Speiss, lead, nickel-contg.

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 24, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to GLP and actual testing guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD testing guideline 437: Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe irritants (September, 2009)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Speiss, lead, nickel containing

IUPAC Name:

Speiss, lead, nickel containing

Constituent 2

Reference substance name:

Speiss, lead, nickel-contg.

EC Number:

308-765-5

EC Name:

Speiss, lead, nickel-contg.

Cas Number:

98246-91-4

IUPAC Name:

Speiss, lead, nickel-contg.

Constituent 3

Reference substance name:

Nickel Speiss

IUPAC Name:

Nickel Speiss

Details on test material:

- Name of test material (as cited in study report): Nickel Speiss
- Analytical purity: Nickel content 41.8%
- Manufacturing date: January 07, 2010
- Lot/batch No.: 52690100309M000000
- Storage condition of test material: at room temperature

Test animals / tissue source

Species:

other: in vitro

Details on test animals or tissues and environmental conditions:

in vitro test with corneae of bovine eyes

Test system

Vehicle:

other: cMEM (complete Minimum Essential Medium), supplemented with sodium bicarbonate and L-glutamine. Immediately before starting the test, cMEM was supplemented with 1% fetal calf serum.

Controls:

other: in vitro: 3 positive and 3 negative controls

Amount / concentration applied:

0.75 mL per cornea

Duration of treatment / exposure:

240 minutes (+/- 5 minutes)

Number of animals or in vitro replicates:

in vitro test:
- 3 cornea for positive control
- 3 cornea for negative control
- 3 cornea with test item

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with 0.9 (w/v) saline
- Time after start of exposure: 240 minutes

SCORING SYSTEM: in vitro irritancy score (IVIS)

TOOL USED TO ASSESS SCORE: a formula was used:
To determine the in vitro score of the negative control the following formula was used:
In vitro score = opacity value + (15 x optical density value at 490 nm)

To determine the in vitro score of the positive control and the test item the following formula was used:
In vitro score = (opacity value - opacity value of the mean negative control) + (15 x corrected optical density[OD] at 490 nm)

The positive control: 10% (w/v) benzalkonium chloride
The negative control: 0.9% NaCl solution

Results and discussion

In vivo

Results

Irritation parameter:

other: in vitro corneal opacity and permeability

Basis:

mean

Time point:

other: 240 minutes

Score:

ca. 10.47

Max. score:

16.09

Reversibility:

other: in vitro: reversibility is not relevant

Irritant / corrosive response data:

Mean in vitro score, calculated from opacity and permeability of the cornea:
- Negative control: 1.94
- Positve control: 194.59
- Nickel Speiss: 10.47

Any other information on results incl. tables

Depending on the score obtained, the test item, positive and negative controls were classified into scaling categories.

Test group	Meanin vitroscore	In vitroirritation scale
Negative control	1.94	Non eye irritant
Positive control	194.59	Very severe eye irritant
Nickel Speiss	10.47	Mild eye irritant

Applicant's summary and conclusion

Interpretation of results:

slightly irritating

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

Nickel speiss was classified as a mild irritant. But according to the OECD 437 guideline, Nickel speiss is considered as a non-irritant.

Executive summary:

Bovine Cornea Opacity and permeability Assay (BCOP)

A key in vitro study for eye irritation was conducted with Nickel speiss according to the GLP guidelines and OECD 437 test guideline. After the background opacity of fresh bovine corneas (t₀) was determined with an opacitometer, a 20% (w/v) suspension in saline [0.9% (w/v) NaCl in deionised water] of Nickel speiss was applied to corneas and incubated for 240 minutes at 32±1°C. The posterior chamber contained MEM medium supplemented with sodium bicarbonate and L-glutamine and 1% fetal calf serum. After the incubation phase, Nickel speiss was rinsed from the corneas and opacity was measured again (t₂₄₀). Thereafter, the permeability of the corneas was determined while application of 1ml of sodium fluorescein for 90 minutes at 32±1°C in a horizontal position. The concentration of sodium fluorescein that crossed into the posterior corneal chamber was measured spectrophotometrically. An in vitro score was calculated on basis of the results obtained on opacity and permeability.

 

The acceptance criteria for the study with negative and positive controls were met and the study was considered reliable, adequate and relevant.

Nickel speiss slightly increased opacity of the corneas but did not cause permeability of the corneas compared with the results of the negative control. The calculation mean in vitro score was 10.47. According to the criteria mentioned in the report, Nickel speiss was classified as a mild irritant. But according to the OECD 437 guideline, Nickel speiss is considered as a non-irritant.