Amines, C12-14-branched alkyl, dodecylbenzenesulfonates (1:1)

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 February - 02 August 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study conducted in accordance with international guidelines and in accordance with GLP. All guideline validity criteria were met.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

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)

Version / remarks:

29 July 2016

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Council Regulation (EC) No 440/2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Amines, C12-14-branched alkyl, octylbenzenesulfonates (1:1)

Molecular formula:

Amines, C12-branched alkyl, octylbenzenesulfonates: C27H51NO3S Amines, C13-branched alkyl, octylbenzenesulfonates: C28H53NO3S Amines, C14-branched alkyl, octylbenzenesulfonates: C29H55NO3S

IUPAC Name:

Amines, C12-14-branched alkyl, octylbenzenesulfonates (1:1)

Constituent 2

Reference substance name:

Amines, C12-14-branched alkyl, nonylbenzenesulfonates (1:1)

Molecular formula:

Amines, C12-branched alkyl, nonylbenzenesulfonates: C28H53NO3S Amines, C13-branched alkyl, nonylbenzenesulfonates: C29H55NO3S Amines, C14-branched alkyl, nonylbenzenesulfonates: C30H57NO3S

IUPAC Name:

Amines, C12-14-branched alkyl, nonylbenzenesulfonates (1:1)

Constituent 3

Reference substance name:

Amines, C12-14-branched alkyl, decylbenzenesulfonates (1:1)

Molecular formula:

Amines, C12-branched alkyl, decylbenzenesulfonates: C29H55NO3S Amines, C13-branched alkyl, decylbenzenesulfonates: C30H57NO3S Amines, C14-branched alkyl, decylbenzenesulfonates: C31H59NO3S

IUPAC Name:

Amines, C12-14-branched alkyl, decylbenzenesulfonates (1:1)

Constituent 4

Reference substance name:

Amines, C12-C14-branched alkyl, benzenesulfonates C12-C15 (1:1)

Molecular formula:

Amines, C12-branched alkyl, benzenesulfonates C12 (1:1) C31H59NO3S Amines, C12-branched alkyl, benzenesulfonates C13 (1:1) C32H61NO3S Amines, C12-branched alkyl, benzenesulfonates C14 (1:1) C33H63NO3S Amines, C12-branched alkyl, benzenesulfonates C15 (1:1) C34H65NO3S Amines, C13-branched alkyl, benzenesulfonates C12 (1:1) C32H61NO3S Amines, C13-branched alkyl, benzenesulfonates C13 (1:1) C33H63NO3S Amines, C13-branched alkyl, benzenesulfonates C14 (1:1) C34H65NO3S Amines, C13-branched alkyl, benzenesulfonates C15 (1:1) C35H67NO3S Amines, C14-branched alkyl, benzenesulfonates C12 (1:1) C33H63NO3S Amines, C14-branched alkyl, benzenesulfonates C13 (1:1) C34H65NO3S Amines, C14-branched alkyl, benzenesulfonates C14 (1:1) C35H67NO3S Amines, C14-branched alkyl, benzenesulfonates C15 (1:1) C36H69NO3S

IUPAC Name:

Amines, C12-C14-branched alkyl, benzenesulfonates C12-C15 (1:1)

Constituent 5

Reference substance name:

Amines, C12-14-branched alkyl, hexadecylbenzenesulfonates (1:1)

Molecular formula:

Amines, C12-branched alkyl, hexadecylbenzenesulfonates (1:1): C35H67NO3S Amines, C13-branched alkyl, hexadecylbenzenesulfonates (1:1): C36H69NO3S Amines, C14-branched alkyl, hexadecylbenzenesulfonates (1:1): C37H71NO3S

IUPAC Name:

Amines, C12-14-branched alkyl, hexadecylbenzenesulfonates (1:1)

Constituent 6

Reference substance name:

Residual Amines, C12-14-tert-alkyl

IUPAC Name:

Residual Amines, C12-14-tert-alkyl

Test material form:

liquid: viscous

Details on test material:

Storage: Room Temperature

Specific details on test material used for the study:

RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, in darkness
- Stability under test conditions: Assumed stable
- Solubility and stability of the test substance in the solvent/vehicle: N/A
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: N/A
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: Applied unchanged.
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material) : Applied as supplied

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable) N/A

OTHER SPECIFICS: N/A

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Three-dimensional human skin model, comprising a reconstructed epidermis with a functional stratum corneum supplied by In Vitro Life Science Laboratories, Bratislava, Slovak Republic

Details on animal used as source of test system:

N/A

Justification for test system used:

