Slags, sponge iron production by coal reduction in retort

Endpoint summary

Description of key information

The corrosivity potential of the test item was assessed in accordance with OECD Guideline 431.  The test item was considered to be non-corrosive to the skin. Furthermore, the skin irritation potential of the test material was assessed in accordance with OECD Guideline 439.  The relative mean viability of the test item treated tissues was 90.9% after the 15-Minute exposure period and 42-Hours post-exposure incubation period.  As such, the test item was classified as non-irritant.

The eye irriation potential of the test item was assessed in accordance with OECD Guideline 437.   The in-vitro irritancy score of the test item was 356.3, therefore the test substance is considered to be classified as causing irreversible effects on the eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Remarks:

in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 June 2015 - 05 June 2015

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)

Deviations:

yes

Remarks:

The deivation was considered not to affect the purpose or integrity of the study.

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

Deviations:

yes

Remarks:

The deivation was considered not to affect the purpose or integrity of the study.

GLP compliance:

yes (incl. QA statement)

Species:

other: Human Epidermis Model EpiDerm™

Details on test animals or test system and environmental conditions:

Source - EpiDerm™ Reconstructed Human Epidermis Model Kit - supplier MatTek. Upon receipt of the tissues the sealed 24-well plate was placed into a refridgerator.

Amount / concentration applied:

Test material used as supplied

Duration of treatment / exposure:

3 minutes & 60 minute exposure periods.

Details on study design:

Pre-Test Procedure - Assessment of direct test item reduction of MTT

-50mg of test item wsa added to 900µL of a freshly prepared 1.0mg/mL MTT solution.
- The solution was incubated in the dark at 37°C, 5% CO2 in air for 3-hours
- 100µL of sterile distilled water was tested concurrently to act as a control
- The test item was found to have the ability to directly reduce MTT.

Main Test - Preincubabtion

- Assay medium was pre-warmed before use.
- 0.9mL of assay medium was pipetted into well of two pre-labelled 6-well plates for the 3 minutes and 60 minute exposure periods.
- The EpiDerm™ tissues were transferred into the 6-well plates containing the assay medium and pre-incubated for approximately 1 hour before dosing at 37°C 5% CO2

Application of Test Item and Rinsing of test item

- After pre-incubation of the EpiDerm™ tissues, the medium was aspirated and replaced with 0.9mL of fresh assay medium
- For the 60 minute exposure 50 μL of sterile distilled water (negative control) was added to the first two tissues. The tissues were dosed at regular intervals to allow for the time taken to rinse each tissue following exposure and to ensure that each tissue gets an equal exposure time. 25 mg of
the test item and 50 μL of 8.0 N Potassium Hydroxide (positive control) were also applied to the corresponding tissues in turn. 25 μL of sterile water was added for wetting of the test item. The plate was returned to the incubator (37 °C, 5% CO2) for the 60-Minute exposure period.
- The procedure was repeated for the 3-minute exposure period.
- Rinsing was done by filling and emptying each tissue under a constant stream of DPBS to remove any residual test item. Excess DPBS was removed by blotting the bottom of the tissue insert with tissue paper.
- Each tissue was transferred to the 24-well plate prepared for MTT loading. The plate was incubated (37 °C, 5% CO2)) for 3 hours.
- Once the 60-minute exposure period was complete the same rinsing and MTT loading procedure was repeated.
- AFter the 3-hour MTT incubation period, the inserts were blotted and transferred to labelled 24-well plates for MTT extraction.
- 2mL of MTT extractant (isopropanol) was used to completely immerse each insert and a sealed to prevent isopropanol evaporation and the plates were placed into a refridgerator overnight to allow extraction to proceed.

Absorbance/Optical Density Measurements

- After extraction each tissue was pierced with a pipette fittled with a 1000µL tip
- The extraction solution was mixed to form a homogenous solution
- Three 200µL aliquots of blue formazan extract was transferred to the pre-labelled plate
- 200µL L of isopropanol was added to the three well designated as blanks.
- Absorbency at 562nm (OD562) was measured using the Anthos 2001 microplate reader

Interpretation of Results
The corrosivity potential of the test item was predicted from the relative mean tissue viabilities obtained after the 3 and 60-Minute exposure periods, compared to the mean of the negative control tissues (n=2) treated with sterile distilled water. The relative mean viabilities were calculated in the following way:

Relative mean viability (%) = (mean OD562 of test item / mean OD562 of negative control) x 1000

Classification of the corrosivity potential is based on relative viabilities for both exposure times accoding to the attached table.

