[cyclohexane-1,2-diylbis[nitrilobis(methylene)]]tetrakisphosphonic acid, sodium salt

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

3 November 2016 - 22 November 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

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

Test material form:

liquid

Test animals / tissue source

Species:

other: in-vitro test

Strain:

other: isolated bovine cornea

Details on test animals or tissues and environmental conditions:

TEST MATERIAL
- Source: slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands)
- Acclimation period: Eyes were collected and transported in physiological saline in a suitable container under cooled conditions. Then, the corneas were dissected and incubated in incubation medium at 32 +/- 1°C in an incubation chamber for 1 hour.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 32 +/- 1°C

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

An amount of 750 µl of either the negative control, positive control or test item was applied directly on the corneas in such a way that the cornea was completely covered.

Duration of treatment / exposure:

The corneas were incubated in a horizontal position for 10 +/- 1 minutes at 32 +/- 1°C.

Duration of post- treatment incubation (in vitro):

120 +/- 10 minutes at 32+/-1°C.

Details on study design:

The aim of this study was to evaluate the ocular irritancy of the test item as measured by its ability to induce opacity and increase permeability in an isolated bovine cornea.

PROTOCOL
- For each treatment group (negative control, positive control and test item), three replicates were used.
- The opacity of each cornea was determined before exposure.
- The corneas were exposed for 10 minutes.
- After the incubation the change in opacity for each individual cornea (including the negative control) was determined.
- Following the final opacity measurement, permeability of the cornea to Na-fluorescein was evaluated.

INTERPRETATION
The assay is considered acceptable if:
- The positive control gives an in vitro irritancy score that falls within two standard deviations of the current historical mean.
- The negative control responses should result in opacity and permeability values that are less than the upper limits of the laboratory historical range.
- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro irritancy score (IVIS)
- A test substance that induces an IVIS > 55 is defined as a corrosive or severe irritant.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

-0.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

In vivo

Irritant / corrosive response data:

The negative control showed no irritation (mean IVIS was 0.443). The positive control induced a very severe irritation on the cornea, mean IVIS was 195.5. It was therefore concluded that the test conditions were adequate and that the test system functioned properly. The mean in vitro irritancy score of the test item was -0.8 after 10 minutes of treatment with the test item.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

ALBRITE CIX(N) is not severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test.

Executive summary:

A GLP compliant in-vitro study was performed according to OECD Guideline 437 in order to evaluate the ocular irritation properties of ALBRITE CIX(N) on an isolated bovine cornea. The possible ocular irritancy of the test item was tested through topical application for approximately 10 minutes. For each treatment group (negative control, positive control and test item), three replicates were used. After removal of the test item and two hours post-incubation, opacity and permeability values were measured. After treatment, the IVIS (in vitro irritancy score) in the controls is within the acceptability criterion. The positive control showed clear irritating effects. ALBRITE CIX(N) showed no effects on the cornea of the bovine eyes. The IVIS score is -0.8 and therefore it can be concluded that ALBRITE CIX(N) is not a severe eye irritant in the BCOP Test.