Guideline specific test system.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: MatTek, In Vitro Life Science Laboratories, Bratislava, Slovak Republic
- Tissue batch number(s): 23395
- Production date: 22 Feb 2017
- Shipping date: 22 Feb 2017
- Delivery date: 22 Feb 2017
- Date of initiation of testing: 22 Feb 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ºC
- Temperature of post-treatment incubation (if applicable): 37 ºC

REMOVAL OF TEST MATERIAL AND CONTROLS - Volume and number of washing steps: the tissues were washed with phosphate buffered saline (PBS) to remove residual material. The rinsed tissues were kept in 24-well plates (holding plates) until all tissues were dosed and rinsed
- Observable damage in the tissue due to washing: No
- Modifications to validated SOP: No

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL
- Incubation time: 3 h
- Spectrophotometer: Not reported
- Wavelength: 570 nm
- Filter: Not reported
- Filter bandwidth: Not reported
- Linear OD range of spectrophotometer: Not reported

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: Pass - details provided in CoA (OD540-570 was between 1.0 and 3.0).
- Barrier function: Pass - details provided in CoA (ET-50 was between 4.77 and 8.72 h).
- Morphology: Not reported
- Contamination: Pass - details provided in CoA (no contamination - tested for HIV-1-virus; Hep B; Hep C; bacteria; yeast and other fungi).
- Reproducibility: Coefficient of Variation between tissue replicates was ≤ 30%.

NUMBER OF REPLICATE TISSUES: 4 per test group (2 each for 3 min and 60 min exposure)

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
The test item did not reduce MTT.

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- 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): 50 µL
- Concentration (if solution): Unchanged/ undiluted

VEHICLE
- Amount(s) applied (volume or weight with unit): N/A
- Concentration (if solution): N/A
- Lot/batch no. (if required): N/A
- Purity: N/A

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL
- Concentration (if solution): Purified water

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL
- Concentration (if solution): 8 M

Duration of treatment / exposure:

3 mins and 60 mins

Duration of post-treatment incubation (if applicable):

3 h MTT incubation followed by overnight isopropnaol extraction.

Number of replicates:

2 per treatment and exposure group

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minute exposure

Value:

86.3

Vehicle controls validity:

not applicable

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:

23.1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: No
- Direct-MTT reduction: No
- Colour interference with MTT: No

DEMONSTRATION OF TECHNICAL PROFICIENCY: Not reported

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD570 for the negative control treated tissues was 1.357 for the 3-Minute exposureperiod and 1.338 for the 60-Minute exposure period. The negative control acceptance criteria were therefore satisfied.
- Acceptance criteria met for positive control: The relative mean tissue viability for the positive control treated tissues was 6.6 and 8.0 % relative to the negative control following the 3 and 60-,minute exposure periods, respectively. The positive control acceptance criterion was therefore satisfied.
- Acceptance criteria met for variability between replicate measurements: In the range 20 to 100% viability the Coefficient of Variation between the two tissue replicates of each treatment group did not exceed 30%. The acceptance criterion was therefore satisfied.
- Range of historical values if different from the ones specified in the test guideline: All values for the positive and negative control were within the historical ranges achieved by the testing facility in previous tests (Feb 2008 - Feb 2017, n=46), thus confirming the acceptable functioning of the test system.

Any other information on results incl. tables

Table 1       Relative mean viabilities for each treatment group

 

Exposure Period	Percentage Viability (% SD / % CoV)^*
	Negative Control^§	Positive Control	Test Item
3 minutes	100 (0.037 / 2.7)	6.6 (0.017 / 4.9)	86.3 (0.035 / 2.9)
60 minutes	100 (0.096 / 7.2)	8.0 (0.017 / 7.4)	23.1 (0.025 / 8.2)

^*mean of 2 replicates

^§negative controls set to 100 %

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

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

Executive summary:

OECD 431 (2017) - The skin corrosivity potential of reaction mass of Amines, C12-14-branched alkyl, dodecylbenzenesulfonates (1:1) was assessed using an EpiDerm Reconstructed Human Epidermis (RHE) Model Kit in accordance with OECD guidance 431 and GLP.

 

Duplicate tissues were treated with the test item for exposure periods of 3 and 60 mins. At the end of the exposure period the test item was rinsed from each tissue before being loaded with MTT. After MTT loading each tissue was placed in 2 mL isopropanol for MTT extraction. After extraction, each tissue was pierced and the extraction solution aliquoted for absorbance measurements. Absorbency at 570 nm of each well was measured using a spectrophotometer.

 

Mean viability of tissues exposed to the test substance after 3 and 60 minutes were 86.3 and 23.1 %, respectively. The quality criteria required for acceptance of the results was met.

 

Under the conditions of this study the test substance is not considered to be corrosive to the skin.