Sulfonic acids, C14-18-alkane hydroxy and C12-20-alkapolyene and C14-18-alkene and C12-20-alkene hydroxy, sodium salts

Endpoint summary

Description of key information

Skin irritation (Reconstructed Human Epidermis Model, 15-minute exposure): non-irritant to the skin.

Skin corrostion (Reconstructed Human Epidermis Model, 3-minute and 60-minute exposure): non-corrosive to the skin.

Eye irritation (Eyes from adult cattle, 240-minute exposure): Category 1. UN GHS or EU CLP Causes serious eye damage

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 August 2017 - 11 August 2017

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 (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

other: Reconstructed Human Epidermis

Cell source:

other: not specified

Source strain:

not specified

Justification for test system used:

This model incorporates several features, which make it advantageous in the study of potential dermal corrosivity. The target cells are epithelial, derived from human skin, and formed into a stratified, cornified epithelium. Test items are applied to the culture surface, at the air interface, so that undiluted and/or end use dilutions can be tested directly.

Vehicle:

not specified

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ Reconstructed Human Epidermis Model Kit
- Tissue batch number(s): 25835
- Delivery date: 08 August 2017
- Date of initiation of testing: 09 August 2017

- Temperature used during treatment / exposure: 37 °C,
- MTT concentration: 1.0 mg/mL MTT solution
- Incubation time: 60 minutes
- Spectrophotometer: Labtech LT-4500 microplate reader
- Wavelength: Absorbency at 570nm (OD570)

- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg of the test item
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL of sterile distilled water
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL of 8.0 N Potassium Hydroxide

Duration of treatment / exposure:

60 minutes and 3 minutes

Duration of post-treatment incubation (if applicable):

3 hours

Number of replicates:

two

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minutes

Value:

ca. 91

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 minutes

Value:

ca. 84.1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- Direct-MTT reduction: test item did not reduce MTT
- Colour interference with MTT: none

- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes

Interpretation of results:

GHS criteria not met

Conclusions:

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:

16 August 2017 - 28 August 2017

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

other: EPISKIN™ Reconstructed Human Epidermis

Cell source:

other: not specified

Source strain:

not specified

Details on animal used as source of test system:

not applicable

Justification for test system used:

Following a full validation study the EpiSkinTM reconstructed human epidermis model showed evidence of being a reliable and relevant stand-alone test for predicting rabbit skin irritation when the endpoint is measured by MTT reduction and for being used as a replacement for the Draize Skin Irritation Test for the purpose of distinguishing between Irritating and Non-Irritating test items.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN™ Reconstructed Human Epidermis
- Tissue batch number(s): 17-EKIN-034
- Delivery date: 22 August 2017
- Maintenance Medium lot number: 17-MAIN3-035
- Assay Medium lot number: 17-ESSC-033
- Date of initiation of testing: 16 August 2017

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

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: DPBS with Ca++ and Mg++.

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 hours
- Spectrophotometer: Labtech LT-4500 microplate reader
- Wavelength: 570 nm
- Filter: no filter

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mg (26.3 mg/cm2)
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 10 μL of DPBS
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 10 μL of SDS 5% w/v

Duration of treatment / exposure:

15 minutes

Duration of post-treatment incubation (if applicable):

42 h

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

test item

Value:

ca. 81.1

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- Direct-MTT reduction: test item did not directly reduce MTT.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes

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.
UN GHS Not classified for Irritation (category 3 can not be determined).

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:

03 October 2017 - 03 October 2017

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 (incl. QA statement)

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

- Source: local abbatoir

Vehicle:

physiological saline

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 20% w/v in sodium chloride 0.9% w/v

Duration of treatment / exposure:

240 minutes

Duration of post- treatment incubation (in vitro):

none

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS : eyes 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.