Quality Criteria : The results of the assay are considered acceptable if the following critera are acheived

Negative Control: The assay establishes the acceptance criterion for an acceptable test if the mean optical density for the negative control treated tissues is ≥ 0.8.
Positive Control: The assay establishes the acceptance criterion for an acceptable test if the relative mean tissue viability for the positive control treated tissues was ≤ 30% relative to the negative control treated tissues following the 3-Minute exposure period

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 minute exposure

Value:

83.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minute exposure

Value:

95.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The relative mean viability of the test item treated tissues was 60 minute exposure 83.9% and 3 minute exposure 95.8%

Direct MTT Reduction

An assessment found the test item was able to directly reduce MTT. Therefore, an additional procedure using freeze-killed tissues was performed during the determination of skin irritation potential. However, the results obtained showed no degree of interference due to direct reduction of MTT occurred. It was therefore considered unnecessary to use the results of the freeze-killed tissues for quantitative correction of results or for reporting purposes.

Test Item, Positive Control Item and Negative Control Item

Mean OD562 values and viabilities for the negative control, positive control and test item are given in the attached table.

The relative mean viability of the test item treated tissues was as follows:

60 minute exposure : 83.9%

3 minute exposure : 95.8%

Quality Criteria

The relative mean tissue viability for the positive control treated tissues was 7.4% relative to the negative control treated tissues following the 3-Minute exposure period. The positive control acceptance criterion was therefore satisfied. The mean OD562 for the negative control treated tissues was 2.127 for the 3-Minute exposure period and 2.304 for the 60-Minute exposure period. The negative control acceptance criteria were therefore satisfied.

Interpretation of results:

GHS criteria not met

Conclusions:

The corrosivity potential of the test item was assessed in accordance with OECD Guideline 431. The test item was considered to be non-corrosive to the skin.

Executive summary:

Summary

The study was performed to evaluate the corrositivity of the test item using the EpiDerm^TMSkin Model after treatment periods of 3 & 60 minutes

^Results

^{Duplicate tissues treated with the test item for exposure periods of 3 and 60 minutes produced relative mean viabilities of 95.8 & 83.9% respectively.}

^Conclusion

^{The test item was considered to be non-corrosive to the skin}

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental Starting Date: 09 June 2015. Experimental Completion Date: 15 June 2015.

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)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN™ Reconstructed Human Epidermis Model Kit
- EpiSkin™ Tissues (0.38cm2) lot number:: 15-EKIN-023
- Maintenance Medium lot number: 15-MAIN3-023
- Assay MEdium lot number: 15-ESSC-023
- Delivery date: 09 June 2015

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

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: At the end of the exposure period, each tissue was removed from the well using forceps and rinsed using a wash bottle containing DPBS with Ca++ and Mg++. Rinsing was achieved by filling and emptying each tissue insert for approximately 40 seconds using a constant soft stream of DPBS to gently remove any residual test item. The rinsed tissues were transferred to the second column of 3 wells containing 2 mL of maintenance medium in each well.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 2 mL of a 0.3 mg/mL MTT solution
- Incubation time: 3 h at 37 °C, 5 % CO2 in air
- Optical density: measured (quantitative viability analysis) at 562 nm (without a reference filter) using the Anthos 2001 microplate reader.

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability:
- Barrier function:
- Morphology:
- Contamination:
- Reproducibility:

NUMBER OF REPLICATE TISSUES: 3

ASSAY ACCEPTANCE CRITERIA
The results of the assay are considered acceptable if the following assay acceptance criteria are achieved:
- Positive Control: The assay establishes the acceptance criterion for an acceptable test if the relative mean tissue viability for the positive control treated tissues was ≤40% relative to the negative control treated tissues, and the standard deviation value of the percentage viability is ≤18%.
- Negative Control: The assay establishes the acceptance criterion for an acceptable test if the mean OD562 for the negative control treated tissues was ≥0.6, and the standard deviation value of the percentage viability is ≤18%.
- Test Item: The assay establishes the acceptance criterion for an acceptable test if the standard deviation calculated from individual percentage tissue viabilities of the three identically treated tissues is ≤18%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Approximately 10 mg of test item was applied to the epidermal surface.

