Hexanamide, N-(2-hydroxyethyl)-3,5,5-trimethyl-

Endpoint summary

Description of key information

Considering the results of the in vitro skin irritation/corrosion studies, the test substance can be considered to be irritating to skin. However, in the absence of data and based on the available weight of evidence from the skin irritation and the surface tension studies, the test substance is considered to be corrosive to the eyes as a conservative approach.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 September 2012 to 08 February 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD (2010), In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method, OECD Guidelines for the Testing of Chemicals No. 439, OECD, Paris.

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: ECVAM Scientific Advisory Committee (ESAC) Statement on the Validity of In Vitro Tests for Skin Irritation (2007).

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: ECVAM Skin Irritation Validation Study. Validation of the EpiSkin® Irritation Test – 42 Hours Assay for the Prediction of Acute Skin Irritation of Chemicals, L’Oreal Recherche, January 2005.

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: ECVAM Skin Irritation Validation Study (SIVS). Performance Standards for Applying Human Skin Models to In Vitro Skin Irritation Testing (2007).

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

OECD (2010), In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method, OECD Guidelines for the Testing of Chemicals No. 439, OECD, Paris.

Deviations:

no

Principles of method if other than guideline:

in vitro skin irritation using the EpiSkin model.

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

TEST SYSTEM
- SkinEthic Episkin

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 µL
- Surface area of the EpiSkin was ca 0.38 cm2.
- Application rate was ca 26.3 μL/cm2.

Duration of treatment / exposure:

15 minutes.

Duration of post-treatment incubation (if applicable):

42 hours

Number of replicates:

3 replicates

Species:

other: in vitro

Details on study design:

TEST SITE
- Area of exposure: 0.38 cm2; RATE OF 26.3 μL/cm2.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Rinsed with 25 mL PBS and blotted dry on tissue paper.
- Time after start of exposure: 15 mins

INITIAL ASSESSMENT OF TOXICITY
Prior to the conduct of the irritation assay, a preliminary test was conducted to assess the intrinsic ability of the test item to reduce methylthiazoldiphenyl-tetrazolium bromide (MTT) to formazan. The substance coating additives did not reduce MTT to formazan.

PROCEDURE OF THE MAIN TEST
The dermal irritation potential was assessed by applying 10 μL of the substance coating additives to the exposed surface of three EpiSkin tissues for 15 min. The surface area of the EpiSkin was 0.38 cm2, therefore the application rate was 26.3 μL/cm2. After the 15 min exposure period, the test item was washed from the surface of the EpiSkin using Dulbecco’s phosphate-buffered saline (PBS). The EpiSkin tissues were then incubated for a recovery period of 42 h in a humidified incubator, set to maintain temperature and CO2 levels of 37°C and 5%, respectively. Following incubation, the tissues were transferred to assay medium containing MTT (0.3 mg/mL) and returned to the incubator for 3 h. Biopsies of the EpiSkin membranes were removed, added to acidified isopropanol, and refrigerated for ca 68 h in order to extract the formazan. The formazan production (cell viability) was assessed by measuring the optical density of the extracts at a wavelength of 550 nm. Three replicates of the positive control, aqueous sodium dodecyl sulphate (SDS, CAS 151-21-3) solution (5%, w/v), and the negative control, PBS, were tested in parallel to demonstrate the efficacy of the assay. The viability of each individual EpiSkin tissue was calculated as a percentage of the mean negative control viability (defined as 100%).

Irritation / corrosion parameter:

% tissue viability

Value:

9.56

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

Basis: mean mean of 3 replicates. Time point: 42 h. Reversibility: no data. Remarks: Reduced viability of cells in the EpiSkin® model.

Interpretation of results:

other: Category 2 (irritant) based on CLP criteria

Conclusions:

Under the conditions of the in vitro EpiSkin® study, the test substance was demonstrated to be an irritant to skin

Executive summary:

