4-Ethoxy-2,3-difluorphenylboronic acid

Endpoint summary

Description of key information

Skin irritation: The test item is not irritating to skin as determined in a study according to OECD Guideline 439.

Eye irritation: A study according to OECD Guideline 492 and OECD Guideline 437 was conducted. Based on the results, no conclusion on classification can be made. Since the substance is an intermediate under strictly controlled conditions and there are no data requirements according to REACH, no further study was initiated.

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:

February 10, 2016 - April 15, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

other: Skinethic skin irritation test -42bis Standard operating procedure (SOP) 2009

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

foreskin from a single donor

Justification for test system used:

standard model

Vehicle:

unchanged (no vehicle)

Remarks:

No vehicle used in this study; The test item was applied neat to the tissues.

Details on test system:

CELL CULTURE
- Supplier: EpiSkin/SkinEthic Laboratories, Lyon, France)
- Source: human keratinocytes cultured on a polycarbonate filter in conditions which permit their terminal differentiation
- Format: 24 well plate
- Batch: 16-RHE-025
- Expires: March 21, 2016

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment: room temperature
- Temperature of post-treatment incubation: 37°C

REMOVAL OF THE TEST MATERIAL AND CONTROL
After the end of the treatment interval, the residual test item was removed immediately by gently rinsing with a minimum volume of 25 mL DPBS using a pipette. Excess DPBS was removed by gently shaking the inserts and blotting the bottom with blotting paper.

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be irritating to skin if the viability is less than or equal to 50%.
- The test substance is considered to be non-irritating to skin if the viability is greater than 50%.

Control samples:

yes, concurrent negative control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 16 mg of solid test material
- Concentration (if solution): n/a

VEHICLE
- Amount(s) applied (volume or weight with unit): n/a
- Concentration (if solution): n/a
- Lot/batch no. (if required): n/a
- Purity: n/a

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 16 µL (Dulbecco`s Phosphate-Buffered Saline)
- Concentration (if solution): n/a

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 16 µL
- Concentration (if solution): 5% aqueous solution of sodium dodecyl sulfate in deionised water

Duration of treatment / exposure:

42 min (± 1 minute)

Duration of post-treatment incubation (if applicable):

42 hours (± 1 hour)

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Experiment 1 / Run 1

Value:

101.18

Vehicle controls validity:

not applicable

Remarks:

The test item was applied neat to the tissues

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: none
- Direct-MTT reduction: yes
- Colour interference with MTT: none

ACCEPTANCE OF RESULTS:
Acceptability of the Quality Control Data of the Skin Model with Reference to Historical Batch Data:
The negative control OD values were 1.575, 1.555 and 1.698 and, thus, in the range of ≥0.8 and ≤3.0.

Acceptability of the Positive and Negative Control:
After treatment with the negative control (DPBS-buffer) the mean OD was 1.609 (standard deviation: 4.84%) and, thus, higher than the historically established boundary of 1.436.
After treatment with the positive control (5% aqueous solution of sodium dodecyl sulfate) the mean viability value was 1.28% (standard deviation: 13.56%) and, thus, lower than the historically established boundary of 3.28%.

The standard deviation of the negative control and the positive control was ≤18%, respectively.

Variability of the Data:
The standard deviation of the three tissues treated with the test item was 1.07% and, thus, ≤18%.
Therefore, the study fulfilled the validity criteria.

Group	Time / [min]	Mean OD	Mean Relative viability / [%]
Negative Control	42	1.609	100
Positive Control	42	0.021	1.28
Test Material	42	1.708*	101.18

*Corrected optical density after true metabolic conversion

Interpretation of results:

GHS criteria not met

Conclusions:

This study was performed according to GLP and the methods applied are fully compliant with OECD TG 439. Under the conditions of the present study, the test item is not considered to possess an irritant potential to skin (UN GHS: No Category).

Executive summary:

Objective

This in vitro study was performed to assess the skin irritation potential of the test item by means of the Reconstructed Human Epidermis (RHE) Test.

 

Study Design

The test consisted of a topical exposure of the test item to a human reconstructed skin model followed by a cell viability test. Cell viability was quantitatively measured by dehydrogenase conversion of MTT into a blue formazan salt after extraction from tissues. The percent reduction of cell viability in comparison to untreated negative controls was used to predict the skin irritation potential.

