Octadecan-1-ol, ethoxylated, phosphates

Endpoint summary

Description of key information

Skin irritation

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.

Eye irritation

Based on the results of the Bovine Corneal Opacity and Permeability (BCOP) test, the test item is not an irritant to the eye.  No category was assigned and the test item does not require classification under UN GHS or EU CLP.

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:

1 October 2018 - 8 July 2019

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)

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

Specific details on test material used for the study:

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

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

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

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.

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.

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.

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:

3 October 2018 to 30 August 2019

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)

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)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

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

Species:

cattle

Details on test animals or tissues and environmental conditions:

Eyes from adult cattle (typically 12 to 60 months old) were obtained from a local abattoir as a by-product from freshly slaughtered animals. The eyes were excised by an abattoir employee after slaughter, and were placed in Hanks’ Balanced Salt Solution (HBSS) supplemented with antibiotics (penicillin at 100 IU/mL and streptomycin at 100 µg/mL). They were transported to the test facility over ice packs on the same day of slaughter. The corneas were prepared immediately on arrival.

Vehicle:

other: For the purpose of this study the test item was prepared as a 10% w/v solution in sodium chloride 0.9% w/v.

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

For the purpose of this study the test item was prepared as a 10% w/v solution in sodium chloride 0.9% w/v.

The test item was formulated within 2 hours of being applied to the test system. It is assumed that the formulation was 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.

Duration of treatment / exposure:

The EMEM was removed from the anterior chamber of the BCOP holder and 0.75 mL of the formulated test item or control items were applied to the appropriate corneas. The holders were gently tilted back and forth to ensure a uniform application of the item over the entire cornea. Each holder was incubated, anterior chamber uppermost, at 32 +/- 1 deg.C for 10 minutes.

Duration of post- treatment incubation (in vitro):

At the end of the exposure period the test item and control items were removed from the anterior chamber and the cornea was rinsed three times with fresh complete MEM containing phenol red before a final rinse with complete EMEM without phenol red. The anterior chamber was refilled with fresh complete EMEM without phenol red. A post treatment opacity reading was taken and each cornea was visually observed.

The holders were incubated, anterior chamber facing forward, at 32 +/- 1 deg.C for 120 minutes.

Number of animals or in vitro replicates:

Three

Details on study design:

The test item, a surfactant solid, was applied at a concentration of 10% w/v in sodium chloride 0.9% w/v for 10 minutes followed by an incubation period of 120 minutes. Negative (Sodium Chloride 0.9% w/v) and positive (Ethanol) 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).

Irritation parameter:

cornea opacity score

Run / experiment:

1

Value:

ca. 2.6

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The corneas treated with the test item were clear post treatment. The corneas treated with the negative control item were clear post treatment. The corneas treated with the positive control item were cloudy post treatment and post incubation.

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the Bovine Corneal Opacity and Permeability (BCOP) test, the test item is not an irritant to the eye. No category was assigned and the test item does not require classification under UN GHS or EU CLP.

Executive summary:

The purpose of this test was to identify the potential for the test item, Octadecan-1-ol, ethoxylated, phosphates (CAS RN 62362-49-6), to induce serious eye damage and to determine whether the test item requires classification for eye irritation or serious eye damage. The Bovine Corneal Opacity and Permeability (BCOP) test method is an organotypic model that provides short‑term maintenance of normal physiological and biochemical function of the bovine corneain vitro. In this test method, damage by the test item is assessed by quantitative measurements of changes in corneal opacity and permeability.

The test method can correctly identify test items (both chemicals and mixtures) inducing serious eye damage as well as those not requiring classification for eye irritation or serious eye damage, as defined by the United Nations (UN) Globally Harmonized System of Classification and Labelling of Items (GHS) and EU Classification, Labelling and Packaging (CLP) of chemicals (Regulation (EC) No 1272/2008), and it was therefore endorsed as scientifically valid for both purposes. Test items inducing serious eye damage are classified as UN GHS and EU CLP Category 1. Items not classified for eye irritation or serious eye damage are defined as those that do not meet the requirements for classification as UN GHS/EU CLP Category 1 or 2 (2A or 2B), i.e. they are referred to as UN GHS/EU CLP No Category.

The test item, a surfactant solid, was applied at a concentration of 10% w/v in sodium chloride 0.9% w/v for 10 minutes followed by an incubation period of 120 minutes. Negative (Sodium Chloride 0.9% w/v) and positive (Ethanol) 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 anIn VitroIrritancy Score (IVIS). 

Based on the results of theBovine Corneal Opacity and Permeability (BCOP)test, the test item,Octadecan-1-ol, ethoxylated, phosphates (CAS RN 62362-49-6), is not an irritant to the eye. No category was assigned and the test item does not require classification under UN GHS or EU CLP.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Skin irritation

The test item was classified as non-irritant.

Eye irritation

Based on the results of the Bovine Corneal Opacity and Permeability (BCOP) test, the test item is not an irritant to the eye.