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

Endpoint summary

Description of key information

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).
The test article was considered to be severely irritating to eyes and was classified as Category 1 (causing irreversible effects on the eye) according to the Globally Harmonised System of Classification and Labelling of Chemicals (GHS).

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:

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

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

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

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.

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)

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%.

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).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation

Remarks:

other: Preliminary study: In vitro, Main test: In vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 September 2012 to 28 March 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

Remarks:

Preliminary study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan UK Ltd., Bicester
- Age at study initiation: 17 to 18 Weeks
- Weight at study initiation: Not reported
- Housing: the rabbit was housed in a cage that conformed to the 'Code of Practice for the Housing and Care of Animals Used in Scientific Procedures' (Home Office, London, 1989).
- Diet (e.g. ad libitum): Global Diet 2930C (Harlan Teklad, Bicester, UK), ad libitum
- Water (e.g. ad libitum): Mains water, ad libitum
- Acclimation period: 9 Weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15 to 22°C
- Humidity (%): > 45%
- Air changes (per hr): The animal room was designed to permit 15 to 20 air changes per hour.
- Photoperiod (hrs dark / hrs light): The rooms were illuminated by fluorescent strip-lights for twelve hours daily.

IN-LIFE DATES: From: 10 December 2012 To: 14 December 2012

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

One dose consisting of 0.1 mL of undiluted test article was instilled into the left conjunctival sac of a single New Zealand White rabbit (the sentinel).

Duration of treatment / exposure:

4 Days

Number of animals or in vitro replicates:

1

Details on study design:

Before the animal could be dosed, the pH of the test article was checked. A 50% w/v dispersion in purified water had a pH of 6. Since this was within the acceptable range of pH 2.0 to 11.5, the study continued.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 72 Hours

Score:

2

Max. score:

2

Reversibility:

not specified

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 72 Hours

Score:

1

Max. score:

1

Reversibility:

not specified

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 72 hours

Score:

> 3 - < 4

Max. score:

4

Reversibility:

not specified

Irritant / corrosive response data:

Preliminary test results (In vitro experiment)
Corneas treated with the test article were noted to be slightly opaque.
Corneas treated with the positive control article were cloudy and blistered.
The mean opacity reading for the test article was 7.0, for the negative control was 0.0 and for the positive control was 65.3.
The mean group corrected optical density for the test article was 0.232 for the negative control was 0.0 and for the positive control was 0.588.
The test article produced an IVIS score of 10.47.

In vivo experiment
Dulling of the normal lustre of the cornea was noted 4 hours after instillation, with easily discernible translucent areas of corneal opacity noted from 24 to 72 hours after treatment.
Iridial inflammation was noted at all time points from 30 minutes to 72 hours after instillation.
Moderate conjunctival irritation was noted 30 minutes and 1 hour after instillation with severe conjunctival irritation noted 4 hours after instillation persisting to the end of the observation period.
Maximal conjunctival inflammation (swelling grade 4 and redness grade 3) was noted 24 after instillation with no evidence of recovery within 48 hours. The animal was therefore humanely killed in accordance with the UK Home Office Guidelines on Eye Irritation Tests (published March 2006).

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Remarks:

Migrated information

Conclusions:

The test article was considered to be severely irritating to eyes and was classified as Category 1 (causing irreversible effects on the eye) according to the Globally Harmonised System of Classification and Labelling of Chemicals (GHS).

Executive summary:

This study was conducted to determine whether the test article has a potential to cause corrosion or irritation to the eye.

Initially the test article was evaluated using an in vitro test system, the bovine corneal opacity and permeability assay (BCOP). A volume of 750 µL of the test article formulation was applied to each of three corneas followed by a 10 minute incubation at 32 ± 1°C. After this incubation, each cornea was washed with media containing phenol red followed by media without phenol red, the opacities measured and then the anterior chamber emptied. For the permeability endpoint, sodium fluorescein solution was added into the anterior chamber and the corneas were incubated in the vertical position for 1.5 hours ± 5 minutes. Following this period, the media in the posterior chamber was removed and three 350 μL aliquots of this media (per cornea) were analysed for optical density at 490 nanometers (OD₄₉₀).

A volume of 750 µL of the negative or positive control was similarly applied to further groups of three corneas. These corneas were subject to the procedures detailed above.

Corneas treated with the test article were noted to be slightly opaque. Corneas treated with the positive control article were cloudy and blistered. The mean opacity reading for the test article was 7.0, for the negative control was 0.0 and for the positive control was 65.3. The mean group corrected optical density for the test article was 0.232 for the negative control was 0.0 and for the positive control was 0.588. The test article produced an IVIS score of 10.47 and was not considered to be corrosive or severely irritating to the eye according to the BCOP assay.

An in vivo eye irritation test was therefore conducted. The undiluted test article (0.1 mL) was instilled into one conjunctival sac of a New Zealand White rabbit on Day 1. Ocular reactions were assessed for three days after treatment.

Dulling of the normal lustre of the cornea was noted 4 hours after instillation, with easily discernible translucent areas of corneal opacity noted from 24 to 72 hours after treatment. Iridial inflammation was noted at all time points from 30 minutes to 72 hours after instillation. Moderate conjunctival irritation was noted 30 minutes and 1 hour after instillation with severe conjunctival irritation noted 4 hours after instillation persisting to the end of the observation period.

Maximal conjunctival inflammation (swelling grade 4 and redness grade 3) was noted 24 after instillation with no evidence of recovery within 48 hours. The animal was therefore humanely killed in accordance with the UK Home Office Guidelines on Eye Irritation Tests (published March 2006).

The test article was considered to be severely irritating to eyes and was classified as Category 1 (causing irreversible effects on the eye) according to the Globally Harmonised System of Classification and Labelling of Chemicals (GHS).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Corrsosion/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^TMSIT (EPI-200)) was conducted.

EpiDerm^TMSIT (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%.

Thus, 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^TMSIT (EPI-200). Consequently, an in vivo study is not required.

Eye Damage/Irritation

Initially the test article was evaluated using an in vitro test system, the bovine corneal opacity and permeability assay (BCOP). The test article produced an IVIS score of 10.47 and was not considered to be corrosive or severely irritating to the eye according to the BCOP assay.

An in vivo eye irritation test was therefore conducted. Maximal conjunctival inflammation (swelling grade 4 and redness grade 3) was noted 24 after instillation with no evidence of recovery within 48 hours. The animal was therefore humanely killed in accordance with the UK Home Office Guidelines on Eye Irritation Tests (published March 2006).

The test article was considered to be severely irritating to eyes.

Respiratory Irritation

Data on respiratory irritation is not available but considering the low vapour pressure of the test substance irritation of respiratory tract is unlikely.

Justification for selection of skin irritation / corrosion endpoint:
Only study available for this endpoint

Justification for selection of eye irritation endpoint:
Only study available for this endpoint

Effects on skin irritation/corrosion: irritating

Effects on eye irritation: highly irritating

Justification for classification or non-classification

Based on available data, the test substance MonoFA_TETA_PAA_BADGE_BGE_Adduct is considered irritant to skin and causing irreversible effects on eyes and consequently is classified as skin irritant Category 2 and Eye Damaging Category 1 according to CLP (Regulation EC No 1272/2008). Accordingly classification according to DSD (Directive 67/548/EEC) is Xi, R41 and Xi, R38.

Data on respiratory irritation are not available and hence classification for respiratory irritation is not possible.