A study was conducted to determine dermal irritation potential of the test substance, in an EpiSkin® in vitro irritation test, according to OECD 439, in compliance with GLP. Prior to the conduct of the irritation assay, a preliminary test was conducted to assess the intrinsic ability of the test substance to reduce methylthiazoldiphenyl-tetrazolium bromide (MTT) to formazan. The test substance did not reduce MTT to formazan. The dermal irritation potential was assessed by applying 10 μL of the test substance to the exposed surface of three EpiSkin tissues for 15 min. The surface area of the EpiSkin was 0.38 cm2, therefore the application rate was 26.3 μL/cm2. After the 15 min exposure period, the test substance was washed from the surface of the EpiSkin using Dulbecco’s phosphate-buffered saline (PBS). The EpiSkin tissues were then incubated for a recovery period of 42 h in a humidified incubator, set to maintain temperature and CO2 levels of 37°C and 5%, respectively. Following incubation, the tissues were transferred to assay medium containing MTT (0.3 mg/mL) and returned to the incubator for 3 h. Biopsies of the EpiSkin membranes were removed, added to acidified isopropanol, and refrigerated for ca 68 h in order to extract the formazan. The formazan production (cell viability) was assessed by measuring the optical density of the extracts at a wavelength of 550 nm. Three replicates of the positive control, aqueous sodium dodecyl sulphate (SDS) solution (5%, w/v), and the negative control, PBS, were tested in parallel to demonstrate the efficacy of the assay. The viability of each individual EpiSkin tissue was calculated as a percentage of the mean negative control viability (defined as 100%). Exposure to the test substance resulted in a mean EpiSkin viability of 9.56% ± 3.26% of the negative control value. Exposure to the positive control, aqueous SDS solution (5%, w/v), resulted in a mean EpiSkin viability of 13.36% of the negative control value. Under the conditions of the in vitro EpiSkin® study, the test substance was demonstrated to be an irritant to skin (Blackstock, 2013).

Reference 2

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From August 25, 2021 to September 9, 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

adopted 18th June, 2019

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Three-dimensional human skin model, comprising of non-transformed, human-derived epidermal keratinocytes was supplied by MatTek In Vitro Life Science Laboratories, Bratislava, Slovak Republic.

Source strain:

not specified

Justification for test system used:

The EpiDermTM tissue consists of human-derived epidermal keratinocytes which have been cultured to form a multi-layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues are cultured on spe-cially prepared cell cultures inserts.

Vehicle:

unchanged (no vehicle)

Details on test system:

- Test system specifications:
Three-dimensional human skin model, comprising of non-transformed, human-derived epidermal keratinocytes was supplied by MatTek In Vitro Life Science Laboratories, Bratislava, Slovak Republic.

- Chemicals and Media:
MTT
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (=MTT), which can be reduced to a blue formazan.
A MTT stock solution of 5 mg/mL in DPBS buffer was prepared and stored in aliquots of 2 mL in the freezer (– 20±5°C).
One aliquot of the stock solution was thawed and diluted with 8 mL of medium. This MTT-solution with the resulting concentration of 1 mg/mL was used in the test.
- For the non-GLP pre-test (testing the ability of direct MTT reduction), the stock solution was thawed and diluted with serum-free MEM directly before use
- For the main test, the stock solution was thawed and diluted with assay medium directly before use
 
DPBS-Buffer
“Dulbecco’s Phosphate-Buffered Saline”; Solution for the rinsing of the tissues and solvent for MTT con-centrate. A subset was procured by MatTek In Vitro Life Science Laboratories, Bratislava; the other subset was prepared by LAUS GmbH.
Composition of the subset from MatTek In Vitro Life Science Laboratories, Bratislava (main test batch no: 081021MSA):
KCL 0.2g
KH2PO4 0.2 g
NaCl 8.0 g
Na2HPO4 * 7H2O 2.16 g
demin. H2O ad 1 L
The buffer which was procured by MatTek Corporation was used for rinsing the test item from the tissues and for preparing the MTT solution.

MEM with Phenol Red for Pre-Test
Serum-free MEM (Minimum Essential Medium). Procured by Life Technologies GmbH, batch no.: 2267701

Assay Medium for Main Test
Serum-free DMEM (Dulbecco’s Modified Eagle’s Medium). Procured by MatTek In Vitro Life Science La-boratories, main test batch no: 090221MJB

Isopropanol
CH3-CH(OH)-CH3, for synthesis, ≥ 99.5 %, batch no.: 190296551 used as extracting solvent for formazan

- Test Vessels
All test vessels used were made of sterilised plastic. The following vessels were used:
• 96-well-plate
• 24-well-plates
• 6-well-plates

- Demonstration of Proficiency
The validity of the skin corrosion study at LAUS GmbH was demonstrated in a proficiency study. For this purpose, 12 proficiency chemicals (indicated by the OECD guideline 431) were tested. All of the 12 proficiency chemicals were correctly categorized . Therefore, the proficiency of the skin corrosion study was demonstrated.


