Reaction product of trimethylhexamethylenediisocyanate and (3-mercaptopropyl)trimethoxysilane

Endpoint summary

Description of key information

An EpiDerm skin corrosion in vitro study according to OECD guideline 431 and GLP was performed. The test item was considered to be not corrosive to skin.

An EpiSkin skin irritation in vitro study according to OECD guideline 439 and GLP was performed. The test item was considered to be not irritating to skin.

A BCOP eye irritation in vitro study according to OECD guideline 437 and GLP was performed. The test item was considered to be not irritating to the eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The experimental start date was 16 April 2018, and the experimental completion date was 20 April 2018.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Version / remarks:

29 July 2016

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EC Guideline No. 440/2008. Part B: Methods for the Determination of Toxicity and other health effects, Guideline B.40 BIS: "In Vitro Skin Corrosion: Human Skin Model Test". Official Journal of the European Union No. L142.

Version / remarks:

31 May 2008

Deviations:

no

GLP compliance:

yes

Remarks:

Analyses conducted to support the information cited in the Certificate of Analysis for the test item were not conducted in compliance with the GLP or GMP regulations but under a sponsor or sponsor subcontractor quality system.

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BZA11014
- Expiration date of the lot/batch: 21 April 2018
- Manufacturing date: 21 March 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature under nitrogen
- Not stable at higher temperatures

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: No correction was made for purity/composition of the test item. The weighed amount of test item was flushed with nitrogen. The liquid item was applied undiluted.

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Human donors. Tissue is obtained by MatTek Corporation from accredited institutions.

Justification for test system used:

The test is based on the experience that corrosive chemicals show cytotoxic effects following short-term exposure to the stratum corneum of the epidermis. The test is designed to predict and classify the skin corrosion potential of a test item by assessment of its effect on a three-dimensional human epidermis model.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm
- Tissue batch number(s): 28326
- Delivery date: 18 April 2018
- Date of initiation of testing: 16 April 2018

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: all incubations, with the excepton of the test item incubation of 3 minutes at room temperature, were carried out at 37 +/- 1ºC.

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: After the exposure period, the tissues were washed with phosphate buffered saline (Invitrogen Corporation, Breda, The Netherlands) to remove residual test item. Due to the characteristics of the test item it was difficult to remove and therefore some residual test item could still be left on the skin. The skin inserts were carefully dried. Rinsed tissues were kept in 24 well plates on 300 μL DMEM until 6 tissues (= one application time) were dosed and rinsed.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: MTT concentrate (5 mg/mL) diluted (1:5) with MTT diluent (supplemented DMEM). Both supplied by MatTek Corporation.
- Incubation time: 3 hours
- Spectrophotometer: TECAN Infinite® M200 Pro Plate Reader
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: Acceptance criteria: OD (540-570 nm) 1.0 - 3.0. Pass (value: 2.124 +/- 0.066)
- Barrier function: Acceptance criteria: ET-50 3.68 - 8.02 h. Pass (value: 6.13 h)
- No contamination

NUMBER OF REPLICATE TISSUES: Two tissues for a 3-minute exposure and 2 tissues for a 1-hour exposure

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µL

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL
- Concentration (if solution): 8N

Duration of treatment / exposure:

3 minutes and 1 hour

Duration of post-treatment incubation (if applicable):

Not applicable

Number of replicates:

2 replicates for each test condition

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3-minutes incubation

Value:

>= 102

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1-hour incubation

Value:

>= 81

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Direct-MTT reduction: Not observed
- Colour interference with MTT: Not observed

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes. The absolute mean OD570 of the two tissues of the negative control is within the laboratory historical control data range.
- Acceptance criteria met for positive control: yes . The mean relative tissue viability following 1-hour exposure to the positive control is < 15% (observed value = 6.8%).
- Acceptance criteria met for variability between replicate measurements: yes. In the range 20-100% viability, the coefficient of variation between tissue replicates is <= 30% (obtained value = <=22%).

Mean tissue viability:

	3 -minute application, % viability	1 -hour application, % viability
Negative control	100	100
Test item	102	81
Positive control	14	6.8

Interpretation of results:

GHS criteria not met

Conclusions:

The Thio Isocyanate Adduct is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this report.

Executive summary:

The study was performed to evaluate Thio Isocyanate Adduct for its ability to induce skin corrosion on a human three dimensional epidermal model (EpiDerm (EPI-200)). The possible corrosive potential of Thio Isocyanate Adduct was tested through topical application for 3 minutes and 1 hour. The study procedures were based on OECD guideline 431 and performed according to GLP. Thio Isocyanate Adduct was applied undiluted (50 μL) directly on top of the skin tissue. The positive control had a mean relative tissue viability of 6.8% after the 1-hour exposure. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the acceptance limits of OECD 431 (lower acceptance limit ≥0.8 and upper acceptance limit <=2.8) and the laboratory historical control data range. In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was <= 22%, indicating that the test system functioned properly.

