Olivine, cobalt silicate blue

Endpoint summary

Description of key information

skin irritation: not irritating (OECD 439; GLP)

eye irritation: not irritating (OECD 492; GLP)

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:

2019-11-06 to 2019-11-29

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:

2019-06-18

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: MatTek Corporation Protocol: In vitro EpiDermTM Skin Irritation Test (EPI-200-SIT) For use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm (EPI-200-SIT)

Version / remarks:

2017-07-11

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

signed 2019-10-23

Test system:

human skin model

Source species:

human

Cell type:

other: normal, human epidermal keratinocytes

Cell source:

other: humans

Source strain:

other: not applicable

Details on animal used as source of test system:

not applicable

Justification for test system used:

This test uses the EpiDerm™ reconstructed human epidermis model (MatTek) which consists of normal human-derived epidermal keratinocytes (NHEK) and therefore represents in vitro the target organ of the species of interest and closely resembles the biochemical and physiological properties of the upper parts of the human skin, i.e. the epidermis.

Vehicle:

other: Dulbecco's Phosphate Buffered Saline (DPBS)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM (EPI-200-SIT; MatTek)
- Tissue lot number: 30837

TEST FOR MTT INTERFERENCE
- To test if a test item directly reduces MTT, 1 ml of a MTT/DMEM solution (1 mg/mL) including 25±2 mg of the test item was incubated for 1 hour (37 ± 1.5°C, 5 ± 0.5% CO2).
- Untreated MTT/DMEM solution (1 mg/mL) was used as negative control.
- After incubation the change of colour was determined by the unaided eye.

TEST FOR COLOUR INTERFERENCE
- to check the colouring potential of the test item 25±2 mg of the test item was added to 300 µl of deionised water and mixed. 330 µl of deionised water was used as control (blank). Both were incubated for 60 min (37 ± 1.5°C, 5 ± 0.5% CO2).
- In parallel, 25±2 mg of the test item was added to 2 mL of isopropanol and mixed. A control (2 mL of isopropanol, blank) was run concurrently. Both were incubated for 1 hour at room temperature.
- After incubation the change of colour was determined by the unaided eye.

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
Since the test item interfered with MTT and reduced it to formazan by itself in the second pre-experiment two additional controls in duplicates run with the main experiment – the freeze-killed tissue controls (killed controls = KC):
- Negative control/ deionised water treated freeze-killed tissues (NC_KC)
- Test item treated freeze-killed tissues (TI_KC)
At the end, the following data correction procedure for viability plus Killed Control tests was performed::
Corrected viability (%) = TI viability – (TI_KC viability – NC_KC viability)

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1.5 °C for 35 minutes followed by incubation at room temperature until the 60-minute treatment period was completed
- Temperature of post-treatment incubation: 37 ± 1.5 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
- At the end of the treatment interval the tissues were gently rinsed with PBS several times in order to remove any residual test material and controls.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL (300 µL/well)
- Incubation time: 3 hours ± 5 minutes (37 ± 1.5°C, 5 ± 0.5% CO2)
- After 3 hour ± 5 minutes incubation the tissues were rinsed with PBS and carefully dried with blotting paper. The tissues were transferred into new 24-well plates containing 2 mL of extractant solution (isopropanol) in each well ensuring that the tissues were completely covered. The 24-well plates were sealed to minimise isopropanol evaporation. The formazan salt was extracted within about 3 hours while shaking at room temperature. At the end of the extraction period the inserts were pierced with an injection needle to allow the extract to run into the well from which the insert was taken, and the insert were discarded.
- Per each tissue, 3 x 200 µL aliquots of the formazan solution were transferred into a 96-well, flat bottom, microtiter plate for OD measurement. 200 µL of isopropanol were added to the wells designated as blanks for 96-well plate.
- Measurement: The optical density (OD570nm) was determined spectrophotometrically in duplicates by a microplate reader (Versamax® Molecular Devices). The absorbance values were determined using the software SoftMax Pro Enterprise (version 4.7.1).

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: tissues pass analysis for tissue viability
- Barrier function: tissues pass analysis for tissue functionality
- Morphology: presence of a functional stratum corneum, a viable basal cell layer, and intermediate spinous and granular layers.
- Contamination: absence of bacteria, yeast, and other fungi (long term antibiotic, antimycotic free culture) as well as absence of HIV1-virus, Hepatitis B virus and Hepatitis C virus
Please also refer to the field "Attached background material" below.