- Preparation of the test system:
MTT concentrate was thawed. The concentrate was diluted with the assay medium directly before use. The assay medium was warmed in the water bath to 37 ± 1°C.
Four 6-well-plates were prepared with 0.9 mL assay medium in each well. The inserts containing the tissues were transferred to the wells using sterile forceps and the 6-well-plates were set into the incubator at 37 ± 1°C and 5.0 ± 1% CO2 for 1 h (pre-incubation). For each experiment (“3 min” and “1 h”), one 24-well-plate was prepared as holding plate. 12 wells of each plate were filled with 300 µL assay medium, the other 12 with 300 µL MTT solution. One additional plate was left empty. The plates were stored in the incubator at 37 ± 1°C and 5.0 ± 1% CO2. For each experiment (“3 minutes” and “1 hour”), two 6-well-plates for the assay were used. After pre-incubation, the assay medium was replaced by fresh assay medium and the test was started, using two wells as negative control with 50 µL demineralised water, two wells as positive controls with 50 µL potassium hydroxide solution and two other wells for testing the test substance. The liquid test substance was applied without preparation (50 µL).
After the respective incubation time (“3 min” and “1 h”) at 37 ± 1°C and 5.0 ±1% CO2, the inserts were removed from the plates using sterile forceps. The inserts were thoroughly rinsed with DPBS, blotted with sterile cellulose tissue and set into the respective holding plate, using the wells containing assay medium. After transfer of all inserts, they were immediately moved to the wells containing MTT medium, blotting the bottom with cellulose tissue again before setting the insert into the MTT well. The tissues were incubated with MTT solution for 3 h at 37 ± 1°C and 5.0 ± 1% CO2. After this time, the MTT solution was aspirated and replaced by DPBS. This was then aspirated, too, and replaced several times. At last, each insert was thoroughly dried and set into the empty, pre-warmed 24-well-plate. Into each well, 2 mL isopropanol were pipetted, taking care to reach the upper rim of the insert. The plate was then shaken for 2 h at room temperature. Afterwards, the inserts were pierced with an injection needle, taking care that all colour was extracted. The inserts were then discarded and the content of each well was thoroughly mixed in order to achieve homogenisation. From each well, three replicates with 200 µL solution (each) were pipetted into a 96-well-plate which was read in a plate spectrophotometer at 570 nm. In addition, twelve wells of the 96-well-plate were filled with 200 µl isopropanol each, serving as blank.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

50 μL

Duration of treatment / exposure:

3 minutes and 1 h

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 min

Value:

ca. 102.1

Vehicle controls validity:

not valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 hour

Value:

ca. 51.2

Vehicle controls validity:

not valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

Corrosivity of the test substance
The mean value of relative tissue viability of the test item was increased to 102.1% after 3 minutes treat-ment. This value is above the threshold for corrosivity (50%). After 1 hour treatment, the mean value of relative tissue viability of the test item was reduced to 51.2%, lying above the threshold for corrosivity (15%). Therefore, the test substance is considered as non-corrosive to skin.

Validity:
- The criterion for optical density of the negative control (≥ 0.8 and ≤ 2.8) was fulfilled: optical density was 1.643 (3 min) resp. 0.862 (1 h).
- The positive control showed clear corrosive effects. The criterion for the viability of the 1 h experiment, expressed as % of the negative control (< 15%), was fulfilled, too.
- The mean value of relative tissue viability was 4.5%.
- The values for negative control and for positive control were within the range of historical data of the test facility.

Therefore, the experiment is considered valid. The variation within the tissue replicates of the controls and the test sustance were within the acceptable range.

Result details:

Measured Values of the Main Test

As blank, the optical density of isopropanol was measured in 12 wells of the 96-well-plate. The measured values and their mean are given in the following table:

Table 1‑a      (Absorbance values blank isopropanol (OD 570 nm)

Replicate	1	2	3	4	5	6	Mean 0.034
Absorbance		0.035	0.034	0.034	0.034	0.034
Replicate	7	8	9	10	11	12
Absorbance	0.034	0.035	0.034	0.034	0.034	0.035

 

The absorbance values of negative control, test substance and positive control are given in the following table:

Table 1‑b      Absorbance Values (OD 570 nm)

Incubation time	Negative Control		Test Item		Positive Control
	Tissue 1	Tissue 2	Tissue 1	Tissue 2	Tissue 1	Tissue 2
3 min	1.677	1.606	1.541	1.815	0.280	0.324
	1.678	1.611	1.544	1.818	0.281	0.325
	1.682	1.606	1.541	1.803	0.278	0.325
1 h	1.776	1.660	0.953	0.839	0.123	0.098
	1.771	1.655	0.955	0.837	0.123	0.099
	1.781	1.650	0.952	0.839	0.123	0.098

 

From the measured absorbances, the mean absorbance of isopropanol (given in table 1-a) was subtracted. The corrected mean and relative standard deviation (RSD) of the two tissues were also calculated.