Skin corrosion is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after 3-minute and 1-hour treatments with Thio Isocyanate Adduct compared to the negative control tissues was 102% and 81%, respectively. Because the mean relative tissue viability for Thio Isocyanate Adduct was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment Thio Isocyanate Adduct is considered to be not corrosive.

In conclusion, Thio Isocyanate Adduct is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this report.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The experimental start date was 29 May 2018 and the experimental completion date was 04 June 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

28 July 2015

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

20 July 2012

Deviations:

no

GLP compliance:

yes

Remarks:

Analyses conducted to support the information in the Certificate of Analysis were not conducted in compliance with the GLP or GMP regulations. The characterization of the test item was conducted under a sponsor or sponsor subcontractor quality system.

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BZA13762
- Expiration date of the lot/batch: 11 June 2018
- Manufacturing date: 11 May 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature under nitrogen

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: No correction was made for the purity/composition of the test item. The liquid test item was applied undiluted directly on top of the tissue.

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Adult donors

Justification for test system used:

In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimize the need of in vivo testing. One of the validated in vitro skin irritation tests is the EPISKIN test, which is recommended in international guidelines (e.g. OECD and EC).

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN
- Tissue batch number(s): 18 EKIN 022
- Expiry date: 04 June 2018
- Date of initiation of testing: 29 May 2018

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: All incubations, with the exception of the test item incubation of 15 minutes at room temperature, were carried out at 37 +/- 1ºC
- Temperature of post-treatment incubation (if applicable): 37ºC

REMOVAL OF TEST MATERIAL AND CONTROLS
After the exposure period of 15 ± 0.5 minutes at room temperature, the tissues were washed with phosphate buffered saline to remove residual test item. After rinsing, the cell culture inserts were each dried carefully and moved to a new well on 2 mL pre-warmed maintenance medium. This process continued until all tissues were dosed and rinsed. Subsequently the skin tissues were incubated for 42 hours at 37°C.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/mL in PBS
- Incubation time: 3h
- Spectrophotometer: TECAN Infinite® M200 Pro Plate Reader
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Histology scoring >= 19.5 (actual result =22.4)
- IC50 determination was not valid due to a technical problem. Batch was nevertheless released

NUMBER OF REPLICATE TISSUES: 3

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be irritating to skin if the relative mean tissue viability of three individual tissues after 15 minutes of exposure to the test item and 42 hours of post incubation is ≤ 50% of the mean viability of the negative controls.
- The test substance is considered to be non-irritating to skin if the relative mean tissue viability of three individual tissues after 15 minutes of exposure to the test item and 42 hours of post incubation is > 50% of the mean viability of the negative
controls.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 µL
- Concentration (if solution): Undiluted

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 25 µL
- Concentration (if solution): Undiluted

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 25 µL
- Concentration (if solution): 5% (stock concentration)

Duration of treatment / exposure:

15 +/- 0.5 minutes

Duration of post-treatment incubation (if applicable):

42 hours

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Value:

114

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Colour interference with MTT: the test item did not interact with the MTT endpoint.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes. The absolute mean OD570 (optical density at 570 nm) of the three tissues of the negative control was within the laboratory historical control data range and the Standard Deviation value (SD) of the % viability was <=18 (actual value = <=16%).
- Acceptance criteria met for positive control: The mean relative tissue viability of the positive control should be <=40% relative to the negative control and the Standard Deviation value (SD) of the % viability should be <=18 (actual values 4.6% and SD <=16%).
- Acceptance criteria met for variability between replicate measurements: The SD calculated from individual % tissue viabilities of the three identically treated replicates should be <=18 (actual value <=16%).

Mean tissue viability:

	Mean tissue viability (% of control)	SD
Negative control	100	3.4
Test item	114	16
Positive control	4.6	0.8

Interpretation of results:

GHS criteria not met

Conclusions:

Thio Isocyanate Adduct is a non-irritant in the in vitro skin irritation test under the study experimental conditions and should not be classified according to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS) of the United Nations.

Executive summary:

The study was carried out to evaluate Thio Isocyanate Adduct for its ability to induce skin irritation on a human three dimensional epidermal model (EPISKIN Small model (EPISKIN-SMTM)). The possible skin irritation potential of the test item was tested through topical application for 15 minutes. The study procedures described in this report were based on OECD guideline 439 and according to GLP. The test item was applied undiluted (25 μL), directly on top of the skin tissue for 15 ± 0.5 minutes. After a 42 hour post-incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Skin irritation is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with the test item compared to the negative control tissues was 114%. Since the mean relative tissue viability for the test item was above 50% after 15 ± 0.5 minutes treatment the test item is considered to be non-irritant. The positive control had a mean cell viability of 4.6% after 15 ± 0.5 minutes exposure. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was ≤ 16%, indicating that the test system functioned properly.