 

Triplicates of the human skin RHE-model were treated with the test item, the negative or the positive control for 42 minutes (± 1 minute). 16 µL of either the negative control (DPBS-buffer) or the positive control (5% aqueous solution of sodium dodecyl sulfate) were applied to the tissues. Before adding the solid test item, 10 µL of deionised water was spread to the epidermis surface to improve the contact between the test item and the epidermis. Afterwards, 16 mg of the test item were applied to the tissues.

 

The test item has the ability to directly reduce MTT. To evaluate the extent of non-specific interaction, three killed tissues were treated with the test item or the negative control, respectively. The treatment and MTT assay of the killed tissues was similar to the handling of the living tissues. The obtained OD for a non-specific reduction was subtracted from OD-values obtained after treatment of living tissues with the test item to calculate the cell viability.

 

Results

After treatment with the negative control (DPBS-buffer) the mean OD was 1.609 (study acceptance criteria: >1.436). Treatment with the positive control (5% aqueous solution of sodium dodecyl sulfate) revealed a mean viability value of 1.28% (study acceptance criteria: <3.28%). Therefore, the study fulfilled the validity criteria.

 

The tissue viability after treatment with the test item was 101.18% and, thus, higher than 50%,i.e.according to UN GHS classification the test item is considered as non-irritant to skin (UN GHS: No Category).

 

Conclusion

Under the conditions of the present study, the test item is not considered to possess an irritant potential to skin (UN GHS: No Category).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

April 18, 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Version / remarks:

October 9, 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

human

Strain:

not specified

Details on test animals or tissues and environmental conditions:

- Justification of the test method and considerations regarding applicability :
The reconstructed human cornea-like epithelium (RhCE) model is an accepted in vitro method to replace animal testing. The human eye EpiOcular-model closely mimics the biochemical and physiological properties of the human eye, i.e. the cornea.

Analysis for tissue functionality and quality: please refer to "any other information on methods and materials"

- Description of the cell system used
Desigantion: EpiOcular Tissue (OCL-200, OCL-212)
Keratinocyte strain: 4F1188

Vehicle:

unchanged (no vehicle)

Remarks:

The test item was applied neat to the tissues.

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

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

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

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 50 µL

Duration of treatment / exposure:

6 hours +/- 15 min

Duration of post- treatment incubation (in vitro):

18 hours +/- 15 min

Number of animals or in vitro replicates:

2 tissues

Details on study design:

- Details of the test procedure used

- RhCE tissue construct used, including batch number: EpiOcular Tissue (OCL-200, OCL-212), Lot No. 27033

- Doses of test chemical and control substances used: 50 mg test material, 50 µL negative control, 50 µL positive control

- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods (where applicable)
Exposure: 6 hours (+/- 15 min) at 37°C
Post-exposure immersion: 25 min (+/- 2 min) at room temperature
Post-exposure incubation: 18 hours (+/- 15 min) at 37°C

- Indication of controls used for direct MTT-reducers and/or colouring test chemicals (if applicable): Pre-experiments to assess direct MTT reduction and colored or staining test item were performed.
- Number of tissue replicates used per test chemical and controls (positive control, negative control, NSMTT, NSCliving and NSCkilled, if applicable): 2 (test item), 2 (negative control), 2 (positive control)
- Wavelength and band pass (if applicable) used for quantifying MTT formazan, and linearity range of measuring device (e.g. spectrophotometer): wavelength = 570 nm
- Description of the method used to quantify MTT formazan
After the post-treatment incubation period, the treated tissues were incubated with 300 µL MTT solution (1.0 mg/mL MTT) and incubated for 180 min (+/- 10 min) at 37°C. The inserts were removed from the 24-well plate after 180 min (+/- 10 min). The bottom of the inserts was blotted on absorbent material, and then transferred to a 6-well plate containing 2 mL isopropanol so that no isopropanol was flowing into the inserts. The plate was sealed with a standard plate sealer. To extract MTT, the plates were placed on an orbital shaker and shaken for 2 to 3 hours at room temperature. The corresponding negative and positive controls were treated identically.
The extracts solution was mixed and 2 x 200 µL were transferred into a 96-well plate. The OD was read using a spectrophotometer at 570 nm wavelength.

