Registration Dossier

Toxicological information

Skin irritation / corrosion

Currently viewing:

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
Principles of method if other than guideline:
EpiDerm-Test
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 4-Nitrotoluene-2-sulphonic acid
- Analytical purity: 84.2 %
- Lot/batch No.: 090330

Test animals

Species:
other: in vitro test
Strain:
other: in vitro test

Test system

Type of coverage:
other: in vitro test
Preparation of test site:
other: in vitro test
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):25 µl of the solid test substance
Duration of treatment / exposure:
3 minutes and 1 hour
Details on study design:
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 Ø) and commercially available as kits (EpiDerm™ 200), containing 24 tissues on shipping agarose.
TEST PROCEDURE
- For the corrosion test two tissue samples were uses for 3 minutes and 1 hour, the irritaion test was perfomed with tree tissue sampels, which were incubated with the test substance for 1 hour followed by a 42-hours post-incubation period.
- After 3 minutes (room temperature) or after 1 hour (incubator) tissue contact with the test material, the tissues were washed with buffer to remove residual test material. 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 buffer and formazan was extracted with Isopropanol over night at room temperature. The optical density at a wavelength of 570 nm (OD570) of extracted formazan was determined spectrophotometrically and cell viability was calculated for each tissue as % of the mean of the negative control tissues. Skin corrosivity potential of the test material was classified according to remaining cell viability obtained after test material treatment with either of the 2 exposure times.


CONTROLS
- Negative control (NC): Doubly distilled water
- Positive control (PC): 8 N potassium hydroxyde (Sigma-Aldrich, Munich, Germany)


ASSAY ACCEPTANCE CRITERIA:
- Negative control (NC ): mean OD570 of the NC per exposure time 2.5
- Positive control (PC ): mean relative tissue viability of the 3 min positive control is <= 30 %
- Tissue variability: The inter tissue variability is considered to be acceptable if the difference of the OD570 values of the two tissues is <= 0.3

Results and discussion

In vivo

Irritant / corrosive response data:
See result tables. Based on the observed results and applying the evaluation criteria cited, it was concluded, that 4-Nitrotoluene-2-sulphonic acid shows a skin irritation potential in the EpiDerm TM skin corrosion/irritation test under the test conditions chosen.

Any other information on results incl. tables

Corrosion test

 

 

Exposure: 3 min

Exposure: 1 hour

Test Substance

 

Tissue 1

Tissue 2

Mean

Tissue 1

Tissue 2

mean

NC

Mean OD570

1.6233

1.8758

1.7496

1.5827

1.8112

1.6969

 

Viability [% of NC]

92.8

107.2

100

93.3

106.7

100

09/0641-1

Mean OD570

1.9028

1.6308

1.7668

0.2857

0.2772

0.2814

 

Viability [% of NC]

108.8

93.2

101

16.8

16.3

17

PC

Mean OD570

0.3493

0.3223

0.3358

0.1902

0.1907

0.1904

 

Viability [% of NC]

20.0

18.4

19

11.2

11.2

11

 

Irritation Test

Test Substance

 

Tissue 1

Tissue 2

Tissue 3

Mean

SD

NC

Mean OD570

1.8535

2.3000

*

2.0768

 

 

Viability [% of NC]

89.3

110.7

*

100

15.20

09/0641-1

Mean OD570

0.1445

0.1385

0.1635

0.1488

 

 

Viability [% of NC]

7.0

6.7

7.9

7

0.63

PC

Mean OD570

0.1005

0.1140

0.1000

0.1048

 

 

Viability [% of NC]

4.8

5.5

4.8

5

0.38

* For the NC two tissues were valid for evaluation, only (details are available with the raw data)

Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Migrated information