reaction product of diphenylmethanediisocyanate, toluenediisocyanate ( reaction mass of isomers: 65 % 2,4- and 35 % 2,6-diisocyanate), octylamine, oleylamine and 4-ethoxyaniline (molar ratio 3.88:1:6.38:0.47:2.91)

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1999-02-22 to 1999-06-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 1995-07-27

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Version / remarks:

Directive 96/54 EEC

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

430-970-4

EC Name:

-

Cas Number:

1266545-66-7

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Reaction product of (C8 – C18) aliphatic primary amines (partly unsaturated) and p-phenetidine with a mixture of aromatic isocyanates comprising of primarily 4,4’-methylenediphenyl diisocyanate and 4-methyl-m-phenylene

Details on test material:

NA

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan, Winkelmann GmbH, D-33178 Borchen, Germany
- Age at study initiation: 7 - 8 weeks
- Weight at study initiation: female 169-183 g (mean: 176 g); male 234-260 g (mean: 246 g)
- Housing: Macrolon cages on Altromin saw fibre bedding
- Diet: ad libitum, Altromin 1324 totally-pathogen-free-TPF
- Water : tap water: drinking water, municipal residue control, microbiol. controlled periodically
- Acclimatisation period: 12-14 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ±3C°
- Humidity: 55 ± 10%
- Air changes: >10 x /hour
- Photoperiod: 12/12 (light 6.30 - 18.30)

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

1% in aqua bidest.

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
4.000 mg of the test item were suspended in 1% CMC Solution to give a total volume of 20 mL.

VEHICLE
- Justification for use and choice of vehicle: The vehicle was chosen due to its non-toxic characteristics
- Concentration in vehicle: CMC, 1% in aqua bidest.
- Amount of vehicle (gavage): 5 mL/kg bw
- Lot no. : 36H0738

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

Technically not feasible. Due to the extremely low solubility in water and organic solvents test item concentrations could not be determined analytically. See section 4.7 and 4.8 for more detail.

Duration of treatment / exposure:

28 days

Frequency of treatment:

7 days per week

Doses / concentrations

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

10 animals (5 females and 5 males)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose selection was based on results in an acute toxicity study (see section 7.2.1).
- Rationale for animal assignment: in accordance with guidelines.

Positive control:

None

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least once a day. Mortality was observed twice a day.

BODY WEIGHT: Yes
- Time schedule for examinations: The animals were weighed prior to first application (day 0) and once a week thereafter (day 7, 14, 21, 28 or day of sacrifice, respectively).

FOOD CONSUMPTION:
- Food consumption for each animal group (females/males) was determined and calculated in g per group: Yes

FOOD EFFICIENCY:
- Body weight gain g per animal: Yes

HAEMATOLOGY AND CLINICAL CHEMISTRY : Yes
- Time schedule for collection of blood: on the day of necropsy (part of necropsy)
- Animals fasted: Yes (over night)
- How many animals: all animals
- Parameters examined: Haematocrit (Hct), Haemoglobin (Hb), Erythrocyte count (RBC), Total leucocyte count (WBC), Differential leucocyte count Platelet count, Blood clotting parameters: activated Partial Thromboplastin Time (aPTT), AST aspartate aminotransferase (GOT), ALT alanine aminotransferase (GPT), Alkaline Phosphatase (AP), Cholesterol (Chol), Total Protein (TP), Glucose (GLU), Urea, Creatinine (CREA), Albumin (ALB), Na, K;

URINALYSIS: Yes
- Time schedule for collection of urine: on the day of necropsy (part of necropsy)
- Metabolism cages used for collection of urine: No
- Animals fasted: Yes (over night)
- Parameters examined: pH, Urine GLU, Urine TP

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: once before first exposure and in the fourth exposure week
- Dose groups that were examined: all animals
- Battery of functions tested: sensory activity / grip strength / motor activity

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (All animals in the study were subjected to a full, detailed gross necropsy which included careful examination of the surface of the body, all orifices and the cranial, thoracic and abdominal cavities and their contents)
HISTOPATHOLOGY: Yes (Full histopathology was carried out on the preserved organs and tissues of all animals)

Statistics:

For statistical analysis one-way analysis of variance (ANOVA) was carried out, followed by Student's t-test to reveal any differences between control and test groups. In the evaluation of laboratory parameters all values within a range of the mean - value ± the two fold Standard deviation (x ± 2s) are considered to be „normal" values within a „normal" population.

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):

not examined

Food efficiency:

no effects observed

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

No toxicological relevant effects were observed in any of the dosed animals in any of the parameters examined.

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects No toxicological relevant effects were observed in any of the dosed animals

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

NOAEL >= 1000 mg/kg bw/day

Executive summary:

The test substance was tested in a repeated dose 28-day oral toxicity study in rats according to EU method B.7 and OECD Guideline 407. The test item was administered in a dose of 1000 mg/kg bw (limit test) once a day, for a period of 28 days, to one group of 5 male and 5 female Wistar rats by oral gavage. A control group of 5 male and 5 female rats was dosed with the vehicle, (CMC) 1% in aqua bidest.

Clinical and behaviour/ functional observations were carried daily and examination of body weight was performed throughout the study. Blood and urine samples, collected during necropsy, were used to perform a detailed haematology analysis and urinalysis. A detailed gross necropsy was performed as well as histopathological and organ weight examination. In comparison to the untreated control group, no toxicologically relevant difference was observed for any of the examined parameters. All rats treated with the test item Urea 6 survived the testing period without showing compound related toxic effects. Thus the NOAEL value determined was >= 1000 mg/kg bw/day.

See cross-ref. for limitations on solubility and analytics.