N-(2-{[C16-18 (even numbered) alkanoyl]amino}ethyl)-N-(2-hydroxyethyl)[C16-18 (even numbered) alkylamide

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31 July 2017 - 27 February 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study was 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:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

July 2015

Deviations:

no

Test guideline 2

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 material

Test material information

Constituent 1

Reference substance name:

Fatty Acids, C14 and C16 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Molecular formula:

C34H68N2O3

IUPAC Name:

Fatty Acids, C14 and C16 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Constituent 2

Reference substance name:

Fatty Acids, C15 and C16 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Molecular formula:

C35H70N2O3

IUPAC Name:

Fatty Acids, C15 and C16 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Constituent 3

Reference substance name:

Fatty Acids, C16 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Molecular formula:

C36H72N2O3

IUPAC Name:

Fatty Acids, C16 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Constituent 4

Reference substance name:

Fatty Acids, C15 and C18 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Molecular formula:

C37H74N2O3

IUPAC Name:

Fatty Acids, C15 and C18 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Constituent 5

Reference substance name:

Fatty Acids, C16 and C18 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Molecular formula:

C38H76N2O3

IUPAC Name:

Fatty Acids, C16 and C18 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Constituent 6

Reference substance name:

Fatty Acids, C17 and C18 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Molecular formula:

C39H78N2O3

IUPAC Name:

Fatty Acids, C17 and C18 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Constituent 7

Reference substance name:

Fatty Acids, C18 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Molecular formula:

C40H80N2O3

IUPAC Name:

Fatty Acids, C18 sat., bisamides with 2-[(2-aminoethyl)amino]ethanol

Constituent 8

Reference substance name:

Stearic acid

EC Number:

200-313-4

EC Name:

Stearic acid

Cas Number:

57-11-4

Molecular formula:

C18H36O2

IUPAC Name:

stearic acid

Constituent 9

Reference substance name:

Palmitic acid

EC Number:

200-312-9

EC Name:

Palmitic acid

Cas Number:

57-10-3

Molecular formula:

C16H32O2

IUPAC Name:

palmitic acid

Constituent 10

Reference substance name:

Myristic acid

EC Number:

208-875-2

EC Name:

Myristic acid

Cas Number:

544-63-8

Molecular formula:

C14H28O2

IUPAC Name:

myristic acid

Constituent 11

Reference substance name:

2-(2-aminoethylamino)ethanol

EC Number:

203-867-5

EC Name:

2-(2-aminoethylamino)ethanol

Cas Number:

111-41-1

Molecular formula:

C4H12N2O

IUPAC Name:

2-(2-aminoethylamino)ethanol

Test material form:

solid

Details on test material:

Identification: N-(2-{[C16-18 (EVEN NUMBERED) ALKANOYL]AMINO}ETHYL)-N-(2-HYDROXYETHYL)[C16-18 (EVEN NUMBERED) ALKYLAMIDE
Storage Conditions: 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: At room temperature (in the dark*)
- Stability under test conditions: Assumed stable
- Solubility and stability of the test substance in the solvent/vehicle: Not applicable - test item applied undiluted.
- 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: None, applied as supplied.
- Preliminary purification step (if any): n/a
- Final dilution of a dissolved solid, stock liquid or gel: n/a
- Final preparation of a solid: n/a

FORM AS APPLIED IN THE TEST (if different from that of starting material): n/a

OTHER SPECIFICS: n/a

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

other: epidermal keratinocytes

Cell source:

other: MatTek Corporation, Bratislava, Slovakia

Source strain:

other: N/A

Details on animal used as source of test system:

N/A

Justification for test system used:

N/A

Vehicle:

unchanged (no vehicle)

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): approximately 25 mg (~ 39 mg/cm2 according to guideline) of the test item were applied to the tissues, wetted with 25 μL DPBS prior to application.
- Concentration (if solution): See above.

VEHICLE
- Amount(s) applied (volume or weight with unit): See above.
- Concentration (if solution): See above.
- Lot/batch no. (if required): 062717
- Purity: Not reported.

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL
- Concentration (if solution): n/a

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL
- Concentration (if solution): 5 % w/v

Duration of treatment / exposure:

60 mins

Duration of post-treatment incubation (if applicable):

42 hours and 20 minutes

Number of replicates:

3

Test system

Type of coverage:

other: Not applicable

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Value:

ca. 86.2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Any other information on results incl. tables

Table 2. Results after treatment with

N-(2-{[C16-18 (even numbered) alkanoyl]amino}ethyl)-N-(2-hydroxyethyl)[C16-18 (even numbered) alkylamide

and the controls.

Dose Group	Tissue No.	Absor-bance 570 nm Well 1	Absor-bance 570 nm Well 2	Absor-bance 570 nm Well 3	Mean Absor-bance of 3 Wells	Mean Absor-bance of three wells blank corrected	Mean Absor-bance of 3 tissues after blank correction	Rel. Absor-bance [%] Tissue 1, 2 + 3*	Relative Standard Deviation [%]	Mean Rel. Absorbance [%]**
Blank		0.039	0.038	0.044	0.040	0.000
Negative Control	1	1.396	1.399	1.398	1.398	1.357	1.495	90.8	8.1	100.0
	2	1.580	1.593	1.581	1.584	1.544		103.3
	3	1.631	1.626	1.612	1.623	1.583		105.9
Positive Control	1	0.103	0.105	0.104	0.104	0.064	0.063	4.3	3.6	4.2
	2	0.104	0.105	0.105	0.104	0.064		4.3
	3	0.100	0.101	0.100	0.100	0.060		4.0
Test Item	1	1.383	1.397	1.384	1.388	1.348	1.288	90.2	5.1	86.2
	2	1.255	1.273	1.249	1.259	1.219		81.5
	3	1.348	1.327	1.342	1.339	1.299		86.9

OD= Optical Density

SD= Standard deviation

*= The mean viability of the negative control tissues is set at 100 %

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study the test substance is not considered to be irritant to the skin and does not meet the criteria for classification in accordance with UN GHS and EU CLP regulation.

Executive summary:

OECD 439 (2017) -The skin irritation potential of

N-(2-{[C16-18 (even numbered) alkanoyl]amino}ethyl)-N-(2-hydroxyethyl)[C16-18 (even numbered) alkylamide

was assessed using an EpiDerm Reconstructed Human Epidermis (RHE) Model Kit in accordance with OECD guidance 439 and GLP.

Triplicate tissues were exposed to the test item for 60 minutes. At the end of the exposure period the test item was rinsed and the tissues incubated for a further 42 h in the presence of maintenance solution which would be used for possible inflammatory mediator determination. Each tissue was then loaded with MTT. After incubation and extraction, the solutions were 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 60 minutes were 86.2 %. It was considered unnecessary to perform IL-1a analysis as the results of the MTT test were unequivocal. The quality criteria required for acceptance of the results was met.

Under the conditions of this study the test substance,

N-(2-{[C16-18 (even numbered) alkanoyl]amino}ethyl)-N-(2-hydroxyethyl)[C16-18 (even numbered) alkylamide

is not irritant to skin according to UN GHS and EU CLP regulation.