4,4'-methylenebis(2,6-diisopropylaniline)

Endpoint summary

Description of key information

SKIN IRRITATION:

A study was performed to determine the primary skin irritation index and corrosive effects of the test item in the New Zealand White rabbit. The study design was based on OECD Guidelines No. 404. The test item was classified as a non-irritant to rabbit skin. No corrosive effects were noted.

EYE IRRITATION:
This in vitro study was performed according to OECD 405 to assess the irritating potential of the test item by detection of damages in blood vessels under the chorioallantoic membrane of 9 day incubated chicken eggs. The test item was tested pure. The observation time was 5 minutes at room temperature. No irritating effects were observed during 5 min incubation with the test item. The calculated mean irritancy index is 0.00.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 1983

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species, strain and supplier:
Three young adult female New Zealand White rabbits obtained from Morton Commerical Rabbits, Parsonage Road, Stansted, Essex, were used for the study.

Specification:
At the start of the study the rabbits weighed between 2.55 and 2.83 kg and were at least 11 weeks old. They were acclimatised to the laboratory environment for at least 3 days and were examined for signs of ill health or injury shortly before the study commenced. All animals appeared healthy and no animals were rejected.

Husbandry:
The animals were individually housed in grid floor cages in a single air conditioned room at a temperature between 18 and 20°C and a relative humidity between 51 and 62%. The animals were exposed to a constant daily photoperiod of 14 hours artificial light (06.00 - 20.00) and 10 hours darkness. Temperature and humidity were recorded twice daily on weekdays and once daily at week-ends.

Diet and drinking water:
The animals were allowed free access to mains water and food (Standard Rabbit Diet, Special Diets Services Ltd., Stepfield, Witham, Essex) throughout the study.

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped the day prior to treatment

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

The test article was melted by warming to approximately 70°C. The test article was allowed to cool to approximately 37°C at
which temperature it was sufficiently liquid to be administered by a 1 ml syringe. The test article concentration applied was 0.5 ml undiluted.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3 female rabbits

Details on study design:

Treatment procedure:
The back of each animal was clipped free of hair the day prior to treatment using electric veterinary clippers. A portion of test article (0.5 ml) was applied to an area of skin approximately 6 cm2 on each animal and covered with a gauze patch. The patch was occluded and secured using a strip of impermeable adhesive tape. A plastic collar was placed around the neck of each animal to
prevent premature removal of the wrappings and ingestion of the test article. Four hours after application the patches were
removed and the skin wiped with a disposable paper towel moistened with water to remove any test article still remaining.

Evaluation of effects:
One hour after removal of the wrappings the skin reactions were scored using the Draize scale. The evaluation was repeated 24,48 and 72 hours after removal of the patches. No irritation was noted at the 72 hour observation and the study was terminated.

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

All treated skin sites appeared normal throughout the study period. A primary irritation index of 0 was obtained. The test item was regarded as non-irritant to rabbit skin. No corrosive effects were noted.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item was classified as a non-irritant to rabbit skin.

Executive summary:

A study was performed to determine the primary skin irritation index and corrosive effects of the test item in the New Zealand White rabbit. The study design was based on OECD Guidelines No. 404. Three female rabbits were tested and 0.5 ml test substance applied undiluted onto the clipped skin area of the animals. A single 4 -hour application produced a primary irritation index of 0. The test item was classified as a non-irritant to rabbit skin. No corrosive effects were noted.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Remarks:

in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Principles of method if other than guideline:

With the HET-Cam test an irritative or corrosive potential is determined by a detection of damages in blood vessels under the chorioallantoic membrane of 9 day incubated chicken eggs.
The Cam (chorioallantoic membrane) is a vital vascular membrane with a closed blood vessel system. After application of a test item on the membrane the underlying blood vessels could become damaged. Particularly three events are observed: haemorrhage, vessel lysis and coagulation of the blood in the vessels.

GLP compliance:

yes (incl. QA statement)

Species:

other: chicken eggs.

Strain:

other: Lohmann Selected Leghorn

Details on test animals or tissues and environmental conditions:

The eggs were obtained from the LSL Rhein-Main Geflügelvermehrungsbetriebe (D-64807 Dieburg, Germany) on day one. Afterwards they were incubated at 37,5 ± 0.5 °C for eight days. The HET-Cam was carried out with Lohmann Selected Leghorn chicken eggs. The Lohmann Selected Leghorn chicken was selected for several reasons: the ability to hatch the eggs of this breed is very consistent and reproducible. There did not appear to be any hereditary defects in this breed.

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

In the HET-Cam assay the test item was tested pure. Six eggs were treated with the test item. An amount of the solid test item was applied to the membrane so that at least 25% of the membrane was covered by the test item

Duration of treatment / exposure:

An amount of the solid test item was applied to the membrane so that at least 25% of the membrane was covered by the test item. During the observation for 300 seconds lesions in close proximity to the covered membrane were noted.

Observation period (in vivo):

The membrane of the eggs were observed for 300 sec. Lesions of the underlying blood vessels were noted. In general there were three endpoints which were observed and the time when that particular endpoint was observed was noted. The three endpoints were:
• haemorrhage
• coagulation
• lysis of the blood vessel

Number of animals or in vitro replicates:

Six eggs were treated with the test item.

Details on study design:

While incubation the eggs were rotated to prevent an attachment of the embryo to one side of the egg. On day eight the eggs were candled and non-viable eggs were discarded. The rest of the eggs were placed upward in the incubator and were incubated for another day without rotation. Six eggs were treated with the test item.
Physiological sodium chloride solution (0.9% (v/v)) was used as negative control. The negative control showed no irritating effect on the blood vessels under the membrane. An 1 % solution of SDS and 0.1 N NaOH were used as positive controls. The positive controls induced a severe irritation on the blood vessels. The calculated mean irritancy indices are 7.83 for 1% SDS and 19.66 for 0.1 N NaOH.

Irritation parameter:

other: mean irritancy index

Run / experiment:

mean of six experiments

Value:

0

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

It can be stated that in this study and under the experimental conditions reported, the test item does not possess any irritating potential.

Executive summary:

This in vitro study was performed according to OECD 405 to assess the irritating potential of the test item by detection of damages in blood vessels under the chorioallantoic membrane of 9 day incubated chicken eggs. The test item was tested pure. The observation time was 5 minutes at room temperature. No irritating effects were observed during 5 min incubation with the test item. The calculated mean irritancy index is 0.00.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the data available the substance is not classified according to Regulation 1272/2008/EEC (CLP) and according to Directive 67/548/EEC (DSD).