Diphosphoric acid, zinc salt, compd. with 1,3,5-triazine-2,4,6-triamine (1:1:2)

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:

other: Study conducted to OECD guidelines and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of 1,3,5-triazine-2,4,6-triamine and zinc bis(dihydrogen phosphate)

EC Number:

690-512-6

Cas Number:

1271172-98-5

Molecular formula:

Zn(C3H7N6)2(P2O7)

IUPAC Name:

Reaction products of 1,3,5-triazine-2,4,6-triamine and zinc bis(dihydrogen phosphate)

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals / tissue source

Species:

other: Bovine eyes

Details on test animals or tissues and environmental conditions:

Preparation of corneas:
Fresh bovine eyes were obtained from the slaughterhouse on the day of the test. They eyes were transported in Hank's balanced salt solution. The corneas were dissected and incubated with with media at 32 +/- 1 °C in an incubation chamber for 1 hour. The corneas were examined and only those free from defects were used. The corneas were excised with a scalpel and cut from the globe with a 2-3 mm ring of sclera around the outside.
After the corneal holders were cleaned and sterilised, they were kept in an incubation chamber at 32°C. On the day of the assay, the minimum essential medium (MEM) without Phenol red was supplemented with sodium bicarbonate, L-glutamine and 1% fetal calf serum and stored in a water bath at 32°C. The same was performed for the MEM with phenol red. Each cornea was transferred to a corneal holder in which pre-warmed cMEM without phenol red was filled. The holders were then incubated for 1 hour at 32°C.

Test system

Vehicle:

unchanged (no vehicle)

Remarks:

tested neat on the corneal surfaces

Controls:

yes

Amount / concentration applied:

On average, 231.4 mg of the test substance was applied directly on the corneal surface (each cornea) using a weight board. The test item was given on the epithelium in such a manner that as much as possible of the cornea was covered with the test item.

Duration of treatment / exposure:

4 hours (+/- 5 minutes) at 32°C.

Number of animals or in vitro replicates:

Three eyes for the test substance, negative controls and positive control substances.

Details on study design:

After the initial incubation, the medium was changed and the baseline opacity was measured. This was performed by placing the holder with the cornea in a spectrophotometer and recording the absorption at 570 nm.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Remarks:

mean value

Value:

0.016

Negative controls validity:

valid

Positive controls validity:

valid

In vivo

Irritant / corrosive response data:

The individual in vitro irritancy scores for the negative controls were 0.2703 to 0.9344. The individual positive control in vitro irritancy scores ranged from 67.8 to 83.6.
The corneas treated with the test substance showed opacity values of 0.38 to 1.2 and permeability values ranging from 0.0035 to 0.02. Hence, the in vitro irritancy scores ranged from -0.2 to 0.7 after 4 hours of treatment (mean 0.0160).

Other effects:

The positive control substance induced an IVIS score of 75.9 (showing severe eye irritancy) and indicating the test was valid.

Any other information on results incl. tables

IVIS values:

Test Group	IVIS	Mean IVIS
Negative control (0.9% saline)	0.9344	0.5930
	0.2703
	0.5748
Test substance	-0.3112	0.0160
	-0.2993
	0.6598
Positive control (20% imidazole)	67.7997	75.8590
	76.1348
	83.6427

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Since the test substance induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.