4-methoxybenzyl alcohol

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:

April 2016 and August 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

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:

28 July 2015

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-methoxybenzyl alcohol

EC Number:

203-273-6

EC Name:

4-methoxybenzyl alcohol

Cas Number:

105-13-5

Molecular formula:

C8H10O2

IUPAC Name:

(4-methoxyphenyl)methanol

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Human Skin Model (EpiDerm Reconstructed Human Epidermis)
- Tissue batch number: 23338
- Delivery date: 24 May 2016
- Date of initiation of testing: 24 May 2016 (pre-incubation phase started)

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature (3 minutes exposure) and 37°C(1 hour exposure)

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: 20 times
- 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: Versamax®, Molecular Devices, SoftMax Pro Enterprise
- Wavelength: 570 nm
- Filter: without reference filter

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: OD = 2.078 +/- 0.070
- Barrier function: ET-50 = 5.64 hrs
- Contamination: sterile

NUMBER OF REPLICATE TISSUES: 3

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 2 (3 min and 60 min exposure)

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

Amount/concentration applied:

TEST MATERIAL
- Amount applied: 50 µL (79.4 µL/cm2)

NEGATIVE CONTROL
- Amount applied: 50 µL

POSITIVE CONTROL
- Amount applied: 50 µl
- Concentration: 0.8 N

Duration of treatment / exposure:

3 minutes and 60 minutes

Number of replicates:

2 tissues (three wells per tissue)

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

#1 (3 minutes exposure)

Value:

95.4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

#2 (60 min exposure)

Value:

15.7

Vehicle controls validity:

not applicable

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:

GHS criteria not met

Conclusions:

The test item is considered to be non corrosive to skin under the test conditions chosen.

Executive summary:

The corrosive potential of the test substance was investigated by means of the in vitro Human Skin Model Test with EpiDerm tissues models according to OECD Guideline 431.

The test item was melted in water bath and afterwards added to independent duplicate tissues of EpiDerm for an incubation time of 3 minutes or 60 minutes, respectively. The negative control (deionised water) and the positive control (0.8 N KOH) were tested in parallel. Subsequently, the test and control items were rinsed off the tissues, and a 3 hour incubation period with MTT followed. MTT solution was then aspirated from the wells and the wells were rinsed with DPBS. Inserts were transferred into new 24 well paltes. The formazan salt was extracted for about 43 hours in the refrigerator. The OD was determined in a microplate reader at 570 nm.

The required acceptability criteria were met. Furhtermore, the test item passed the MTT- and the Colour Interference pre-tests.

The relative absorbance was decreased to 95.4% after 3 minutes exposure to the test item. After an exposure period of 60 the relative absorbance was reduced to 15.7%.

Based on the results, the test item was not considered to be corrosive under the test conditions chosen.