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Toxicological information

Eye irritation

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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: Modern guideline study conducted according to GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
There are no official national or international guidelines for the EpiOcularTM test yet; however, the study was performed according to the methods described in the following publications:

MatTek Corporation, Ashland, MA 01721, USA: EpiOcularTM human cell construct: Procedure details, Version 3.1a of February 10, 2010.

Harbell J.W. et al. (2009): COLIPA Program on Optimization of Existing In Vitro Eye Irritation Assays for Entry into Formal Validation: Technology

Transfer and Intra/Inter Laboratory Evaluation of EpiOcular Assay for Chemicals. Poster # 378, Society of Toxicology, March 2009.

In addition the study follows the testing strategy for determination of eye irritation/corrosion as given in the following OECD guideline:
OECD Guideline for Testing of Chemicals No. 405, April 24, 2002 (“Acute Eye Irritation/Corrosion”)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
2-morpholinoethanol
EC Number:
210-734-5
EC Name:
2-morpholinoethanol
Cas Number:
622-40-2
Molecular formula:
C6H13NO2
IUPAC Name:
2-(morpholin-4-yl)ethan-1-ol
Test material form:
other: liquid
Details on test material:
Name of test substance:
2-Morpholinoethanol

Test-substance No.:
11/0386-1

Batch identification:
B 1262 vom 21.09.2011

CAS No.:
622-40-2

Purity:
99.7 area-% (for details see report No.: 11L00465)

Homogeneity:
The test substance was homogeneous by visual inspection.

Storage stability:
The stability under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.

Test animals / tissue source

Species:
other: non-keratinized tissue construct composed of normal human derived epidermal keratinozytes

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: negative and positive control
Duration of treatment / exposure:
undiluted liquid test substance was applied covering the whole tissue surface
Observation period (in vivo):
after application, the tissues were placed into the incubator until the total exposure time of 30 minutes was completed

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: optical density [wavelength 570 nm]
Basis:
mean
Time point:
other: after 30 minutes exposure
Score:
0.326
Remarks on result:
other: test substance
Irritation parameter:
other: optical density [wavelength 570 nm]
Basis:
mean
Time point:
other: after 30 minutes exposure
Score:
0.534
Remarks on result:
other: positive control
Irritation parameter:
other: optical density [wavelength 570 nm]
Basis:
mean
Time point:
other: after 30 minutes exposure
Score:
1.856
Remarks on result:
other: negative control
Irritation parameter:
other: tissue viability [% of negative control]
Basis:
mean
Time point:
other: after 30 minutes exposure
Score:
18
Remarks on result:
other: test substance
Irritation parameter:
other: tissue viability [% of negative control]
Time point:
other: after 30 minutes exposure
Score:
29
Remarks on result:
other: positive control
Irritation parameter:
other: tissue viability [% of negative control]
Time point:
other: after 30 minutes exposure
Score:
100
Remarks on result:
other: negative control

Applicant's summary and conclusion

Conclusions:
Based on the observed results and applying the evaluation criteria it was concluded, that 2-Morpholinoethanol shows an eye irritation potential in the EpiOcular™ eye irritation test under the test conditions chosen.
Executive summary:

The potential of 2-Morpholinoethanol to cause ocular irritation was assessed by a single topical application of 50 µL of the undiluted test substance to a reconstructed three dimensional human cornea model (EpiOcular™). Two EpiOcular™ tissue samples were incubated with the test substance for 30 minutes followed by a 2-hours post-incubation period. Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test. The reduction of mitochondrial dehydrogenase activity, measured by reduced formazan production after incubation with a tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the test-substance treated epidermal tissues is compared to that of negative control tissues. The quotient of the values indicates the relative tissue viability. The EpiOcular™ eye irritation test showed the following results: The test substance is able to reduce MTT directly. Therefore an additional MTT reduction control was introduced. However, this ability of direct MTT reduction did not impair the study result as demonstrated by the concurrently performed exposure of control tissues inactivated by freezing. The mean viability of the test-substance treated tissues was 18 %. Based on the observed results it was concluded, that 2-Morpholinoethanol shows an eye irritation potential in the EpiOcular™ eye irritation test under the test conditions chosen. The test method does not yet allow for the evaluation of serious eye damage. The result does not exclude a serious eye irritation potential of the test substance. For final assignment of a risk phrase at present, results from another study would be needed (see 63V0386/11A583).