NUMBER OF REPLICATE TISSUES:
- Three tissue replicates for test item and controls

PREDICTION MODEL / DECISION CRITERIA
The mean optical density (OD) of the three negative control tissues was calculated after blank correction. This value corresponded to 100 % tissue viability in the current test. For each individual tissue treated with the test item or the positive control, the individual relative tissue viability is calculated according to the following formula:
Relative viability (%) = [(mean ODtest item / positive control) / ODmean of negative control] * 100
For the test item and the positive control the mean relative viability ± relative standard deviation of the three individual tissues were calculated.
Irritant potential of the test item was predicted from the relative mean tissue viabilities compared to the negative control tissues concurrently treated with DPBS. The test item is considered to be irritant to skin in accordance with Regulation EC No. 1272/2008 (UN GHS “Category 2”), if the tissue viability after exposure and post-incubation is less or equal to 50%. Further testing is required to resolve between UN GHS categories 1 and 2 and decide on the final classification of the test substance. The test substance may be considered as non-irritant to skin in accordance with regulation EC 1272/2008 and UN GHS “No Category” if the tissue viability after exposure and post-treatment incubation is more than 50%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: The test item was tested neat. Three tissue replicates were wetted with 25 µL of DPBS prior to application and then 25 mg (39 mg/cm²) of the test item were applied uniformly to the epidermis surface.

VEHICLE
- Amount(s) applied: 25 µL of DPBS

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL DPBS

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL of 5 % SDS solution

Duration of treatment / exposure:

60 minutes

Duration of post-treatment incubation (if applicable):

approx. 42 hours

Number of replicates:

triplicates

Irritation / corrosion parameter:

% tissue viability

Remarks:

(mean)

Value:

93.39

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

TEST FOR MTT INTERFERENCE
Optical evaluation of the MTT-reducing capacity of the test item with MTT-reagent showed purple colour. Therefore, an additional test with freeze-killed tissues was necessary for the quantitative correction of the test item viability.

TEST FOR COLOUR INTERFERENCE
The test item did not change colour when mixed with deionised water or isopropanol. No colouring was detectable by unaided eye-assessment. Therefore, an additional test with viable tissues was not necessary.

ACCEPTANCE OF RESULTS:
- the mean OD570 of the tissue replicates treated with the negative control is ≥ 0.8 and ≤ 2.8 (1.953) in accordance with OECD TG 439.
- the viability of the tissue replicates treated with the positive control is ≤ 20% (4.35%) .
- the standard deviation calculated from 3 identically treated replicates is ≤ 18 (0.6 – 8.1).
- Concurrent negative controls (NC) and positive controls (PC) were used in each run to demonstrate that viability (with the NC), barrier function and resulting tissue sensitivity (with the PC) of the tissues were within a defined historical acceptance range.

Please also refer to the field "Any other information on results incl. tables" below.

Results after treatment with Cobalt silicate olivine and the controls

Treatment Group	Tissue No.	OD 570 nm			Mean OD of 3 Wells	Mean OD of 3 Wells blank corrected	Mean OD of 3 tissues	Rel. Viability [%] Tissue 1, 2 + 3	Standard Deviation	Mean Rel. Viability [%]
		Well 1	Well 2	Well 3
Blank		0.040	0.041	0.053	0.044
Negative Control	1	2.041	1.986	2.000	2.009	1.965	1.953	100.591	1.0	100.0
	2	2.019	1.953	1.951	1.974	1.930		98.803
	3	2.063	1.995	1.970	2.009	1.965		100.606
Positive Control	1	0.154	0.143	0.132	0.143	0.098	0.085	5.036	0.6	4.35
	2	0.124	0.122	0.121	0.122	0.078		3.988
	3	0.123	0.122	0.123	0.123	0.078		4.014
Test Item (TI)	1	1.713	1.681	1.674	1.689	1.645	1.809	84.211	8.1	93.39*
	2	1.918	1.843	1.841	1.867	1.823		93.317
	3	2.033	1.995	1.981	2.003	1.959		100.280
Blank		0.038	0.039	0.039	0.039
Negative Control Freeze killed Tissues (NC_KC)	1	0.115	0.113	0.113	0.114	0.075	0.072	3.835	0.2	3.69
	2	0.109	0.108	0.108	0.108	0.069		3.548
Test Item Freeze killed Tissues (TI_KC)	1	0.097	0.096	0.097	0.097	0.058	0.057	2.963	0.1	2.90
	2	0.095	0.094	0.095	0.094	0.056		2.847

* Corrected viability (%) = TI viability – (TI_KC viability – NC_KC viability)

