bis(2-ethylhexyl) 4,4’-{6-[4-tert-butylcarbamoyl)anilino]-1,3,5-triazine-2,4-diyldiimino}dibenzoate

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

GLP compliance:

yes (incl. QA statement)

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

421-450-8

EC Name:

-

Cas Number:

154702-15-5

Molecular formula:

C44H59N7O5

IUPAC Name:

2-ethylhexyl 4-[(4-{[4-(tert-butylcarbamoyl)phenyl]amino}-6-[(4-{[(2-ethylhexyl)oxy]carbonyl}phenyl)amino]-1,3,5-triazin-2-yl)amino]benzoate

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Male (46) and female (46) Wistar rats were obtained from a colony maintained under SPF-conditions at Charles River Wiga GmbH, Sulzfeld Germany
- Age at study initiation: about 4 weeks old
- Weight at study initiation: 157.4 g - 189.1 g and 118.2 g - 145.2 g for males for females, respectively
- Housing: Housing conditions were conventional. From the start of the treatment period, the animals were housed in groups of five, separated by sex, in suspended stainless steel cages with wire mesh floor and front. During the acclimatisation period they were housed in similar cages in groups of five or six .
- Diet: feed (in powder) was provided ad libitum, from the arrival of the rats until the end of the study.
- Water: tap water was available, ad libitum, from the arrival of the rats until the end of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 - 24. The temperature exceeded the upper value twice
- Humidity (%): 50 - 70. The humidity was lower than 50 % for two periods of a few hours and higher than 70 % for three such short periods. In addition, the humidity exceeded 70 % for about ten longer periods.
- Air changes (per hr): about 10
- Photoperiod (hrs dark / hrs light): lighting was artificial by fluorescent tubes, time switch controlled at a sequence of 12 h light (7.30-19.30), 12 h dark

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The oral route was used because this route is recommended in the guidelines for testing of cosmetic ingredients for their safety evaluation. The TNO rodent diet was used as the carrier. The test substance was incorporated into this diet by mixing in a mechanical blender. Because the test substance contained particles of different sizes, the substance was ground in an electric coffee mill prior to diet mixing. Fresh batches of experimental diets were prepared at 2 - 3 week intervals and stored at about 20 °C in a freezer until use.
The doses were selected on the basis of the results of the 14 -day range-finding study.

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

13 consecutive weeks.

Frequency of treatment:

Continuously

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0.15 other: % w/w

Remarks:

Basis: nominal in diet. Group 2

Doses / concentrations 2

Dose / conc.:

0.5 other: % w/w

Remarks:

Basis: nominal in diet. Group 3

Doses / concentrations 3

Dose / conc.:

1.5 other: % w/w

Remarks:

Basis: nominal in diet. Group 4: dietary intake corresponding to 831 and 963 mg/kg bw/day for male and female respectively

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:

- DETAILED CLINICAL OBSERVATIONS: Yes
- BODY WEIGHT: Yes
- FOOD CONSUMPTION AND COMPOUND INTAKE:: Yes
- FOOD EFFICIENCY: Yes
- WATER CONSUMPTION: Yes
- OPHTHALMOSCOPIC EXAMINATION: Yes
- HAEMATOLOGY: Yes
- CLINICAL CHEMISTRY: Yes
- URINALYSIS: Yes
- OTHER: Renal concentration test

Sacrifice and pathology:

- GROSS PATHOLOGY: Yes
- HISTOPATHOLOGY: Yes

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

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

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Endocrine findings:

not examined

Urinalysis findings:

no effects observed

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:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Effect levels

1

Key result

Dose descriptor:

NOAEL

Effect level:

> 831 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Remarks on result:

not determinable due to absence of adverse toxic effects

2

Key result

Dose descriptor:

NOAEL

Effect level:

> 963 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity

Key result

Critical effects observed:

no

Any other information on results incl. tables

RESULTS

STABILITY, HOMOGENEITY AND ANALYIS OF THE TEST SUBSTANCE

The concentrations of the test substance found in diet samples analysed after storage in the animal room for 7 days or in a freezer for five weeks showed that the substance was stable under simulated experimental conditions. Analysis of 5 samples per diet, taken at different locations, showed that the test substance was homogeneously distributed in the diet at all dose levels. The content of the test substance was close to the intended level in all diets analysed.

GENERAL FINDINGS
The ingestion of the test substance was well tolerated, as evidenced by the absence of treatment-related changes in the appearance, general condition and behaviour of the animals, their growth, food and water consumption, red blood cell and clotting potential values, total white blood cell counts, clinical chemistry values, renal concentrating ability, composition of the urine, organ weights, gross necropsy findings and histopathological findings
In comparison with the controls, the mean percentage of neutrophils was higher and that of lymphocytes lower in all groups of female rats given test substance. The differences showed no dose-response relationship and probably resulted from the relatively low neutrophil and high lymphocyte count in the control group. Most importantly, the differences were not reflected in dose-related or significant changes in the absolute numbers of these cell types and are, therefore, regarded as chance findings, unrelated to treatment.

Applicant's summary and conclusion

Conclusions:

Since the ingestion of RA 3643 at dietary levels up to 1.5 % for 13 consecutive weeks was tolerated without signs of toxicity, the dietary concentration of 1.5 % was a no-observed-adverse-effect level of test substance under the conditions of this study. This dietary level provided a mean intake of 831 and 963 mg of test substance/kg bw/day in male and female rats, respectively.

Executive summary:

The objective of this GLP compliant OECD TG 408 study was to examine the possible sub chronic oral toxicity of the test substance RA 3643 in rats. This substance was administered in the diet, at concentrations of 0, 0.15, 0.5 or 1.5 % w/w, to groups of 10 males and 10 female Wistar rats for 13 consecutive weeks. The ingestion of the test substance was well tolerated, as evidenced by the absence of treatment-related changes in the appearance, general condition and behaviour of the animals, their growth, food and water consumption, red blood cell and clotting potential values, total white blood cell counts, clinical chemistry values, renal concentrating ability, composition of the urine, organ weights, gross necropsy findings and histopathological findings
In comparison with the controls, the mean percentage of neutrophils was higher and that of lymphocytes lower in all groups of female rats given test substanct. The differences showed no dose-response relationship and probably resulted from the relatively low neutrophil and high lymphocyte count in the control group. Most importantly, the differences were not reflected in dose-related or significant changes in the absolute numbers of these cell types and are, therefore, regarded as chance findings, unrelated to treatment. The NOAEL was set at 831 and 963 mg/kg bw/day for male and female rats, respectively.