Registration Dossier

Administrative data

Description of key information

The test article, TOFA_TETA_PAA_BADGE_CGE_Adduct, was considered not to be corrosive to the in vitro skin model EpiDermTM, but was considered to be an irritant to the in vitro skin model EpiDermTM SIT (EPI-200).
The test article was classified as causing irreversible effects on the eye (Category 1) according to the Globally Harmonised System of Classification and Labelling of Chemicals (GHS) based on results of an in vivo study.

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:
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
Species:
other: Not applicable
Strain:
other: Not applicable
Details on test animals and environmental conditions:
Three-dimensional human skin model, comprising a reconstructed epidermis with a functional stratum corneum supplied by MatTek Corporation, Ashland, Massachusetts, USA. Human keratinocytes are used to construct the epithelium. Multiple layers of viable epithelial cells are present under a functional stratum corneum. The stratum corneum is multi-layered with the necessary lipid profile to produce a functional barrier. The containment properties of the model prevent the passage of material around the stratum corneum to the viable model tissue. The skin model was supplied free of contamination with bacteria, mycoplasma and fungi.
Type of coverage:
not specified
Preparation of test site:
not specified
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
Three tissues per test article, negative control and positive control were treated by application of 30 µL of the negative control, 30 µL of the positive control and approximately 70 mg of test article onto the surface of the tissues.
Details on study design:
The tissues were incubated at 37 ± 1ºC, 5 ± 1% Carbon Dioxide for a 35 ± 1 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.
Irritation / corrosion parameter:
other: other: relative group mean viability
Value:
7.2
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 7.2%.
The relative group mean viability for the positive control was 4.9%.
Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: not specified
Conclusions:
The test article, TOFA_TETA_PAA_BADGE_CGE_Adduct, was considered not to be corrosive to the in vitro skin model EpiDermTM, but was considered to be an irritant to the in vitro skin model EpiDermTM SIT (EPI-200).
Executive summary:

The purpose of the study was to determine whether the test article, TOFA_TETA_PAA_BADGE_CGE_Adduct, caused dermal corrosion or irritation. Dermal corrosion refers to the production of irreversible tissue damage in the skin following the application of a test material. Dermal irritation refers to the production of reversible inflammatory changes in the skin following the application of a test material.

An in vitro skin corrosivity assay (EpiDermTM) 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 (EpiDermTM SIT (EPI-200)) was conducted.

EpiDermTM 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 7.2% and for the positive control was 4.9%.

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

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation
Remarks:
other: In vitro preliminary test and in vivo main test
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:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
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:
Bovine corneas supplied by a local abattoir. The eyes were removed after slaughter, completely immersed in Hanks’ Balanced Salt Solution (containing penicillin at 100 IU/mL and streptomycin at 100 µg/mL) in a suitably sized container and transported on the same day to the testing facility. All corneas were preserved in 10% Neutral Buffered Formalin.

Main test details
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: 7 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)
Amount / concentration applied:
Preliminary test: A volume of 750 µL of the test article was applied to each of three corneas
Main test: 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).
Details on study design:
Preliminary test: A volume of 750 µL of the test article was applied to each of three corneas followed by a 10 minute incubation at 32°C ± 1°C. After this incubation, each cornea was washed with media containing phenol red (as a pH indicator) until this indicator showed no pH effect occurring (and demonstrating that the test article had been removed successfully). The corneas were then washed once in media without phenol red and incubated at 32°C ± 1°C for 120 ± 10 minutes, the opacities measured and the the anterior chamber emptied. For the permeability endpoint, 1 mL of sodium fluorescein (4 mg/mL 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 held in a labelled tube. Three 350 μL aliqots 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 groups were subject to the procedures detailed above.
Main test: Before the first animal could be dosed, the pH of the test article was checked. Since this was within the acceptable range of pH 2.0 to 11.5, the study continued.
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). The lower eyelid was gently prised away from the eyeball to create a receptacle for the dose. After instillation the eyelids were held closed for a few seconds to prevent loss of the dose. The right eye remained untreated and served as a control to the treated eye. The day of dosing was designated as Day 1.
The condition of the treated eye of the sentinel was assessed for a period of three days. As severe ocular responses were observed, the animal was humanely killed and no further animals were treated.
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 72 Hours
Score:
2
Max. score:
2
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 72 hours
Score:
1
Max. score:
1
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
mean
Time point:
other: 72 hours
Score:
3
Max. score:
3
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Remarks:
Chemosis
Basis:
mean
Time point:
other: 72 hours
Score:
4
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Remarks:
Discharge
Basis:
mean
Time point:
other: 72 hours
Score:
2
Max. score:
2
Reversibility:
not reversible
Irritant / corrosive response data:
Scattered or diffuse areas of cornea opacity were 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, 1, 4 and 24 hours after instillation with severe conjunctival irritation noted 28, 48 and 72 hours after instillation.
Maximal conjunctival inflammation (swelling grade 4) was noted 24 hours after instillation and was associated with redness grade 3 at 48 hours after instillation. As there was no evidence of recovery by the 72-hour observation, the animal was humanely killed after the 72-hour observation 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 classified as causing irreversible effects on the eye (Category 1) 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 four hour incubation at 32°C ± 1°C. After this incubation, each cornea was washed with media containing phenol red followed by media without phenol red and then incubated at 32°C ± 1°C for 120 ± 10 minutes. The opacities were then 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 8.3, for the negative control was 0.0 and for the positive control was 75.7. The mean group corrected optical density for the test article was 0.226 for the negative control was 0.0 and for the positive control was 0.595. The test article produced an IVIS score of 11.73 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.

Scattered or diffuse areas of cornea opacity were 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, 1, 4 and 24 hours after instillation with severe conjunctival irritation noted 24, 48 and 72 hours after instillation.

Maximal conjunctival inflammation (swelling grade 4) was noted 24 hours after instillation and was associated with redness grade 3 at 48 hours after instillation. As there was no evidence of recovery by the 72-hour observation, the animal was humanely killed after the 72-hour observation in accordance with the UK Home Office Guidelines on Eye Irritation Tests (published March 2006).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Corrosion/Irritation

An in vitro skin corrosivity assay (EpiDermTM) was initially conducted. This demonstrated that the test article did not have the potential to cause corrosion to the skin and therefore an in vitro skin irritation assay (EpiDermTMSIT (EPI-200)) was further conducted.

EpiDermTMSIT (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 7.2% and for the positive control was 4.9%.

Thus, the test article, TOFA_TETA_PAA_BADGE_CGE_Adduct, was considered not to be corrosive to the in vitro skin model EpiDermTM, but was considered to be an irritant to the in vitro skin model EpiDermTMSIT (EPI-200).

Eye Damage/Irritation

A 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). 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 8.3, for the negative control was 0.0 and for the positive control was 75.7. The mean group corrected optical density for the test article was 0.226 for the negative control was 0.0 and for the positive control was 0.595. The test article produced an IVIS score of 11.73 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.

Scattered or diffuse areas of cornea opacity were 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, 1, 4 and 24 hours after instillation with severe conjunctival irritation noted 24, 48 and 72 hours after instillation.

Maximal conjunctival inflammation (swelling grade 4) was noted 24 hours after instillation and was associated with redness grade 3 at 48 hours after instillation. As there was no evidence of recovery by the 72-hour observation, the animal was humanely killed after the 72-hour observation in accordance with the UK Home Office Guidelines on Eye Irritation Tests (published March 2006). Thus, the substance is considered causing irreversible effects to the eyes.


Justification for selection of skin irritation / corrosion 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 TOFA_TETA_PAA_BADGE_CGE_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.