Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
November 10th, 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)
Qualifier:
according to
Guideline:
EU method B.48 (Isolated chicken eye test method for identifying occular corrosives and severe irritants)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Specific details on test material used for the study:
Test material name (as stated in the report): Cetonal
Batch No.: SC00011028
Expiry date: 23 June, 2016

Test animals / tissue source

Species:
chicken
Strain:
not specified
Details on test animals or tissues and environmental conditions:
The eyes collected from Chickens obtained from a slaughterhouse where they are killed for human consumption have been used for this assay. The age and weight of the chickens used in this test method are that of spring chickens traditionally processed by a poultry slaughterhouse. (approximately 7 weeks old 1.5 - 2.5 Kg).
Heads have been removed immediately after sedation of the chickens by electric shock, and incision of the neck for bleeding . Because eyes were dissected in the laboratory , the intact heads were transported from the slaughterhouse at ambient temperature in polystyrene boxes humidified with towels moistened with physiological saline.

The eyelids are carefully excised, taking care not to damage the cornea. Then, the eye was further dissected from the skull taking care not to damage the cornea. The eyeball is pulled from the orbit by holding the nictitating membrane firmly with surgical forceps, and the eye muscles are cut with a bent, blunt-tipped scissor. When the eye is removed from the orbit, a visible portion of the optic nerve should be left attached. Once removed from the orbit, the eye is placed on an absorbent pad and the nictitating membrane and other connective tissue are cut away.

Test system

Controls:
yes, concurrent vehicle
yes, concurrent positive control
Amount / concentration applied:
30 microL of test item was applied as supplied to the cornea such taht the entire surface of the cornea is evenly covered with the test item.
Duration of treatment / exposure:
Test item was applied for 10 seconds and then rinsed from the eye with 20 mL of physiological saline at ambient temperature. As residual test item was observed on the cornea, four rinses of 10 mL of physiological saline were added.
Duration of post- treatment incubation (in vitro):
Treated corneas are evaluated pretreatment and starting at 30, 75, 120, 180 and 240 minutes after the post treatment rinse.
Number of animals or in vitro replicates:
3 enucleated chicken eyes
Details on study design:
Damages by the test substance were assessed by determination of corneal swilling, opacity, and fluorescein retention at 30, 75, 120, 180 and 240 minutes post-dose.

Results and discussion

Results of ex vivo / in vitro studyopen allclose all
Irritation parameter:
cornea opacity score
Run / experiment:
maximal mean score
Value:
1
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: corresponding to ICE class II
Irritation parameter:
fluorescein retention score
Run / experiment:
mean score
Value:
0.7
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: corresponding to ICE class II
Irritation parameter:
percent corneal swelling
Run / experiment:
maximal mean score
Value:
32
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: at 240 post dose, corresponding to ICE class III
Other effects / acceptance of results:
The combination of the three endpoints for the positive control, 5% Benzalconium chloride was 3 x IV. Therefore, the positive controle is classified as "Corrosive/Severe irritant" as expected.
The combination of the three endpoints for the negative control, physiological saline was 3 x I. Therefore, the positive controle is classified as "No category" as expected.

Any other information on results incl. tables

The ocular reactions observed in eyes treated with the test item were:

- maximal mean score of corneal opacity: 1.0 corresponding to ICE class II

- mean score of fluorescein retention: 0.7 corresponding to ICE class II

- maximal mean corneal swelling : + 32 at 240 minutes post dose corresponding to ICE class III

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The results obtained, under this experimental conditions enable to conclude that the test item Cetonal does not have to be classified in category 1 "irreversible effects on eye".
Executive summary:

This experience on Cetonal has been performed according to the OECD guideline No 438 adopted 26 July 2013.

The eyes collected from Chickens obtained from a slaughterhouse where they are killed for human consumption have been used for this assay. The age and weight of the chickens used in this test method are that of spring chickens traditionally processed by a poultry slaughterhouse. (approximately 7 weeks old 1.5 - 2.5 Kg). Heads have been removed immediately after sedation of the chickens by electric shock, and incision of the neck for bleeding . Because eyes were dissected in the laboratory , the intact heads were transported from the slaughterhouse at ambient temperature in polystyrene boxes humidified with towels moistened with physiological saline. The eyelids are carefully excised, taking care not to damage the cornea. Then, the eye was further dissected from the skull taking care not to damage the cornea. The eyeball is pulled from the orbit by holding the nictitating membrane firmly with surgical forceps, and the eye muscles are cut with a bent, blunt-tipped scissor. When the eye is removed from the orbit, a visible portion of the optic nerve should be left attached. Once removed from the orbit, the eye is placed on an absorbent pad and the nictitating membrane and other connective tissue are cut away. 30 microL of test item was applied as supplied to the cornea such taht the entire surface of the cornea is evenly covered with the test item. Test item was applied for 10 seconds and then rinsed from the eye with 20 mL of physiological saline at ambient temperature. As residual test item was observed on the cornea, four rinses of 10 mL of physiological saline were added. Damages by the test substance were assessed by determination of corneal swilling, opacity, and fluorescein retention at 30, 75, 120, 180 and 240 minutes post-dose. The ocular reactions observed in eyes treated with the test item were: - maximal mean score of corneal opacity: 1.0 corresponding to ICE class II - mean score of fluorescein retention: 0.7 corresponding to ICE class II - maximal mean corneal swelling : + 32 at 240 minutes post dose corresponding to ICE class III The combination of the three endpoints for the positive control, 5% Benzalconium chloride was 3 x IV. Therefore, the positive controle is classified as "Corrosive/Severe irritant" as expected. The combination of the three endpoints for the negative control, physiological saline was 3 x I. Therefore, the positive controle is classified as "No category" as expected. The results obtained, under this experimental conditions enable to conclude that the test item Cetonal does not have to be classified in category 1 "irreversible effects on eye" according to the CLP Regulation (EC) No. 1272/2008.