VEHICLE
- Amount(s) applied (volume or weight with unit): 5 µL of sterile distilled water was topically applied to the epidermal surface in order to improve contact between the test item and the epidermis.
- Concentration (if solution):

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 10 µL

POSITIVE CONTROL
- Concentration (if solution): 10 µL of 5 % w/v

Duration of treatment / exposure:

15 m

Duration of post-treatment incubation (if applicable):

42 h, 5 % CO2 in air

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Value:

90.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other:

Remarks:

It was considered unnecessary to perform IL-1α analysis as the results of the MTT test were unequivocal.

Other effects / acceptance of results:

OTHER EFFECTS:
- Direct-MTT reduction: An assessment found the test item was able to directly reduce MTT. Therefore, an additional procedure using water-killed tissues was performed during the determination of skin irritation potential. However, the results obtained showed no degree of interference due to direct reduction of MTT occurred. It was therefore considered unnecessary to use the results of the water-killed tissues for quantitative correction of results or for reporting purposes.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD562 for the negative control treated tissues was 0.789 and the standard deviation value of the percentage viability was 15.6%. The negative control acceptance criterion was therefore satisfied.
- Acceptance criteria met for positive control: The relative mean tissue viability for the positive control treated tissues was 21.5% relative to the negative control treated tissues and the standard deviation value of the percentage viability was 11.3%. The positive control acceptance criterion was therefore satisfied.
- Acceptance criteria met for variability between replicate measurements: The standard deviation calculated from individual percentage tissue viabilities of the three identically test item treated tissues was 6.1%. The test item acceptance criterion was therefore satisfied.

Mean OD₅₆₂Values and Percentage Viabilities for the Negative Control Item, Positive Control Item and Test Item

Item	OD562of tissues	Mean OD562of triplicate tissues	± SD of OD562	Relative individual tissue viability (%)	Relative mean viability (%)	± SD of Relative mean viability (%)
Negative Control Item	0.823	0.789	0.124	104.3	100*	15.6
	0.651			82.5
	0.892			113.1
Positive Control Item	0.266	0.170	0.089	33.7	21.5	11.3
	0.091			11.5
	0.153			19.4
Test Item	0.662	0.717	0.048	83.9	90.9	6.1
	0.753			95.4
	0.736			93.3

OD= Optical Density

SD= Standard Deviation

*= The mean viability of the negative control tissues is set at 100 %

Interpretation of results:

GHS criteria not met

Conclusions:

The skin irritation potential of the test material was assessed in accordance with OECD Guideline 439. The relative mean viability of the test item treated tissues was 90.9% after the 15-Minute exposure period and 42-Hours post-exposure incubation period. As such, the test item was classified as non-irritant.

Executive summary:

Introduction

The purpose of this test was to evaluate the skin irritation potential of the test item using the EPISKIN™ reconstructed human epidermis model after a treatment period of 15 minutes followed by a post-exposure incubation period of 42 hours. The principle of the assay was based on the measurement of cytotoxicity in reconstructed human epidermal cultures following topical exposure to the test item by means of the colorimetric MTT reduction assay. Cell viability is measured by enzymatic reduction of the yellow MTT tetrazolium salt (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to a blue formazan salt (within the mitochondria of viable cells) in the test item treated tissues relative to the negative controls.

Method

Triplicate tissues were treated with the test item for an exposure period of 15 minutes. At the end of the exposure period each tissue was rinsed before incubating for 42 hours. At the end of the post-exposure incubation period each tissue was taken for MTT-loading. The maintenance medium from beneath each tissue was transferred to pre-labeled micro tubes and stored in a freezer for possible inflammatory mediator determination. After MTT-loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT-loaded tissues.

At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200 μL samples were transferred to the appropriate wells of a pre-labeled 96-well plate. The optical density was measured at 562 nm.

Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues).

Results

The relative mean viability of the test item treated tissues was 90.9% after the 15-Minute exposure period and 42-Hours post-exposure incubation period.

Quality criteria:The quality criteria required for acceptance of results in the test were satisfied.

Conclusion

The test item was classified as non-irritant.

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:

09 July 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Deviations not considered to affect the purpose or the integrity of the study (see below for further explanation)

Principles of method if other than guideline:

Deviation No. 1
Opacitometer Calibration: The reading recorded from calibrator number 2 was out of range. However, the values obtained for the positive control and the test item were closest to calibrators 1 (75) and 3 (225) respectively and therefore the results of this study are considered to be unaffected by calibrator number 2.

