumption after allowance for body weight showed a similar trend. No treatment-related changes in food consumption before or after allowance for body weight were noted in rats treated up to 100 mg/kg bw/day.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Only data from animals in Groups 1, 2 and 3 (0, 50 and 100 mg/kg bw/day, respectively) were available as the high dose animals (200 mg/kg bw/day; Group 4) had to be euthanized preterm.

Haematology parameters were considered not to be affected by treatment up to 100 mg/kg bw/day. The mean number of total white blood cells was lower at 50 and 100 mg/kg bw/day in both sexes. This was statistically significant for males at 50 mg/kg bw/day and females at 100 mg/kg bw/day. However, as all values remained in the historical control range and in the absence of a dose-response relationship, it was not considered treatment-related.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

Only data from animals in Groups 1, 2 and 3 (0, 50 and 100 mg/kg bw/day, respectively) were available as the high dose animals (200 mg/kg bw/day; Group 4) had to be euthanized preterm.

The mean concentration of urea was statistically significantly lower in males at 100 mg/kg compared to controls (4.2 mmol/L versus 5.2 mmol/L; relative difference: 19%). Group mean value was below the available historical control range. The statistically significant change noted for calcium level in males at 50 mg/kg bw/day was considered unrelated to treatment due to the absence of a dose-related trend.

Urinalysis findings:

no effects observed

Description (incidence and severity):

Only data from animals in Groups 1, 2 and 3 (0, 50 and 100 mg/kg bw/day, respectively) were available as the high dose animals (200 mg/kg bw/day; Group 4) had to be euthanized preterm. Urinalysis parameters were considered not to be affected by treatment up to 100 mg/kg bw/day.

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

Only data from animals of Groups 1, 2 and 3 (0, 50 and 100 mg/kg, respectively) were available as the high dose animals (200 mg/kg; Group 4) had to be euthanized preterm.
Hearing ability, pupillary reflex and static righting reflex were normal in all examined animals. Grip strength was not affected by treatment.
The variation in motor activity did not indicate a relation with treatment. All groups showed a similar habituation profile with a decreasing trend in activity over the duration of the test period.
It was noted that the mean values for total movements and ambulations at 50 mg/kg in males were about 1.5-fold higher compared to the control values. In the absence of a dose-related response, these differences were considered to be unrelated to treatment with the test item.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Only data from animals in Groups 1, 2 and 3 (0, 50 and 100 mg/kg bw/day, respectively) were available as the high dose animals (200 mg/kg bw/day; Group 4) had to be euthanized preterm. There were no test item-related alterations in organ weights up to 100 mg/kg bw/day.

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were no test item-related gross observations for any animal at 50 or 100 mg/kg bw/day. Principal macroscopic findings at 200 mg/kg bw/day related to moribundity consisted of emaciation (one female), distension with gas of (parts of) the gastro-intestinal tract (four males and two females), enlarged or thickened lung (one male and one female) and pale discoloration of the lung (two males and one female). In addition, one female at 200 mg/kg bw/day (no. 113) was noted with discoloration and dark red gelatinous contents of the small intestine. This necropsy observation together with the microscopic findings of esophagus, trachea and/or glandular stomach erosions were indicative for an irritating property of the test item.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Only data from animals in Groups 1, 2 and 3 (0, 50 and 100 mg/kg bw/day, respectively) were available as the high dose animals (200 mg/kg bw/day; Group 4) had to be euthanized preterm. At microscopic examination of the scheduled sacrifices, test item-related findings were noted in the thymus of males of Group 3 (100 mg/kg bw/day), consisting of minimal degrees of lymphoid atrophy (2/5 examined tissues) or increased lymphocytolysis (1/5 examined tissues). The remainder of the recorded microscopic findings of the scheduled sacrifices were within the range of background pathology encountered in rats of this age and strain. There was no test item related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Description (incidence and severity):

Thyroid hormone analyses:
Serum levels of T4 in F0 males were considered not to be affected by treatment up to 100 mg/kg bw/day

Details on results:

