3-[(2-acetamidoacetyl)amino]propanoic acid

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March 2012

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

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)

Version / remarks:

Official journal of the European union dated august, 24'h, 2009

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: ECVAM protocol

Version / remarks:

November 5th, 2008

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3-[(2-acetamidoacetyl)amino]propanoic acid

EC Number:

816-146-0

Cas Number:

1016788-34-3

Molecular formula:

C7H12N2O4

IUPAC Name:

3-[(2-acetamidoacetyl)amino]propanoic acid

Test material form:

solid: particulate/powder

In vitro test system

Test system:

human skin model

Details on test system:

Human reconstructed epidermis, SkinEthic model 0.5 cm', maintained according to the supplier instructions.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Series definition:
16 microL of the test item diluted at 5 %, as well as the reference items, are tested on three epidermis.
Test protocol:
16 microL ± 0.5 microL of the test item have been deposited with a positive displacement micropipette on the surface of the epidermis and a 7.5 mm diameter nylon mesh is gently applied on the surface of the
epidermis with tweezers.
16 microL ± 0.5 microL of reference item are deposited with a micropipette positive displacement on the surface of the tissue. A 7.5 mm diameter nylon mesh is gently applied on the surface of the epidermis in with
tweezers.

Duration of treatment / exposure:

The epidermis are incubated in 0.3 ml of maintenance medium (24 wells plate) at room temperature for 42 minutes ± 1 minute.

Duration of post-treatment incubation (if applicable):

Nylon meshes are removed and the epidermis are rinsed with 25 ml of PBS by epidemlis (25 times 1 ml using a dispenser). The residual PBS is eliminated on absorbent paper. If necessary the epidermis
can be gently swept with a cotton tip. The epidermis are incubated in 2 ml of growth medium in 6 wells plate at 37 °C, 5% CO, for 42 hours ± 1 hour.
All epidermis (living and dead) are incubated in 0.3 ml of maintenance medium at 1 mg/ml MIT in 24 well plate.
After 3 hours ± 5 minutes incubation at 37°C, 5% CO2 outside of inserts is rinsed with1I to 2 ml of PBS.
Extraction is performed by placing the epidermis into wells filled with 0.8 ml of isopropanol and covered with 0.7 ml isopropanol for 2 hours ± 5 minutes under gentle agitation protected from light.
The absorbance is measured in triplicat on 200 microL extract in 96-wells plates. Absorbances are measured at 540 nm against a blank consisting with isopropanol.

Number of replicates:

3 replicates for test item, 3 replicates for positive control and 3 replicates for negative control

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Tissue Viability (%)

Value:

ca. 100

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

		ASSESMENT OF VIABILITY				CONCLUSION
		Abs.	mean	Standard deviation	Viability %
Negative control PBS	Ep.1 Ep.2 Ep.3	1.904 1.894 1.892 1.951 1.946 1.905 1.778 2.124 1.777	1.908	0.103	100%	Non Irritant
Positive control SDS 5%	Ep.1 Ep.2 Ep.3	0.020 0.020 0.018 0.021 0.021 0.019 0.022 0.022 0.020	0.020	0.001	1.1%	Irritant
Test item	Ep.1 Ep.2 Ep.3	2.160 2.148 2.108 1.862 1.907 1.971 1.707 1.680 1.620	1.907	0.207	100%	Non Irritant

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In accordance with the Regulation EC No. 127212008 on classification, labelling and packaging of substances and mixtures, the test item must not be classified.