Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
impurity

Test animals / tissue source

Species:
cattle

Test system

Vehicle:
water
Controls:
yes, concurrent vehicle
yes, concurrent positive control
Amount / concentration applied:
Application of volume of 750 µL of 20% w/v test item (in water)
Duration of treatment / exposure:
3 hours and 55 minutes
Duration of post- treatment incubation (in vitro):
Post incubation of 85 minutes
Number of animals or in vitro replicates:
three in vitro replicates
Details on study design:
A volume of 750 µL of 20% w/v test item,vehicle (distilled water) and positive control (20% w/v imidazole)was introduced into anterior chamber in triplicates to the designated cornea holders and incubated at 32±1 ºC for 3 hours and 55 minutes. Treated corneas were washed till no visual evidence of test item observed with EMEM containing phenol red and finally with EMEM without phenol red. The anterior chamber was then refilled with fresh EMEM without phenol red. Opacity was measured with the aid of opacitometer and permeability was determined spectrophotometrically at 490 nm (OD490) using 5 mg/mL sodium fluorescein, post incubation of 85 minutes at 32±1 ºC.

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Value:
38.6
Vehicle controls validity:
valid
Remarks:
The vehicle control and test item resulted in mean change in opacity value after treatment of 2.584 and 18.241 respectively. The mean corrected opacity and mean corrected permeability values of test item are 15.66 and 1.532 respectively.
Positive controls validity:
valid
Remarks:
Positive control resulted in mean change in opacity of 86.93. The mean corrected opacity and mean corrected permeability values of positive control are 84.33 and 1.549 respectively with an in vitro Irritancy Score (IVIS) of 107.6.

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Conclusions:
Based on the results obtained in the Bovine Corneal Opacity and Permeability Test, the test item Cocoyl proline CAS: 960241-19-4 induced an IVIS of 38.6 after 3 hours and 55 minutes of treatment. As the test item induced an IVIS > 3 and ≤ 55, the results are equivocal and no prediction can be made regarding the corrosivity or severe irritancy to Bovine corneas.