Slags, phosphorus-manufg.

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 January 2010 - 28 January 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test was conducted according to OECD Test Guideline No. 437, 2009, under GLP Standards, and QA.

Justification for data waiving:

other:

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD Guideline for Testing of Chemicals 437: Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants (September, 2009)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: INVITTOX (UK) protocol no. 124 "Bovine Opacity and Permeability Assay", 1999

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: OTWG/ ICCVAM / NICEATM (BRD): current status of in vitro test methods for identifying ocular corrosives and severe irritants: The Bovine Corneal Opacity and Permeability (BCOP) Test Method, March 2006

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Remarks:

Statement of Compliance

Test material

Test material information

Constituent 1

Reference substance name:

Slags, phosphorus-manufg.

EC Number:

273-732-3

EC Name:

Slags, phosphorus-manufg.

Cas Number:

69012-32-4

Molecular formula:

not applicable due to the multi constituent mineral structure of the substance

IUPAC Name:

aluminium(3+) heptacalcium magnesium(2+) λ²-iron(2+) disodium tris([(trioxidosilyl)oxy]silanetris(olate)) oxosilanebis(olate) hydroxysilanoylolate difluoride

Details on test material:

- Name of test material (as cited in study report): Phosphorous slag
- Physical state: powder
- Analytical purity: confidential
- Lot/batch No.: Not indicated
- Expiration date of the lot/batch: confidential
- Stability under test conditions: stable
- Storage condition of test material: At room temperature in the dark

Test animals / tissue source

Species:

other: in vitro test with bovine cornea

Strain:

other: not applicable

Details on test animals or tissues and environmental conditions:

BOVINE EYES:
- Source: Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands)
- Age at study initiation: young cattle - Bovine eyes were used as soon as possible after slaughter on the same day.

Test system

Vehicle:

physiological saline

Remarks:

20% (w/v)

Controls:

yes, concurrent vehicle

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 ml

VEHICLE
- Concentration (if solution): 20% (w/v)

POSITIVE CONTROL: 20% (w/v) Imidazole

Duration of treatment / exposure:

Incubation 240 ± 10 minutes (actual treatment period 230 minutes)

Observation period (in vivo):

Not applicable

Number of animals or in vitro replicates:

9 freshly isolated bovine cornea: negative control, positive control and test item: 3 cornea each

Details on study design:

EXPOSURE OF THE CORNEA: in cornea holder

PRE-INCUBATION: 1 hour

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The test item or control items, respectively, were rinsed off from the application side by changing cMEM (complete Eagle's Minimum Essential Medium), at least three times.
- Time after start of exposure: 240 ± 10 minutes

SCORING SYSTEM:
The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).
Additionally the opacity and permeability values were evaluated independently to determine whether the test substance induced irritation through only one of the two endpoints.
A test substance that induces an IVIS ≥ 55.1 is defined as a corrosive or severe irritant.

TOOL USED TO ASSESS SCORE:
Cornea opacity value: opacitometer (OP_KiT)
Permeability (optical density value) at 490 nm (OD490): fluorescein; 96-well microplate reader

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

Opacity

Basis:

mean

Remarks:

Cornea 1, 2, and 3

Score:

0

Reversibility:

other: not applicable

Results 2

Irritation parameter:

cornea opacity score

Remarks:

Permeability

Basis:

mean

Remarks:

Cornea 1, 2, and 3

Score:

0

Reversibility:

other: not applicable

Results 3

Irritation parameter:

other: In vitro irritation Score

Basis:

mean

Remarks:

Calculated using the negative control mean opacity and mean permeability values

Score:

0

Reversibility:

other: not applicable

Remarks on result:

other: In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value)

Any other information on results incl. tables

The individual in vitro irritancy scores for the negative controls ranged from -0.1 to 0.1. The individual positive control in vitro irritancy scores ranged from 87 to 109. The corneas treated with the positive control were turbid after the 240 minutes of treatment. Negative and positive controls were within historical ranges. The corneas treated with Phosphorous slag showed an opacity value of 0 for all three corneas and permeability values ranging from -0.005 to 0.006. The corneas were clear after the 240 minutes of treatment with Phosphorous slag. Hence, the in vitro irritancy scores were 0 for all three corneas after 240 minutes of treatment with Phosphorous slag.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Phosphorous slag did not induce ocular irritation through both endpoints, opacity and permeability, resulting in a mean in vitro irritancy score of 0 after 240 minutes of treatment. It is concluded that this test is valid and that Phosphorous slag is a non irritant in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.

Executive summary:

This in vitro study was performed according to OECD Guideline 437, 2009 to assess the corneal irritation and damage potential of Phosphorous slag by means of the Bovine Corneal Opacity and Permeability Assay (BCOP) using fresh isolated bovine corneae. The possible ocular irritancy of Phosphorous slag was tested through topical application for 240 ± 10 minutes. The test substance was applied as a 20% (w/v) suspension (750 ul) directly on top of the corneas.

The negative control responses of the opacity and permeability values were less than the upper limits of the laboratory historical range, indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (20% w/v Imidazole) was 98 and was within the historical positive control data range.

Phosphorous slag did not induce ocular irritation through both endpoints, opacity and permeability, resulting in a mean in vitro irritancy score of 0 after 240 minutes of treatment.

Finally, it is concluded that this test is valid and that Phosphorous slag is a non irritant in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.