Alkanol reaction products with bis(aminoalkyl(C=1~6))carbomonocycle, alkoxy(C=3~8)alkanol(C=2~7) and 2,4-TDI

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

17 march 2016 - 06 april 2016

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

GLP compliance:

no

Remarks:

Range-finding toxicity study

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Details on test material:

Name of test material (as cited in study report): 20231250
- Physical state: white powder
- Storage condition of test material: room temperature (ca. 20°C) in the dark

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories Italia, Calco, Italy.
- Age at study initiation: approximately 10 weeks old on the day of treatment
- Mean body weight at study initiation: for the males, 347 g (range: 326 g to 369 g) and the females 249 g (range: 234 g to 268 g).
- Fasting period before study: no
- Housing: in polycarbonate cages with stainless steel lids
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: a period of 6 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12

IN-LIFE DATES: 17 March 2016 to 06 April 2016

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION
The test item was administered as a suspension in the vehicle. The test item was ground to a fine powder, using a mortar and pestle, and then mixed with the required quantity of vehicle.
The test item dose formulations were prepared daily and were delivered to the study room at room temperature and protected from light.


VEHICLE
- Justification for use and choice of vehicle (if other than water): homogeneous suspensions were obtained with corn oil as a vehicle
- Concentration in vehicle: 20, 60 and 200 mg/ml
- Amount of vehicle (if gavage): 5 ml/kg bw

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

14 days

Frequency of treatment:

7 days/week

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

5 males and 5 females per dose

Control animals:

yes, concurrent vehicle

Details on study design:

Dose selection rationale:
The dose-levels used in this study (0, 100, 300 and 1000 mg/kg bw/day) were selected on the basis of the results of an acute oral toxicity study. In this study, two groups of three Sprague-Dawley females were treated once with the test item at 2000 mg/kg bw and one group of three animals were treated once with the test item at 300 mg/kg bw in corn oil and then observed for fourteen days. There were no mortalities and no clinical signs in any animals.
Based on these results, a dose-level of 1000 mg/kg bw/day was selected for the high dose. The dose levels for the mid and low doses (300 and 100 mg/kg bw/day respectively) were selected to explore any possible dose relationship.

Positive control:

no

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once a day.

BODY WEIGHT: Yes
The body weight of each animal was recorded once before the beginning of the treatment period, on the first day of treatment, and then on days 4, 8, 11 and 14 before necropsy.

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
The quantity of food consumed by the animals in each cage was measured twice weekly (days 1-4, 4-8, 8-11 and 11-14) during the treatment period.
Food consumption was calculated per animal and per day.When one of the animals in the same cage died, the number of days for which that animal had been present in the cage was taken into consideration for the calculation of food consumption.

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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER: ORGAN WEIGHTS
The body weight of each animal was recorded before sacrifice at the end of the treatment period. Kidneys, liver, heart, ovaries with oviducts, spleen and testes were weighed wet as soon as possible after dissection.
The ratio of organ weight to body weight was calculated.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
A complete macroscopic post-mortem examination was performed on all study animals. This included
examination of the external surfaces, all orifices, the cranial cavity, the external surfaces of the brain, the
thoracic, abdominal and pelvic cavities with their associated organs and tissues and the neck with its
associated organs and tissues.

HISTOPATHOLOGY: No
No microscopic examination was performed in first instance.

Other examinations:

The heterogeneity of variance between groups were checked by Bartlett’s homogeneity of variance test. Where no significant heterogeneity was detected, a one-way analysis of variance was carried out. If the obtained result was positive, Dunnett test was used to assess the significance of inter-group differences. Where significant heterogeneity was found, the normal distribution of data was examined by Kolmogorov-Smirnov test. If the data was not normally distributed, the non-parametric method of Kruskal-Wallis One-Way analysis of variance was used. If there was a positive result, the inter-group comparisons was performed using Dunn test.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Description (incidence):

There were no clinical signs that could be ascribed to the treatment with the test item. Clinical signs were observed in females given 1000 mg/kg/day and in all test item-treated groups for males.
Signs of alopecia, chromodacryorrhea, loud breathing, scabs and thinning of hair were isolated and/or commonly encountered in rats of this age and strain. As such, they were considered to be incidental and therefore unrelated to the test item treatment.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

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

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not specified

Details on results:

MORTALITY:
No deaths occurred during the study.

CLINICAL SIGNS:
There were no clinical signs that could be ascribed to the treatment with the test item. Clinical signs were observed in females given 1000 mg/kg/day and in all test item-treated groups for males. Signs of alopecia, chromodacryorrhea, loud breathing, scabs and thinning of hair were isolated and/or commonly encountered in rats of this age and strain. As such, they were considered to be incidental and therefore unrelated to the test item treatment.

BODY WEIGHT:
There were no test item-related changes in body weight and body weight gain during the study at any dose level.

FOOD CONSUMPTION:
There were no test item-related effects on food consumption during the study.

ORGAN WEIGHTS:
There were no changes in the mean organ weights which suggested a test item-related effect.
The lower absolute and relative-to-body heart weights in females treated at 300 or 1000 mg/kg/day were of low magnitude (up to -12% in relative-to-body weights at 1000 mg/kg/day; p<0.05), not associated with any macroscopic correlates and not seen in the test item-treated males. Consequently, it was considered that these differences were probably unrelated to test item administration.

GROSS PATHOLOGY:
There were no changes at necropsy which indicated a test item-related effect.

Effect levels

Dose descriptor:

other: maximum tolerated dose

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Target system / organ toxicity

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

The administration of the test item daily by the oral route to male and female Sprague-Dawley rats at dose-levels of 100, 300 or 1000 mg/kg bw/day for two weeks did not elicit any signs of toxicity.
Consequently, the same dose-levels were chosen for the subsequent reproduction/developmental toxicity screening test in this species.

Executive summary:

The objective of this dose-range finding study was to evaluate the potential toxicity of the test item following daily oral administration by gavage to rats for 14 days in order to select the dose-levels for a further OECD 421 reproductive/developmental toxicity screening in this species.

Three groups of five male and five female Sprague-Dawley rats received the test item daily, by gavage at dose-levels of 100, 300 or 1000 mg/kg bw/day for 14 days. A group of five males and five females received the vehicle alone,corn oil under the same experimental conditions and acted as a control group. The dose formulations were administered under a constant dosage-volume of 5 mL/kg/day.

The animals were checked daily for mortality and clinical signs. Body weights were recorded once during the pre-treatment period, on the first day of treatment and then on days 4, 8, 11 and 14 before necropsy. Food consumption was recorded twice weekly during the treatment period.

On completion of the treatment period, animals were sacrificed. Kidneys, liver, heart, ovaries, spleen and testes were weighed and a full macroscopic post-mortem examination was performed.

 No unscheduled deaths related to the test item treatment occurred during the study. No clinical signs were observed in animals.

There were no test item-related changes in body weight and body weight gain during the study at any dose-level, and no relevant effects were recorded on food consumption.

No test item-related changes were observed in the mean organ weights or at the macroscopic post-mortem examination.

The administration of the test item daily by the oral route to male and female Sprague-Dawley rats at dose-levels of 100, 300 or 1000 mg/kg/day for 2 weeks did not elicit any signs of toxicity.

Consequently, the same dose-levels were chosen for a further reproduction/developmental toxicity screening test in this species.