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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:
15 January 2015 - 16 January 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
This method is approved by international regulatory agencies as a replacement for the identification of corrosives in the in vivo Rabbit skin assay (OECD No. 404) and is specifically approved (OECD 431) as a replacement for the in vivo skin corrosivity test.

Data source

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

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:
Due to technical reason (upgrade of the plate reader), plate was measured at 540 and 570 nm, instead of 540 nm as indicated in the Study Plan. As the maximum of the absorption peak is at 570 nm, those results are reported, however all the data will be kept and archived in the raw data binder. The measurements on the two different wavelength resulted identical conclusion.

Due to unscheduled delay of reporting, the Draft Report was issued two days later than indicated in the Study Plan. However, this fact had no impact on the results or integrity of the study.
Deviations:
yes
Remarks:
This fact was considered not to adversely affect the results or integrity of the study.
Qualifier:
according to
Guideline:
EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
Version / remarks:
Due to technical reason (upgrade of the plate reader), plate was measured at 540 and 570 nm, instead of 540 nm as indicated in the Study Plan. As the maximum of the absorption peak is at 570 nm, those results are reported, however all the data will be kept and archived in the raw data binder. The measurements on the two different wavelength resulted identical conclusion.

Due to unscheduled delay of reporting, the Draft Report was issued two days later than indicated in the Study Plan. However, this fact had no impact on the results or integrity of the study.
Deviations:
yes
Remarks:
This fact was considered not to adversely affect the results or integrity 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
- Batch number: 141016
- Appearance: Blue powder
- Purity: ≥85.5%
- Expiry date: 19 November 2018
- Storage conditions: Controlled Room Temperature (15-25°C, below 70 RH%)

In vitro test system

Test system:
human skin model
Cell type:
non-transformed keratinocytes
Cell source:
other: Main basal, supra basal, spinous and granular layers and a functional stratum corneum
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN-SM
- Tissue batch number(s): 15 MAIN3 002
- Expiration date: 21 Jan 2015

KILLED EPIDERMIS
- Tissue batch number(s): 14-EKIN-043
- Expiration date: 17 November 2014

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C
- Temperature of post-treatment incubation (if applicable): 25.1-26.4°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 25 mL PBS solution
- Observable damage in the tissue due to washing: N/A
- Modifications to validated SOP:

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/mL
- Batch no: 10175554
- Expiration date: 02 May 2023
- Wavelength:

NUMBER OF REPLICATE TISSUES:
In this assay, two replicates per test item per time point were used. Two negative controls and two positive controls were also run in the assay. As the test item was coloured, two additional test item-treated tissues were used for the non specific OD evaluation. Furthermore, as the test item had an MTT interacting potential, two additional test item-treated killed epidermis and two negative control treated killed epidermis were used in the study.
Control samples:
yes, concurrent negative control
yes, concurrent no treatment
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 20mg
- applied as is, but 100 μL physiological saline was added to the test item to ensure good contact with the epidermis.

NEGATIVE CONTROL
Physiological saline
- Amount(s) applied (volume or weight): 50 μL
- Concentration (if solution): 0.9% (w/v) NaCl solution
- Batch no: 19GI12GF
- Expiry Date: 31 Aug 2018


POSITIVE CONTROL
Glacial acetic acid
- Amount(s) applied (volume or weight): 50 μL
- Batch no: 12D120031
- Expiry date: 30 Apr 2015
Duration of treatment / exposure:
4h at room temperature
Duration of post-treatment incubation (if applicable):
3h
Number of replicates:
2 per control group (plus 2 positive and negative control on killed epidermis)

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
mean
Value:
94.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Following exposure with test item, the mean cell viability was 94.7% compared to the negative control (after adjustment for non-specific MTT reduction).
This is above the threshold of 35%, therefore the test item was considered as being non-corrosive.

The experiment met the validity criteria, therefore the study was considered to be valid.

In conclusion, in this in vitro EPISKIN model test with test item, the results indicate that the test item is not corrosive to the skin.