Registration Dossier

Administrative data

Description of key information

Non skin irritant
Eye irritant

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study was conducted according to GLP and valid methods and is considered relevant and reliable for classification. It is an in vitro study predicting skin irritation, which is considered adequate in combination with the in vitro corrosivity testing.
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
other: a three-dimensional human skin model
Strain:
other: The Skin model EST-1000 was used.
Details on test animals and environmental conditions:
Not applicable
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 300 mg of the supplied test item were dissolved in 1 mL water for injection. 30 µL Butanedioic acid, sulfo-, 4-(2-((2-hydroxyethyl)amino)ethyl) ester, N-C18-unsatd. acyl derivs., disodium salts were applied to the skin model with a surface area of 0.6 cm2
Irritation / corrosion parameter:
other: other: cell viability test group
Value:
110.5
Remarks on result:
other:
Remarks:
Basis: mean. Time point: 20 minutes, followed by 42h incubation . Remarks: % versus negative control group . (migrated information)
Irritation / corrosion parameter:
other: other: cell viability positive control group
Value:
1.3
Remarks on result:
other:
Remarks:
Basis: mean. Time point: 20 minutes, followed by 42h incubation . Remarks: % versus negative control group . (migrated information)

The cell viability was measured by determining the optical density (OD) at a wavelength of 540 nm. An exposure time of 20 minutes was employed.

The test item, Butanedioic acid, sulfo-, 4 -(2 -((2 -hydrocyethyl)amino)ethyl)ester, N-C18 -unsatd. acyl derivs., disodium salts, was applied to the model skin surface. Water for injection was used as the negative control. 5% aqueous sodium dodecyl sulphate (SS) was used as the positive reference item.

The mean viability of the cells exposed to the test item was 110.5% of the mean negative control value. The OD540values were well above the cut-off percentage cell viability value that distinguishes irritant from non-irritant test items of >50% for a 20-minute exposure.

The test item was considered to be non-cytotoxic and predicted to be not irritant to skin.

The viability of cells treated with the positive reference item, 5% SDS,was 1.3% of the negative controls and below the 50% cut-off value. Hence,5% SDS is predicted to cause pronounced skin irritation.

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the present test conditions Butanedioic acid, sulfo-, 4-(2-((2-hydroxyethyl)amino)ethyl) ester, N-C18-unsatd. acyl derivs., disodium salts, tested at an exposure time of 20 minutes, was non-cytotoxic and not irritant to skin in an experiment with an artificial three-dimensional model of human skin.
Executive summary:

The purpose of this study was to determine cytotoxic properties to skin cells, which might lead to irritation by Butanedioic acid, sulfo-, 4 -(2 -((2 -hydroxyethyl)amino)ethyl) ester, N-C-18 -unsatd. acyl derivs., disodium salts to human skin, in an experiment with an artificial three-dimensional model of human skin.

The EST1000 model was employed.

The cell viability was measured by determining the optical density (OD) at 540 nm. An exposure time of 20 minutes was employed.

The test item, Butanedioic acid, sulfo-, 4 -(2 -((2 -hydroxyethyl)amino)ethyl) ester, N-C18 -unsatd. acyl derivs., disodium salts, was applied to model skin surface. Water for injection was used as the negative control. 5% aqueous sodium dodecyl sulphate (SDS) was used as the positive reference item.

The mean viability of the cells exposed to the test item was 110.5% of the mean negative control value. The OD540values were well above the cut-off percentage cell viability value that distinguishes irritant from non-irritant test items of >50% for a 20-minute exposure.

The test item was considered to be non-cytotoxic and predicted to be not irritant to skin.

The viability of cells treated with the positive reference item, 5% SDS,was 1.3% of the negative controls and below the 50% cut-off value. Hence, 5% SDS is predicted to cause pronounced skin irritation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
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)
Qualifier:
according to
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. certificate)
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).
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
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
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

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

- In a first in vitro study, the corrosive properties were studied in an experiment with a three-dimensional EST-100 human skin model (Flügge, 2012a) on the similar substance N-C18-unsatd. acyl derivs. sulphosuccinate, disodium salts. The test item was non-corrosive in this human skin model and is predicted to be non-corrosive to human skin.

- In a second in vitro study, the irritant properties to skin cells were also studied in an experiment with a three-dimensional EST-100 human skin model (Flügge, 2012b) on the same substance. The test item was considered to be non-cytotoxic and predicted to be not irritant to skin.

- According to ECHA progress report 2010 (p. 32), it is accepted that in vitro methods for skin irritation represent a full replacement of the in vivo method in a tiered testing strategy and in conjunction with in vitro skin corrosivity tests, if necessary. A negative result in the human skin model for irritation does not need to be confirmed by additional testing. - In a third in vitro study the potential of Sulfosuccinate of Lanolin Alcohol solid to be irritant to the skin was investigated through an in vitro skin irritation study using a commercial reconstructed human epidermis (RhE) model named EPISKINTM The test item did not induce cell death, and good homogeneity was observed between replicates. According to the established criteria (cell viability less than 50%), the test item is considered to have no irritant effect on the skin under the reported experimental conditions.

The registered substance has been tested for eye irritation and only a very slight irritant potential has been assessed. Based on the studies on the similar substances and the outcome of the eye irritation study the registered substance can be considered as a non irritant for the skin

Eye irritation:

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

Also two in vitro study on a similar substances are reported:

In a first in vitro study the eye irritancy potential of test item containing 29.9% active ingredient was tested by means of the chorioallantoic membrane of hens' eggs (HET-CAM) method (Haferkorn, 2012). Three eggs each were treated with 300 µL/egg. After administration of the test item, blood vessels including the capillary system and the albumen were examined and scored for irritant effects (haemorrhage, coagulation and lysis) during 5 minutes. The test item treated eggs revealed an effect with an irritation index (IS) of 4.0, compared to IS of 19.0 or 9.3 for 0.1 N NaOH and 1% SDS positive controls and no effects in the negative control 0.9% NaCl solution. The test item was considered to be a slight irritant. - In a second in vitro study severe eye irritancy potenital and corrosivity potential of the test item containing 29.9% active ingredient was tested by means of the BCOP test method (Leuschner, 2013) with negative results.

Those results are confirming the eye irritation potential of the registered substance


Justification for selection of skin irritation / corrosion endpoint:
Although the in vitro study for irritation was selected, the corrosion study was equally valuable in a weight-of-evidence approach.

Justification for selection of eye irritation endpoint:
The study has been performed on the registered substance, with a scientifically accepted method, supported by several in vitro tests on similar substances

Effects on eye irritation: irritating

Justification for classification or non-classification

Skin irritation/corrosion:

The substance is not classified a skin irritant

Eye irritation:

The substance is classified an eye irritant cat 2: H319