2-Naphthalenol, 1-[2-[2-ethyl-4-[2-(2-ethylphenyl)diazenyl]phenyl]diazenyl]-, ar-heptyl derivs.

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

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)

GLP compliance:

yes

Test material

Test material information

1

Reference substance name:

Reaction products of 2-Ethylaniline with heptyl naphthalen-2-ol

Cas Number:

1401000-07-4

Molecular formula:

Not applicable (UVCB Substance)

IUPAC Name:

Reaction products of 2-Ethylaniline with heptyl naphthalen-2-ol

Test material form:

solid

Specific details on test material used for the study:

Batch No.: 80-47-16
Storage Conditions: Room temperature, protected from light
Expiry Date: August 2017

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

The assay uses isolated corneas obtained as a by-product from animals freshly slaughtered.
On the test day, fresh eyes were collected from the slaughterhouse and were transported in HBSS containing Pen/Strep on ice to the laboratories. Immediately after arrival of the eyes, cornea preparation was initiated.
The eyes were carefully examined for defects and any defective eyes were discarded.
The tissue surrounding the eyeball was carefully pulled away and the cornea was excised leaving a 2 to 3 mm rim of sclera. The isolated corneas were stored in a petri dish containing HBSS. Before the corneas were mounted in corneal holders (Duratec GmbH) with the endothelial side against the O-ring of the posterior chamber, they had been visually examined for defects and any defective cornea had been discarded. The anterior chamber was then positioned on top of the cornea and tightened with screws. The chambers of the corneal holder were then filled with RPMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete RPMI). The posterior chamber was always filled first. The corneas were incubated for one hour at 32 +/- 1 °C.

Test system

Vehicle:

physiological saline

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750 mg of the test item preparation or the control substance was introduced into the anterior chamber (open-chamber method).

Duration of treatment / exposure:

4 hours ± 5 minutes incubation at 32 +/- 1 °C

Number of animals or in vitro replicates:

3 corneas for the test item
3 corneas as negative controls treated with physiological saline 0.9% NaCl
3 corneas as positive controls treated with imidazole 20% in physiological saline 0.9% NaCl

Details on study design:

The test item was moistened with one drop of physiological saline 0.9% NaCl (B. Braun Melsungen, lot no. 17031412, expiry date: 12/2019) before direct application.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

Test item Cornea 1

Value:

3.12

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

in vitro irritation score

Run / experiment:

test item Cornea 2

Value:

2.14

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

in vitro irritation score

Run / experiment:

test item Cornea 3

Value:

0.68

Negative controls validity:

valid

Positive controls validity:

valid

Results 4

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean Value

Value:

1.96

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to the evaluation criteria the test item RED 2735 is classified into UN GHS No Category.

Executive summary:

The eye irritancy potential of RED 2735 was investigated in the bovine corneal opacity and permeability assay.

Preparation of the test item:                      The test item was moistened with physiological saline 0.9% NaCl.

Visual Observation after treatement:         All 3 corneas treated with RED 2735 showed slight yellow discolouration of the tissue.

Mean in vitro irritation score:                   1.96

The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.

The negative control responses should result in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.