Registration Dossier

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011-06-29 to 2011-07-01
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
Deviations:
no
Qualifier:
according to
Guideline:
other: OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid

Test animals

Species:
other: EpiDerm human epidermis model
Strain:
other: not applicable
Details on test animals and environmental conditions:
organotypic reconstituted three-dimensional model of the human epidermis

Test system

Type of coverage:
other: direct application
Preparation of test site:
not specified
Vehicle:
unchanged (no vehicle)
Controls:
other: two tissues per treatment period for negative control (A. dest.) and positive control (8 N KOH)
Amount / concentration applied:
50 µL
Duration of treatment / exposure:
3 minutes and 60 minutes
Number of animals:
two replicate tissues per treatment period
Details on study design:
2 replicate tissues per treatment period are dosed with the neat test item, the negative control (A. dest.) and the positive control (8 N KOH), repectively. After each treatment period the test item and controls are rinsed off and the tissues are stained via MTT for 3 hours. Isopropanol extracts are measured photometrically at 550 nm.

Results and discussion

In vitro

Resultsopen allclose all
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: 3 min. Remarks: corrosive. (migrated information)
Irritation / corrosion parameter:
other: other: mean relative tissue viability
Value:
< 15
Remarks on result:
other:
Remarks:
Basis: other: mean tissue viability of the negative control tissues. Time point: 60 min. Remarks: corrosive. (migrated information)

In vivo

Irritant / corrosive response data:
if mean tissue viability after 3 min is >= 50% and after 60 min is >= 15% of the respective negative control, the test item is classified as non-corrosive
Other effects:
The mixture of 100 µl test item per 1 ml MTT medium showed reduction of MTT compared to the solvent. The mixture turned blue/purple. Treatment of killed tissues with the test item showed that the test item was washed away before the addition of the MTT solution. Therefore, an influence on the assay was not detectable and a quantitative correction of results was not necessary.

Any other information on results incl. tables

3 min.

Name

Negative Control

Test Item

Positive Control

Tissue

1

2

1

2

1

2

absolute OD550 -values

1.823

1.801

1.732

1.702

0.216

0.223

1.880

1.869

1.801

1.742

0.219

0.216

1.839

1.800

1.782

1.739

0.219

0.224

OD550(mean of 3 aliquots)

1.847

1.823

1.772

1.728

0.218

0.221

sd

0.029

0.040

0.036

0.022

0.002

0.004

OD550(mean of 2 replicate tissues)

1.835*

1.750

0.220

mean relative tissue viability [%]

100

95

12**

mean inter tissue viability difference [%]***

1.3

2.5

1.3

*       mean OD550 >= 0.8

**      mean relative tissue viability of the 3 min. positive control <= 30%

***     inter tissue viability difference <= 30%

60 min.

Name

Negative Control

Test Item

Positive Control

Tissue

1

2

1

2

1

2

absolute OD550 -values

1.915

1.898

0.265

0.243

0.181

0.157

1.920

1.927

0.271

0.246

0.182

0.160

1.956

1.824

0.263

0.245

0.182

0.153

OD550(mean of 3 aliquots)

1.930

1.883

0.266

0.245

0.182

0.157

sd

0.022

0.053

0.004

0.002

0.001

0.003

OD550(mean of 2 replicate tissues)

1.907*

0.255

0.169

mean relative tissue viability [%]

100

13

9

mean inter tissue viability difference [%]***

2.5

8.5

14.8

*       mean OD550 >= 0.8

***     inter tissue viability difference <= 30%

Applicant's summary and conclusion

Interpretation of results:
corrosive
Remarks:
Migrated information Criteria used for interpretation of results: other: OECD 431
Conclusions:
The test item is classified as "corrosive".
Executive summary:

The potential of the test item to induce skin corrosion was analysed by using the three-dimensional human skin model EpiDermTM, comprising a reconstructed epidermis with a functional stratum corneum. The undiluted substance was applied topically to the EpiDermTM tissue for 3 minutes and 60 minutes respectively, followed by immediate determination of cytotoxic effects via the MTT reduction assay. Corrosivity potential was predicted from the relative mean tissue viabilities obtained after both treatment periods compared to the corresponding negative control tissues concurrently treated with A. dest (distilled water).

The test item showed corrosive effects in that the mean relative tissue viability (% negative control) was > 50% (95%) after3 minute treatment and < 15% (13%) after 60 minute treatment. The controls confirmed the validity of the study. The mean OD550 of the two negative control tissues was > 0.8 for each exposure period. The mean relative tissue viability (% negative control) of the positive control was < 30% (12%) after 3 minute treatment. The maximum inter tissue difference of replicate tissues of all dose groups was < 30% (1.3% - 14.8%).

Based on the results of this study, the test item showed corrosive effects. The test item therefore is classified as "corrosive".