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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:
weight of evidence
Study period:
2014-11-25 to 2015-05-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Version / remarks:
26 July 2013
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: 003
- Expiration date of the lot/batch: 01 Jan 2020

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, exclusion of direct sunlight, test substance is hygroscopic
- Stability under test conditions: stable

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
water
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™
- Tissue batch number: 21604

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature

REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: once after treatment and once after MTT incubation
- Observable damage in the tissue due to washing: no
- Modifications to validated SOP: no

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 hours
- Spectrophotometer: SunriseTM Absorbance Reader
- Wavelength: 570 nm
- Filter: without reference filter

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: 1.821±0.108 (Acceptance criteria: OD (540-570 nm) [1.0 - 3.0])
- Barrier function: 5.32 hrs (Acceptance criteria: ET-SO [4.77 - 8.72 hrs])
- Contamination: sterile (acceptance criteria: no contamination)

NUMBER OF REPLICATE TISSUES: 1

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- killed tissues
- Procedure used to prepare the killed tissues: tissues were freeze killed
- No. of replicates: 2
- Method of calculation used: The formazan production of the test substance treated epidermal tissues is compared to that of negative control tissues. The quotient of the values indicates the relative tissue viability.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 2

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50 %, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15 %
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is greater than or equal to 15 %.
Control samples:
yes, concurrent vehicle
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 25 µL

VEHICLE
- Amount applied: 25 µL

NEGATIVE CONTROL
- Amount applied: 50 µL de-ionized water

POSITIVE CONTROL
- Amount applied: 50 µL
- Concentration: 8 N potassium hydroxide
Duration of treatment / exposure:
3 min and 1 hour
Number of replicates:
2

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 min exposure
Value:
102.4
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1 hours exposure
Value:
107.6
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: No
- Direct-MTT reduction: Test substance is not able to reduce MTT directly.
- Colour interference with MTT: Not applicable

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

Table 1: 3 min exposure

Test substance

Tissue 1

Tissue 2

Mean

SD

CV [%]

NC

Mean OD570

2.007

1.967

1.987

 

 

Viability [% of NC]

101.0

99.0

100.0

1.4

1.4

Test substance

Mean OD570

1.963

2.107

2.035

 

 

Viability [% of NC]

98.8

106.1

102.4

5.1

5.0

PC

Mean OD570

0.378

-

0.378

 

 

Viability [% of NC]

19.0

-

19.0

 

 

Table 2: 1 hour exposure

Test substance

Tissue 1

Tissue 2

Mean

SD

CV [%]

NC

Mean OD570

2.031

2.032

2.031

 

 

Viability [% of NC]

100.0

100.0

100.0

0.02

0.02

Test substance

Mean OD570

2.093

2.279

2.186

 

 

Viability [% of NC]

103.0

112.2

107.6

6.5

6.0

PC

Mean OD570

0.113

0.089

0.101

 

 

Viability [% of NC]

5.5

4.4

5.0

0.8

16.5

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met