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

Eye irritation

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

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)

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

243-421-7

EC Name:

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

Cas Number:

19900-69-7

Molecular formula:

C25 H38 N2

IUPAC Name:

4-{[4-amino-3,5-bis(propan-2-yl)phenyl]methyl}-2,6-bis(propan-2-yl)aniline

Constituent 2

Reference substance name:

Lonzacure M-DIPA

IUPAC Name:

Lonzacure M-DIPA

Test material form:

solid: compact

Details on test material:

- Identity: Lonzacure M-DIPA
- Molecular Weight: 366.6 g/mol
- Appearance: Solid, white to brown
- Solubility: In EtOH, acetone, acetonitrile, and DMSO > 1 g/L
- Stability in Solvent: In H2O, EtOH, acetone, acetonitrile, and DMSO > 96 hours
- Storage: At room temperature protected from light
- Expiration Date: December 31, 2009

Test animals / tissue source

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.

Test system

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.

Results and discussion

In vitro

Results

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

Applicant's summary and conclusion

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.