Fatty acids, C16 and C18-unsatd., polymers with bisphenol A, Bu glycidyl ether, epichlorohydrin and triethylenetetramine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 September 2012 to 25 March 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Fully GLP compliant and in accordance with current test guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Test material form:

other: Viscous semi-solid

Details on test material:

Name: MonoFA_TETA_PAA_BADGE_BGE_Adduct
CAS name: 105839-18-7
Batch number: WA521
Purity: 100%
Expiry date: 28 Feb 2014
Receipt date: 24 Aug 2012
Storage: stored in a sealed container, at 15 to 25ºC, in the dark.

Test animals

Species:

other: Not applicable

Strain:

other: Not applicable

Details on test animals or test system and environmental conditions:

TEST SYSTEM
Three-dimensional human skin model, comprising a reconstructed epidermis with a functional stratum corneum was supplied by MatTek Corporation, Ashland, Massachusetts, USA.

ENVIRONMENTAL CONDITIONS
The tissues were incubated at 37°C, 5 % carbon dioxide for a 35 minute period. The plates were then removed from the incubator and placed into a sterile hood until the 60 minute treatment period was complete for each tissue.
Following treatment, substances were removed by washing the tissues. The tissues were then placed on the appropriate medium and incubated for 42 hours. The protocol stated that the incubation period would be 24 hours. This deviation from protocol was considered not to have affected the integrity or outcome of the study as this was an error in the protocol and the correct incubation period was used for this test system.
To evaluate whether residual test article was binding to the tissue, a functional check (using freeze-killed control tissue) was performed. Freeze-killed tissues were prepared by placing untreated EpiDerm constructs in the -20°C freezer overnight. The freeze-killed tissues were allowed to thaw once at room temperature and placed back in the freezer until required.

IN-LIFE DATES: From: 08 October 2012 To: 17 November 2012

Test system

Type of coverage:

not specified

Preparation of test site:

not specified

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

Three tissues per test article, negative control and positive control were treated by application of 30 µL of either negative/positive control or test article.

Duration of treatment / exposure:

The tissues were incubated at 37°C, 5% carbon dioxide for a 35 minute period. The plates were then removed from the incubator and placed into a sterile hood for the remaining 60 minutes.

Observation period:

Following treatment, the substances were removed by washing the tissues. The tissues were then placed on the appropriate medium and incubated for 42 hours.

Number of animals:

Not applicable

Details on study design:

The test article was positive in the MTT interaction test causing a change in colour intensity. Therefore 0.5mL of isopropanol was instead added into each well so that the MTT was extracted through the bottom of the tissue insert during shaking for two hours at 150 rpm.
Upon completion of the extraction, each tissue was pierced using a hypodermic needle so that the extract could run through the tissue. Once drained, the tissue was discarded and the extract was made up to 2 mL with isopropanol and mixed by pipette. Two x 200 µL aliquots of each resultant extract were placed into a 96-well plate for spectrophotometric determination of optical density at 570 nm using extraction solution as a blank. Tissue viability was calculated for each tissue as a percentage of the mean of the negative control tissues.

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

other: other: relative group mean viability

Value:

14.2

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 60 minutes treatment period. Max. score: 100.0. Reversibility: no data. (migrated information)

In vivo

Irritant / corrosive response data:

The relative group mean viability for the test article was 14.2%.
The relative group mean viability for the positive control was 5.7%.

Applicant's summary and conclusion

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: not specified

Conclusions:

The test article, MonoFA_TETA_PAA_BADGE_BGE_Adduct, was considered not to be corrosive to the in vitro skin model EpiDerm, but was considered to be an irritant to the in vitro skin model EpiDerm SIT (EPI-200).

Executive summary:

The purpose of the study was to determine whether the test article, MonoFA_TETA_PAA_BADGE_BGE_Adduct, caused dermal corrosion or irritation.

An in vitro skin corrosivity assay (EpiDerm^TM) was initially conducted. This demonstrated that the test article did not have the potential to cause corrosion to the skin therefore an in vitro skin irritation assay (EpiDerm^TM SIT (EPI-200)) was conducted.

EpiDerm^TM SIT (EPI-200) inserts were treated with test article, negative control (phosphate buffered saline (PBS)) and positive control (5% w/v sodium dodecyl sulphate (SDS)) for 60 minutes. At the end of the treatment period, the tissues were washed with PBS and cell viability was assessed using the MTT assay. The skin irritation potential was classified according to the remaining cell viability obtained after test article treatment.

The relative group mean viability for the test article was 14.2% and for the positive control was 5.7%.

The test article, MonoFA_TETA_PAA_BADGE_BGE_Adduct, was considered not to be corrosive to the in vitro skin model EpiDerm^TM, but was considered to be an irritant to the in vitro skin model EpiDerm^TM SIT (EPI-200).