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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
Study period:
2013-12-18 until 2013-12-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant, guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: UN (2009), United Nations Globally Harmonized System of Classification and Labelling of Chemicals (GHS), third revised edition, UN New York and Geneva.
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EpiSkin™ SOP, Version 1.8 (February 2009), ECVAM Skin Irritation Validation Study: Validation of the EpiSkin™ test method 15 min - 42 hours for the prediction of acute skin irritation of chemicals.
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
2,4,6-trichloroaniline
EC Number:
211-219-8
EC Name:
2,4,6-trichloroaniline
Cas Number:
634-93-5
Molecular formula:
C6H4Cl3N
IUPAC Name:
2,4,6-trichloroaniline
Test material form:
solid: crystalline
Details on test material:
- Physical state: Off white crystals
- Expiration date of the lot/batch: Recertification date end November 2015
- Storage condition of test material: Room temperature (<30°C)

Test animals

Species:
other: Reconstructed human epidermis model EPISKIN (SM)
Strain:
other: Not applicable
Details on test animals or test system and environmental conditions:
HUMAN SKIN: EPISKIN (SM) (Manufacturer: SkinEthic, France, Batch No.:13-EKIN-045, Expiry Date: 23 December 2013) is a three-dimensional human epidermis model.

EXPERIMENTAL DATES: From: 18 December 2013 To: 20 December 2013

Test system

Type of coverage:
other: Not applicable
Preparation of test site:
other: Not applicable
Vehicle:
unchanged (no vehicle)
Controls:
other: Not applicable
Duration of treatment / exposure:
15 mins
Observation period:
Not applicable
Number of animals:
Not applicable

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
other: other: optical density
Value:
0.731
Remarks on result:
other:
Remarks:
Basis: mean. (migrated information)
Irritation / corrosion parameter:
other: other: viability
Value:
99
Remarks on result:
other:
Remarks:
Basis: mean. Remarks: %. (migrated information)

In vivo

Irritant / corrosive response data:
The optical density for the test item treated skin was 0.731 and showed a viability of 99%.
The optical density for the negative and positive controls were 0.740 and 0.088, respectively and showed viabilities of 100% and 12% respectively.

The test item is, therefore, considered not to be irritant to skin, as the mean relative viability after 15 minutes exposure and 42 hours post incubation, is greater than 50% of the negative control. The positive control was irritant.

Any other information on results incl. tables

Table 1: Optical density and calculated % viability of cells

Substance

Optical density

Viability (%)

Negative control

PBS

1

0.770

104

2

0.728

98

3

0.721

97

mean

0.740

100

Positive control

SDS 5%

1

0.100

14

2

0.095

13

3

0.070

9.5

mean

0.088

12

Test item

1

0.746

101

2

0.698

94

3

0.748

102

mean

0.731

99

 

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an in vitro EPISKIN (SM) model test, the results indicate that the test item is Non Irritant (UN GHS: No Category).
Executive summary:

The reconstructed human epidermis model EPISKIN (SM) is designed to predict and classify the skin irritant potential of chemicals, by measuring its cytotoxic effect, as reflected in the MTT assay, on the EPISKIN reconstituted human epidermis. This method is approved by international regulatory agencies as a replacement for the identification of irritants / corrosives in the in vivo rabbit skin assay (OECD 404).

Disks of EPISKIN (three units / chemical) were treated with the test item and incubated for 15 minutes at room temperature. Exposure of test material was terminated by rinsing with phosphate buffered saline (PBS). Epidermis units were then incubated at 37°C for 42 hours in an incubator with 5% CO2. The viability of the epidermis on each disk was assessed by incubating the tissues for 3 hours with MTT solution at 37°C in 5% CO2 protected from light. The formazan extract in acidified isopropanol was then spectrophotometrically evaluated for optical density (OD) and quantified.

SDS 5% and PBS treated epidermis were used as positive and negative controls, respectively. An additional disk was used to provide an estimate of colour contribution from the test item. For each treated tissue viability was expressed as a % relative to negative control. If the mean relative viability after 15 minutes exposure and 42 hours post incubation is less or equal (≤) to 50% of the negative control, the test substance is considered to be irritant to skin.

Following exposure with the test item, the mean cell viability was 99% compared to the negative control and, therefore, non irritant. All validity criteria were within acceptable limits and therefore the study can be considered as valid.

In the in vitro EPISKIN (SM) model test, the results indicate that the test item is Non Irritant [UN GHS: No category].