Alcohols, C18-22, reaction products with maleic anhydride and sodium bisulfite

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012, November

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP study, well described, according to the HCE. The Human Corneal Epithelium 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)

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: Human Corneal Epithelium model

Principles of method if other than guideline:

The method allows to evaluate a toxic event and related cytotoxicity by a colorimetric assay. Solution of MTT in balanced saline is yellowish in color. Mitochondrial 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:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Disodium C18-C22 sulphosuccinate

IUPAC Name:

Disodium C18-C22 sulphosuccinate

Test material form:

other: liquid

Details on test material:

PRODUCT NAME: Sulfosuccinate of C18-22 alcohol 30% AS (RTC No.13349)
BATCH: FL21-20-RS
MANUFACTURER : TFL Ledertechnik GmbH
PHYSICAL FORM: Liquid
STORAGE: Room temperature
EXPIRATION DATE 31.03.2014

Test animals / tissue source

Species:

other: The SkinEthic© Reconstituted Human Corneal Epithelium

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.
The model has been fully characterized (Ref. Beuermann and Nguyen) and it presents the same feature and morphology of human epithelium in vivo. The sensitivity, specificity and concordance of the model versus the Draize test (OECD TG 405) has been evaluated by using the cytotoxicity parameter for discriminating irritant and non irritant reference products from the ECETOC list (Van Goethem et al. 2006).

Test system

Amount / concentration applied:

The test was performed on triplicate tissues.
The positive control was Ethanol (99,99%) and the negative control was saline solution (NaCl 0,9%).
30 μL of controls (liquids) were directly and uniformly applied topically.
Sulfosuccinate of C18-22 alcohol 30% AS 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 1h followed by product washing according to the relevant procedure and a post incubation period of 16h.

Duration of treatment / exposure:

The experimental design is reported in the following scheme:
1h TREATMENT + 16h RECOVERY


DAY TIME OF TREATMENT PROCEDURES and METHODS

Tuesday 20.11.2012 OVERNIGHT ARRIVAL and PLACEMENT of TISSUES
in 12-WELL PLATE in MAINTENANCE MEDIUM
Wednesday 21.11.2012 MAINTENANCE MEDIUM CHANGE
1h TOPICAL APPLICATION of TEST ITEM,
POSITIVE CONTROL (ETHANOL) and
NEGATIVE CONTROL (SALINE SOLUTION)
PRODUCT WASHING WITH SALINE SOLUTION
16h RECOVERY INCUBATION IN FRESH MEDIUM
Thursday 22.11.2012 3h MTT TEST
(10 min QUALITATIVE EVALUATION)
2h FORMAZAN EXTRACTION IN ISOPROPANOL
SPECTRO-PHOTOMETRICAL MEASUREME

Results and discussion

In vivo

Results

Irritation parameter:

other: viability

Basis:

mean

Time point:

other: 16h

Score:

ca. 50

Max. score:

100

Reversibility:

not specified

Irritant / corrosive response data:

A cell viability < 50% corresponding to the cut-off value for eye irritant classification
has been quantified for positive control (0,65%). For Sulfosuccinate of C18-22 alcohol 30% AS, a residual viability lower than 50%
(49,89%) has been calculated: this value classifies the test item as potential irritant for
the eye

Applicant's summary and conclusion

Interpretation of results:

slightly irritating

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

For Sulfosuccinate of C18-22 alcohol 30% AS, a residual viability lower than 50% (49,89%) has been calculated: this value classifies the test item as potential slightly irritant for the eye

Executive summary:

Sulfosuccinate of C18-22 alcohol 30% AS has been tested for eye irritation according to the HCE. The Human Corneal Epithelium 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). A residual viability lower than 50% (49,89%) has been calculated: this value classifies the test item as potential slightly irritant for the eye