Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 5th, 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test conducted according to internationally accepted Guideline and in according to the GLP Principles

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Principles of method if other than guideline:
This study design is in agreement with the procedures described in the OECD series on testing and assessment no. 160 – Guidance Document on the bovine corneal opacity and permeability (BCOP) and isolated chicken eye (ICE) test methods: collection of tissues for histological evaluation and collection of data on non-severe irritants.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder

Test system

Controls:
yes
Amount / concentration applied:
A suspension of the test item at 20 % w/v was prepared in physiological saline, with correction for the displacement due to the volume of the test item. Concentrations were expressed in terms of material as received. The suspension was sonicated and maintained in magnetic agitation until aliquots preparation. Three syringes were filled in with the suspension and maintained at room temperature until use.
Duration of treatment / exposure:
4-hour exposure period
Observation period (in vivo):
There was no post-exposure period.
Number of animals or in vitro replicates:
n.a.
Details on study design:
REPLICANTS: three corneas per Test Item, Negative and Positive control respectively

CONTROLS: positive and negative control
Positive control: 20 % (w/v) Imidazole (Sigma, batch no. 0001422794), in physiological saline (0.9 % NaCl) (Baxter, batch no. 13D0406).
Negative control: Physiological saline (0.9 % NaCl) (Baxter, batch no. 13D0406).

REMOVAL OF TEST SUBSTANCE
- Washing and time after start of exposure:
After exposure, corneas were rinsed thoroughly with complete EMEM with phenol red. For the test item treated group, additional washings were performed and the glass windows of the anterior chambers were removed to allow a more effective removal of the test item from the corneas. A final wash with prewarmed complete EMEM without phenol red was carried out. Finally, the anterior chamber was re-filled with prewarmed complete EMEM without phenol red.

SCORING SYSTEM: in vitro irritacy score (IVIS)

TOOL USED TO ASSESS SCORE: opacitometer

Results and discussion

In vivo

Results
Irritation parameter:
other: IVIS (in vitroirritation score)
Basis:
mean
Time point:
other: 4 hours
Score:
ca. 0
Reversibility:
other: No effects observed under test condition
Remarks on result:
other: in vitro test
Irritant / corrosive response data:
No relevant increases in opacity or permeability values were recorded after treatment with the test item, when compared to those of negative control.
The calculated in vitro irritancy score (IVIS ) was 0.0.

Negative and positive controls gave the expected results. The test is therefore considered as valid.

Any other information on results incl. tables

Assessment of ocular corrosion/irritation bovine corneal opacity/permeability (BCOP) assay.

Treatment code Pre-test corna number Basal opacity Mean group basal opacity
N1 10 0 0.00
N2 6 0
N3 9 0
P1 2 1 0.00
P2 5 0
P3 7 0
A1 1 1 0.00
A2 4 0
A3 8 0

Mean of basal opacity: 0.00; the mean refers to the total number of selected pre-test corn

The IVIS cut-off values for identifying test chemicals as inducing serious eye damage (UN GHS Category 1) and test chemicals not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are:

- IVIS ≤ 3 –No Classification(UN GHS)

- IVIS > 3; ≤ 55 – No prediction can be made

- IVIS > 55 –Category 1(UN GHS)

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Calculated in vitro irritancy score (IVIS ) = 0.0.
No relevant increases in opacity or permeability values were recorded after treatment with the test item, when compared to those of negative control.
Executive summary:

Method

The potential of the test item to cause corrosion/severe irritation by using the Bovine Corneal Opacity and Permeability (BCOP) assay was examined, in agreement with OECD Guideline no. 437 and the Guidance Document OECD series on testing and assessment no. 160.

The test item, a non surfactant powder, was dissolved at 20 % (w/v) in physiological saline and tested on the epithelial surface of three idoneous bovine corneas, for an exposure period of 4 hours.

Observations

The opacity of two of the treated corneas, detected with an opacimeter at the end of the test item exposure period, was 0.0; residual test item was observed on the surface of the third replicate thus the opacity value obtained (18) was not considered reliable. This was confirmed by macroscopic observation, where no alterations of corneas surfaces were noted. After the determination of opacity, the epithelial surface was treated with a 0.5 % solution of sodium fluorescein in DPBS for approximately 90 minutes to investigate alteration in cornea permeability. The mean value of permeability (OD490) of the corneas treated with the test item was 0.0022. The calculated in vitro irritancy score (IVIS) of the test item was 0.0.

Positive and negative controls [a 20 % (w/v) Imidazole solution in physiological saline and physiological saline alone, respectively] were concurrently tested in similar conditions and gave the expected results.

Result

According to the OECD Guideline no. 437, the test item can be classified as Not irritant.