Amines, C16-18-alkyl, acetates

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-05-09 to 2012-05-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany

Test material

Test material information

Constituent 1

Reference substance name:

C16-18-(even numbered)-alkylamines acetates

EC Number:

800-526-8

Cas Number:

1273322-45-4

Molecular formula:

R-NH2xHOOCCH3 R = alkyl, mainly C16-18-(even numbered)

IUPAC Name:

C16-18-(even numbered)-alkylamines acetates

Details on test material:

Name: C16-18-(even numbered)-alkylamines acetates
CAS No: 1273322-45-4
Physical State at RT: solid
Colour: white-yellow
pH: 5.7 (1% a.i. in water)
Melting Point: 70-72 °C
Storage Conditions: at room temperature

In vitro test system

Test system:

human skin model

Amount/concentration applied:

solids: 25 µL DPBS + 25 mg

Duration of treatment / exposure:

60 +/- 1 minutes

Duration of post-treatment incubation (if applicable):

42 +/- 4 h

Number of replicates:

3

Test animals

Species:

other: EpiDerm reconstructed human epidermis model (MatTek Corp.)

Details on test animals or test system and environmental conditions:

organotypic reconstructed three-dimensional model of the human epidermis

Test system

Type of coverage:

other: direct application

Controls:

other: three tissues for negative control (30µL DPBS) and positive control (30µL 5% SDS)

Details on study design:

3 replicate tissues are dosed with the test item, the negative control (DPBS) and the positive control (5% SDS), respectively. The tissues are incubated for 35 +/-1 minutes in a humidified incubator (37+/- 1 °C, 5 +/- 1% CO2, 95% RH) and further 25 minutes at room temperature. Then the test item and the controls are rinsed off with DPBS and the tissues are post-incubated for 24 +- 2 h and, after medium renewal, for further 18 +/- 2 h. Then the tissues are stained via MTT for 3 hours. Isopropanol extracts are measured photometrically at 550 nm.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: mean relative tissue viability

Value:

> 50

Remarks on result:

other:

Remarks:

Basis: other: mean tissue viability of the negative control tissues. Time point: 60 min treatment 42 h post-incubation. Remarks: non-irritant; EU CLP and UN GHS: No Category. (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: mean relative tissue viability

Value:

<= 50

Remarks on result:

other:

Remarks:

Basis: other: mean tissue viability of the negative control tissues. Time point: 60 min treatment 42 h post-incubation. Remarks: irritant; EU CLP and UN GHS Category 2. (migrated information)

Applicant's summary and conclusion

Interpretation of results:

other: irritant

Remarks:

Criteria used for interpretation of results: other: OECD 439

Conclusions:

The test item is classified as "irritant" (Category 2).

Executive summary:

In the in vitro skin irritation test using the EpiDerm human epidermis model 25 mg test item + 25 µL DPBS were applied topically for 60 minutes.

After 42 h post-incubation cytotoxic effects were determined via MTT reduction assay.

The mean relative tissue viability (% negative control) was <= 50%.

The test item is therefore classified as "irritant" (EU CLP and UN GHS: Category 2).

The test item showed irritant effects. The mean relative tissue viability (% negative control) was< 50% (30.6%) after 60 min treatment and 42 h post incubation. The test item could not be applied evenly on the tissue surface due to its waxy/jellylike consistence which formed clots after application. Therefore, the three tissues treated identically with the test item showed varying viabilities, whereas two replicates showed consistent values (17.2% and 22.0% versus 52.6%) indicating irritant potential. The standard deviation between the three tissues was therefore exceeding the 18% cut-off (19.2%). This is accepted due to the properties of the test item as described above.

This study is classified as acceptable:

Mean OD550 of the three negative control tissues is >=1 and <=2.5.

Mean relative tissue viability of the three positive control tissues is <= 20%.

Standard deviation (SD) of relative tissue viablity obtained from each three concurrently tested control tissues is < 18%.

This study satisfies the requirement for Test Guideline OECD 439 for in vitro skin irritation.