Hexanedioic acid, polymer with 2,2'-oxybis[ethanol]

Eye irritation

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:

other: Study conducted according to scientifically valid methods and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

Assessment of ocular irritation by a single topical application of the test substance to a reconstructed 3D human cornea model (EpiOcularTm eye irritation test).

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol]

EC Number:

618-460-1

Cas Number:

9010-89-3

Molecular formula:

(C6 H10 O4 . C4 H10 O3)x

IUPAC Name:

Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol]

Test material form:

other: liquid

Details on test material:

ca. 99% (w/w) Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol] / Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol] / 9010-89-3

IMPURITIES
ca. 2% (w/w) 2,3,7,8-tetrahydro-2,5,8-benzotrioxacycloundecine-1,9-dione / 1,4,7-Trioxacyclotridecane-8,13-dione / 1,4,7-Trioxacyclotridecane-8,13-dione / 6607-34-7

ca. 10 ppm tin bis(2-ethylhexanoate) / Bis(2-ethylhexanoyloxy)tin / Hexanoic acid, 2-ethyl, tin(2+) salt (2:1) / 301-10-0

ca. 10 ppm titanium tetrabutanolate / titanium tetrabutanolate / tetrabutyl titanate / 5593-70-4

Test animals / tissue source

Species:

other: 3D human cornea model

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

50 ul of undiluted test substance

Duration of treatment / exposure:

30 minutes

Observation period (in vivo):

2 hours post-incubation

Details on study design:

The EpiOcularTM assay is designed to predict an eye irritation potential of a chemical based on the observation that irritant chemicals produce cytotoxicity in human reconstructed cornea after a short-term topical exposure. Cytotoxicity is expressed as a reductionof the activity of mitochondrial dehydrogenase, which reduces the yellow water-soluble 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolim bromide (MTT) to the insoluble blue formazan in the test medium. Thus,the loss of viability of the tissues is measured by a colorimetric assay after administration of the substance.
For the EpiOcularTM assay, the OCL-200 model supplied by MatTEK Corp. (Ashland MA, USA) was used. Methyl acetate was used as a positive control.
Two tissues were treated with the test compound, the positive control and negative control, respecitvely. Tissues were incubated in sterile 6-well plates with 1 ml assay medium at standard culture conditions for 16-24 hours. After that, tissues were treated with 20 ul of PBS at standard culture conditions for 30 minutes to wet the tissue surface. 50 ul of the undiluted test substance were then applied covering the whole tissue surface, . Control tissues were concurrently treated with 50 ul of de-ionized water (negative control) or methyl acetate (positive control). After that, the tissues were incubated until the total exposure time of 30 minutes was completed. To remove the test substance, the tissues were washed with PBS and transferred to pre-warmed medium for 12 minutes to remove residual test substance. The tissues were then dried, transferred to 6-well-plates filled with pre-warmed medium and incubated at standard culture conditions for 2 hours of postincubation. After that, the assay medium was replaced by 0.3 ml MTT solution (1 mg/ml in assay medium) and the tissues were placed in the incubator for 3 hours. After incubation, tissues were washed with PBS and subsequently incubated in isopropanol at room temperature overnight or for at least 2 hours on a plate shaker to extract the metabolically produced formazan. The OD570 of the extracts was then determined spectrophotometrically.

Results and discussion

In vivo

Results

Irritation parameter:

other: tissue viability, presented as the quotient of the mean OD570 divided by the respective OD570 NC value in %.

Score:

> 60

Reversibility:

not specified

Remarks on result:

other: A substance is considered to be non-irritant in the EpiOcular test if the mean tissue viability is >60% of the negative control.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: The irritation potential is predicted from the mean relative tissue viabilities as compared to the negative control tissues according to the following criteria: <=50% --> irritant; >50 <=60% --> no prediction; >60% --> non-irritant.

Conclusions:

Under the test conditions chosen, Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol] does not show an eye irritation potential in the EpiOcular eye irritation test.

Executive summary:

Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol] was tested for eye irritation potential in the EpiOcularTM assay. 50 ul of the undiluted test substance were applied to a reconstructed 3D human cornea model (two tissue samples per treatment group). Negative (de-ionized water) and positive controls (methyl acetate) were run concurrently. The tissues were incubated with the test or control substances for 30 minutes, followed by a 2 hour post-incubation period. Formazan production after incubation with MTT was measured as indicator for cytotoxicity. The ratio of formazan production in the test substance-treated tissues and that in the negative control tissues indicates relative tissue viability. The test substance was not able to reduce MTT, and the mean viability of the test substance-treated tissues was 102%. Based on these results, it was concluded that the test substance does not show an eye irritation potential in the EpiOcularTM test under the test conditions chosen.