Registration Dossier

Diss Factsheets

Administrative data

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

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes (incl. QA statement)
Remarks:
testing lab.

Test material

Constituent 1
Chemical structure
Reference substance name:
m-terphenyl-2'-ol
EC Number:
219-401-9
EC Name:
m-terphenyl-2'-ol
Cas Number:
2432-11-3
Molecular formula:
C18H14O
IUPAC Name:
1,1':3',1''-terphenyl-2'-ol
Details on test material:
- CAS name: [1,1':3',1''-Terphenyl]-2'-ol
- Name of the test substance used in the study report: 2,6-Diphenylphenol
- Test substance No.: 10/0493-1
- Batch No.: LV 83/2010
- pH-value: ca. 4 (undiluted test substance, moistened with water)
- Homogeneity: homogenous by visual inspection
- Stability under test conditions: The stability under storage conditions over the study period was guaranteed by the sponsor.

Test animals

Species:
other: not applicable (in vitro test)
Strain:
other: not applicable (in vitro test)

Test system

Controls:
other: not applicable (in vitro test)
Amount / concentration applied:
Approximately 25 μL bulk volume of the test substance was added to 0.9 mL of the MTT solution.
Duration of treatment / exposure:
3 minutes and 1 hour
Number of animals:
not applicable (in vitro test)
Details on study design:
The EpiDermTM tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs, 10 mm diameter) and commercially available as kits (EpiDerm™ 200), containing 24 tissues on shipping agarose.

Negative control (NC):
- highly de-ionized water (corrosion test)
- PBS, sterile (irrritation test)

Positive control (PC):
- 8-n potassium hydroxide solution (Sigma-Aldrich, Munich, Germany) (corrosion test)
- 5% (w/v) sodium dodecyl sulfate (SDS, Sigma, Germany) in highly de-ionized water, sterile (irritation test)

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. 25 μL highly de-ionized water was applied first. Thereafter, a bulk volume of 25 μL of the solid ground test material was applied with a sharp spoon and homogeneously distributed with the water. Control tissues were concurrently applied with 50 μL of highly de-ionized water (negative control, NC) or with 50 μL of 8-n potassium hydroxide (positive control, PC). 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:
Three tissues were treated with the test substance, the PC and NC, respectively. 25 μL sterile PBS was applied first. Thereafter, a bulk volume of 25 μL of the solid ground test material was applied with a sharp spoon and homogeneously distributed together with the fluid. 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 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

Any other information on results incl. tables

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 103%, and it was 109% 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 106%.

Applicant's summary and conclusion