2,4,6-trichloroaniline

Skin irritation / corrosion

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other: GLP compliant, guideline study, available as unpublished report, no restrictions, fully adequate for assessment.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 guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

Test guideline 3

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

Test guideline 4

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

Test material information

Constituent 1

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

Results

Results 1

Irritation / corrosion parameter:

other: other: optical density

Value:

0.731

Remarks on result:

other:

Remarks:

Basis: mean. (migrated information)

Results 2

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].