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Diss Factsheets

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 March 2012 to 30 March 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.G

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

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
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
sodium (2S)-5-oxopyrrolidine-2-carboxylate
Cas Number:
28874-51-3
Molecular formula:
C5H6NO3. Na
IUPAC Name:
sodium (2S)-5-oxopyrrolidine-2-carboxylate
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Physical state: White to pale yellow powder.
- Storage condition of test material: Controlled Room temperature (15-25°C, below 70 RH %), Protected from humidity.

Test animals

Details on test animals or test system and environmental conditions:
EPISKIN-SM
Supplier: SkinEthic, France

Test system

Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 20 mg
Duration of treatment / exposure:
15 ± 0.5 minutes
Number of animals:
Not applicable

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Value:
95
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid

In vivo

Other effects:
CELL VIABILITY
- Cell viability of treated disks was 95 %, see Table 1 for results.

VALIDITY OF THE TEST
The mean OD value of the three negative control tissues was 0.641. The positive control result showed 26 % viability. Each standard deviation value (SD) of the % viability was below 18. All validity criteria were within acceptable limits and therefore the study can be considered as valid.

INDICATOR FOR POTENTIAL FALSE VIABILITY
> Possible direct MTT reduction with the test material: No colour change was observed after three hours of incubation of the test item in MTT solution. The test material did not interact with the MTT, therefore additional controls and data calculations were not necessary. A false estimation of viability can be precluded.
> Colouring potential of the test material: As the test material has an intrinsic colour, one additional chemical-treated tissue was used for the non specific OD evaluation. Mean OD (measured at 540 nm) of this tissue was determined as 0.025, Non Specific Colour % was calculated as 4 %. Therefore additional data calculation was not necessary.

Any other information on results incl. tables

Table 1: Optical Density (OD) and the Calculated % Viability

Substance

Optical Density (OD)

Viability (%)

Negative Control

1

0.639

100

2

0.607

95

3

0.676

105

Mean

0.641

100

Standard Deviation (SD)

5.00

Positive Control

1

0.150

23

2

0.191

30

3

0.155

24

Mean

0.165

26

Standard Deviation (SD)

3.79

Test Material

1

0.555

87

2

0.627

98

3

0.633

99

Mean

0.605

95

Standard Deviation (SD)

6.66

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the study, exposure to the test material resulted in a mean relative cell viability of 95 %. Since the cell viability was determined to be > 50 % of the negative control the test material was considered to be non-irritating.
Executive summary:

The potential for the test material to cause skin irritation was predicted in an in vitro study using the EPISKIN Model. The study was conducted under GLP conditions and in line with OECD 439 and EU Method B.46. The test material was applied topically to the surface of the skin for 15 minutes, which was terminated by rinsing with PBS 1 x solution (0.9 %). Epidermis units were then incubated at 37°C for 42 hours in an incubator with 5 % CO2. The viability of 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. Positive and negative controls were run concurrently and tissue viability was expressed as a % relative to the negative control.

Under the conditions of the test, exposure to the test material resulted in a mean relative cell viability of 95 %. Since the cell viability was determined to be > 50% of the negative control the test material was considered to be non-irritating.