Registration Dossier

Diss Factsheets

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:
2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and OECD testing guideline compliant study with well-characterized material

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report date:
2011

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: OECD Guideline for Testing of Chemieals No. 437, September 07,2009 (BCOP)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction products of diazotized 2-amino-4-(methylsulfonyl)-phenol with 2-(3-Chlorophenyl)-2,4-dihydro-5-methyl-3H-pyrazol-3-one and Methyl-7-hydroxy-1-naphthylcarbamate – and chromium (III) 2:1, salted by ammonium, sodium and acid
EC Number:
936-609-1
Molecular formula:
C40H32CrN4O12S2-R+] . [C36H28ClCrN7O10S2-R+] . [C34H26Cl2CrN8O8S2-R+]; R+ = NH4+, Na+ or H+
IUPAC Name:
Reaction products of diazotized 2-amino-4-(methylsulfonyl)-phenol with 2-(3-Chlorophenyl)-2,4-dihydro-5-methyl-3H-pyrazol-3-one and Methyl-7-hydroxy-1-naphthylcarbamate – and chromium (III) 2:1, salted by ammonium, sodium and acid
Details on test material:
Expiry date: 02 September 2012
purity: > 80% (see certificate of analysis)
Physical state: solid

Test animals / tissue source

Species:
other: Bovine
Strain:
not specified
Details on test animals or tissues and environmental conditions:
The test system (target tissue) is the isolated bovine cornea. Bovine eyes are obtained as a by-product of freshly
slaughtered cattle (age of the animals: minimum 12 months, maximum 60 months).

Test system

Vehicle:
water
Controls:
yes, concurrent vehicle
Amount / concentration applied:
20% (w/v) suspension
Duration of treatment / exposure:
4 hours
Observation period (in vivo):
none
Number of animals or in vitro replicates:
6 corneas per group
Details on study design:
Corneas free of defects (opacity, scratches, pigmentation etc.) are dissected with a 2 to 3 mm rim of sclera. Isolated corneas are mounted in specially designed cornea I holders that consists of anterior and posterior chambers. 80th chambers are filled to excess with prewarmed Eagles's MEM (without phenol red) and then equilibrated in a ver1ical position at about 32°C far at least 1 hour. After the equilibration period the medium in both chambers is replaced with fresh pre-warmed medium and initial cornea I opacity readings are taken for each cornea with an opacitometer.

Before application the medium in the anterior chamber is removed using a syringe. 0.750 mL of the 20% (w/v) test-substance preparation is applied directly to the epithelial surface of the cornea using a pipette (open chamber method). Control tissues are concurrently applied into the anterior chamber with 0.750 mL of highly deionized water (negative control, NC) or with 0.750 mL of 20% (w/v) solution of Imidazole in
highly de-ionized water (positive control, PC) using a pipette. The corneas will be incubated in a horizontal position at about 32°C for approximately 4 hours. The test substance, NC and PC is then be removed from the anterior chamber using a syringe and the epithelium is washed at least 3 times with Eagle's MEM (containing phenol red) and once with Eagle's MEM (without phenol red). Both chambers are then refilled with fresh Eagle's MEM (without phenol red). Before measurement, each cornea is observed visually and observations are recorded.
Final corneal opacity readings are taken for each cornea with an opacitometer.

For determination of permeability the medium in the anterior chamber is replaced by 1 mL sodium fluorescein solution (5 mg/mL) and incubated for 90 ± 5 min in a horizontal position at about 32 °C.
The amount of sodium fluorescein that permeates through the corneas into the posterior chamber is measured spectrophotometrically. Three aliquots per cornea are transferred 10 a 96-well microtiter plale and the optical density (OD 490 nm) is determined.

Results and discussion

In vivo

Results
Irritation parameter:
other: IVIS
Basis:
mean
Time point:
other: 4h
Score:
33
Reversibility:
other: not relevant for BCOP assay
Remarks on result:
other: not "highly irritating". Two of six corneas were not scorable because of strong staining by the test item.

Applicant's summary and conclusion

Interpretation of results:
other: not higly irritating
Remarks:
Criteria used for interpretation of results: EU