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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
Study period:
from 20th November 2012 to 22nd November 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented study conducted according to a recognized method.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: SOP M04
Deviations:
not specified
Principles of method if other than guideline:
The method allows to evaluate a toxic event and related cyotoxicity by a colorimetrtc assay. Solution of MTT in balanced saline is yeliowish in color. Mitocondrial dehydrogenase of viable cells cleaves the tetrazolium ring yielding blue/purple MTT crystals which are insoluble in aqueous solutions. Crystals formed by viable cells are retained in the polycarbonate filter used as substrate for the 3D construct. Intense purple colour of the cultures indicates the viability of the cell, whereas colour remains white when necrosis occurs. Negative controls are of a dark blue colour and positive controls are white-yellow.
MTT crystals are extracted by Isopropanol and optical density Is measured at 570 nm using the TECAN INFINITE M-200 spectrophotometer. Results are expressed as percentage of cell viability compared to the negative control.
GLP compliance:
no

Test material

Constituent 1
Reference substance name:
Alkenes, C11-14, hydroformylation products, distn. residues, reaction products with maleic anhydride and sodium bisulfite, sodium salts
EC Number:
938-654-2
Molecular formula:
not applicable
IUPAC Name:
Alkenes, C11-14, hydroformylation products, distn. residues, reaction products with maleic anhydride and sodium bisulfite, sodium salts
Test material form:
other: liquid
Details on test material:
Name of the substance as cited in the report: Alkenes, C 11-14, hydroformylation products, distn. residues, reaction products with maleic anhydride and sodium bisulfite, sodium salts
Physical form: liquid
Storage: -20°C

Test animals / tissue source

Species:
other: not applicable
Strain:
other: not applicable
Details on test animals or tissues and environmental conditions:
the SkinEthic Reconstituted Human Corneal Epithelium of 0.5 cm2 is used for the evaluation Epithelial human immortalized cells (HICEC) are deposed on a polycarbonate filter and cultured at the air-liquid interface for 5 days in a chemically defined medium In order to form a structured epithelium.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
30 µl
Duration of treatment / exposure:
Eye Irritation potential of the test item was assessed at 1 h followed by product washing and a post Incubation period of 16h.
Number of animals or in vitro replicates:
not applicable
Details on study design:
Immediately after the arrival in the laboratory the HCE was removed from the agarose nutrient solution under a sterile air flow cabin The inserts were rapidly placed in a 12- well plate previously filled with 0.75 mL of the SkinEthic maintenance medium at room temperature. The wells were placed in an incubator at 37°C 5% CO2 and saturated humidity overnight. The test was started the day after the arrival.

The test was performed on triplicate tissues
The positive control was Ethanol (99,99%) and the negative control was saline solution (NaCI 0,9%) 30 µl of controls (liquids) were directly and uniformly applied topically
Alkenes, C11-l4 hydroformylation products, distn. residues reaction products with maleic anhydride and sodium bisulfite sodium salts (C11— 14) is a liquid and it was tested at the dose defined for liquid (30 µl) according to SOP M 04
Eye Irritation potential of the test item was assessed at lh followed by product washing and a post Incubation period of 16h.

Results and discussion

In vivo

Results
Irritation parameter:
other: cell viability
Basis:
mean
Time point:
other: 1h + 16h exposure
Reversibility:
other: not applicable
Remarks on result:
other: mean tissue viability is < 50%.
Irritant / corrosive response data:
The prediction model for eye irritation classification is based on cell viability (MTT - modified protocol) after 1h + 16h exposure.
A cut-off of 50% is accepted for dicriminating between potential irritant (≤ 50%) and not classified (> 50%). Positive control (ethanol) satisies acceptance criteria if it is classified as irritant (≤ 50%).

Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: expert judgment
Conclusions:
According to the adopted prediction model, based on MTT results, the test item has been classified as reported irritant for the eye.
Executive summary:

An in-vitro on Human Corneal Epithelium model (HCE) was applied in order to assess the eye irritation potential of Alkenes, C11-14, hydroformylation products, distn. residues, reaction products with maleic anhydride and sodium bisulfite, sodium salts.

The HCE, Human Corneal Epithalium model, is currently under a cosmetic Europe (CosEU, x COLIPA)-ECVAM validation study (2009 -2012) as test method replacement alternative for regulatory classification (irritant/non irritant for the eye) by using the MTT test after exposure of 1h+16h of post incubation. According to the adopted prediction model, based on MTT results, the test item has been classified as reported irritant for the eye.

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