Historical Control Data

Positive Control; OD at 570 nm after exposition to 5% SDS solution in deionized water (MatTek)		Negative Control OD at 570 nm DPBS (MatTek)
Tissue Viability [%]	3.98	Mean OD	1.69
Standard Deviation	1.01 p.p.	Standard Deviation	0.19
Range of Viabilities	2.24% - 6.19%	Range of OD*	1.28 - 2
Mean OD	0.07	* should be ≥0.8 and ≤ 2.8 (OECD 439/ MatTek)
Standard Deviation	0.02
Range of OD	0.03 - 0.11
Data of 51 sets of controls shared between 197 studies performed from August 2015 until June 2019 (p.p.: percentage points)

 

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, in this study and under the experimental conditions reported, Cobalt silicate olivine is non-irritant to skin and does not require classification and labelling for skin irritation according to UN GHS and EU CLP.

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:

2019-11-06 to 2019-11-28

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:

2019-06-18

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: MatTek Corporation Protocol: EpiOcular™ Eye Irritation Test (OCL-200-EIT) for the prediction of acute ocular irritation of chemicals; for use with MatTek Corporation’s Reconstructed Human EpiOcular™ Model

Version / remarks:

2015-06-29

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

signed 2019-10-23

Details on test animals or tissues and environmental conditions:

JUSTIFICATION OF THE TEST METHODS AND CONSIDERATIONS REGARDING APPLICABILITY:
This in vitro method is recommended to identify chemicals that do not require classification for eye irritation or serious eye damage according to UN GHS (UN GHS “No Category”) without further testing within a tiered testing strategy from those requiring classification and labelling (UN GHS categories 1 and 2). Therefore, it can be used for regulatory purposes as an initial step in the bottom-up approach or as one of the last steps in a top-down approach to test eye irritation/corrosion potential. It is not intended to differentiate between UN GHS “Category 1” (serious eye damage) and UN GHS “Category 2” (eye irritation) which would require additional testing. Ocular irritation potential is predicted by the relative viability of the tissue after a single exposure to the test substance. Relative viability is determined by measuring the MTT dye to formazan conversion by the EpiOcular™ tissue construct after topical exposure to the test substance.

RhCE TISSUE CONSTRUCT USED: EpiOcular™ (Lot No.: 30636; Standard Assay Kit and MTT-100 kit; source: MatTek Corporation (82105 Bratislava, Slovakia))
The test was carried out with the EpiOcular™ reconstructed human cornea-line epithelium (RhCE) model (MatTek). The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a stratified, highly differentiated squamous epithelium morphologically similar to that found in a human cornea. The EpiOcular™ RhCE tissue construct consists of at least 3 viable layers of cells and a non-keratinized surface, showing a cornea-like structure analogous to that found in vivo.

Please also refer to the field "Attached background material " below.

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): 50 ± 2 mg were applied to the tissue surface. The test item was tested neat.

Duration of treatment / exposure:

6 hours

Observation period (in vivo):

not applicable

Duration of post- treatment incubation (in vitro):

about 18 hours

Number of animals or in vitro replicates:

Number of EpiOcular tissues:
Test item: duplicates
Negative control: duplicates
Positive control: duplicates

Details on study design:

PRE-TEST FOR MTT INTERFERENCE
- To test if a test item directly reduces MTT, 1 ml of a MTT solution (1 mg/mL) including 50 ± 2 mg of the test item was incubated for 180 min (37 ± 1.5°C, 5 ± 0.5% CO2).
- 50 µL deionised water in MTT solution was used as negative control.
- After incubation the change of colour was determined by the unaided eye.

PRE-TEST FOR COLOUR INTERFERENCE
- To check the colouring potential of the test item, 50± 2 mg of the test item was added to 1 mL of deionised water and mixed. 1 mL of deionised water was used as control (blank). Both were incubated for 60 min (37 ± 1.5°C, 5 ± 0.5% CO2).
- In parallel, 50 ± 2mg of the test item was added to 2 mL of isopropanol and mixed. A control (2 mL of isopropanol, blank) was run concurrently. Both were incubated for 3 hours at room temperature.
- After incubation the presence of the staining was evaluated by OD measurement.

