4,4'-thiodiethylene hydrogen -2-octadecenylsuccinate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

7th August 2014 - 17th September 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

GLP compliance:

yes

Test material

Test material information

1

Reference substance name:

4,4'-thiodiethylene hydrogen -2-octadecenylsuccinate

EC Number:

299-434-3

EC Name:

4,4'-thiodiethylene hydrogen -2-octadecenylsuccinate

Cas Number:

93882-40-7

Molecular formula:

C48H86O8S

IUPAC Name:

2-octadecenylsuccinic anhydride, thiodiethanol esterification products

Test material form:

solid

Remarks:

Waxy Solid

Specific details on test material used for the study:

Lot Number 00031-353/ E00298-234-001
Physical State: Brown Solid
Sotrage Conditions: Ambient Temerpature in the dark, sealed container

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: MatTek

Justification for test system used:

The EpiDerm™ model closely mimics the human epidermis and thus provides an important in vitro approach in the evaluation of dermal corrosion and toxicity.

Vehicle:

water

Details on test system:

The 3-dimensional Human Dermal Epithelial Model (EpiDerm™, MatTek Corp., Ashland, MA) is made up of normal human keratinocytes in serum free medium. The cells form an epithelial tissue that consists of organized basal, spinous, granular, and cornified layers analogous to those found in vivo. The EpiDerm™ model also contains epidermis-specific differentiation markers such as profilaggrin, the K1/K10 cytokeratin pair, involucrin, and type I epidermal transglutaminase, as well as keratohyalin granules, tonofilament bundles, desmosomes, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns characteristic of in vivo epidermis. This model has been used with several common tests of cytotoxicity, corrosion and irritation including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), release of lactate dehydrogenase (LDH), expression and release of interleukin 1-alpha (IL-1a), release of prostaglandin E2 (PGE2) and sodium fluorescein permeability. A growing body of evidence indicates that EpiDerm™ effectively provides a non-animal means to assess dermal corrosion and other toxicological issues.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

25 mg of test article

Duration of treatment / exposure:

3 minutes at room temperature and 60 minutes at approximately 37°C, approximately 5% CO2 in a humidified incubator.

Duration of post-treatment incubation (if applicable):

The MTT assay was performed by transferring the tissues to 24- well plates containing 300 µL MTT medium (1.0 mg/mL). After 3 hours MTT incubation at approximately 37°C, approximately 5% CO2 in a humidified incubator, the tissues were rinsed
twice with DPBS. The blue formazan salt was extracted by submerging tissues in 2 mL isopropanol in a 24-well plate. The extraction time was overnight protected from light, followed by a 15 minute shaking period the following day.

Number of replicates:

n=6

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 Minute Mean

Value:

ca. 107.9

Negative controls validity:

valid

Remarks:

100

Positive controls validity:

valid

Remarks:

30.6

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 minute Mean

Value:

ca. 81.2

Negative controls validity:

valid

Remarks:

100

Positive controls validity:

valid

Remarks:

5.7

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

Negative Control (NC)
The assay meets the acceptance criterion if the Optical Density (OD570) of the NC tissues (treated with sterile ultrapure water) in the MTT assay is =0.8 and =2.8. This is an indicator of tissue viability following shipping and conditions under use. In this assay, the acceptance
criterion was met with each negative control OD between 1.870 and 2.798.

Positive Control (PC)
8N KOH (potassium hydroxide) was used as a PC and tested concurrently with the test article. The assay is meeting the acceptance criteria if the mean viability of the positive control after 1 hour exposure, expressed as mean % of the negative control, is <15%. In this
assay, this acceptance criterion was met with mean % viability of 5.7% viable after 1 hour exposure, resulting in the classification as a corrosive.

Tested Samples
The assay is meeting the acceptance criteria if the tested samples, when in the range of 20 - 100% viability, the Coefficient of Variation (CV) between tissue replicates is =30%. This is an indicator of tissue reproducibility and proper performance of the assay. In this assay, this
acceptance criterion was met with a percent CV of 7.7% and 7.8% for the 3 minute and 1 hour exposures, respectively.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Substance was demonstrated to be non-corrosive in vitro corrosion assay.

Executive summary:

The MatTek EpiDerm™ model was used to assess the potential dermal corrosivity of a test article by determining the cell viability of the tissues after exposure to the test article. The pre-testing (color and MTT pre-test) was performed on August 20, 2014 while the dosing and MTT assay were performed on August 26th. The objective of this study was to assess the dermal corrosion potential of the test article. Tissues were exposed to the test article and controls for three minutes and one hour. The viability of each tissue was determined by MTT assay. The positive control, 8N potassium hydroxide (KOH), reduced cell viability to 30.6% of control after 3 minute exposure and to 5.7 of control after 1 hour exposure. The test article had no effect on the cell viability after 3 minutes exposure (>100% viable) and only a mild effect at 60 minutes exposure (>81% viable). These data suggest that the test material is non-corrosive according to the OECD test guideline using MatTek EpiDerm™ tissues.