- Description of evaluation criteria used including the justification for the selection of the cut-off point for the prediction model
No categeory: Mean tissue viability > 60%
No prediction can be made: Mean tissue viability: <= 60%

- Reference to historical positive and negative control results demonstrating suitable run acceptance criteria
Negative control: OD > 0.8 and < 2.5
Positive control: mean relative viability is a) 30 min exposure: below 50% of control viability and b) 6 hour exposure: below 50% of control viability
- Demonstration of proficiency in performing the test method before routine use by testing of the proficiency chemicals
- Positive and negative control means and acceptance ranges based on historical data
1. Negative control is OD > 0.8 and < 2.5 (1.577 and 1.795)
2. The mean relative viability of the positive control is below 50% of the negative control viability (17.8%)
- Acceptable variability between tissue replicates for positive and negative controls: < 20%
- Acceptable variability between tissue replicates for the test chemical: < 20%

Irritation parameter:

other: viability (%)

Run / experiment:

1

Value:

13.7

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

other: viability (%)

Run / experiment:

2

Value:

13.9

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: no

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes (OD > 0.8 and < 2.5 (1.577 and 1.795))
- Acceptance criteria met for positive control: Yes (below 50% of the negative control (17.8%))

The pre-test for direct MMT-reducing capacity of the test item did not result in blue color, i.e. the test item is not a direct MTT reducer and the test item has no colorant properties.

Interpretation of results:

study cannot be used for classification

Conclusions:

Following treatment with the test item, the tissue viability was 13.8% and, thus, lower than 60%, i.e. according to OECD 492 no prediction can be made regarding the eye hazard potential of the test item.

Executive summary:

The objective of the present study was to investigate the potential of the test item to induce eye irritation in an in vitro human cornea model. The test item was applied topically to a reconstructed human cornea-like epithelium model (EpiOcular) followed by determination of the cell viability. Cell viability was determined by enzymatic conversion of vital dye MTT into a blue formazan salt and measurement of the formazan salt after extraction from tissues. The percent reduction of cell viability in comparison to untreated negative controls was used to predict the eye irritation potential.

Duplicates of the EpiOcular-model were treated with the test item, the negative or the positive control for 6 hours (+/- 15 min). 50 mg of the test item and 50 µL of either the negative control (sterile deionized water) or the positive control (methyl acetate) were applied to the tissues.

After treatment with the negative control (sterile deionized water) the mean OD was 1.686 (study acceptance criterion: > 0.8 and < 2.5). Treatment with the positive control (methyl acetate) revealed a mean viability value of 17.8% (study acceptance criterion: < 50%). Thus, the acceptance criteria were met. Following treatment with the test item, the tissue viability was 13.8% and, thus. lower than 60%, i.e. according to OECD 492 no prediction can be made regarding the eye hazard potential of the test item.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2016-02-10 to 2017-01-18

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 i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Version / remarks:

2013

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Version / remarks:

2010

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Odenwaldschlachthof Brensbach, 64395 Brensbach, Germany
- Characteristics of donor animals (e.g. age, sex, weight): 15 - 33 months
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): The eyes were kept and transported in transport medium cooled on ice.
- indication of any existing defects or lesions in ocular tissue samples: All eyes were carefully examined macroscopically for defects. Those presenting defects such as vascularization, pigmentation, opacity or scratches were discarded.

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

yes, concurrent positive control

Amount / concentration applied:

750 μL of the suspended test item (i.e. 150 mg/750 μL), positive or negative control were applied on the corneas

Duration of treatment / exposure:

4 hrs

Number of animals or in vitro replicates:

3 replicates

Details on study design:

SELECTION AND PREPARATION OF CORNEAS : Freshly isolated bovine eyes of cattle were collected from the slaughterhouse. Excess tissue was removed from the eyes. The eyes were kept and transported in transport medium cooled on ice. The corneas were prepared immediately after delivery of the eyes to the laboratory. The corneas were carefully removed from the eyes using scalpel and rounded scissors. A rim of about 2 to 3 mm of tissue (sclera) was left for stability and handling of the isolated cornea. All corneas used in the study were collected in incubation medium (pre-warmed at 32 +/- 1°C) and the corneal diameter of each cornea was measured and recorded. Each cornea was mounted in a cornea holder with the endothelial side against the sealing ring (O-ring) of the posterior part of the holder. The cornea was gently flattened over the O-ring without stretching the cornea. Afterwards, the anterior part of the holder was positioned on top of the cornea and fixed in place with screws. Both compartments of the holder were filled with incubation medium. The posterior compartment was filled first to return the cornea to its natural convex form.

