1,1'-(methylenedi-4,1-phenylene)bis(3-butylurea)

Endpoint summary

Description of key information

HAT ISO was tested in a 28 Day repeated dose toxicity study by oral application in rats. The oral administration at dose levels up to 1000 mg/kg bw/day was not associated with any toxic change. The no-observed-adverse-effect level (NOAEL) can be considered to be greater than 1000 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:

1993-08-23 to 1994-09-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Limit test:

no

Specific details on test material used for the study:

Analytical purity: >= 99.65 %
Lot/batch No.: 93.166

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: IFFA CREDO; BP 0109; 69592 L'Arbresle, Cedex; France;
- Age at study initiation: 6 weeks;
- Weight at study initiation: males: 178 to 203 g; females: 156 to 179 g;
- Fasting period before study: not indicated,
- Housing: groups of 5 of the same sex and dose group;
- Diet (e.g. ad libitum): not indicated;
- Water (e.g. ad libitum): not indicated;
- Acclimation period: not indicated;


ENVIRONMENTAL CONDITIONS
- Temperature: 22 +/- 2 °C;
- Humidity: 55 +/- 15 % R.H.;
- Air changes: minimum 15 air changes per hour;
- Photoperiod: 12 hours light (artificial) / 12 hours dark;

Route of administration:

oral: gavage

Vehicle:

other: carboxymethylcellulose 1 % in water for injection

Details on oral exposure:

Method of administration:
Gavage

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Method:
Measurement of the UV absorption at 290 nm of solutions subjected to high performance liquid chromatography (HPLC).
Mobile phase:
Mixture of water (25 parts) with dimethylformamide (75 parts), degassed by an online degasser.
Calibration:
Approximately 100 mg of test substance was accurately weighed, dissolved in mobile phase and made up to 100 mL. Appropriate volumes of test substance stock solutions were diluted with mobile phase to produce a set of standard calibrants in the nominal range 1 to 30 µg/mL.
HPLC-UV analysis:
Pump: Jasco PU-980;
Auto sampler programme: 100 µL of sample was rinsed through a 20 µL sample loop for injection;
Column: 256 cm x 2.0 mm 3 micron Phenomenex Hypersil;
Temperature: Ambient;
Mobile phase: cf. above;
Detector: Jasco type 875 UV absorption detector, wavelength 290 nm;
Cycle time: Typically 26 min.;
Calculations:
The response from calibration solutions (R) and the amount of test substance (A) were used to generate a straight line (R = Bo + B1A).
Concentrations of test substance (A) in samples were calculated from the resulting equation: A = (R - Bo)/B1.
All peak height measurements and calibrations were performed on VG Data Systems Multichrom 2.
Validation:
The method was validated using the test substance, and shown to be acceptable in accordance with HE departmental standard operating procedures.

Duration of treatment / exposure:

Test duration: 28 days

Frequency of treatment:

Dosing regime: 7 days/week

Dose / conc.:

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

Remarks:

vehicle control

Dose / conc.:

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

Dose / conc.:

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

Dose / conc.:

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

No. of animals per sex per dose:

Male: 5 animals at 0 mg/kg bw/day
Male: 5 animals at 150 mg/kg bw/day
Male: 5 animals at 600 mg/kg bw/day
Male: 5 animals at 1000 mg/kg bw/day
Female: 5 animals at 0 mg/kg bw/day
Female: 5 animals at 150 mg/kg bw/day
Female: 5 animals at 600 mg/kg bw/day
Female: 5 animals at 1000 mg/kg bw/day

Control animals:

no

Details on study design:

Four groups of each 5 male and 5 female animals were scheduled for the test:
Group 1 - Control group - receiving the control article (carboxymethylcellulose 1 %);
Group 2 - Low dose group - receiving 150 mg/kg bw/day;
Group 3 - Intermediate dose group - receiving 600 mg/kg bw/day;
Group 4 - High dose group - receiving 1000 mg/kg bw/day;
The test article was administered daily by oral gavage in a dose volume of 10 mL/kg/bw/day; the corresponding dose concentrations were 0; 15; 60; 100 mg/mL.

