reaction product of diphenylmethanediisocyanate, octylamine, 4-ethoxyaniline and ethylenediamine (1:0,37:1,53:0,05)

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:

1998-12-01 to 1999-01-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to other study

Reference

Endpoint:

water solubility

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1999-02-10 to 1999-02-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 105 (Water Solubility)

Version / remarks:

July 1995

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method A.6 (Water Solubility)

Version / remarks:

December 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 830.7840 (Water Solubility)

Version / remarks:

August 1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 830.7860 (Water Solubility Generator Column Method)

Version / remarks:

August 1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: DIN EN 1484, "Anleitung zur Bestimmung des gesamten organischen Kohlenstoffs (TOC) und des gelösten organischen Kohlenstoffs (DOC)", Normungsausschuss Wasserwesen im Deutschen Institut für Normung e. V., August 1997.

Principles of method if other than guideline:

NA

GLP compliance:

yes (incl. QA statement)

Type of method:

other: DOC (Dissolved Organic Carbon)

Key result

Water solubility:

< 1 mg/L

Conc. based on:

test mat. (dissolved fraction)

Incubation duration:

3 d

Temp.:

20 °C

pH:

6

Details on results:

The water solubility of Urea 4 at room temperature was estimated to be < 1 g/L by visual judgement in the preliminary test. The saturation concentration of Urea 4 was analytically estimated by a simplified flask method in the preliminary test at 20 °C to be in the range of the quantification limit for the DOC-measurements (1 mg DOC/L).

Conclusions:

Interpretation of results: slightly soluble (0.1-100 mg/L)
The water solubility of Urea 4 was estimated to be in the range of < 1 mg/L (quantification limit of the DOC measurements) at 20 °C +/- 1 °C. Thus, Urea 4 was considered to be slightly soluble or insoluble.

Executive summary:

The water solubility of Urea 4 was determined according to EU method A.6, OECD Guideline 105, and EPA OPPTS 830.7840, EPA OPPTS 830.7860 guideline and DIN EN 1484 guideline. Because of the low solubility of the test substance in water and the low sensitivity of the analytical method available the experimental determination of the water solubility of Urea 4 was technically not feasible. Therefore, the water solubility of Urea 4 was estimated to be in the range of < 1 mg/L (quantification limit of the DOC measurements) at 20 °C +/- 1 °C. Thus, Urea 4 was considered to be slightly soluble or insoluble.

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Reference

Endpoint:

partition coefficient

Type of information:

calculation (if not (Q)SAR)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

EU Method A.8 (Partition Coefficient)

Version / remarks:

December 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 117 (Partition Coefficient (n-octanol / water), HPLC Method)

Version / remarks:

March 1989

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 830.7570 (Partition Coefficient, n-octanol / H2O, Estimation by Liquid Chromatography)

Version / remarks:

August 1996

Deviations:

no

Principles of method if other than guideline:

NA

GLP compliance:

no

Type of method:

other: calculation (Leo-Hansch method)

Partition coefficient type:

octanol-water

Analytical method:

other: theoretical calculation

Key result

Type:

log Pow

Partition coefficient:

> 6

Temp.:

20 °C

Remarks on result:

other: calculation

As Urea 4 consists of several constituents, for each constituent a partition coefficient was calculated. The partition coefficients (log Pow values) of all possible constituents are > 6.

Conclusions:

The partition coefficient of Urea 4 was estimated to be log Pow > 6 using the Leo-Hansch method.

Executive summary:

The partition coefficient of Urea 4 was estimated according to EU method A.8 (Leo-Hansch method), OECD Guideline 117, and EPA OPPTS 830.7570 guideline. The partition coefficient of Urea 4 was estimated to be log Pow > 6 using the Leo-Hansch method. Urea 4 was not soluble in n-octanol and only slightly soluble in water. Therefore, due to the low solubility of Urea 4 in water and n-octanol and the low sensitivity of the analytical methods available the experimental determination of the partition coefficient by any analytical technique was technically not feasible.

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:

430-750-8

EC Name:

-

Cas Number:

1266545-86-1

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Reaction product of (C8 – C18) aliphatic primary amines, ethylene diamine, p-phenetidine with 4,4’-methylenediphenyl diisocyanate

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 150-185 g (mean: 167 g); male 201-246 g (mean: 241 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 ±3 C°
- 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:
600 mg of the test item were suspended in 1% CMC Solution to give a total volume of 20 mL.
2000 mg of the test item were suspended in 1% CMC Solution to give a total volume of 20 mL.
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 (if 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

Doses / concentrations 1

Dose / conc.:

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

Remarks:

Basis: actual ingested

Doses / concentrations 2

Dose / conc.:

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

Remarks:

Basis: actual ingested

Doses / concentrations 3

Dose / conc.:

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

Remarks:

Basis: actual ingested

No. of animals per sex per dose:

10 animals per dose (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). 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 perspective to demonstrate any dosage related response and no-observed-adverse effects at a certain dose level.

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

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Other effects:

effects observed, non-treatment-related

Description (incidence and severity):

Some borderline deviations were found (weight gain, organ weight, Na - values) without clear dose dependency or toxicological Compound related relevance.

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

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The NOAEL of the test item was determined to be >= 1000 mg/kg bw/day.

Executive summary:

The test substance Urea 4 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 doses of 150, 500 and 1000 mg/kg bw once a day, for a period of 28 days, to three groups 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, several borderline deviations were observed for few parameters examined in the treated groups. However, a clear dose dependency or toxicological compound related relevance could not be established. All rats treated with the test item Urea 4 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.