Deviation No. 2
The IVIS obtained for the positive control also fell below the mean for 2014 as given in the Study Plan. However, it is considered that the significant in vitro irritancy score obtained for the test item (356.3) and the condition of the test item treated corneas (opaque/white) is indicative of severe ocular irritation and therefore the lower then expect positive control score is considered

GLP compliance:

yes (incl. QA statement)

Species:

other: Cattle

Details on test animals or tissues and environmental conditions:

TEST SYSTEM
- Eyes from adult cattle (typically 12-60 months old) were obtained from a local abattoir
- The eyes were excised by an abattoir employee after slaughter and placed in Hanks Balanced Salt Solution supplement with antibiotics (pencillin at 100IU/mL and streptomycin at 100µg/mL)
- Eye were transported over ice pack on the same day of slaughter to the test facility
- Corneas were prepared immediately on arrival

Vehicle:

physiological saline

Controls:

other: Negative Control: 0.9% w/v sodium chloride solution. Positive Control: Imadazole used as a 20% w/v solution in 0.9% w/v sodium chloride solution

Amount / concentration applied:

TEST ITEM FORMLATION & EXPERIMENTAL PREPARATION

- The test item was prepared as a 20% w/v solution in 0.9% w/v sodium chloride solution
- The test item was formulated within 2 hours of being applied to the test system and assumed stable for this duration
- No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.
- Positive control : Imadazole at 20%w/v solution in 0.9% w/v sodium chloride solution
- Negative control : 20% solution in 0.9%w/v sodium chloride solution

TEST ITEM
- 0.75mL of test item was applied on each cornea
- Concentration : 20% w/v solution in 0.9% w/v sodium chloride solution
- Positive Control 20%w/v solution in 0.9% w/v sodium chloride solution
- Negative Control : Used as supplied

Duration of treatment / exposure:

-Test item was applied to the corneas, the holder were gently tilted back and forth to ensure uniform application.
- Each holder was incubated at 32±1°C for 240 minutes.

Observation period (in vivo):

Not applicable

Number of animals or in vitro replicates:

Total: 9 corneas - 3 corneas/group for test item, negative and positive controls

Details on study design:

PREPARATION OF CORNEAS
- Only corneas free of damage were used
- The cornea from each eye was removed and immersed (epithelial side uppermost) in a dish containing HBSS.
- Corneas were mounted into cornea holders
- The anterior and posterior chambers of each holder were filled with Eagle's minimum essential mdium (MEM) without phenol red and plugged.
- The holders were incubated at 32±1°C for 1 hour.
- At the end of the incubation period each cornea was examined for defects and only corneas free of damage were used.

SELECTION OF CORNEAS
- Pre-treatment opacity reading was taken using a calibrated opacitometer
- Average opacilty for all corneas was calculated.

TREATMENT OF CORNEAS
- MEM was removed from the anterior chamber of the holder
- 0.75mL of the test item or control items were applied to the corneas and gently tilted back and forth before being incubated at 32±1°C for 240 minutes.
- At the end of the exposure period the test item and control items were removed from the anterior chamber and rinsed 3 times with MEM containing phenol red and once with MEM withough phenol red.
- an opacity reading and a visual assessment of the cornea was taken.

APPLICATION OF SODIUM FLUORESCEIN
- Permeability of the corneas to sodium flurescein was evaulated.
- Sodium fluorescein solution (5mg/mL) replaced the medium from the anterior chamber, the dosing holes were plugged and the holder incubated at 32±1°C for 90 minutes.

PERMEABILITY DETERMINATIONS
- Following incubation the medium was decanted and retained.
- 360µL of medium representing each cornea was applied to a designated well on a 96-well plate and the optical density at 792nm (OD492) was measured using the Anthos 2001 microplate reader.

HISTOPATHOLOGY
- Corneas were retained for possible conduct of histopathology.

Irritation parameter:

in vitro irritation score

Value:

356.3

Negative controls validity:

valid

Positive controls validity:

other:

Remarks:

The positive control in-vitro irritation score narrowly missed the lower acceptable value of 66.9 but is considered insignificant.

Individual and mean corneal opacity measurements and individual and mean corneal permeability measurements are given in Table 1.