QUALITY CHECK OF THE ISOLATED CORNEAS: macroscopically examined

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED : yes

POSITIVE CONTROL USED : yes

APPLICATION DOSE AND EXPOSURE TIME : 20% w/v in sodium chloride 0.9% w/v for 240 minutes

R insed three times with fresh complete EMEM 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.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: yes
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of [UV/VIS spectrophotometry / microtiter plate reader] (OD490) yes

SCORING SYSTEM: In Vitro Irritancy Score (IVIS) : yes

Irritation parameter:

cornea opacity score

Run / experiment:

Mean corrected value

Value:

ca. 22.7

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

other: Permeability (OD)

Run / experiment:

Mean permeability

Value:

ca. 2.903

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Value:

ca. 66.2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: The corneas treated with the test item were cloudy 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



ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

Interpretation of results:

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

Conclusions:

Category 1. UN GHS or EU CLP Causes serious eye damage.

Executive summary:

The opacity and in Vitro Irritancy Score were found to be 22.7 and 66.2, respectively. In accordance with the Regulation (EC) 1272/2008 Appendix 1 (table 3.3.1), the substance will be classified as causing serious eye damage Category 1. The applicable hazard phrase is “H318: Causes serious eye damage”.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

A Skin Irritation study (Envigo Research Limited, 2018, study code FF02HX) was conducted to determine the skin irritation potential of Sulfonic acids, C14-18-alkane hydroxy and C12-20-alkapolyene and C14-18-alkene and C12-20-alkene hydroxy, sodium salts. The study was performed according to OECD test guideline 439 and EC method B46; the study was performed in compliance with GLP.

The effects of 15-minute dermal exposure followed by a post-exposure incubation period of 42 hours to Sulfonic acids, C14-18-alkane hydroxy and C12-20-alkapolyene and C14-18-alkene and C12-20-alkene hydroxy, sodium salts were investigated in EPISKIN reconstructed human epidermis model.

On the basis that test item did not directly reduce MTT, Sulfonic acids, C14-18-alkane hydroxy and C12-20-alkapolyene and C14-18-alkene and C12-20-alkene hydroxy, sodium salts was classified as non-irritant to skin.

 

A Skin Corrosion study (Envigo Research Limited, 2018, study code HF48BF) was conducted to determine the skin corrosion potential of Sulfonic acids, C14-18-alkane hydroxy and C12-20-alkapolyene and C14-18-alkene and C12-20-alkene hydroxy, sodium salts. The study was performed according to OECD test guideline 431 and EC method B.40bis; the study was performed in compliance with GLP.

The effects of 3-minute and 60-minute dermal exposure to Sulfonic acids, C14-18-alkane hydroxy and C12-20-alkapolyene and C14-18-alkene and C12-20-alkene hydroxy, sodium salts were investigated in EPISKIN reconstructed human epidermis model.

On the basis that test item did not directly reduce MTT, Sulfonic acids, C14-18-alkane hydroxy and C12-20-alkapolyene and C14-18-alkene and C12-20-alkene hydroxy, sodium salts was classified as non-corrosive to skin.

 

An Eye Irritation study (Envigo Research Limited, 2018, XD76KK) was conducted to assess the effect of the instillation of Sulfonic acids, C14-18-alkane hydroxy and C12-20-alkapolyene and C14-18-alkene and C12-20-alkene hydroxy, sodium salts into the eyes from adult cattle. The study was performed according to OECD test guideline 437 and EC method B47; the study was performed in compliance with GLP.

Sulfonic acids, C14-18-alkane hydroxy and C12-20-alkapolyene and C14-18-alkene and C12-20-alkene hydroxy, sodium salts (concentration 20% w/v in sodium chloride 0.9% w/v) was applied to three corneas for 240 minutes.

 

On vitro irritation score 66.2;

Permeability (OD) (mean permeability) score 2.903;

Cornea opacity score (mean corrected value) 22.7

 

Effects on skin irritation/corrosion: non-irritant/ non-corrosive to the skin

Effects on eye irritation: irritating (Category 1. UN GHS or EU CLP Causes serious eye damage)

Justification for classification or non-classification

The opacity and in Vitro Irritancy Score were found to be 22.7 and 66.2, respectively. In accordance with the Regulation (EC) 1272/2008 Appendix 1 (table 3.3.1), the substance will be classified as causing serious eye damage Category 1. The applicable hazard phrase is “H318: Causes serious eye damage”.

The substance was predicted to be non-corrosive and non-irritant to the skin; on this basis no classification for skin corrosion or irritation will be applied.