Octadecan-1-ol, ethoxylated, phosphates

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1 October 2018 - 8 July 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Octadecan-1-ol, ethoxylated, phosphates

EC Number:

500-155-9

EC Name:

Octadecan-1-ol, ethoxylated, phosphates

Cas Number:

62362-49-6

Molecular formula:

Variable

IUPAC Name:

Octadecan-1-ol, ethoxylated, phosphates

Specific details on test material used for the study:

Purity: 100% (UVCB)
Appearance/Physical state: Solid, white pastille
Storage: Room temperature in the dark

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

other: reconstructed human epidermis tissues

Cell source:

other: Not specified

Vehicle:

unchanged (no vehicle)

Details on test system:

Duplicate tissues were treated with the test item for exposure periods of 3 and 60 minutes. Negative and positive control groups were treated for each exposure period. At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTT loading. After MTT-loading each tissue was placed in 2 mL of Isopropanol for MTT extraction.

At the end of the formazan extraction period each well was mixed thoroughly and triplicate 200 L samples were transferred to the appropriate wells of a pre-labeled 96 well plate. The optical density (OD) was measured at 570 nm (OD570).

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

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

25 mg of the test item was applied to the corresponding tissues in duplicate.

Duration of treatment / exposure:

3 and 60 minutes

Duration of post-treatment incubation (if applicable):

3 hours

Number of replicates:

Two

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minute exposure

Value:

ca. 98.3

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 minute exposure

Value:

ca. 90.8

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The mean OD570 for the negative control treated tissues was 1.962 for the 3 mMinute exposure period and 1.828 for the 60 mMinute exposure period. The negative control acceptance criteria were therefore satisfied.

The relative mean tissue viability for the positive control treated tissues was 3.5% following the 3 minute exposure and 4.2% following the 60-minute exposure relative to the negative control. The positive control acceptance criterion was therefore satisfied.

The coefficients of variation between the two tissue replicates of the test item and negative control groups were 0.2 – 10.3%. The acceptance criterion was therefore satisfied. In the range 20 to 100% viability, coefficient of variation between the two tissue replicates of each treatment group did not exceed was less than 30%. The acceptance criterion was therefore satisfied.

Any other information on results incl. tables

The MTT solution containing the test item did not turn blue/purple. This was taken to indicate the test item did not reduce MTT.

The solution containing the test item did not become colored. This was taken to indicate the test item did not have the potential to cause color interference.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was classified as non-irritant. The following classification criteria apply: EU CLP Not classified for Irritation and UN GHS Not classified for Irritation.

Executive summary:

The purpose of this test is to evaluate the corrosivity potential of the test item using the EpiDerm™ Human Skin Model after treatment periods of 3 and 60 minutes.

Corrosion is directly related to cytotoxicity in the EpiDerm™ tissue. Cytotoxicity is determined by the reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to formazan by viable cells in the test item treated tissues relative to the corresponding negative control. The results are used to make a prediction of the corrosivity potential of the test item.

Duplicate tissues were treated with the test item for exposure periods of 3 and 60 minutes. Negative and positive control groups were treated for each exposure period. At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTT‑loading. After MTT-loading each tissue was placed in 2 mL of Isopropanol for MTT extraction.

At the end of the formazan extraction period each well was mixed thoroughly and triplicate 200 mL samples were transferred to the appropriate wells of a pre-labeled 96‑well plate. The optical density (OD) was measured at 570 nm (OD₅₇₀).

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

The test item did not reduce MTT and did not have the potential to cause color interference.

The relative mean percentage viability for the test item treated tissues was 98.3% following the 3 minute exposure period and 90.8% following the 60 minute exposure period relative to the negative control. The criterion for non-corrosive classification (relative mean percentage tissue viability ≥ 50% after 3 minutes and ≥ 15% after 60 minutes of exposure) was, therefore, met.

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

The test item was considered to be non-corrosive to the skin.