Mortality:
At 100 mg/kg, one male (no. 30) was euthanized in extremis on Day 14 of treatment and one female (no. 94) was found dead on Day 11 of treatment.
At 200 mg/kg, there were eight animals euthanized in extremis (male nos. 38, 42, 43 and 51, female nos. 111, 113, 119 and 123, sacrificed between Days 3-13 of treatment). Due to this toxicity, the remaining rats of the 200 mg/kg group were no longer dosed from Study Day 11 and were sacrificed a few days later (Study Days 13-15).
Respiratory difficulties (rales, laboured respiration and/or gasping) were noted in all decedents. This was accompanied by hunched posture and/or piloerection in most of them, while a flat posture or chromodacryorrhoea (snout) were noted in one or two decedents.
At macroscopic examination of the 100 mg/kg decedents, no abnormalities were seen in the male while the female found dead showed advanced autolysis. Principal macroscopic findings at 200 mg/kg related to moribundity consisted of emaciation (one female), distension with gas of (parts of) the gastro-intestinal tract (four males and two females), enlarged or thickened lung (one male and one female) and pale discoloration of the lung (two males and one female). In addition, one female at 200 mg/kg (no. 113) was noted with discoloration and dark red gelatinous contents of the small intestine. This necropsy observation together with the microscopic findings of esophagus, trachea and/or glandular stomach erosions were indicative for an irritating property of the test item.
At 100 mg/kg, principal microscopic findings for the two unscheduled deaths were seen in the larynx and consisted of:
- Erosion/necrosis of the epithelium with extension to the underlying tissues, in the male (massive) and the female (moderate).
- Inflammatory cell infiltrate, mainly lymphogranulocytic, in the female (slight).
Principal microscopic findings at 200 mg/kg for the eight unscheduled deaths were seen in the larynx and nasal tissues (in general most obvious in the most caudal levels of the nose):
Larynx:
- Erosion/necrosis of the epithelium with extension to the underlying tissues, in one male (minimal) and three females (marked).
- Inflammatory cell infiltrate, mainly lymphogranulocytic, in three males and four females (up to moderate).
- Luminal exudate, in three females (up to moderate).
- Epithelial hyperplasia, in four males and one female (up to slight).
Nasal tissues:
- Erosions/necrosis of the olfactory epithelium, in four males and one female (up to marked).
- Erosions/necrosis of the respiratory epithelium in four males and one female (up to massive).
- Inflammation, in four males and one female (up to moderate).
- Luminal exudate, granulocytic and/or necrotic and fibrin-like in four males and one female (up to moderate).
- Epithelial metaplasia, in two males and one female (up to moderate).
One female of the control group (no. 67) died at blood sampling prior to scheduled necropsy. As this was a control animal it was not due to treatment with the test item.

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical signs

gross pathology

histopathology: non-neoplastic

mortality

Critical effects observed:

no

Conclusions:

A combined repeated 28-Day repeated dose toxicity study with the reproduction/developmental toxicity screening according to OECD 422 and in compliance with GLP is available for the test substance. Rats were exposed to 200, 100 and 50 mg/kg bw/day by gavage. Due to high toxicity at 200 mg/kg bw/day the test was terminated for the high-dose group within 10 days. As no relevant effects were observed at 100 mg/kg bw/day a systemic NOAEL of 100 mg/kg bw/day was determined.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

100 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.6, of Regulation (EC) No 1907/2006.

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

A dose-range finding study was conducted with 1,2 -ethanediamine, N-{3-(trimethoxysilyl)propyl}-,N-{(ethenylphenyl)methyl}derivs,hydrochlorides (CAS 171869-89-9) prior to the commencement of the OECD 422 study. Three male and three female rats were dosed with water, 100, 300, or 1,000 mg/kg bw/day for 14 days via gavage, respectively. Mortalities were observed in the high-dose (2/3 female) and mid-dose (1/3 male) groups. All animals were sacrificed moribund. The main clinical signs seen in the decedents were salivation, laboured respiration, gasping and piloerection. The survivors mostly showed slight rales and salivation in the mid and high-dose groups. No abnormal clinical signs were observed in the low-dose group. At necropsy of the decedents in the mid and high-dose groups, it was found that parts of the gastrointestinal tract were distended with gas and several reddish foci in the glandular mucosa stomach were observed. In contrast, the survivors showed no macroscopic abnormalities in the low and mid-dose groups. In the high-dose group, several reddish/dark red foci in the glandular mucosa of the stomach of two males were observed. Based on the results of this dose-range finding study, dose levels for the primary study were determined to be: 50, 100 and 200 mg/kg bw/day. The peak effect of the occurrence of salivation occurred immediately after dosing. The other clinical signs noted at 300 mg/kg bw/day (i.e., the dose closest to the highest dose level selected for the main study) generally started later. Based on this, detailed daily clinical observations in the main study were conducted immediately after dosing and 1 hour after that.