Following treatment with the test substance, the corneas were opaque/white post treatment. The corneas treated with the negative control were clear post treatment and those treated with the positive control were cloudy post treatment (see attached Table 2)

Criteria for an Acceptable Test

The positive control In Vitro Irritancy Score narrowly missed the lower acceptable value of 66.9. Therefore the positive control acceptance criterion was marginally outside the positive control range. However, it is considered that the significant in vitro irritancy score obtained for the test item (356.3) and the condition of the test item treated corneas (opaque/white) is indicative of severe ocular irritation and therefore the lower then expect positive control score is considered insignificant.

The negative control gave opacity of ≤4.1 and permeability ≤0.105. The negative control acceptance criteria were therefore satisfied.

Interpretation of results:

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

Conclusions:

The eye irriation potential of the test item was assessed in accordance with OECD Guideline 437. The in-vitro irritancy score of the test item was 356.3, therefore the test substance is considered to be classified as causing irreversible effects on the eye according to the GHS criteria.

Executive summary:

An in vitro eye irritation study was performed according to the OECD Guideline 437 and in compliance with GLP to assess the corneal damage potential of the test substance by means of the BCOP assay using fresh bovine corneae. 

The test item was applied at a concentration of 20% w/v in 0.9% w/v sodium chloride solution for 240 minutes. Negative and positive control items were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS).

 

Following treatment with test substance, the corneas were noted as opaque/white. The corneas treated with the positive control, 20% w/v Imidazole, were opaque and the corneas treated with the negative control, 0.9w/v% saline, were clear post treatment

 

The negative and positive controls met the acceptance criteria for this assay.

The In Vitro irritancy scores are summarized as follows:

Treatment	In Vitro Irritancy Score
Test Item	356.3
Negative Control	2.1
Positive Control	54.3

 

The test substance is predicted to have a classification of Category 1 (H318: Causes serious eye damage) according to the Regulation (EC) No 1272/2008(CLP).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Read-across has been used for several endpoints in this dossier (see section 13 for the complete justification). It has to be assessed whether a common underlying mechanism leads to the same quantitative outcome (for Source and Target) with regard to the prediction of the property under consideration; and the provided evidence supports the explanation. The Source substance has very low intrinsic toxicological properties, as evidenced by the study data. The available data for the Target substance indicate minimal toxicity. But, it is noted that there is a minor difference in terms of eye irritation in that the Source substance is not an eye irritant whereas the Target substance is classified as an eye irritant. However, this is considered to be caused by the different methodologies used to examine the eye irritation potential of the two substances. For the Source the methods were the HET-Cam ex-vivo assay and the in vivo rabbit eye assay, whereas the BCOP assay was used for the Target substance. The BCOP assay had a 4-hour exposure period whereas the HET-Cam and rabbit study use only 30 minutes and 1 hour respectively. The HET-Cam assay showed no irritant effects whereas the rabbit study showed effects but insufficient for classification. Blast furnace slags in solution have an alkaline pH in the range of 8 to 10 and given the extended exposure period of the BCOP assay then this is the likely explanation for the difference in classification.

Justification for classification or non-classification

In accordance with the prediction model for corrosivity and CLP Regulation, No 1272/2008, a test item should be classified as non-corrosive if the relative mean tiossue viability (% of negative control) greater or equal to 50 after a 3 minute exposure and greater or equal to 15 after a 1 hour exposure. The relative mean viability of the test item treated tissues was 83.9 % after a 1 hour exposure and 95.8 % after a 3 minute exposure. Therefore, the test item does not meet the criteria for classification for corrosivity and can be considered to be non-corrosive.

In accordance with the prediction model for skin irriation and CLP Regulation, No 1272/2008, the test item should be classified as non-irritant if the relative mean tissue viability is greater than 50 %. The relative mean viability of the test item treated issues was 90.9 % after a 15 -Minute exposure period and 42 -Hour post-exposure incubation period. Therefore, the test item does not meet the classification criteria as a skin irritant and can be considered to be non-irritant.

In acordance with the prediction model for eye irritancy and the CLP Regulation, No 1272/2008, a test item should be classified as a Category 1, irreversible effects on the eye, if the in-vitro irritancy score is greater than 55. The in-vitro irritancy score of the test item was 356.3, therefore the test substance meets the criteria for classification.