lithium nickel dioxide

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

other: part of a testing strategy

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Test guideline 1

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)

Version / remarks:

26 July 2013

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Version / remarks:

09.12.2010

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Lithium; oxido(oxo)nickel

Cas Number:

12031-65-1

Molecular formula:

LiNiO2

IUPAC Name:

Lithium; oxido(oxo)nickel

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No. of test material: 8800315
- Content: approx. 99.8 g/100 g
- Homogeneity: the test item appeared to be homogeneous
- Appearance: solid, anthracite
- Expiry Date: 12 August 2021

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, under N2

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item (non-surfactant) was tested as suspension in deionised water using sonication for 10 minutes. Before each removal the test item was resuspended.
- Form of application: 20% (w/v) suspention in deionized water

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Freshly isolated bovine cornea; supplier: AB Schlachthof GmbH & Co. KG, 63739 Aschaffenburg, Germany
- Characteristics of donor animals: age of the animals: 14 month old
- indication of any existing defects or lesions in ocular tissue samples: Corneas free of defects (opacity, scratches, pigmentation etc.)

Test system

Vehicle:

water

Remarks:

deionized

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 20% (w/v) solution in de-ionized water

NEGATIVE CONTROL
- Amount(s) applied (volume or weight with unit): 750 µL of de-ionized water

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 750 μL of 20% (w/v) solution of imidazole in deionized wateror

Duration of treatment / exposure:

4 hours

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS: Corneas free of defects (opacity, scratches, pigmentation etc.) were dissected with a 2 to 3 mm rim of sclera. Isolated corneas were mounted in cornea holders that consists of anterior and posterior chambers. Both chambers were filled to excess with pre-warmed Eagle’s MEM (without phenol red) and then equilibrated in a vertical position at about 32°C for at least 1 hour. After the equilibration period, the medium in both chambers was replaced by fresh pre-warmed medium and initial corneal opacity readings were taken for each cornea with an opacitometer.

QUALITY CHECK OF THE ISOLATED CORNEAS: Any corneas that showed macroscopic tissue damage or an opacity value < 550 opacity units were discarded. The remaining corneas were then distributed into negative control, positive control and treatment groups.(According to OECD TG 437, corneas that have an opacity value >7 or equivalent for the opacitometer and cornea holders used after an initial one-hour equilibration period are to be discarded. In the test facility´s opacitometer in combination with the used cornea holder set 2013-22 and 2013-24, the maximal initial opacity of >7 arises from I= 550 lux with Io= 641 lux).

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED: Yes

POSITIVE CONTROL USED: Yes

APPLICATION DOSE AND EXPOSURE TIME: Application of 750 µL of the 20% (w/v) test-substance preparation, 4 hours exposure time

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: at least 3 times.

- POST-EXPOSURE INCUBATION: no

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Yes.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader: Yes, OD490, OP_KiT opacitometer (Electro Design, 63-Riom France).

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA:
IVIS ≤ 3, Prediction: No Category
IVIS > 3; ≤ 55, Prediction: No prediction can be made
IVIS > 55, Prediction: Category 1

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Remarks:

IVIS

Run / experiment:

Run 1, mean of three single corneas

Value:

355.13

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: Cathegory 1, serious eye damage

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: No.

DEMONSTRATION OF TECHNICAL PROFICIENCY: Proficiency given

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

Any other information on results incl. tables

Table 1: Results of the BCOP Test, in vitro irritancy score (IVIS) of the test substance, the NC and the PC.

	Opacity value per cornea	Permeability at 490 nm (OD490)		IVIS
substance identification			per cornea	per group
				mean	SD
	400.00*	0.889*	413.34
Test substance	326.00*	1.201*	344.02	355.13	53.52
	291.00*	1.136*	308.04
	0	0.066	0.99
NC (Saline)	0	0.078	1.17	1.08	0.09
	0	0.072	1.08
	**	**	**
NC (deionised water)	2.00	0.078*	2.00	1.00	1.41
	0.00	0.061*	0.00
	81.00*	0.605*	90.08
PC (10% (w/v) Benzalkonium chloride in saline)	68.00*	0.471*	75.07	80.11	8.63
	67.00*	0.545*	75.18
	88.00*	0.771*	99.57
PC (20 % Imidazole in saline)	97.00*	1.239*	115.59	97.04	19.93
	65.00*	0.731*	75.97

*Correction was performed with the values of the Negative Control Saline.
** not taken into the evaluation since this value is declared as an outlier.

 

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria