Hydrogen peroxide-urea

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

23 October 2017 - 07 February 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

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

2016

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EU Methode B.40 bis (In vitro Skin Corrosion: Human Skin Model Test)

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Hydrogen peroxide-urea

EC Number:

204-701-4

EC Name:

Hydrogen peroxide-urea

Cas Number:

124-43-6

Molecular formula:

CH4N2O.H2O2

IUPAC Name:

carbamimidic acid; peroxol

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Reconstructed Human Epidermis (RHE)

Justification for test system used:

The use of the EpiDerm Kit is state of the art.

Vehicle:

water

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM (EPI-212-SCT) from MatTek In Vitro Life Science Laboratories, Bratislava
- Tissue batch number(s): 25850
- Delivery date: 24 Oct. 2017
- Date of initiation of testing: 26 Oct. 2017

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

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: thoroughly rinsed with DPBS
- Modifications to validated SOP: no

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 h
- Spectrophotometer: Anthos Reader 2010 Flexi (Anthos Microsysteme GmbH)
- Wavelength: 570 nm

NUMBER OF REPLICATE TISSUES: 2

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

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 negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount applied to the tissues: ca. 25 mg

NEGATIVE CONTROL
- Amount applied: 50 µL water

POSITIVE CONTROL
- Amount applied: 50 µL potassium hydroxide
- Concentration: 8 M

Duration of treatment / exposure:

3 and 60 minutes

Number of replicates:

2 tissues per exposure period, 3 replicates each

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

exposure period 3 minutes

Value:

72.8

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

exposure period 60 minutes

Value:

19.3

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Direct-MTT reduction: No
- Colour interference with MTT: No

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

Applicant's summary and conclusion

Interpretation of results:

other: not Category 1 (corrosive) based on GHS criteria

Conclusions:

The test item is considered to be non-corrosive to skin.

Executive summary:

The test substance was tested for skin corrosion in vitro using the EpiDermTM test kit according to the OECD guideline 431 under GLP conditions. In brief, reconstructed human epidermis was exposed to either water (negative control), 8 N potassium hydroxide (positive control), or the test material (25 mg with 25 µL distilled water) for 3 and for 60 minutes. Three replicates were run per dose and exposure period. Thereafter, the tissues were incubated with MTT medium. MTT is reduced to a blue formazan dye which is extracted and measured at 570 nm as a parameter for cell viability.

Following 3 minutes treatment with the test item, the mean value of relative tissue viability was reduced to 72.8 %.This value is above the threshold for corrosion potential (50%). After 1 hour treatment, mean value of relative tissue viability was reduced to 19.3 %.This value, too, is above the threshold for corrosion potential (15%). Therefore, the test item is considered to be non-corrosive to skin in the Reconstructed Human Epidermis (RhE) Test Method.