Aluminium tris(dihydrogen phosphate)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was performed on 29 May 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study conducted to GLP and in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do no effect the quality of the relevant results. This study is conducted according to an appropriate guideline and under the conditions of GLP. In accordance with Regulation (EC) No. 1272/2008 and as detailed in the Guidance on the application of the CLP criteria, a positive BCOP assay is sufficient for the purposes of classification and labelling and as such the study is considered to be acceptable and to adequately satisfy both the guideline requirement and the regulatory requirement as a key study for this endpoint.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Version / remarks:

Study performed in 2010

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of inspection: 15-09-2009 Date of Signature: 26-11-2009

Test material

Test material information

Constituent 1

Reference substance name:

Aluminium tris(dihydrogen phosphate)

EC Number:

236-875-2

EC Name:

Aluminium tris(dihydrogen phosphate)

Cas Number:

13530-50-2

Molecular formula:

H6AlO12P3

IUPAC Name:

aluminium(3+) tridihydrogen phosphate

Test animals / tissue source

Species:

other: Excised Bovine Cornea

Strain:

other: Not Applicable

Details on test animals or tissues and environmental conditions:

Not applicable

Test system

Vehicle:

physiological saline

Controls:

no

Amount / concentration applied:

TEST MATERIAL

-Amounts(s) applied (volume or weight with unit):
Triplicate corneas were treated with 0.75 ml of the test material.

-Concentration (if solution):
The test material was used as a 20% dilution.

VEHICLE
0.9% w/v sodium chloride solution.

Duration of treatment / exposure:

240 Minutes.

Observation period (in vivo):

Not Applicable.

Number of animals or in vitro replicates:

Not Applicable.

Details on study design:

TEST SITE:
-Area of exposure:
Triplicate corneas were treated with 0.75 ml of the test material.

-% coverage:
The test material preparation was topically applied to the cornea. The corneal holders were gently tilted back and forth to ensure a uniform application of the material over the entire cornea.

-Type of wrap used:
None used.

REMOVAL OF TEST SUBSTANCE:
-Washing (if done):
At the end of the exposure period the test material preparation was removed from the anterior chamber and the cornea was rinsed three times with fresh complete MEM containing phenol red before a final rinse with complete MEM.

-Time after start of exposure:
240 Minutes post exposure.

SCORING SYSTEM:
Results from the two test method endpoints, opacity and permeability, were combined in an empirically derived formula to generate an In Vitro Irritancy Score.
Opacity Measurement: The change in opacity for each cornea (including the negative control) was calculated by subtracting the initial opacity reading from the final opacity reading. These values were then corrected by subtracting from each the average change in opacity observed for the negative control corneas. The mean opacity value of each treatment group was then calculated by averaging the corrected opacity values of each cornea for that treatment group.
Permeability Measurement: The corrected OD492 was calculated by subtracting the mean OD492 of the negative control corneas from the OD492 value of each treated cornea. The OD492 value of each treatment group was calculated by averaging the corrected OD492 values of the treated corneas for the treatment group.

The following formula was used to determine the in vitro score:
In Vitro Irritancy Score = mean opacity value + (15 x mean OD492 value)

Data Interpretation:
A test material that induces an in vitro irritancy score >55.1 is defined as an ocular corrosive or severe irritant.

Additionally, the opacity and permeability values were evaluated independently to determine whether the test material induced a response through only one of the two endpoints.

Visual Observation: The condition of the cornea was visually assessed immediately after rinsing and at the final opacity measurement.

Results and discussion

In vivo

Results

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

mean viability of opacity & Permeability (%)

Time point:

other: 240 Minutes post exposure

Score:

86.7

Max. score:

100

Reversibility:

other: not investigated

Remarks on result:

other: See below

Irritant / corrosive response data:

This BCOP is a short term assay which does not assess reversibility of reactions.

Other effects:

Not applicable.

Any other information on results incl. tables

RESULTS

Corneal Opacity and Permeability Measurement

Individual and mean corneal opacity measurements and individual and mean corneal permeability measurements are given in Table 1. 

Corneal Epithelium Condition

The condition of the cornea immediately after rinsing is given in Table 2.

The corneas treated with the test material or positive control material were cloudy post treatment. The corneas treated with the negative control material were clear post treatment.

In Vitro Irritancy Score

The results are summarised as follows:

Treatment	In VitroIrritancy Score
Test Material	86.7
Negative Control	5.2
Positive Control	115.4

Criteria for an Acceptable Test

The positive contro lIn Vitro irritancy Score was within the range of 55.8 to 126.1. The positive control acceptance criterion was therefore satisfied.

Table 1          Individual and Mean Corneal Opacity and Permeability Measurements

Treatment	Cornea Number	Opacity				Permeability (OD)		In vitroIrritancy Score
		Pre-Treatment	Post-Treatment	Post-Treatment-Pre‑Treatment	Corrected Value		Corrected Value
Negative Control	1	4	8	4		0.083
	2	3	7	4		0.065
	3	3	7	4		0.090
				4.0*		0.079^		5.2
Positive Control	4	4	96	92	88.0	2.715	2.636
	5	3	69	66	62.0	3.175	3.096
	6	3	69	66	62.0	3.295	3.216
					70.7·		2.982·	115.4
Test Material	7	3	102	99	95.0	0.396	0.317
	8	4	88	84	80.0	0.346	0.267
	9	1	77	76	72.0	0.364	0.285
					82.3·		0.289·	86.7

 

OD= Optical density                 * = Mean of the post treatment-pre‑treatment values      ^= Mean permeability             ·= Mean corrected value

Table 2          Corneal Epithelium Condition

Treatment	Cornea Number	Observation
		Immediately after Rinsing
Negative Control	1	clear
	2	clear
	3	clear
Positive Control	4	cloudy
	5	cloudy
	6	cloudy
Test Material	7	cloudy
	8	cloudy
	9	cloudy

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

The test material was considered to be an ocular corrosive or severe irritant.

Study conducted to GLP and in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do no effect the quality of the relevant results.
This study is conducted according to an appropriate guideline and under the conditions of GLP. In accordance with Regulation (EC) No. 1272/2008 and as detailed in the Guidance on the application of the CLP criteria, a positive BCOP assay is sufficient for the purposes of classification and labelling and as such the study is considered to be acceptable and to adequately satisfy both the guideline requirement and the regulatory requirement as a key study for this endpoint.

Executive summary:

Introduction. A study was performed to assess the ocular irritancy potential of the test material to the isolated bovine cornea.

Method. The test material was applied at a concentration of 20% w/v for 240 minutes. Negative and positive control materials were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS). 

Results. The in vitro Irritancy scores are summarised as follows:

Treatment	In Vitro Irritancy Score
Test Material	86.7
Negative Control	5.2
Positive Control	115.4

Conclusion.  The test material was considered to be an ocular corrosive or severe irritant.