In conclusion, Thio Isocyanate Adduct is a non-irritant in the in vitro skin irritation test under the study experimental conditions and should not be classified according to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS) of the United Nations.

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:

Experimental start date: 17 April 2018. Experimental completion date: 17 April 2018.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

October 09, 2017

Deviations:

no

GLP compliance:

yes

Remarks:

Analyses conducted to support the information cited in the Certificate of Analysis for the test item were not conducted in compliance with the GLP or GMP regulations. The characterization of the test item was conducted under a Sponsor quality system.

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BZA11014
- Expiration date of the lot/batch: 21 April 2018
- Manufacturing date: 21 March 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature under nitrogen
- Not stable at higher temperatures

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: No correction was made for the purity/composition of the test item. The test item was tested neat.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL

Duration of treatment / exposure:

10 +/- 1 minutes

Duration of post- treatment incubation (in vitro):

120 +/- 10 minutes

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
The eyes were checked for unacceptable defects, such as opacity, scratches, pigmentation and neovascularization by removing them from the physiological saline and holding them in the light. Those exhibiting defects were discarded. The isolated corneas were stored in a petri dish with cMEM (Earle’s Minimum Essential Medium (Life Technologies, Bleiswijk, The Netherlands) containing 1% (v/v) L-glutamine (Life Technologies) and 1% (v/v) Foetal Bovine Serum (Life Technologies)). The isolated corneas were mounted in a corneal holder (one cornea per holder) of BASF (Ludwigshafen, Germany) with the endothelial side against the O-ring of the posterior half of the holder. The anterior half of the holder was positioned on top of the cornea and tightened with screws. The compartments of the corneal holder were filled with cMEM of 32 +/- 1ºC. The corneas were incubated for the minimum of 1 hour at 32 +/- 1ºC.

QUALITY CHECK OF THE ISOLATED CORNEAS
After the incubation period, the medium was removed from both compartments and replaced with fresh cMEM. Opacity determinations were performed on each of the corneas using an opacitometer (BASF-OP3.0, BASF, Ludwigshafen, Germany). The opacity of each cornea was read against a cMEM filled chamber, and the initial opacity reading thus determined was recorded. Corneas that had an initial opacity reading higher than 7 were not used.

NUMBER OF REPLICATES
Three corneas were selected at random for each treatment group.

NEGATIVE CONTROL USED
A negative control, physiological saline (Eurovet Animal Health, Bladel, The Netherlands) was included to detect non-specific changes in the test system and to provide a baseline for the assay endpoints.

POSITIVE CONTROL USED
Ethanol.

APPLICATION DOSE AND EXPOSURE TIME
The test item was tested neat (colourless liquid) for 10 +/- 1 minutes.

TREATMENT METHOD: closed chamber

POST-INCUBATION PERIOD: yes, 120 +/- 10 minutes

REMOVAL OF TEST SUBSTANCE
After the incubation the solutions were removed and the epithelium was washed with MEM with phenol red (Earle’s Minimum Essential Medium, Life Technologies) and thereafter with cMEM. Possible pH effects of the test item on the corneas were recorded. The medium in the posterior compartment was removed and both compartments were refilled with fresh cMEM.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: The opacity of a cornea was measured by the diminution of light passing through the cornea. The light was measured as illuminance (I = luminous flux per area, unit: lux) by a light meter. The opacity value (measured with the device OP-KIT) was calculated according to:
Opacity = ((I0/I) - 0.9894) / 0.0251
With I0 the empirically determined illuminance through a cornea holder but with windows and medium, and I the measured illuminance through a holder with cornea.
The change in opacity for each individual cornea (including the negative control) was calculated by subtracting the initial opacity reading from the final post-treatment reading. The corrected opacity for each treated cornea with the test item or positive control was calculated by subtracting the average change in opacity of the negative control corneas from the change in opacity of each test item or positive control treated cornea. The mean opacity value of each treatment group was calculated by averaging the corrected opacity values of the treated corneas for each treatment group.
- Each cornea was inspected visually for dissimilar opacity patterns.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of [UV/VIS spectrophotometry / microtiter plate reader] (OD490) . Following the final opacity measurement, permeability of the cornea to Na-fluorescein (Sigma-Aldrich, Germany) was evaluated. The medium of both compartments (anterior compartment first) was removed. The posterior compartment was refilled with fresh cMEM. The anterior compartment was filled with 1 mL of 4 mg Na-fluorescein (Sigma-Aldrich Chemie GmbH, Germany)/mL cMEM solution. The holders were slightly rotated, with the corneas maintained in a horizontal position, to ensure uniform distribution of the sodium-fluorescein solution over the entire cornea. Corneas were incubated in a horizontal position for 90 +/- 5 minutes at 32 +/- 1ºC. After the incubation period, the medium in the posterior compartment of each holder was removed and placed into a sampling tube labelled according to holder number. 360 μL of the medium from each sampling tube was transferred to a 96-well plate. The optical density at 490 nm (OD490) of each sampling tube was measured in triplicate using a microplate reader (TECAN Infinite® M200 Pro Plate Reader). Any OD490 that was 1.500 or higher was diluted to bring the OD490 into the acceptable range (linearity up to OD490 of 1.500 was verified before the start of the experiment). OD490 values of less than 1.500 were used in the permeability calculation. The mean OD490 for each treatment was calculated using cMEM corrected OD490 values. If a dilution has been performed, the OD490 of each reading of the positive control and the test item was corrected for the mean negative control OD490 before the dilution factor was applied to the reading.