QUALITY CHECK OF THE ISOLATED CORNEAS : All eyes were carefully examined macroscopically for defects. Those presenting defects such as vascularization, pigmentation, opacity or scratches or an opacity > 7 opacity units were discarded.

NUMBER OF REPLICATES : 3 (in three independent experiments)

NEGATIVE CONTROL USED : 0.9% sodium chloride solution

POSITIVE CONTROL USED : Imidazole (20 %)

APPLICATION DOSE AND EXPOSURE TIME : 750 µL of the suspended test item (i.e. 150 mg/750 µL) and an exposure time of 240 minutes

TREATMENT METHOD: closed chamber (run 1), open chamber (run 2 and 3)

POST-INCUBATION PERIOD: no.

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: three times

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: For equilibration, the corneas in the holder were incubated in a vertical position at 32 +/- 1°C for about one hour. At the end of the incubation period, the incubation medium was replaced by fresh pre-warmed (32 +/- 1°C) incubation medium in both compartments. The baseline opacity was determined with a calibrated opacitometer. The light transmission through the corneas, given as lux value, was recorded in a table and thereafter converted into an opacity value (baseline opacity values).
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD490)


SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: IVIS <= 3 (No category); IVIS > 3, <= 55 (No prediction can be made); IVIS > 55 (Category 1)

Irritation parameter:

in vitro irritation score

Run / experiment:

1

Value:

52.4

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

2

Value:

55.1

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

3

Value:

4.1

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Other effects / acceptance of results:

Definition of Study Acceptance Criteria
A test is considered acceptable if the positive control gives an IVIS that falls within two standard deviations of the current historical mean (IVIS positive control run 1: 76.5 - 136.8; run 2: 76.9 - 136.6; run 3: 77.4 - 136.3).
The negative control responses should result in an IVIS that falls within three standard deviations of the current historical mean (IVIS negative control run 1: -1.4 to -3.5; run 2: -1.3 to -3.4; run 3: -1.4 to -3.4).

A single test run with three corneas should be sufficient for a test item when the resulting classification is unequivocal. In cases of the following borderline results in the first testing run, a second test run should be considered.
-2 of the 3 corneas give discordant predictions from the mean of all 3 corneas or
-1 of the 3 corneas give discordant predictions from the mean of all 3 corneas, and the discordant result is > 10 IVIS units from the cut-off threshold of 55.

Interpretation of results:

study cannot be used for classification

Conclusions:

The resulting classifications of the test item in the different runs were equivocal and, in addition, borderline results were obtained. Therefore, it was considered that the BCOP assay is not applicable to examine the potential of the test item to induce serious eye damage.

Executive summary:

The objective of the present study was to examine the potential of the test item to induce serious eye damage in the BCOP assay. The BCOP assay with isolated fresh bovine corneas is an accepted in vitro model for ocular hazard assessment.

To determine the eye hazard potential the induced opacity and increased permeability was investigated in isolated bovine corneas after exposure to the test item as a 20 % (w/v) suspension in a 0.9 % sodium chloride solution. As negative control 0.9 % sodium chloride solution and as positive control 20 % (w/v) Imidazole was used.

Three corneas were used per group (negative control, positive control or test item group).

After a first opacity measurement of the untreated bovine corneas, 750 μL of the dissolved test item, positive or negative control were applied on the corneas and incubated for 240 minutes. After the incubation phase the test item, the positive, and the negative control were rinsed from the corneas and the opacity was measured again.

After the opacity measurements, the permeability of the corneas was determined by application of a fluorescein solution for 90 minutes. The amount of fluorescein solution that crossed the cornea was measured spectrophotometrically.The opacity and permeability assessments were combined to determine an In Vitro Irritancy Score (IVIS).

