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Administrative data

Endpoint:
skin irritation / corrosion
Remarks:
other: in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 February 2013 - 25 February 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
other: EU Method B.46 (In vitro dermal irritation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 439 (In vitro dermal irritation)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
N-(2-hydroxypropyl)oleamide
EC Number:
203-828-2
EC Name:
N-(2-hydroxypropyl)oleamide
Cas Number:
111-05-7
Molecular formula:
C21H41NO2
IUPAC Name:
N-(2-hydroxypropyl)oleamide
Test material form:
other: waxy solid
Details on test material:
- Name of test material: N-(2-hydroxypropyl) Oleamide
- Physical state: beige waxy solid
- Lot/batch No.: T22221 without solvent
- Analytical purity: 100% dry matter
- Storage condition: at room temperature.

Test animals

Species:
other: reconstructed human epidermis
Strain:
other: not applicable
Details on test animals or test system and environmental conditions:
Episkin TM Model Kit (0.38 cm2 tissues) supplied by SkinEthic Laboratories, Lyon, France.
Medium and Incubation T°C: 37°C
Dates of experimental phase: from 12 February 2013 to 20 February 2013.

Test system

Type of coverage:
other: not applicable (in vitro)
Preparation of test site:
other: not applicable (in vitro)
Vehicle:
unchanged (no vehicle)
Controls:
other: in vitro negative and positive controls
Amount / concentration applied:
10 mg (± 2 mg) spread on tissues
Duration of treatment / exposure:
Exposure period of 15 minutes, followed by rinsing.
Observation period:
MTT-loading after a 42h-incubation period following rinsing. Observation of MTT-> formazan transformation by viable cells.
Number of animals:
Not applicable
Triplicate for tested substance (test item, negative control, positive control)
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Rinsing: At the end of the treatment period, each tissue was removed from the well of the treatment plate, and rinsed with D-PBS. Rinsing was achieved by gently filling and emptying several times each tissue with D PBS to gently remove any residual test or control items. Excess D-PBS was removed by blotting the bottom of the tissue culture insert with absorbent paper.
The rinsed tissues were transferred to the second column of 3 wells containing 2 mL of maintenance medium in each well and the plates were incubated at +37°C, 5% CO2 in a humidified incubator for 42 (± 1) hours.

POSITIVE CONTROL
Name: Sodium Dodecyl Sulphate (SDS) at a 5% (w/v) aqueous solution.

NEGATIVE CONTROL
Name: Dulbecco’s Phosphate-Buffered Saline (D-PBS).

SCORING SYSTEM:
- Optical density (OD) was measured at 570 nm:
Relative mean viability (%) = 100 x mean cOD(test item) / mean cOD(negative control)
where:
- mean cOD Negative Control = mean ODNC – mean ODblank
- mean cOD Test Item = mean ODTI – mean ODblank

Interpretation: see below

Results and discussion

In vivo

Results
Irritation parameter:
other: relative mean viability
Basis:
mean
Time point:
other: 15 min exposure + 42h expression
Reversibility:
no data
Remarks:
not applicable
Remarks on result:
other: 100% = control

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test item is considered to be non-irritant to skin.
According to the results of this study, the classification of the test item should be:
. not classified (Directive 67/548/EEC) and no category (Regulation (EC) No. 1272/2008).
Executive summary:

The objective of this study was to evaluate the skin irritation potential of the test item using the EpiskinTM reconstructed human epidermis model.

The study design was based upon international guidelines (OECD Guideline No. 439 and Commission Regulation (EC) No. 761/2009, B.46) and thestudy was conducted in compliance with CiToxLAB’s standard operating procedures and the principles of Good Laboratory Practice.

 

Methods

Preliminary tests were performed to detect the ability of the test item to directly reduce MTT as well as its colouring potential.

Following the preliminary tests, the skin irritation potential of the test item was tested in the main test. The test item and both the negative and positive controls were topically applied on triplicate tissues and incubated at room temperature for 15 (± 1) minutes. At the end of the treatment period, each tissue was rinsed with D-PBS and incubated for 42 (± 1) hours at, 5% CO2in a humidified incubator. The cell viability was then assessed by means of the colourimetric MTT reduction assay.

Relative viability values were calculated for each tissue and expressed as a percentage of the mean viability of the negative control tissues which was set at 100% (reference viability).

 

Results

Preliminary tests

In the preliminary test, thetest item was found not to have direct MTT reducing properties.

 

The test item was found not to have a colouring potential in the preliminary test.

Main test

All acceptance criteria for the negative and positive controls were fulfilled. The study was therefore considered to be valid.

 

Following a 15 -minute exposure period and a 42-hour recovery period, the mean relative viability of the test item-treated tissues was 102% with a standard deviation of 9%.

 

Conclusion

The test item is considered to be non-irritant to skin.

According to the results of this study, the classification of the test item should be:

.  not classified (Directive 67/548/EEC) and no category (Regulation (EC) No. 1272/2008).