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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD 431: In vitro Skin Corrosion: Human Skin Model Test
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test substance: Golpanol ALS wasserfrei
- Test-substance No.: 09/0505-1
- Batch identification: 04749356P0
- Purity: Golpanols ALS = 71.3%; NaCI = 26.0%
- Homogeneity: The homogeneity of the test substance was provided after shaking the test-substance container.
- Storage stability: The stability under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.

ADDITIONAL TEST-SUBSTANCE INFORMATION
-pH-value: Ca. 5.5 (undiluted test substance, moistened with water)

Test animals

Species:
other: EpiDermTM (reconstructed three dimensional human epidermis model)

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: Negative control (NC): Highly de-ionized water (corrosion test); PBS, sterile (irrritation test); Positive control (PC): 8-n potassium hydroxide solution
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 μL bulk volume (about 12 mg)
Duration of treatment / exposure:
corrosion test: 3 minutes and 1 hour
irritation test: 1 hour
Observation period:
corrosion test: 3 minutes and 1 hour
irritation test: 1 hour incubation followed by a 42-hours post-incubation period
Number of animals:
corrosion test :Two tissue samlpes per exposure time and test group (test material, negative control and positive control; 12 tissues per test)
irritation test: three EpiDerm™ tissue samples with the test substance, the PC and NC
Details on study design:
Corrosion test:

For the corrosion test two EpiDerm™ tissue samples were incubated with the test substance for 3 minutes or 1 hour, respectively. 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 over night at room temperature. The optical density at a wavelength of 570 nm (OD570) of the extracts was determined spectrophotometrically.

Irritation test:

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 for at least 2 hours at room temperature on a plate shaker. The optical density at a wavelength of 570 nm (OD570) of the extracts was determined spectrophotometrically.

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 min corrosion experiment
Value:
ca. 114
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1h corrosion experiment
Value:
ca. 89
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3min irritation experiment (42 post incubation)
Value:
ca. 95

Any other information on results incl. tables

The EpiDerm skin corrosivity/irritation test showed the following results:

The test substance is not able to reduce MTT directly.

The mean viability of the test-substance treated tissues determined after an exposure period of 3 minutes was 114%, and it was 89% after an exposure period of 1 hour.

The mean viability of the test-substance treated tissues determined after an exposure period of 1 hour with about 42 hours post-incubation was 95%.

Based on the observed results it was concluded, that Golpanol ALS wasserfrei does not show a skin irritation potential in the EpiDerm skin corrosion/irritation test under the test conditions chosen.

Corrosion Test

 

     Exposure time 3 min           

Test-Article

OD570

tissue 1

OD570

tissue 2

mean  

OD570

Viability

(% of NC)

NC

1.7627

1.7522

1.7574

100

09/0505 -1

1.8622

2.1477

2.0049

114

PC_C

0.4438

0.3498

0.3968

23

 

     Exposure time 1 h           

Test-Article

OD570

 tissue 1

OD570

tissue 2

mean  

OD570

Viability

(% of NC)

NC

1.7657

1.6962

1.7309

100

09/0505 -1

1.5547

1.5312

1.5429

89

PC_C

0.1597

0.1867

0.1732

10

 

Irritation Test   

Test-Article

OD570

tissue 1

OD570

tissue 2

OD570

tissue 3

mean  

OD570

Viability

(% of NC)

NC

1.6795

1.6125

1.5725

1.6215

100

09/0505 -1

1.4935

1.5625

1.5745

1.5435

95

PC_C

0.1165

0.1320

0.1215

0.1233

8

 

NC: Negative control

PC_C: Positive control

OD570: Optical density (wavelength 570 nm)

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: other: CLP (1272/2008/EC)