Chromium (III) complexes with 2-[{2,4-dihydroxy-3-[(5-sulfo-1-naphthyl)diazenyl]phenyl}diazenyl]benzoic acid (1:2), sodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 2015

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Chromium (III) complexes with 2-[{2,4-dihydroxy-3-[(5-sulfo-1-naphthyl)diazenyl]phenyl}diazenyl]benzoic acid (1:2), sodium salts

EC Number:

700-953-9

Molecular formula:

C46H26CrN8O14S2.3Na

IUPAC Name:

Chromium (III) complexes with 2-[{2,4-dihydroxy-3-[(5-sulfo-1-naphthyl)diazenyl]phenyl}diazenyl]benzoic acid (1:2), sodium salts

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals / tissue source

Species:

other: Bovine eye

Strain:

other: Bos primigenius Taurus (fresh bovine corneas)

Details on test animals or tissues and environmental conditions:

Fresh bovine eyes were obtained from the slaughterhouse Müller Fleisch GmbH, Enzstr. 2-4, 75217 Birkenfeld, Germany, on the day of the test. The cattle were between 12 and 60 months old. The eyes were transported to the test facility in Hank’s balanced salt solu-tion (supplemented with 0.01% streptomycin and 0.01% penicillin). Then, the corneas were dissected and incubated in medium at 32 ± 1 °C in an incubation chamber for 1 hour.

Test system

Vehicle:

other: MEM

Amount / concentration applied:

The following amounts of the test item were tested neat and applied directly on the cornea using a weight board:
98.6-102.3-104.4 mg

Duration of treatment / exposure:

The test item was given on the epithelium in such a manner that as much as possible of the cornea was covered with test item.
Exposition time on the corneas was 4 h. at 32 ± 1°C. After thorough rinsing with cMEM with phenol red and final rinsing with cMEM without phenol red, both chambers were filled with cMEM without phenol red, and the final opacity value of each cornea was rec-orded at once (again by measurement at 570 nm). The cMEM without phenol red was then removed from the front chamber, and 1 mL sodium fluorescein solution (concentra-tion: 5 mg/mL) was added to the front chamber.
The chambers were then closed again and incubated for 90 min at 32 ± 1 °C. After incu-bation, the content of the posterior chamber was thoroughly mixed. Then, the permeability of the cornea was measured as optical density of the liquid at 490 nm.

Details on study design:

7.2 Experimental Parameters
Date of treatment 14. Apr. 2015
Incubation time 4 hours
Negative control 0.9 % sodium chloride solution
Positive control imidazole, 20 % solution in 0.9 % sodium chloride solu-tion

Results and discussion

In vivo

Results

Irritation parameter:

other: IVIS

Basis:

mean

Time point:

other: 4h

Score:

ca. 38.5

Max. score:

51.9

Reversibility:

other: Not applicable

Irritant / corrosive response data:

According to the guideline, the test is considered as valid if the positive control causes an IVIS that falls within two standard deviations of the current historical mean.
The negative control has to show an IVIS ≤ 3.
The validity criteria and findings are given in the following table:
Table 9.3 a Validity
Parameter Criterion Found Assessment
IVIS of negative control 0.9% NaCl ≤ 3 0.7 ok
IVIS of positive control
20% imidazole solution 35.1 – 134.7 63.0 ok

Values for negative and positive controls were within the range of historical data of the test facility (see chapter 14, page 21). Therefore, the test system was acceptable.
9.4 Assessment
According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS > 3 and ≤ 55 induces effects on the cornea, that cannot be classified in an UN GHS Category.
Table 9.4 a Classification Scheme
IVIS UN GHS Category Eye Damage
≤ 3 No category
> 3; ≤ 55 No prediction can be made
> 55 Eye damage Category I

In the negative control, no signs of eye irritation were observed.
Two replicates of the positive control induced serious eye damage, which would be clas-sified as GHS category 1. The third replicate showed effects on the cornea of the bovine eye.
The test item Acid Brown 432 showed effects on the cornea of the bovine eye. The calcu-lated IVIS (in vitro irritancy score) is 38.5 and cannot be classified in an UN GHS Catego-ry.
The experiment is considered as sufficient for the classification of the test item, because all three replicates of the test item lead to the same assessment for the test item.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

A BCOP in vitro study has been performed to evaluate the risk of severe eye lesion. According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS > 3 and ≤ 55 induces effects on the cornea, that cannot be classified in an UN GHS Category. (IVIS: 38.5)

Executive summary:

This in vitro study was performed to assess corneal damage potential of Acid Brown 432 by quantitative measurements of changes in opacity and permeability in a bovine cornea.

The test item Acid Brown 432was brought onto the cornea of a bovine eye which previously had been incubated with cMEM without phenol red at 32 ± 1 °C for one hour and whose opacity had been determined. The test item was incubated on the cornea for 4 hoursat32 ± 1 °C. After removal of the test item, opacity and permeability values were measured.

 

The test item was tested pure. Under the conditions of this test, the test item Acid Brown 432 showed effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 38.5.

According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS > 3 and ≤ 55 induces effects on the cornea, that cannot be classified in an UN GHS Category for eye damage.

 

The negative control (physiological sodium chloride solution) and the positive control (20% imidazole solution) have met the validity criteria. No observations were made which might cause doubts concerning the validity of the study outcome. The study is considered valid.