Three runs were performed with the test item.

Run 1: After treatment with the negative control (0.9 % sodium chloride solution) the calculated IVIS was 1.1 (study acceptance criteria range: -1.4 to -3.5). Treatment with the positive control (20 % Imidazole) revealed an IVIS of 111.9 (study acceptance criteria range: 76.5 to 136.8). Therefore, this run fulfilled the acceptance criteria for the negative and positive control. The IVIS obtained after treatment with the test item was 52.4.

Run 2: After treatment with the negative control (0.9 % sodium chloride solution) the calculated IVIS was -0.1 (study acceptance criteria range: -1.3 to -3.4). Treatment with the positive control (20 % Imidazole) revealed an IVIS of 111.5 (study acceptance criteria range: 76.9 to 136.6). Therefore, this run fulfilled the acceptance criteria for the negative and positive control. The IVIS obtained after treatment with the test item was 55.1.

Run 3: After treatment with the negative control (0.9 % sodium chloride solution) the calculated IVIS was 0.1 (study acceptance criteria range: -1.4 to -3.4). Treatment with the positive control (20 % Imidazole) revealed an IVIS of 111.2 (study acceptance criteria range: 77.4 to 136.3). Therefore, this run fulfilled the acceptance criteria for the negative and positive control. The IVIS obtained after treatment with the test item was 4.1.

The resulting classifications of the test item in the different runs were equivocal and, in addition, borderline results were obtained. Therefore, it was considered that the BCOP assay is not applicable to examine the potential of the test item to induce serious eye damage.

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation/corrosion

In vitro Skin Irritation study (OECD 439)

This in vitro study was performed to assess the skin irritation potential of the test item by means of the Reconstructed Human Epidermis (RHE) Test.

After treatment with the negative control (DPBS-buffer) the mean OD was 1.609 (study acceptance criteria: >1.436). Treatment with the positive control (5% aqueous solution of sodium dodecyl sulfate) revealed a mean viability value of 1.28% (study acceptance criteria: <3.28%). Therefore, the study fulfilled the validity criteria.

The tissue viability after treatment with the test item was 101.18% and, thus, higher than 50%,i.e.according to UN GHS classification the test item is considered as non-irritant to skin (UN GHS: No Category)

Eye irritation/corrosion

In vitro Eye Irritation study (OECD 492)

The objective of the present study was to investigate the potential of the test item to induce eye irritation in an in vitro human cornea model. The test item was applied topically to a reconstructed human cornea-like epithelium model (EpiOcular) followed by determination of the cell viability. Cell viability was determined by enzymatic conversion of vital dye MTT into a blue formazan salt and measurement of the formazan salt after extraction from tissues. The percent reduction of cell viability in comparison to untreated negative controls was used to predict the eye irritation potential.

After treatment with the negative control (sterile deionized water) the mean OD was 1.686 (study acceptance criterion: > 0.8 and < 2.5). Treatment with the positive control (methyl acetate) revealed a mean viability value of 17.8% (study acceptance criterion: < 50%). Thus, the acceptance criteria were met. Following treatment with the test item, the tissue viability was 13.8% and, thus lower than 60%, i.e. according to OECD 492 no prediction can be made regarding the eye hazard potential of the test item.

In vitro Eye irritation/corrosion study (OECD 437)

The objective of this GLP study performed according to OECD GL 437 was to examine the potential of the test item to induce serious eye damage in the BCOP assay. The BCOP assay with isolated fresh bovine corneas is an accepted in vitro model for ocular hazard assessment. The resulting classifications of the test item in the different runs were equivocal and, in addition, borderline results were obtained. Therefore, it was considered that the BCOP assay is not applicable to examine the potential of the test item to induce serious eye damage.

No conclusion on classification für eye irritation/corrosion can be made based on the available studies.

Justification for classification or non-classification

Based on data provided the test item is not classified for skin irritation according to Regulation (EC) No 1272/2008. Based on the two in vitro eye irritation/corrosion studies, no prediction can be made on classification and labelling regarding eye irritation/corrosion potential. However, no further studies were initiated as the substance is registered transported isolated intermediate according to Article 18 of REACH Regulation and therefore no data requirements are needed.