Reaction mass of hexadecyl acrylate and octadecyl acrylate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Remarks:

and skin corrosion

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-04-17 to 2012-05-31

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study OECD guidelines were followed.

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:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Version / remarks:

April 13, 2004

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

July 22, 2010

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Part B: Methods for the determination of toxicity and other health effects: In Vitro Skin Corrosion: Human Skin Model Test

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

BASF SE

Test material

Test material information

Constituent 1

Reference substance name:

Stearylacrylat, Synative MM SA, Stearyl 1618

IUPAC Name:

Stearylacrylat, Synative MM SA, Stearyl 1618

Test material form:

other: solid

Test animals

Species:

human

Strain:

other: human epidermis model

Details on test animals or test system and environmental conditions:

Test system:
- Model: Three dimensional human epidermis model
The EpiDermTM model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multi layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs, 10 mm ∅) and commercially available as kits (EpiDerm™ 200), containing 24 tissues on shipping agarose.

Test system

Type of coverage:

open

Preparation of test site:

other: in vitro test

Vehicle:

unchanged (no vehicle)

Controls:

other: Negative control: De-ionized water (corrosion test); PBS, sterile (irritation test); positive control: 8-n potassium hydroxide solution for the corrosion test 5% (w/v) sodium dodecyl sulfate; MTT-reduction control: De-ionized water or test substance

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: Corrosion test: 50 µL of the undiluted liquid test substance (liquide after heating at ca. 30°C)
irritatin test: 30 µL of the undiluted liquid test substance (liquide after heating at ca. 30°C)

Duration of treatment / exposure:

- Corrsosion test: 3 min and 1 hour
- Irritation test: 1 hour with about 42 hours post-incubation

Observation period:

42-hour post-incubation period

Number of animals:

2 samples per application time

Details on study design:

TEST SYSTEM
The EpiDermTM model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multi layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs, 10 mm ∅) and commercially available as kits (EpiDerm™ 200), containing 24 tissues on shipping agarose.
-Tissue model: Epi-200
-Origin: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia

TEST PROCDURE
- Corrosion test: Two tissues per exposure time (3 minutes at room temperature or 1 hour in the incubator, as a rule) and test group (test material, negative control and positive control; 12 tissues per test) were used.
Fifty microliter (50 μL) of the undiluted liquid test substance (after heating at ca. 30 °C) was applied using a pipette.
Control tissues were concurrently applied with 50 μL of de-ionized water (negative control, NC) or with 50 μL of 8 n potassium hydroxide (positive control, PC) or test substance (killed tissue control, KC).
The tissues were washed with PBS to remove residual test material 3 minutes or 1 hour after start of the application treatment. Rinsed tissues were kept in 24-well plates (holding plates) at room temperature on assay medium until all tissues per application time were dosed and rinsed. The assay medium was then replaced by MTT solution and tissues were incubated for 3 hours.
After incubation, tissues were washed with PBS and the formazan produced by the tissues was extracted with isopropanol. The optical density at a wave length of 570 nm (OD570) of the extracts was determined spectrophotometrically. Blank values were established of 6 microtiter wells filled with isopropanol for each microtiter plate.
- Irritation test: Three tissues were treated with the test substance, the PC and NC, respectively. Thirty microliter (30 μL) of the undiluted liquid test substance (after heating at ca. 30 °C) was applied using a pipette.
Control tissues were concurrently applied with 30 μL of sterile PBS (negative control, NC) or with 30 μL of 5% SDS (positive control, PC). A nylon mesh was placed carefully onto the tissue surface afterwards. The tissues were kept under the laminar flow hood at room temperature for 25 minutes overall and for 35 minutes in the incubator. The tissues were washed with sterile PBS to remove residual test material 1 hour after start of application. Rinsed tissues were blotted on sterile absorbent paper and transferred into new 6-well plates, pre-filled with 0.9 mL fresh medium. When all tissues were rinsed, the surface of each tissue was carefully dried with a sterile cotton swab. Subsequently, the tissues were incubated in the incubator at 37°C for 24 ± 2 hours. After 24 ± 2 hours the tissues were transferred into new 6-well plates pre-filled with 0.9 mL of fresh medium and placed into the incubator for additional 18 ± 2 hours post-incubation period. After the post-incubation period, the assay medium was replaced by 0.3 mL MTT solution and the tissues were incubated in the incubator for 3 hours. After incubation, the tissues were washed with PBS to stop the MTT-incubation. The formazan that was metabolically produced by the tissues was extracted by incubation of the tissues in isopropanol. The optical density at a wavelength of 570 nm (OD570) of the extracts was determined spectrophotometrically. Blank values were established of 6 microtiter wells filled with isopropanol for each microtiter plate.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: Corrosion test

Value:

98

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: Viability [% of NC]

Results 2

Irritation / corrosion parameter:

other: Irritation test

Value:

117

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: Viability [% of NC]

Any other information on results incl. tables

- Corrosion test: The mean viability of the test-substance treated tissues determined after an exposure period of 3 minutes was 99%, and it was 98% after an exposure period of 1 hour.

- Irritation test: The mean viability of the test-substance treated tissues determined after an exposure period of 1 hour with about 42 hours post-incubation was 117%.

Based on the observed results and applying the evaluation criteria it was concluded, that Stearylacrylate, Synative MM SA, Stearyl 1618 does not show a skin irritation potential in the EpiDerm™ skin corrosion/irritation test under the test conditions chosen.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information