Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The experiment was performed on 10 April 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Test material form:
liquid

Test animals / tissue source

Species:
chicken
Details on test animals or tissues and environmental conditions:
Historically, eyes collected from chickens obtained from a slaughterhouse where they are killed for human consumption have been used for this assay, eliminating the need for laboratory animals. Only the eyes of healthy animals considered suitable for entry into the human food chain are used.

Although a controlled study to evaluate the optimum chicken age has not been conducted, the age and weight of the chickens used historically in this test method are that of spring chickens traditionally processed by a poultry slaughterhouse (i.e., approximately 7 weeks old, 1.5 - 2.5 kg).

Test system

Vehicle:
physiological saline
Controls:
yes
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
Test item was applied at the dose of 30 µl, to 3 enucleated chicken eyes, during 10 seconds.
Duration of treatment / exposure:
Test item was applied at the dose of 30 µl, to 3 enucleated chicken eyes, during 10 seconds. Treated corneas were evaluated pretreatment and starting at 30, 75, 120, 180, and 240 minutes after the post-treatment rinse.
Observation period (in vivo):
Treated corneas were evaluated pretreatment and starting at 30, 75, 120, 180, and 240 minutes after the post-treatment rinse.
Duration of post- treatment incubation (in vitro):
Treated corneas were evaluated pretreatment and starting at 30, 75, 120, 180, and 240 minutes after the post-treatment rinse.
Number of animals or in vitro replicates:
3
Details on study design:
All observations of the cornea and measurement of corneal thickness are performed using a Haag-Streit BP900 slit-lamp microscope with depth-measuring device no. I. For the measurement of corneal thickness, the slit-width is set at 9½, equalling 0.095 mm.

Treated corneas are evaluated prior to treatment and at 30, 75, 120, 180, and 240 minutes (± 5 minutes) after the post-treatment rinse. These time points provide an adequate number of measurements over the four-hour post-treatment period, while leaving sufficient time between measurements for the requisite observations to be made for all eyes.

The endpoints evaluated are corneal opacity, swelling, fluorescein retention, and morphological effects (e.g., pitting or loosening of the epithelium). All of the endpoints, with the exception of fluorescein retention (which is determined only at prior to treatment and at 30 minutes after test item exposure) are determined at each of the above time points.

Results and discussion

In vitro

Results
Irritation parameter:
cornea opacity score
Value:
ca. 2.7
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
In accordance with the Regulation (EC) No. 1272/008, the results obtained under these esperimental conditions enable to conclude that the test item Pine tar / Tar, Pinus sylvestris has to be classified in Category 1 "Irreversible effects on the eye". The signal word "Danger" and hazard statements H318 "Causes serious eye damage" are required.
Executive summary:

The test item Pine tar / Tar, Pinus sylvestris was applied at the dose of 30 µl, to 3 enucleated chicken eyes, during 10 seconds. A positive and a negative control was included. Damages of the test item were assessed by determination of corneal swelling, opacity, and fluorescein retention at 30, 75, 120, 180 and 240 minutes post-dose. In accordance with the Regulation (EC) No. 1272/008, the results obtained under these esperimental conditions enable to conclude that the test item Pine tar / Tar, Pinus sylvestris has to be classified in Category 1 "Irreversible effects on the eye". The signal word "Danger" and hazard statements H318 "Causes serious eye damage" are required.