Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental start date 15 August 2017. Experimental completion date 15 August 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Deviations:
yes
Remarks:
incorrect version of the study plan was followed in error. This deviation was considered to have not affected the integrity or validity of the study.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
Identification: 1-(methylamino)-4-[(3-methylphenyl)amino] anthraquinone
Physical state/Appearance: dark red powder
Batch: 702W01
Purity: 99.4%
Expiry Date: 08 February 2020
Storage Conditions: room temperature in the dark
Intended use/Application: industrial chemical

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
Source of Bovine Eyes
Eyes from adult cattle (typically 12 to 60 months old) were obtained from a local abattoir as a by-product from freshly slaughtered animals. The eyes were excised by an abattoir employee after slaughter, and were placed in Hanks’ Balanced Salt Solution (HBSS) supplemented with antibiotics (penicillin at 100 IU/mL and streptomycin at 100 µg/mL). They were transported to the test facility over ice packs on the same day of slaughter. The corneas were prepared immediately on arrival.

Test system

Vehicle:
other: 0.9% w/v sodium chloride solution
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
For the purpose of this study the test item was prepared as a 20% w/v solution in sodium chloride 0.9% w/v.

The negative control item, sodium chloride 0.9% w/v, was used as supplied.
The positive control item, Imidazole, was used as a 20% w/v solution in sodium chloride 0.9% w/v.
Duration of post- treatment incubation (in vitro):
240 minutes
Number of animals or in vitro replicates:
test substance: 3
negative control: 3
positive control: 3

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Run / experiment:
Test item
Value:
6.6
Negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

Corneal Epithelium Condition

The corneas treated with the test item were partly cloudy post treatment. The corneas treated with the negative control item were clear post treatment. The corneas treated with the

positive control item were cloudy post treatment.

6.3 In Vitro Irritancy Score

The In Vitro irritancy scores are summarized as follows:

Treatment In Vitro Irritancy Score

Test Item 6.6

Negative Control 1.6

Positive Control 99.4

6.4 Criteria for an Acceptable Test

The positive control In Vitro Irritancy Score was within the range of 65.1 to 123.3. The

positive control acceptance criterion was therefore satisfied.

The negative control gave opacity of ≤2.4 and permeability ≤0.072. The negative control

acceptance criteria were therefore satisfied.

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Conclusions:
No prediction of eye irritation can be made.