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Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation / corrosion, other
Remarks:
other: in vitro (Epiderm)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

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:
other: EU Method b.46 (Skin Irritation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany

Test material

Constituent 1
Chemical structure
Reference substance name:
Fatty acids, C16-18, reaction products with diethanolamine
EC Number:
293-014-3
EC Name:
Fatty acids, C16-18, reaction products with diethanolamine
Cas Number:
91032-08-5
Molecular formula:
C22H45NO3 to C52H101NO5
IUPAC Name:
Fatty acids, C16-18, reaction products with diethanolamine

Test animals

Species:
other: EpiDerm reconstructed human epidermis model (MatTek Corp.)
Details on test animals or test system and environmental conditions:
organotypic reconstructed three-dimensional model of the human epidermis

Test system

Type of coverage:
other: direct application
Controls:
other: three tissues for negative control (30µL DPBS) and positive control (30µL 5% SDS)
Amount / concentration applied:
solids: 25 µL DPBS + 25 mg
Duration of treatment / exposure:
60 +/- 1 minutes
Observation period:
42 +/- 4 h
Number of animals:
three replicate tissues per dose group
Details on study design:
3 replicate tissues are dosed with the test item, the negative control (DPBS) and the positive control (5% SDS), respectively. The tissues are incubated for 35 +/-1 minutes in a humidified incubator (37+/- 1 °C, 5 +/- 1% CO2, 95% RH) and further 25 minutes at room temperature. Then the test item and the controls are rinsed off with DPBS and the tissues are post-incubated for 24 +/- 2 h and, after medium renewal, for further 18 +/- 2 h. Then the tissues are stained via MTT for 3 hours. Isopropanol extracts are measured photometrically at 550 nm.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: mean relative tissue viability
Basis:
other: mean tissue viability of the negative control tissues
Time point:
other: 60 min treatment 42 h post-incubation
Score:
> 50
Remarks on result:
other: non-irritant; EU CLP and UN GHS: No Category
Irritation parameter:
other: mean relative tissue viability
Basis:
other: mean tissue viability of the negative control tissues
Time point:
other: 60 min treatment 42 h post-incubation
Score:
<= 50
Remarks on result:
other: irritant; EU CLP and UN GHS Category 2
Irritant / corrosive response data:
If mean tissue viability is > 50% relative to the mean negative control, the test item is classified as non-irritant (EU CLP and UN GHS: No Category).
If mean tissue viability is =< 50% relative to the mean negative control, the test item is classified as irritant (EU CLP and UN GHS: Category 2).
Other effects:
The test item showed no direct MTT reducing capability and no colouring potential.

Any other information on results incl. tables

Name

NC

PC

TM

Tissue

1

2

3

1

2

3

1

2

3

OD550(blank-corrected)

1.320

2.141

1.843

0.123

0.117

0.117

1.961

2.564

2.449

1.626

1.912

1.814

0.137

0.089

0.121

2.566

2.601

2.443

mean OD550of the replicates (blank-corrected)

1.473

2.026

1.828

0.130

0.103

0.119

2.263

2.582

2.446

total mean OD550of 3 replicate tissues (blank-corrected)

1.776

0.117

2.430

SD OD550

 

0.279

 

 

0.016

 

 

0.239

 

relative tissue viability [%]

82.9

114.1

103.0

7.3

5.8

6.7

127.5

145.4

137.7

SD tissue viability [%]***

15.8

0.8

9.0

CV [% viability]

 

15.8

 

 

11.5

 

 

6.6

 

mean relative tissue viability [%]

100.0

6.6

136.9

*          Corrected mean OD550of the negative control corresponds to 100% absolute tissue viability.

**      mean relative tissue viability of the three positive control tissues is < 20%

***     standard deviation (SD) obtained from the three concurrently tested tissues is < 18%

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test item is classified as "non-irritant" (EU CLP and UN GHS: No Category).
Executive summary:

In the in vitro skin irritation test using the EpiDerm human epidermis model 25 mg test item + 25 µL DPBS were applied topically for 60 minutes. After 42 h post-incubation cytotoxic effects were determined via MTT reduction assay.

The test item showed no irritant effects. The mean relative tissue viability (% negative control) was> 50% (137%) after 60 min treatment and 42 h post incubation.

This study is classified as acceptable:

Mean OD550 of the three negative control tissues is >=1 and <=2.5.

Mean relative tissue viability of the three positive control tissues is <= 20%.

Standard deviation (SD) of relative tissue viablity obtained from each three concurrently tested tissues is < 18%.

This study satisfies the requirement for Test Guideline OECD 439 for in vitro skin irritation.