Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
25 November 2014 to 03 February 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to GLP and testing guidelines; coherence between data, results and conclusion.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
Qualifier:
according to
Guideline:
other: OECD series on testing and assesment no. 160
Qualifier:
according to
Guideline:
other: Use of an alternate testing framework for classification of eye irritation potential of EPA pesticide products - Office of Pesticide Programs, U.S. Environmental Protection Agency (31 May 2013)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Test item: Disperse Blue CVG


Test animals / tissue source

Strain:
not specified
Details on test animals or tissues and environmental conditions:
Test system: isolated corneas from the eyes of freshly slaughtered bovine.
Age of animals: 6-12 months

Transport solution: Hanks balanced salt solution (HBSS) Modified supplemented with Penicillin sulphate and Streptomycin sulphate.
Transport conditions: in transport solution at approximately 4°C.

Test system

Vehicle:
physiological saline
Controls:
yes
Amount / concentration applied:
The test item was spread on each epithelial surface of three idoneous bovine corneas as supplied (150 mg/cornea); 0.75 mL of physiological saline was then added on each cornea in order to better distribute the substance on the cornea surface (final concentration on cornea = 200 mg/mL, approximately 20% w/v)
Number of replicates: 3
Duration of treatment / exposure:
Corneas were exposed in horizontal position for 4 hours ± 10 minutes, incubated in a liquid bath at 32 ± 1°C.
There was no post-exposure period.
Observation period (in vivo):
exposure period: 4 hours --> determination of opacity
fluorescein staining: 90 minutes --> corneal permeability
Number of animals or in vitro replicates:
three idoneous bovine corneas
Details on study design:
Positive control item: corneas treated 20% (w/v) Imidazole (Sigma, batch no. 0001422794) in Physiological saline (0.9% NaCl) (Baxter, batch no. 13D0406).
Negative control item: corneas treated with Physiological saline (0.9% NaCl) (Baxter, batch no. 13D0406).
Volume of treatment: 0.75 mL
Number of replicates: 3

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: mean opacity score
Basis:
mean
Time point:
other: at the end of the test item exposure period
Score:
4
Remarks on result:
other: 4 h exposure time
Irritation parameter:
other: mean permeability OD490 score
Basis:
mean
Time point:
other: 90 min
Score:
0
Remarks on result:
other: staining with a 0.5% solution of sodium fluorescein in DPBS for 90 minutes
Irritation parameter:
other: In vitro irritancy score (IVIS)
Basis:
mean
Time point:
other: 4 h exposure + 90 min staining
Score:
4
Max. score:
55
Remarks on result:
other: In vitro irritancy score (IVIS) = mean opacity score + (15 x mean permeability OD490 score)

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Remarks:
Migrated information
Conclusions:
The test item Disperse Blue CVG cannot be clearly classified.
Executive summary:

The potential of the test item, Disperse Blue CVG, to cause corrosion/severe irritation by using the Bovine Corneal Opacity and Permeability (BCOP) assay, was examined in agreement with OECD Guideline no. 437 (adopted on 26 July 2013), the Guidance Document OECD series on testing and assessment no. 160 and the method described in the document issued by U.S. EPA/OPP (31 May 2013). The test item was spread on each epithelial surface of three idoneous bovine corneas as supplied (150 mg/cornea); 0.75 mL of physiological saline was then added on each cornea in order to better distribute the substance on the cornea surface (final concentration on cornea = 200 mg/mL, approximately 20% w/v); an exposure period of 4 hours was used. The mean opacity detected with an opacitometer at the end of the test item exposure period was 4.0. At macroscopic observation, no visible change of the treated corneas was noted. After the determination of opacity, the epithelial surface was treated with a 0.5% solution of sodium fluorescein in DPBS for 90 minutes, to investigate alteration in cornea permeability. The calculated mean permeability OD490 value of the corneas treated with the test item was 0.00. The calculated in vitro irritancy score (IVIS) for the test item was 4.0. 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. According to the OECD Guideline no. 437, the test item cannot be clearly classified.