Positive control:

NA

Observations and examinations performed and frequency:

Morbidity/mortality checks were performed twice daily.
Clinical examinations were performed daily.
A full clinical examination was performed weekly.
Individual body weights were recorded weekly.
Food consumption was measured weekly for each cage of animals.

Sacrifice and pathology:

Clinical pathology investigations were performed at termination.
All animals were killed and necropsied after week 4; selected organs were weighed and tissue samples were fixed and preserved at necropsy.
Selected tissues from group 1 and 4 animals killed after week 4 were examined histopathologically.

Other examinations:

NA

Statistics:

None

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):

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not specified

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 specified

Histopathological findings: neoplastic:

not specified

Other effects:

no effects observed

Details on results:

There were no unscheduled deaths.
There were no remarkable clinical signs.
There were no differences in body weight gain between the groups.
There were no differences in food consumption between the groups.
There were no differences in haematology profiles.
There were no differences in clinical parameters between the groups which were sufficient in magnitude to indicate an effect of treatment.
There was no difference in urine volume between the groups.
There were no differences in the organ weights between the groups which were sufficient in magnitude to indicate an effect of treatment.
There was no macroscopic or microscopic evidence of change associated with treatment.

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: original NCD unit is mg/kg/day.

Remarks on result:

other: no adverse effects observed at the highest dose tested, i. e. limit dose

Key result

Critical effects observed:

no

In conclusion, oral (gavage) administration of PATE HAT to the rat at dose levels up to 1000 mg/kg bw/day was not associated with any toxic change. Under the defined experimental conditions, the no-effect level of PATE HAT in the rat be considered to be greater than 1000 mg/kg bw/day.

Conclusions:

PATE HAT was tested in an 28 Day repeated dose toxicity test in the rat. The oral administration at dose levels up to 1000 mg/kg bw/day was not associated with any toxic change. The no-effect level (NOEL) can be considered to be greater than 1000 mg/kg bw/day. No classification and labelling is necessary according to Regulation 1272/2008/EC (CLP).

Executive summary:

PATE HAT was tested in an 28 Day repeated dose toxicity test in the rat.

The oral administration at dose levels up to 1000 mg/kg bw/day was not associated with any toxic change.

The no-effect level (NOEL) of PATE HAT can be considered to be greater than 1000 mg/kg bw/day.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

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

HAT ISO was tested in a 28 day repeated dose toxicity study by oral application in rats. The oral administration at dose levels up to 1000 mg/kg bw/day (limit dose) was not associated with any toxic change. The no-observed-adverse-effect level (NOAEL) was greater than 1000 mg/kg bw/day.

Repeated dose toxicity testing by the dermal and inhalation route was waived. Exposure to HAT ISO by the dermal and inhalation route is unlikely, as:

1) HAT-ISO is manufactured and formulated in closed systems. Thus, exposure via the dermal route or inhalation during manufacturing and formulation is unlikely.

2) Use of formulated HAT-ISO is also unlikely to result in exposure via the dermal route or inhalation. During use HAT-ISO is included in three-dimensional matrices of polymeric, resinous material and completely retained. Therefore, exposure via the dermal route or inhalation is practically negligible.

For more details on exposure assessment see the risk assessment report in section 13.

The physico-chemical properties of HAT ISO suggest also no evidence of a significant absorption by the dermal and inhalation route. No signs of systemic effects were observed in the acute oral and dermal toxicity test. Further, neither skin nor dermal irritation tests showed any signs of systemic effects by absorption through skin or mucous membranes. In two one generation studies and one teratotoxicity study with test item administered via oral application, parent animals were treated with up to 1000 mg/kg bw/day (limit dose). No signs of toxicity were reported in any of these studies.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data the test item does not require classification according to Regulation (EC) No 1272/2008 (CLP), as amended for the tenth time in Regulation (EU) No 2017/776.