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
Since the test item interfered with MTT and reduced it to formazan by itself in the pre-experiment two additional controls in duplicates run with the main experiment – the freeze-killed tissue controls (killed controls = KC):
- Negative control/ deionised water treated freeze-killed tissues (NC_KC)
- Test item treated freeze-killed tissues (TI_KC)
At the end, the following data correction procedure for viability plus Killed Control tests was performed:
Corrected viability (%) = TI viability – (TI_KC viability – NC_KC viability)

DETAILS OF THE TEST PROCEDURE USED
Pre-warming:
- EpiOcularTM tissues were equilibrated at room temperature for 15 min. The inserts with the tissues were transferred into 6-well-plates containing 1.0 ml assay medium and incubated for 60 minutes (37 ± 1.5°C, 5 ± 0.5% CO2). Afterwards, the medium was changed and a further pre-incubation for approx. 17 hours (37 ± 1.5°C, 5 ± 0.5% CO2) follows.
- Treatment:
After pre-warming and prior to application of the test item respectively the controls, all tissues were pre-wetted with 20 µL Ca2+Mg2+free-DPBS and incubated for 30 min (37 ± 1.5°C, 5 ± 0.5% CO2).
Concurrent negative and positive control were applied at a volume of 50 µL and for the test item 50 ± 2 mg to the tissue surface and incubated for 6 h in assay medium (37 ± 1.5°C, 5 ± 0.5% CO2)
Afterwards all tissues were rinsed 3 times in 100 mL PBS and incubated for 25 min in 5 ml assay medium at room temperature in a 12-well plate (post-exposure immersion). At the end of this incubation the tissues were transferred into a 6-well plate with 1 ml assay medium and were incubated for a post exposure incubation for about 18 h (37 ± 1.5°C, 5 ± 0.5% CO2).
- MTT Assay:
Each insert was removed from the 6-well plate and gently blotted on absorbent material. The tissues were placed into the 24-well plate containing 300 µL of MTT solution and were incubated for 180 min (37 ± 1.5°C, 5 ± 0.5% CO2).
The inserts were removed from the 24-well plates after 180 minutes. The bottom of each insert was blotted on absorbent material, and then transferred into a pre-labelled 24-well plate containing 2 mL isopropanol so that no isopropanol was flowing into the insert on the tissue surface. For this, tissues treated with the test item were extracted from the bottom of the tissue only, to minimise any potential contamination of the isopropanol extraction solution with any test item that may have remained on the tissue. The concurrently tested negative and positive control substances were treated similarly to the tested item. Each tissue was extracted with isopropanol within approx. 2.5 h while shaking at room temperature. At the end of the extraction period, the tissues were not pierced.
Then, the extracts were mixed and two 200 µL aliquots were transferred to a 96-well plate for OD measurement. 200 µL of isopropanol were added to the wells designated as blanks for 96-well plate.
- Measurement:
The optical density (OD570nm) was determined spectrophotometrically in duplicates by a microplate reader (Versamax® Molecular Devices). The absorbance values were determined using the software SoftMax Pro Enterprise (version 4.7.1).

TEST ACCEPTANCE CRITERIA
The test meets acceptance criteria:
- mean absolute OD570 nm of the negative control is > 0.8 and < 2.5
- mean relative tissue viability of the positive control is < 50%
- relative tissue viability difference of replicate tissues is < 20%
- OD control values are not be below historically established boundaries/ positive and negative control mean values and acceptance ranges based on historical data.

DEMOSTRATING OF PROFICIENCY IN PERFORMING THE TEST METHOD BEFORE ROUTINE USE BY TESTING OF THE PROFICIENCY CHEMICALS
Prior to routine use of EpiOcularTM EIT for regulatory purposes, the laboratory demonstrated technical proficiency by correctly predicting the eye irritation potential of fifteen proficiency chemicals listed in Table 1 of OECD TG 492. The respective proficiency certificate given by MatTek is attached in the field "Attached background material" below.

DESCRIPTION OF DATA EVALUATION
1) The mean OD value of the two wells for each tissue and the blank control (ODBlk) was calculated (Mean [OD570] (well 1 and well 2).
2) The mean ODBlk was subtracted from each mean OD value of the two wells.
(Mean [OD570] blank corr. (well 1 and well 2)). These values were used for all further calculations below.
3) The mean OD of the two relating tissues for each test group (negative control (NC), positive control (PC)) and the test item (TI) was calculated with the blank corrected mean OD (Mean [OD570] of T1 and T2)
4) The percent viability of each test group relative to the negative control (= 100%) was calculated:
Viability (%) =100 × (mean OD_TI/PC/NC) / mean OD_NC)
5) The relative OD of each tissue per test group was calculated. 100 divided by the mean ODNC T1 and T2 x mean OD of each test group.
6) The difference of the viability values between duplicate tissues was calculated: The relative OD of T2 was subtracted from T1.

