Ethyl 2-chloroacetoacetate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: 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: According to OECD Guideline for Testing of Chemicals 431: In vitro Skin Corrosion: Human Skin Model Test (Original Guideline adopted April 13, 2004).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Guideline for Testing of Chemicals 431: In vitro Skin Corrosion: Human Skin Model Test (Original Guideline adopted April 13, 2004).

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

EACA

IUPAC Name:

EACA

Details on test material:

Identity: EACA
Batch-No.: 001008
Purity: 93.9%
Stability in Solvent: Not relevant
Storage: At 2 8 °C
Retest Date: July 26, 2010

Test system

Amount / concentration applied:

Each two tissues of the human skin Model EST1000 were treated with either the test item, the negative or the positive control for 3 minutes and 1 hour
50 µL of the test item and the controls were applied to each tissue
the test item was tessted neat
the negative control was deionised water
the positive control was 8.0 N potassium hydroxide

Duration of treatment / exposure:

the tissues were tretaed for 3 minutes and 1 hour, respectively

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Value:

ca. 10.7

Any other information on results incl. tables

Table 1: Results after treatment with EACA

 

Dose Group	Ex-posure Inter-val	Absorbance 570 nm Tissue 1*	Absorbance 570 nm Tissue 2*	Mean Absorbance of 2 Tissues	Rel. Absorbance  [% of Negative Control]**
Negative Control	3 min	2.036	2.066	2.051	100.0
Positive Control	3 min	0.118	0.092	0.105	5.1
EACA	3 min	0.506	0.411	0.458	22.3
Negative Control	1 hour	1.646	1.939	1.792	100.0
Positive Control	1 hour	0.030	0.025	0.028	1.5
EACA	1 hour	0.187	0.195	0.191	10.7

*       Mean of three replicate wells after blank correction (0.037 for the 3 minutes treatment, 0.036

        for the 1 hour treatment)
**      relative absorbance [rounded values]:

The optical evaluation of the MTT-reducing capacity of the test item after a 1 hour incubation with MTT-reagent did not show evidence of a blue colour and thereby was not considered to be an MTT reducer.

The test item is considered to be corrosiveto skin, since

·       the viability after 3 minutes exposure is less than 50%, and

·       the viability after 1 hour exposure is less than 15%.

Applicant's summary and conclusion

Interpretation of results:

corrosive

Remarks:

Migrated information Criteria used for interpretation of results: other: OECD Guideline for Testing of Chemicals 431: In vitro Skin Corrosion: Human Skin Model Test (Original Guideline adopted April 13, 2004).

Conclusions:

In conclusion, it can be stated that in this study and under the reported experimental conditions, the test item EACA was corrosive to skin. According to EC Regulation 1272/2008 the test item is classified as corrosive (category 1).

Executive summary:

This in vitro study was performed to assess the corrosive potential of EACA by means of the Human Skin Model Test. This test consists of topical application of the test material to the skin tissue for 3 minutes and 1 hour, followed by immediate determination of the cytotoxic effect. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the exposure period. Produced amount of formazan is measured photometrical at 570 nm.

Independent duplicate tissues of the human skin model EST-1000 were exposed to either the test item, the negative control or the positive control for 3 minutes and 1 hour, respectively.

The liquid test item (50 µL) was applied to each tissue and spread evenly over the surface of the tissue.

A volume of 50 µL of either the negative control (deionised water) or the positive control (8.0 N KOH) was applied to each tissue.

After exposure to the negative control the absorbance values exceeded the required acceptability criterion of mean OD₅₇₀= 0.8 for both treatment intervals thereby confirming the acceptable quality of the tissues. Both absorbance values exceeded the minimum range of at least 20 times blank value(based on blank values of 0.037 for the 3 minutes treatment and 0.036 for the 60 minutes treatment the threshold for the 3 minutes treatment is 0.74 and 0.72 for the 60 minutes treatment).

Exposure to the positive control induced a decrease in the relative absorbance as compared to the negative control, both for the 3 minutes exposure period and for the 1 hour exposure period thus confirming the validity of the test system.

After exposure to the test item EACA the relative absorbance values decreased to 22.3% after 3 minutes. After the 1 hour exposure relative absorbance values were reduced to 10.7%. Both values exceeded the threshold for corrosivity of 50% for the 3 minutes exposure and 15% for the 1 hour exposure. Therefore, the test item was considered to be corrosive.

In conclusion, it can be stated that in this study and under the reported experimental conditions, the test item EACA was corrosive to skin. According to EC Regulation 1272/2008 the test item is classified as corrosive (category 1).