Registration Dossier

Administrative data

Endpoint:
skin irritation / corrosion
Remarks:
in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12-14 February 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Modern guideline study performed in a GLP certified laboratory

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014

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
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Batch number: RW 2013-10-22
Form: Powder
Color: Orange
Storage conditions: Room temperature
Expiring date: 22. 10. 2018

Test system

Controls:
other: SDS (5% aq.) and PBS treated epidermis were used as positive and negative controls respectively (three units / control).
Amount / concentration applied:
10 mg test item was applied evenly to each disks of EPISKIN (three units)
Duration of treatment / exposure:
15 minutes
Observation period:
2 days
Number of animals:
3 replicates
Details on study design:
Disks of EPISKIN (three units) were treated with test item and incubated for 15 minutes at room temperature. Exposure of test material was terminated by rinsing with PBS 1x solution. 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 precipitated formazan was then extracted using acidified isopropanol and quantified spectrophotometrically.
The test item has an intrinsic colour (orange), one additional chemical-treated tissue was used for the non-specific OD evaluation.
SDS (5% aq.) and PBS treated epidermis were used as positive and negative controls respectively (three units / control). For each treated tissue viability was expressed as a percentage relative to negative control.

Results and discussion

In vivo

Results
Irritation parameter:
other: mean tissue viabilityin-vitro
Basis:
mean
Remarks:
%
Time point:
other: day 2
Score:
67
Max. score:
77
Reversibility:
no data
Remarks:
not applicable
Irritant / corrosive response data:
All obtained test item viability results (mean OD: 0.550) were above 50 % when compared to the viability values obtained from the negative control, therefore the test item was considered to be non-irritant to skin

Any other information on results incl. tables

The results of the optical density (OD) measured at 570 nm of each replicate and the calculated % viability of the cells is presented below:

 

OD values and viability percentages of the controls

Controls

Optical Density (OD)

Viability (%)

Negative Control:
1x PBS

1

0.764

93

2

0.860

105

3

0.833

102

mean

0.819

100

standard deviation (SD)

6.02

Positive Control:
SDS (5 % aq.)

1

0.067

8

2

0.045

6

3

0.037

5

mean

0.050

6

standard deviation (SD)

1.91

 


OD values and viability percentages of the test item

Test Item

Optical Density (OD)

Viability (%)

YELLOW LF 6881

1

0.593

72

2

0.629

77

3

0.428

52

mean

0.550

67

standard deviation (SD)

13.07

 

OD values and NSC % of additional control

Additional colour control

Optical Density (OD)

Non Specific Colour %(NSC %)

YELLOW LF 6881
(
test item treated tissueswithout MTT incubation)

1

0.024

3.0

 

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In this in vitro skin irritation test in the EPISKIN model with test item YELLOW LF 6881 the results indicated that the test item is Non Irritant (NI) [UN GHS: No Category].
Executive summary:

Disks of EPISKIN (three units) were treated with test item and incubated for 15 minutes at room temperature. Exposure of test material was terminated by rinsing with PBS 1x solution. 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% CO2protected from light. The precipitated formazan was then extracted using acidified isopropanol and quantified spectrophotometrically.

The test item has an intrinsic colour (orange), one additional chemical-treated tissue was used for the non-specific OD evaluation.

SDS (5% aq.) and PBS treated epidermis were used as positive and negative controls respectively (three units / control). For each treated tissue viability was expressed as a percentage relative to negative control.

The test item is considered to be irritant to skin, if the mean relative viability after 15 minutes exposure and 42 hours post incubation is less or equal (≤) to 50% of the negative control.

 

The test item showed reduced cell viability in comparison to the negative control (mean value: 67 %). However all obtained test item viability results were above 50 % when compared to the viability values obtained from the negative control, therefore the test item was considered to be non-irritant to skin.

Positive and negative controls showed the expected cell viability values within acceptable limits.The experiment was considered to be valid.

In this in vitro skin irritation test in the EPISKIN model withtest itemYELLOW LF 6881the results indicated that the test itemis Non Irritant (NI) [UN GHS: No Category].