Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16.03.2018 - 29.03.2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
Adopted: 29th July 2016
Deviations:
yes
Remarks:
In the MTT test there were used only two tissues per each time interval for the positive control (PC) and negative control (NC).
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
2-methylhydroquinone
EC Number:
202-443-7
EC Name:
2-methylhydroquinone
Cas Number:
95-71-6
Molecular formula:
C7H8O2
IUPAC Name:
2-methylbenzene-1,4-diol
Test material form:
solid: particulate/powder

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
foreskin from a single donor
Source strain:
not specified
Justification for test system used:
non-animal test
Vehicle:
water
Details on test system:
MTT Test
Test item application: 25 mg of the test item was placed directly atop to the moistened tissue and it was spread to match size of the tissue.
Controls:
Negative control - 50 μL H2O tested with every exposure time
Positive control - 50 μL 8N KOH tested with every exposure time

Procedure: After delivery, tissues were pre-incubated overnight to release transport stress related compounds and debris. After overnight pre-incubation, medium was replaced to fresh one and tissues were topically exposed to the test item for 3 (room temperature) and 60 minutes at culture conditions (37±1°C, 5±1 % CO2, humidified). In each time interval three tissues were used per test item (C1), two tissues for the positive control (PC) and two tissues for negative control (NC). In additional freeze-killed tissues were used. In each time interval two tissues were used for the test item (C1) and two tissues for negative control (NC). After exposition, tissues were thoroughly rinsed and blotted to remove the test item/controls.
After that, tissues were transferred to 24-well plates containing MTT medium (1 mg/mL) and incubated at culture conditions for 3 hours. After MTT incubation, the blue formazan salt (formed by cellular mitochondria) was extracted with 2.0 mL/tissue of isopropyl alcohol for 2 hours at room temperature and shaking. Optical density of the extracted formazan was determined using a spectrophotometer at 570 nm.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
25 mg
Duration of treatment / exposure:
3 min./ 60 min.

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
after 3 min.
Value:
7.2
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
after 60 min.
Value:
-31.6
Negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

Direct MTT reduction - functional check in tubes

25 mgof the test item was addedto 1.0 mL of MTT medium. Suspension was incubated for 1 hour (37±1 °C, 5±1 % CO2, humidified). The test item changed colour of MTT medium from red to dark purple.

The test item reduces MTT directly.

Colour interference

After addition of test item the water changed colour to light pink and in isopropyl alcohol there were not observed any change of colour. The change of colour in water was not significant so it was no need for additional controls (colorant control).

Applicant's summary and conclusion

Interpretation of results:
Category 1A (corrosive) based on GHS criteria
Conclusions:
The test item methylhydroquinone should be regarded as corrosive, sub-category 1A since it was corrosive in an In-vitro Skin Corrosion test on EpiDermTM tissues (OECD Test No. 431: In vitro skin corrosion: reconstructed human epidermis (RHE) test method).
Executive summary:

In an In-vitro Skin Corrosion test on EpiDerm tissues (OECD Test No. 431: In vitro skin corrosion: reconstructed human epidermis (RHE) test method) the average viability of affected tissues was 7.2% of negative control average value after 3 minutes of treatment. The viability after 3-minute exposure is < 50 % and < 25 %.