Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental starting date: 2015-07-08; Experimantal completion date: 2015-07-08
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Version / remarks:
resp. EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Deviations:
yes
Remarks:
The temperature during storage was outside the range of 20+/-5°C. The temperature was 14.4°C and 25.8°C for one day - marginal deviation- uncritical; Instead of an opacitometer a photometer was used for mesurement of the opacity - uncritical
GLP compliance:
yes (incl. certificate)
Remarks:
Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht; Kaiser-Friedrich-Straße 7, D-55116 Mainz

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: viscous liquid
Details on test material:
Stabilized EMIM BF4:
Appearance: clear lightly yellow viscous liquid
Purity: 93.36% (EMIM+, HPLC); 86.68% (BF4-, titration); 4.89% (Stabilizer, HPLC)

Test animals / tissue source

Species:
other: fresh bovine corneas (Bos primigenius Taurus)
Details on test animals or tissues and environmental conditions:
The BCOP test method uses isolated corneas from the eyes of cattle which are freshly slaughtered in the slaughterhouse Müller Fleisch GmbH, Enzstr. 2-4, 75217 Birkenfeld, Germany. The cattle were between 12 and 60 months old.
The eyes are removed as soon as possible after death of the cattle and immersed in Hanks’ Balanced Salt Solution in a suitable and cool container. Like that, they are transported to the laboratory within 4 h where they are used at once.

Test system

Vehicle:
unchanged (no vehicle)
Duration of treatment / exposure:
Exposition time on the corneas: 10 min +/- 30 sec at 32 +/- 1 °C
Details on study design:
Closed Chamber Method:
The “closed chamber-method” is used for liquid substances.
The respective substance (negative control solution, positive control or test item) is applied by pipetting 750 µL of the appropriate liquid throug h the refill hole in the chamber on the cornea. Exposition time on the corneas is 10 min ± 30 sec at 32 ± 1 °C. After thorough rinsing with cMEM with phenol red and final rinsing with cMEM without phenol red, the chamber is filled with cMEM without phenol red, and the corneas are stored for additional 2 h ± 5 min at 32 ± 1 °C (post-incubation). Then, the final opacity value of each cornea is recorded (again by measurement at 57 0 nm).

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Run / experiment:
1 valid experiment
Value:
ca. 1.23
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
The test item was tested pure. Under the conditions of this test, the test item stabilized EMIM BF4 showed no effects on the cornea of the bovine eye. The calculated IVIS (invitro irritancy score) is 1.23. According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS <= 3 requires no classification for eye irritation or serious eye damage.
Executive summary:

One valid experiment was performed.

Bovine corneas were used. They were collected from slaughtered cattle wich were between 12 and 60 month old.

The test item stabilized EMIM BF4 was brought onto the cornea of a bovine eye which had been incubated with CMEM without phenol red at 32 +/- 1 °C for 1 h and whose opacity had been measured. The test item was incubated on the cornes for 10 min at 32 +/- 1 °C. After removal of the test item and 2 h post incubation, opacity and permeability values were measured.

Physiological sodium chloride solution was used as negative control. The negative control showed no irritating effect on the cornea and the calculated IVIS (in vitro irritancy score) is -0.75.

Dimethylformamide (DMF) undiluted was used as positive control. The positive control induced serious eye damge on the cornea and falls within two standard deviations of the current historical mean. The calculated IVIS (in vitro irritancy score) is 105.62.

Under the conditions of this study, the test item stabilized EMIM BF4 showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 1.23.

According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS <=3 requires no classification for eye irritation or serious eye damage.