PREDICTION MODEL
If the test item-treated tissue viability is > 60% after exposure and post-exposure incubation relative to the negative control treated tissue viability, the test item is identified as not requiring classification and labelling according to UN GHS (No Category).
If the test item-treated tissue viability is ≤ 60% after exposure and post-exposure incubation relative to negative control treated tissue viability, no prediction can be made for this test item.

Irritation parameter:

other: % tissue viability (mean)

Value:

66.45

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

TEST FOR MTT INTERFERENCE
Optical evaluation of the MTT-reducing capacity of the test item with MTT-reagent showed purple colour. Therefore, an additional test with freeze-killed tissues was necessary in the main experimet.

TEST FOR COLOUR INTERFERENCE
The test item did not change colour when mixed with deionised water or isopropanol and the mean OD of the test item in deionised water or isopropanol was < 0.08. Therefore, an additional test with viable tissues without MTT addition was not necessary in the main experiment.

ACCEPTANCE OF RESULTS:
- mean absolute OD570 nm of the negative control is > 0.8 and < 2.8 (2.033)
- mean relative tissue viability of the positive control is < 50% (33.63%)
- relative tissue viability difference of replicate tissues is < 20% (0.17 p.p. to 14.83 p.p.)
- OD control values are not be below historically established boundaries/ positive and negative control mean values and acceptance ranges based on historical data.

The acceptance criteria were met. Regarding the reproducibility of the data, the absorbance of the negative controls and positive controls is within the historical range of absorbance.

Please also refer to the field "Another information on results incl. tables" below.

Results after treatment with Cobalt silicate olivine and the controls for 6 hours:

Test Group	Tissue No.	Well 1 [OD570]	Well 2 [OD570]	Mean [OD570] (Well 1 and well 2)	Mean [OD570] blank corr. (Well 1 and well 2)	Mean [OD570] of T1 and T2	Tissue viabil. [%]		Viabil. of T1 and T2 [%]		Diff. of viabil. between T1 and T2 [p.p.]	Cor. Mean viability of test item [%]
Blank		0.037	0.037	0.037
Negative Control	1	2.125	2.039	2.082	2.045	2.033		100.00		100.6	1.21	66.5*
	2	2.076	2.039	2.057	2.021					99.4
Positive Control	1	0.833	0.803	0.818	0.781	0.684		33.63		38.4	9.57
	2	0.622	0.624	0.623	0.586					28.8
Test Item (TI)	1	1.271	1.214	1.243	1.206	1.357		66.74		59.3	14.83
	2	1.571	1.518	1.544	1.508					74.2
Negative Control Freeze killed Tissues (NC_KC)	1	0.094	0.095	0.094	0.058	0.055		2.69		2.8	0.30
	2	0.089	0.088	0.088	0.052					2.5
Test Item Freeze killed Tissues (TI_KC)	1	0.097	0.094	0.096	0.059	0.061		2.98		2.9	0.17
	2	0.099	0.099	0.099	0.062					3.1

* Corrected viability (%) = TI viability – (TI_KC viability – NC_KC viability)

Historical Control Data

Positive Control (Methyl acetate)		Negative Control OD at 570 nm (Deionised water)
Mean Viability	27.04%	Mean Absorption	1.54
Standard Deviation	10.39 p.p.	Standard Deviation	0.248
Range of Viabilities	6.73% - 42.54%	Range of Absorbance	1.02 - 2.09
Mean Absorption	0.463
Standard Deviation	0.169
Range of Absorbance	0.078 – 0.776
Data of 51 studies performed from July 2015 until March 2019. (p.p.: percentage points)

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, it can be stated that in this study and under the experimental conditions reported, Cobalt silicate olivine does not require classification and labelling for eye irritation or serious eye damage according UN GHS.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

The substance was not observed to be a skin irritant in a reliable in vitro skin irritation study according to OECD 439.

Eye irritation:

The substance was not observed to be an eye irritant in a reliable in vitro eye irritation study according to OECD 492.

Justification for classification or non-classification

Skin irritation:

The substance does not possess a skin irritation potential based on an in vitro OECD 439 test and does not require classification as skin irritant according to Regulation (EC) No 1272/2008 and its subsequent adaptations.

Eye irritation:

The substance does not possess an eye irritation potential based on an in vitro OECD 492 test and does not require classification as eye irritant according to Regulation (EC) No 1272/2008 and its subsequent adaptations.