Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
available as unpublished report, no restrictions, fully adequate for assessment

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)
GLP compliance:
yes (incl. certificate)
Remarks:
TNO Triskelion, Utrechtseweg 48, 3700 AV, Zeist

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Celitement
- Chemical name: Calcium hydrosilicate
- Physical state: White powder
- pH-value: 10.5
- Expiration date of the lot/batch: 12/2015
- Stability under test conditions: Hydration reaction with water
- Storage condition of test material: Room temperature protected from light, in original container in evacuated desiccator

Test animals / tissue source

Species:
other: eyes of male or female chickens (ROSS, spring chickens)
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: slaughterhouse, v.d. Bor, Nijkerkerveen, The Netherlands
- Age at study initiation: approximately 7 weeks
- Weight at study initiation: approximately 1.5 - 2.5 kg
- Heads of the animals were cut off immediately after sedation of the animals by electric shock and incision of the neck for bleeding, and before they reached the next station on the process line. The heads were placed in small plastic boxes on a bedding of paper tissues moistened with isotonic saline. Next they were transported to the testing facility. During transportation, the heads were kept at ambient temperature.
- The preparation and validation of the eyes prior to the ICE-test were all according to OECD guideline 438.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 30 mg

Duration of treatment / exposure:
10 seconds
Observation period (in vivo):
0, 30, 75, 120, 180, and 240 minutes
Number of animals or in vitro replicates:
3 eyes
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: The eyes were rinsed with 20 mL saline
- Time after start of exposure: 10 seconds

SCORING SYSTEM: According to OECD 438 guideline.

TOOL USED TO ASSESS SCORE: All examinations were carried out with the hand-slit lamp microscope. Fluorescein retention was only scored at approximately 30 minutes after treatment.

CONTROLS: A negative control (30 µL physiological saline) and 3 positive controls (30 mg Imidazole) were included.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Remarks:
swelling (%)
Basis:
mean
Time point:
other: 240 min
Score:
7
Remarks on result:
other: Maximum value during the test
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
mean
Time point:
other: 120, 180, and 240 min
Score:
1.8
Max. score:
4
Remarks on result:
other: Maximum value during the test
Irritation parameter:
cornea opacity score
Remarks:
fluorescein retention
Basis:
mean
Time point:
other: 30 min
Score:
2.2
Max. score:
3
Irritant / corrosive response data:
Slit-lamp examination: After rinsing of the cornea, remnants of the test substance were present on the cornea and could not be removed. The test substance caused slight corneal swelling (7%), slight to moderate or moderate corneal opactiy (1.8) and moderate or moderate to severe fluorescein retention (2.2). In addition, spots with erosion of epithelium was observed in one eye. The negative control eye did not show any corneal effect and demonstrated that the general conditions during the test were adequate. The positive control eyes showed severe corneal effects and demonstrated the ICE test valid to detect severe eye irritants.

MIcroscopic examination: Microscopic examination of the corneas treated with the test substance revealed very slight erosion of the epithelium. Microscopic examination of the cornea treated with the negative control (saline) did not reveal any abnormalities. The positive control Imidazole caused severe erosion and necrosis of the epithelium.

Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the test conditions (OECD 438 and GLP) the test substance is considered to be an eye irritant.
Executive summary:

In accordance to OECD guideline 438 and GLP the test substance was examined for its in vitro eye irritating potential using the Isolated Chicken Eye (ICE) Test. In the ICE test, 3 eyes were exposed to 30 mg test substance for 10 seconds. In addition, one negative control eye (30 µL saline) and three positive control eyes (30 mg imidazole) were tested. After the exposure the eyes were rinsed with 20 mL saline and were examined at approximately 0, 30, 75, 120, 180, and 240 minutes after treatment. The test substance caused slight corneal swelling (7%), slight to moderate or moderate corneal opactiy (1.8) and moderate or moderate to severe fluorescein retention (2.2). In addition, spots with erosion of epithelium was observed in one eye. The negative control eye did not show any corneal effect while the positive control eyes showed severe corneal effects. Microscopic examination of the corneas treated with the test substance revealed very slight erosion of the epithelium. Microscopic examination of the cornea treated with the negative control did not reveal any abnormalities. The positive control caused severe erosion and necrosis of the epithelium. Based on the results obtained in the present study, the test substance is considered to be irritating to eyes. Based on GHS and CLP, the test substance can be classified as category 2A and category 2 substance, respectively.