Table 1‑c      Mean Absorbance Values of the 3 Minutes Experiment

Designation	Negative Control	Test Item	Positive Control
Mean – blank (tissue 1)	1.645	1.508	0.245
Mean – blank (tissue 2)	1.573	1.778	0.290
Mean of the two tissues	1.609	1.643	0.268
RSD	3.1%	11.6%	11.9%

Table.1‑d      Mean Absorbance Values of the 1 h Experiment

Designation	Negative Control	Test Item	Positive Control
Mean – blank (tissue 1)	1.742	0.919	0.089
Mean – blank (tissue 2)	1.621	0.804	0.064
Mean of the two tissues	1.681	0.862	0.076%
RSD	5.1%	9.4%	22.8%

Comparison of Tissue Viability of the Main Test

For the test item and the positive control, the following percentage values of mean tissue viability were calculated in comparison to the mean of the negative controls:

Table 2‑a      % Tissue Viability

Incubation	Test Item	Positive Control
3 min	102.1%	16.7%
1 h	51.2%	4.5%

Additional tables:

Historical data:

In the following table, the means of the negative controls and positive controls of all performed experiments up to 16. Jun. 2021 are stated and compared with the values which were found in this study.

Table 3-a       Historical Data

Parameter	Optical Density Negative Control	Optical Density Negative Control	% Tissue viability Positive Control	% Tissue viability Positive Control
Incubation time	3 min.	1 h	3 min.	1 h
Mean	1.827	1.784	23.2 %	9.8 %
Standard Deviation	0.275	0.240	6.4 %	4 %
Range min-max	1.197 - 3.077	1.215 - 2.571	9.6 - 57.3 %	4.1 - 24.2 %
Range ± 2 SD	1.282 - 2.376	1.311 - 2.262	10.5 - 35.9 %	1.8 - 17.8 %
Study21062801G820	1.609	1.681	16.7%	4.5%

List of Proficiency Chemicals

In the following table the outcome of the proficiency chemical testing is stated.

All 12 proficiency chemicals were correctly predicted.

 

The demonstration of proficiency was performed under non-GLP conditions but within the GLP-environment at LAUS GmbH.

 

Table 4-a       Results of Proficiency Testing

Chemical Name	CAS No.	Prediction OECD 431 UN GHS Category	Findings LAUS GmbH
Bromoacetic acid	79-08-3	1A	1A
Boron trifluoride dihydrate	13319-75-0	1A	1A
Phenol	108-95-2	1A	1A
Dichloroacetyl chloride	79-36-7	1A	1A
Glyoxylic acid monohydrate	563-96-2	1B and 1C	1B and 1C
Lactic acid	598-82-3	1B and 1C	1B and 1C
Ethanolamine	141-43-5	1B and 1C	1B and 1C
Hydrochloric acid (14.4%)	7647-01-0	1B and 1C	1B and 1C
Phenethyl bromide	103-63-9	not corrosive	not corrosive
4-Amino-1,2,4-triazole	584-13-4	not corrosive	not corrosive
4-(methylthio)-benzaldehyde	3446-89-7	not corrosive	not corrosive
Lauric acid	143-07-7	not corrosive	not corrosive

Interpretation of results:

other: CLP criteria not met

Remarks:

non-corrosive

Conclusions:

Under the conditions of the RHE study, the test substance was considered to be non-corrosive to skin

Executive summary:

A study was conducted to determine the skin corrosion potential of the test substance in an in vitroReconstructed Human Epidermis (RHE) test,according to OECD Guideline 431 and EU Method B.40 bis in compliance with GLP.In the study, two tissues of the human skin model EpiDerm^TMwere treated with the test substancefor 3 minutes and 1 hour, respectively. The test substance was applied to each tissue and spread to match the tissue size. Demineralised water was used as negative control and 8 M KOH was used as positive control. After treatment, the respective substance was rinsed from the tissues. Then, cell viability of the tissues was evaluated by addition of MTT, which can be reduced to a blue formazan. Formazan production was evaluated by measuring the optical density (OD) of the resulting solution. Skin corrosion was expressed as the remaining cell viability after exposure to the test substance. After treatment with the negative control, the absorbance values were within the required acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for both treatment intervals thus showing that the quality of the tissues was adequate for the testing. The OD was 1.609 (3 minutes experiment) and 1.681 (1 h experiment). The positive control showed clear corrosive effects for both treatment intervals. The mean relative tissue viability value was reduced to 4.5% for the 1 h treatment. After 3 minutes and 1 h treatment with the test substance, the mean value of relative tissue viability was increased to 102.1 and 51.2% respectively. These values were above the respective thresholds for corrosion potential (i.e., 50 and 15%). Therefore, under the conditions of the RHE study, the test substance was considered to be non-corrosive to skin (Himmelsbach, 2021).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Reason / purpose for cross-reference:

