Registration Dossier

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Remarks:
source of read across
Adequacy of study:
key study
Study period:
15 November 2016 to 15 December 2016
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
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
2016
Qualifier:
according to guideline
Guideline:
other: Method B.40bis of Commission Regulation (EC) No 440/2008, of 30 May 2008,
Version / remarks:
laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Similar Substance 01
IUPAC Name:
Similar Substance 01

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
other: EpiDerm™ Human Skin Mode
Cell source:
other: EpiDerm™ Human Skin Mode
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ Human Skin Mode (MatTek)
- Tissue batch number(s):23385
- Delivery date: 13 December 2016

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ˚C, 5% CO2
- Temperature of post-treatment incubation (if applicable): 37 ˚C, 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS: rinsed under a constant soft stream of DPBS and blotted dry with a tissue

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL MTT
- Incubation time: 3 h
- Spectrophotometer: Anthos 2001 microplate reader
- Wavelength: 562 nM

NUMBER OF REPLICATE TISSUES: 2/treatment

TISSUES:
- Fresh tissues: for treatment, negative control and positive control and color interference
- Killed tissues: for MTT interference and correction for color interference
- Procedure used to prepare the killed tissues (if applicable): freezing
- N. of replicates : 2/treatment
- Method of calculation used:
(TKT-UKT) + (CCT-CCU) – (ktCCT-ktCCU)
UKT Treated killed
TKT Untreated killed
CCT Color correction treated
CCU Color correction untreated
ktCCU Killed color correction treated
ktCCT Killed color correction untreated

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1 experiment

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if [complete, e.g. the viability after 3 minutes exposure is less than 50% (H314 1A), 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% (H314 1B or 1C)
- 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% (not classified as corrosive)
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
other: MTT reduction controls and killed color correction
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg as such

VEHICLE
- Amount(s) applied (volume or weight with unit): 50 µL distilled water

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL 8.0 N Potassium Hydroxide
Duration of treatment / exposure:
3 min and 60 min
Duration of post-treatment incubation (if applicable):
The plates was incubated (37 ˚C, 5% CO2) for 3 hours in presence of MTT. Thereafter tissues were placed in isopropanol for MTT extraction, which was measured in triplicate samples as optical density at 562 nm.
Number of replicates:
2/treatment

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
A 3 minutes exposure
Value:
102.6
Vehicle controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: non-corrosive
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
B 60 minutes exposure
Value:
96.3
Vehicle controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: non-corrosive
Other effects / acceptance of results:
The results were in agreement with the quality criteria:

Negative Control
The absolute OD562 of the negative control treated tissues in the MTT-test is an indicator of tissue viability obtained in the testing laboratory after the shipping and storing procedure and under specific conditions of the assay. The mean OD562 of the two negative control tissues should be ≥ 0.8 and ≤ 2.8 for each exposure time, which ensures that the tissue viability meets the acceptance criteria.
Positive Control
Potassium Hydroxide 8.0N solution is used as a positive control. An assay meets the acceptance criterion if mean relative tissue viability of the 60 minute positive control is < 15%.
Coefficient of Variation
In the range 20 and 100% viability, the Coefficient of Variation between tissue replicates should be ≤ 30%.

Any other information on results incl. tables

 

 

Exposure time

Mean OD562

Relative mean viability

Negative control

3 min

1.694

100%

 

60 min

1.773

100%

Test substance

3 min

1.783*

102.6%

 

60 min

1.707**

96.3%

Positive control

3 min

0.058

3.4%

 

60 min

0.051

2.9%

*Correction factor 0.020

** Correction factor 0.151

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Remarks:
according to the criteria set up in the OECD guideline 431
Conclusions:
The substance is considered non-corrosive
Executive summary:

The substance was tested in the EpiDerm™ Human Skin Model using duplicate tissues during 3 and 60 minutes. Additional controls were included to account for direct MTT interference and color interference. The relative mean viability of the test substance treated tissues was 102.6 % for the 3 minutes exposure and 96.3% for the 60 minutes exposure. Therefore it can be concluded that the substance is not corrosive to the skin.