SCORING SYSTEM: The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score: In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value). Additionally the opacity and permeability values were evaluated independently to determine whether the test item induced irritation through only one of the two endpoints.

DECISION CRITERIA: The IVIS cut-off values for identifying the test items as inducing serious eye damage (UN GHS Category 1) and test items not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are given hereafter:
<=3 No Category
>3 <= 55 No prediction can be made
>55 Category 1
The assay is considered acceptable if:
- The positive control gives an in vitro irritancy score that falls within two standard deviations of the current historical mean.
- The negative control responses should result in opacity and permeability values that are less than the upper limits of the laboratory historical range.

Irritation parameter:

cornea opacity score

Value:

>= -3 - <= 1

Negative controls validity:

valid

Remarks:

Opacity and permeability values less than the upper limits of the laboratory historical range

Positive controls validity:

valid

Remarks:

Value was 53 and within two sandard deviations of the current historical positive control mean

Remarks on result:

no indication of irritation

Irritation parameter:

fluorescein leakage

Remarks:

Permeability values

Value:

>= 0.001 - <= 0.004

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The corneas treated with Thio Isocyanate Adduct showed opacity values ranging from -3.0 to 1.0 and permeability values ranging from 0.001 to 0.004. The corneas were translucent after the 10 minutes of treatment with Thio Isocyanate Adduct. No pH effect of the test item was
observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from -2.9 to 1.1 after 10 minutes of treatment with Thio Isocyanate Adduct.

OTHER EFFECTS:
- The corneas treated with the negative control item were translucent after the 10 minutes of treatment.
- The corneas treated with the positive control item were turbid after the 10 minutes of treatment.
- The corneas treated with the test item were translucent after the 10 minutes of treatment.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes

Summary of opacity, permeability and in vitro scores:

Treatment	Mean opacity	Mean permeability	Mean in vitro irritation score
Negative control	1.8	0.000	1.8
Positive control	18	2.319	53
Test item	-1.1	0.003	-1.1

Interpretation of results:

GHS criteria not met

Conclusions:

Since the Thio Isocyanate Adduct induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.

Executive summary:

The study was performed to evaluate the eye hazard potential of Thio Isocyanate Adduct as measured by its ability to induce opacity and increase permeability in an isolated bovine cornea using the Bovine Corneal Opacity and Permeability test (BCOP test). The eye damage of Thio Isocyanate Adduct was tested through topical application for 10 minutes. The study procedures were based on OECD guideline 437 and according to GLP. The test item was applied as it is (750 μL) directly on top of the corneas. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (Ethanol) was 53 and was within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

Thio Isocyanate Adduct did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of -1.1 after 10 minutes of treatment. In conclusion, since Thio Isocyanate Adduct induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In the EpiDerm skin corrosion study, the relative mean tissue viability obtained after 3-minute and 1-hour treatments with the test substance compared to the negative control tissues was 102% and 81%, respectively. Because the mean relative tissue viability was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment, the substance is considered to be not corrosive.

In the EpiDerm skin irritation study, the relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with the test item compared to the negative control tissues was 114%. Since the mean relative tissue viability for the test item was above 50% after 15 ± 0.5 minutes treatment the test item is considered to be non-irritant.

In the BCOP eye irritation study, the substance did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of -1.1 after 10 minutes of treatment. Since the substance induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.

Justification for classification or non-classification

Based on the available data for Thio Isocyanate Adduct, no classification for skin irritation and eye irritation is required according to Regulation (EC) No 1272/2008.