data waiving: supporting information

Reason / purpose for cross-reference:

data waiving: supporting information

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin

Study1: A study was conducted to determine dermal irritation potential of the test substance, in anEpiSkin®in vitro irritation test, according to OECD 439, in compliance with GLP. Prior to the conduct of the irritation assay, a preliminary test was conducted to assess the intrinsic ability of the test substance to reduce methylthiazoldiphenyl-tetrazolium bromide (MTT) to formazan. The test substance did not reduce MTT to formazan. The dermal irritation potential was assessed by applying 10 μL of the test substance to the exposed surface of three EpiSkin tissues for 15 min. The surface area of the EpiSkin was 0.38 cm2, therefore the application rate was 26.3 μL/cm2. After the 15 min exposure period, the test substance was washed from the surface of the EpiSkin using Dulbecco’s phosphate-buffered saline (PBS). The EpiSkin tissues were then incubated for a recovery period of 42 h in a humidified incubator, set to maintain temperature and CO2 levels of 37°C and 5%, respectively. Following incubation, the tissues were transferred to assay medium containing MTT (0.3 mg/mL) and returned to the incubator for 3 h. Biopsies of the EpiSkin membranes were removed, added to acidified isopropanol, and refrigerated for ca 68 h in order to extract the formazan. The formazan production (cell viability) was assessed by measuring the optical density of the extracts at a wavelength of 550 nm. Three replicates of the positive control, aqueous sodium dodecyl sulphate (SDS) solution (5%, w/v), and the negative control, PBS, were tested in parallel to demonstrate the efficacy of the assay. The viability of each individual EpiSkin tissue was calculated as a percentage of the mean negative control viability (defined as 100%). Exposure to the test substance resulted in a mean EpiSkin viability of 9.56% ± 3.26% of the negative control value. Exposure to the positive control, aqueous SDS solution (5%, w/v), resulted in a mean EpiSkin viability of 13.36% of the negative control value. Under the conditions of the in vitroEpiSkin®study, the test substance was demonstrated to be an irritant to skin (Blackstock, 2013).

Study 2:

A study was conducted to determine the skin corrosion potential of the test substance in an in vitro Reconstructed Human Epidermis (RHE) test,according to OECD Guideline 431 and EU Method B.40 bis in compliance with GLP. In the study, two tissues of the human skin model EpiDerm^TM were treated with the test substancefor 3 minutes and 1 hour, respectively. The test substance was applied to each tissue and spread to match the tissue size. Demineralised water was used as negative control and 8 M KOH was used as positive control. After treatment, the respective substance was rinsed from the tissues. Then, cell viability of the tissues was evaluated by addition of MTT, which can be reduced to a blue formazan. Formazan production was evaluated by measuring the optical density (OD) of the resulting solution. Skin corrosion was expressed as the remaining cell viability after exposure to the test substance. After treatment with the negative control, the absorbance values were within the required acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for both treatment intervals thus showing that the quality of the tissues was adequate for the testing. The OD was 1.609 (3 minutes experiment) and 1.681 (1 h experiment). The positive control showed clear corrosive effects for both treatment intervals. The mean relative tissue viability value was reduced to 4.5% for the 1 h treatment. After 3 minutes and 1 h treatment with the test substance, the mean value of relative tissue viability was increased to 102.1 and 51.2% respectively. These values were above the respective thresholds for corrosion potential (i.e., 50 and 15%). Therefore, under the conditions of the RHE study, the test substance was considered to be non-corrosive to skin (Himmelsbach, 2021)

Eye:

No study was available for eye irritation. However, considering the surface-active nature of the test substance and the skin irritation results, the test substance can be considered to cause serious eye damage as a conservative approach and in accordance with Annex XI, Section 1.2 (weight of evidence) of the REACH regulation.

Justification for classification or non-classification

Based on the results of the in vitro skin irritation/corrosion studies, the test substance warrants a Skin Irrit. 2; H315: ‘Causes skin irritation’ classifications according to EU CLP (EC 1272/2008) criteria. Further based on the available weight of evidence and as a conservative approach, the test substance has been classified as Eye Damage 1; H318: 'Causes serious eye damage'.