A key study (combined repeated dose toxicity study with reproductive/developmental toxicity screening test (oral)) with 1,2 -ethanediamine, N-{3-(trimethoxysilyl)propyl}-,N-{(ethenylphenyl)methyl}derivs,hydrochlorides (CAS 171869-89-9) is available and was performed according to OECD TG 422 and in compliance with GLP (CRL, 2018). The test substance was administered daily in graduate doses (50, 100, or 200 mg/kg bw/day) to 3 groups of test animals per gavage, one dose level per group for a treatment period of up to 55 days, i.e. during two weeks prior to mating, during mating, during post-coitum, and during 13-15 days of lactation. Males were dosed after the mating period until the minimum total dosing period of 29 days was completed. Animals of an additional control group were handled identically as the dose groups but received water, as the test substance was administered undiluted. Due to toxicity in the highest dose group, resulting in eight unscheduled sacrifices during the pre-mating period, treatment was terminated after 10 days and the surviving animals were sacrificed a few days later.

At 100 and 200 mg/kg, several animals became moribund in the course of the pre-mating period, resulting in the premature death of two animals (one female and one male, euthanized on Days 11 and 14 of treatment, respectively) at 100 mg/kg and eight animals (four/sex; euthanized between Days 3-13 of treatment) at 200 mg/kg. These animals showed respiratory difficulties (rales, laboured respiration and/or gasping), often accompanied by hunched posture and/or piloerection, and occasionally flat posture or chromodacryorrhoea at the snout. Related morphological findings included dilation of (parts of) the gastro-intestinal tract with gas, pale discoloration and/or enlargement of the lungs and microscopic lesions in the larynx (up to massive erosion/necrosis, inflammatory cell infiltrate, luminal exudate and/or epithelial hyperplasia) and nasal tissues (up to massive erosion/necrosis of the olfactory/respiratory epithelium, inflammation, luminal exudate, and/or epithelial metaplasia). Due to this severe toxicity at 200 mg/kg, treatment was terminated after 10 days and the surviving animals were sacrificed a few days later. A single incidence of rales was observed in one female at 50 mg/kg; no signs of respiratory difficulties were noted in males at 50 mg/kg. Based on a combination of factors, it was considered that observed findings were indicative of reflux.

At 200 mg/kg bw/day, treatment-related decreases in body weights and food consumption were noted; however, these effects correspond to the severe toxicity of the test material.

At 100 mg/kg bw/day, microscopic test item-related findings were noted in the thymus of males consisting of minimal degrees of lymphoid atrophy or increased lymphocytolysis. Based on their low severity (minimal), the thymic findings in 100 mg/kg males were also regarded as non-adverse. The remainder of the recorded microscopic findings of the scheduled sacrifices was within the range of background pathology encountered in rats of this age and strain. All other effects noted at 100 and 50 mg/kg bw/day were not considered treatment related and were incidental in nature. These changes included alterations in hematological and clinical chemistry endpoints. With regard to the hematological parameters, the mean number of total white blood cells was lower at 50 and 100 mg/kg bw/day in both sexes. This was statistically significant for males at 50 mg/kg bw/day and females at 100 mg/kg bw/day. However, as all values remained in the historical control range and in the absence of a dose-response relationship, it was not considered treatment-related. Concerning the clinical chemistry parameters, the mean concentration of urea was statistically significantly lower in males at 100 mg/kg bw/day compared to controls (4.2 mmol/L versus 5.2 mmol/L; relative difference: 19%). Group mean value was below the available historical control range. The statistically significant change noted for calcium level in males at 50 mg/kg bw/day was considered unrelated to treatment due to the absence of a dose-related trend.

 

Based on these results, the systemic NOAEL was 100 mg/kg bw/day. Moribundity/mortality due to respiratory effects (breathing difficulties, adverse microscopic changes in the larynx) in two animals at 100 mg/kg bw/day was considered a local, reflux-related effect and as such considered local rather than systemic effects of the test item.

Justification for classification or non-classification

According to CLP (1272/2008/EC) classification criteria for repeated dose toxicity, 1,2 -ethanediamine,N-{3-(trimethoxysilyl)propyl}-,N-{(ethenylphenyl)methyl}derivs,hydrochlorides does not fulfill the criteria for classification and thus a non-classification is warranted for this endpoint.