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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04 Jul - 06 Jul 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
The test item was found to have the potential to cause color interference with the MTT endpoint, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred.
Deviations:
yes
Remarks:
This deviation was considered to have not affected the integrity or validity of the study.
Qualifier:
according to
Guideline:
EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
Version / remarks:
The test item was found to have the potential to cause color interference with the MTT endpoint, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred.
Deviations:
yes
Remarks:
This deviation was considered to have not affected the integrity or validity of the study.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
Identification: Lumière Pink S.M. 8135N
Batch: 021340
Purity: >90%
Physical state/Appearance: Red powder
Expiry Date: 01 July 2022
Storage Conditions: Room temperature in the dark

Negative Control
Identification: sterile distilled water
Batch 3012436
Purity: Not supplied
Expiry Date: 01 October 2018
Storage Conditions: Room temperature

Positive Control
Identification: 8.0N Potassium Hydroxide
Batch: SLBM9898V
Purity: 7.92N
Expiry Date: 07 April 2020
Storage Conditions: Room temperature

EpiDerm™ Reconstructed Human Epidermis Model Kit
Supplier: MatTek
Date received: 04 July 2017
EpiDermTM Tissues (0.63cm2) lot number: 25828
Assay Medium lot number: 062917TMA

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Source strain:
other: epithelial, derived from human skin, and formed into a stratified, cornified epithelium
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
Supplier: MatTek
Date received: 04 July 2017
EpiDermTM Tissues (0.63cm2) lot number: 25828
Assay Medium lot number: 062917TMA

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C
- Temperature of post-treatment incubation: 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Rinsing was achieved by filling and emptying each tissue under a constant soft stream of DPBS to gently remove any residual test item.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL
- Incubation time: 60
- Optical density – 570nm
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):25 mg

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL


POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL
- Concentration (if solution): 8.0 N

25 μL of sterile water was added for wetting of the test item to increase tissue surface contact.
Duration of treatment / exposure:
3 and 60 minutes
Duration of post-treatment incubation (if applicable):
3 h at 37 °C, 5% CO2
Number of replicates:
2

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Nean Negative Control 3 min
Value:
100
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Negative Control 30 min
Value:
100
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Positive Control 3 min
Value:
5.6
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Positive Control 60 min
Value:
4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Test Item 3 min
Value:
120.4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Test Item 60 min
Value:
100.1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
- OTHER EFFECTS:
- Direct-MTT reduction:
- Colour interference with MTT:
The solution containing the test item was a purple color, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred.


ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements:yes

Any other information on results incl. tables

The relative mean viabilities for each treatment group :

Exposure Period

Percentage Viability

Negative Control

Positive Control

Test Item

3 minute

100

5.6

120.4

60 minute

100

4

100.1

 

Mean OD570Values and Viabilities for the Negative Control Item, Positive Control Item and Test Item

Tissue

Exposure Period

Mean OD570 of individual tissues

Mean OD570 of duplicate tissues

Standard Deviation

Coefficient of Variation (%)

Relative Mean Viability (%)

Negative Control

3 Minutes

1.524

1.568

0.062

4

100

1.612

60 Minutes

1.759

1.795

0.051

2.8

1.831

Positive Control

3 Minutes

0.097

0.088

0.013

N/A

5.6

0.079

60 Minutes

0.071

0.072

0.001

N/A

4

0.073

Test Item

3 Minutes

1.796

1.888

0.13

6.9

120.4

1.98

60 Minutes

1.777

1.796

0.026

1.5

100.1

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test item was considered to be non-corrosive to the skin.