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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
supporting study
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well documented study according to international accepted guidelines and GLP compliant.

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
Cas Number:
Molecular formula:
Test material form:
solid: crystalline
Details on test material:
Test item: Ethisterone-ketal
Batch No.: 252123N
Physical state: powder
Colour: white
Storage: at 15-30 °C, protected from light and moisture

Test animals / tissue source

other: chicken eyes
Details on test animals or tissues and environmental conditions:
Chicken heads collection and transport:
Species of chicken: ROSS 308. Source: TARAVIS KFT. Heads were removed immediately after sedation of the chickens (sedation was happened by electric current). The heads were transported to TOXI-COOP ZRT. at the earliest convenience for use approximately within 2 hours from collection. The ambient temperature was optimal (19.6 ºC to 20.2 ºC) during the transport. All eyes used in the assay were from the same groups of eyes collected on one specific day.
After collection, the heads were inspected for appropriate quality and wrapped with paper moistened with saline, then placed in a plastic box that can be closed (4-5 heads/box).
Eyes selection:
After removing the head from the plastic box, it was put on soft paper. The eyelids were carefully cut away with scissors, avoiding damaging the cornea. If the cornea was in good condition, the eyeball was carefully removed from the orbit.

Test system

unchanged (no vehicle)
Amount / concentration applied:
The three test item were treated with 0.03 g/eye Ethisterone-ketal.
The three positive control eyes were treated in a similar way with 0.03 g Imidazole.
One negative control eye was treated with saline solution (9 g/L).
Duration of treatment / exposure:
The time of application was monitored, then after an exposure period of 10 seconds from the end of the application the cornea surface was rinsed thoroughly with 20 mL saline solution at ambient temperature, while taking care not to damage the cornea but attempting to remove all the residual test item if possible.
Observation period (in vivo):
The control and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse. Minor variations within ±5 minutes were considered acceptable.
The cornea thickness and cornea opacity were measured at all time points. Fluorescein retention was determined at baseline (t=0) and 30 minutes after the post-treatment rinse.
Number of animals or in vitro replicates:
Three test item treated eyes, three positive control eyes and one negetaive control eye were used in this study.
Details on study design:
The purpose of this Isolated Chicken Eye Test (ICET) was to evaluate the potential ocular corrosivity or severe irritancy of the test item ETHISTERONE-KETAL by its ability to induce toxicity in enucleated chicken eyes. Each eye which was used in this study was collected in the slaughterhouse, eliminating the need for laboratory animals. The test compound was applied in a single dose onto the cornea of isolated chicken eyes in order to potentially classify the test compound as ocular corrosive and/or severe irritant. The damage by the test substance was assessed by determination of corneal swelling, opacity, fluorescein retention and morphological effects. These parameters were evaluated pre-treatment and starting at approximately 30, 75, 120, 180, and 240 minutes after the post-treatment rinse. The endpoints evaluated were corneal opacity, swelling, fluorescein retention, and morphological effects. All of the endpoints, with the exception of fluorescein retention (which was determined only at pre-treatment and 30 minutes after test substance exposure) were determined at each of the above time points.

Results and discussion

In vitro

Other effects / acceptance of results:
In this ICET, ETHISTERONE-KETAL did not cause ocular corrosion or severe irritation in the enucleated chicken eyes.
Positive and negative controls showed the expected results. The experiment was considered to be valid.

Any other information on results incl. tables



ICE Class

Mean maximum corneal swelling at up to 75 min

3 %


Mean maximum corneal swelling at up to 240 min

3 %


Mean maximum corneal opacity



Mean fluorescein retention



Other Observation


Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Migrated information
In this in vitro eye corrosives and severe irritants study, using the Isolated Chicken Eye model with ETHISTERONE-KETAL, no ocular corrosion or severe irritation potential was observed. According to the guideline OECD 438, ETHISTERONE-KETAL has been categorized as “No Category”.