Amides, tall-oil fatty, N,N'-(iminodi-2,1-ethanediyl)bis-, maleated

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12-06-2013 to 17-06-2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study performed according to the guideline and under GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

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)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Principles of method if other than guideline:

exposure period 15 minutes with post-exposure incubation period of 42 hours

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

(2Z)-3-({2-[(8E)-9-(1,3-dioxo-7-pentyl-1,3,3a,4,7,7a-hexahydro-2-benzofuran-4-yl)non-8-enamido]ethyl}({2-[(9E)-octadec-9-enamido]ethyl})carbamoyl)prop-2-enoic acid; (2Z)-3-[bis({2-[(8E)-9-(1,3-dioxo-7-pentyl-1,3,3a,4,7,7a-hexahydro-2-benzofuran-4-yl)non-8-enamido]ethyl})carbamoyl]prop-2-enoic acid; (2Z)-3-[bis({2-[(9E)-octadec-9-enamido]ethyl})carbamoyl]prop-2-enoic acid

EC Number:

800-430-6

Cas Number:

1419212-76-2

Molecular formula:

ca. C44H75N3O5 - C48H81N305

IUPAC Name:

(2Z)-3-({2-[(8E)-9-(1,3-dioxo-7-pentyl-1,3,3a,4,7,7a-hexahydro-2-benzofuran-4-yl)non-8-enamido]ethyl}({2-[(9E)-octadec-9-enamido]ethyl})carbamoyl)prop-2-enoic acid; (2Z)-3-[bis({2-[(8E)-9-(1,3-dioxo-7-pentyl-1,3,3a,4,7,7a-hexahydro-2-benzofuran-4-yl)non-8-enamido]ethyl})carbamoyl]prop-2-enoic acid; (2Z)-3-[bis({2-[(9E)-octadec-9-enamido]ethyl})carbamoyl]prop-2-enoic acid

Test material form:

other: gel

Details on test material:

- Substance type: UVCB
- Physical state: paste
- Purity: 100% (UVCB)
- Storage condition of test material: room temperature in dark

In vitro test system

Test system:

human skin model

Control samples:

yes, concurrent negative control

Test system

Details on study design:

The EpiSKIN assay uses reconstructed human epidermis to assess skin irritation. For the viability test enzymatic conversion of MTT to formazan is used.

In a pre-test direct interaction of the test substance with the detection chemical MTT is assessed.
In the main study triplicate tissues were treated with 30 uL of the test substance during 15 minutes. In addition a negative control (DPBS) and a positive control (5% SDS) were included. After treatment tissues were rinsed, incubated in maintenance fluid for 42 hours at 37 °C and treated with MTT during 3 hours (at 37 °C). After MTT loading a total biopsy of each epidermis (epidermis and collagen) was made, that was suspended in acidified iso-propanol for 3 days and kept in the refrigerator at 1-10°C. Thereafter the optical density of the extracted MTT solutions was measured at 562 nm. Viability was expressed as percentage MTT conversion versus negative control.
Tissue viability > 50% means non-irritant
Tissue viability ≤ 50% means irritant

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Value:

81.8

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

rel viability 10.6%

In vivo

Other effects:

The test substance did not interfere with the reduction of MTT to blue formazan salt.

Any other information on results incl. tables

	Mean OD562 ± SD	Relative mean viability ± SD
Negative control:	0.946 ± 0.119	100 ± 12.6%
Test substance:	0.774 ± 0.021	81.8 ± 2.2%
SDS:	0.100 ± 0.014	10.6 ± 1.5%

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance is non-irritant in the EPISKIN reconstructed human epidermal model.
Criteria used for interpretation of results: other: tissue viability > 50%: non-irritant

Executive summary:

In this assay triplicate reconstructed human skin tissues were treated with 30 uL of the test substance during 15 minutes. After treatment tissues were rinsed, kept for 42 hours and treated with MTT. Optical density of extracted MTT solutions was measured at 562 nm. Viability was expressed as percentage MTT conversion versus negative control. The viability was 81.8%, which is indicative for non-irritant substances.

The test substance is non-irritant in the EPISKIN reconstructed human epidermal model.