Phenyl 4-hydroxybenzoate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-02-20 to 2018-02-21

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: 28th July 2015

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Phenyl 4-hydroxybenzoate

EC Number:

241-698-9

EC Name:

Phenyl 4-hydroxybenzoate

Cas Number:

17696-62-7

Molecular formula:

C13H10O3

IUPAC Name:

phenyl 4-hydroxybenzoate

Specific details on test material used for the study:

- Sponsor’s identification: PHENYL 4-HYDROXYBENZOATE
- Batch No.: 018964K19K
- CAS No.:17696-62-7
- Storage: room temperature
- Form: powder
- Colour: white
- Expiry date: 2018-10-01
- Purity: 99.1%

In vitro test system

Test system:

human skin model

Source species:

human

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: The 0.6 cm² reconstituted epidermis (epiCS, Cell Systems – Batch No. 100-AH0252-1) were received on 20 February 2018. The insert (filter + epidermis) was gently removed from the agarose while avoiding leaving agarose on the polycarbonate filter. The insert was placed in a 6 wells culture plate which had been previously filled with 1 mL of culture medium (Cell Systems Batch No. 305-AH-0432). The culture dishes were incubated at 37 ± 2 °C, 5% CO2 during 21 hours and 23 minutes before treatment. Just before the treatment, the culture medium was replaced by a new culture medium (Cell Systems, Batch No. 305-AH-0432).

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature (3 min), 37 °C ± 1 °C, 5% ± 1% CO2 (1 hour)
- Temperature of post-treatment incubation (MTT incubation): 37 ± 1 °C (for 3 h, 5.0% CO2)

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: 3 minutes and 1 hour after the test item application the human epidermis were washed 20 times with 1 mL of DPBS (DPBS – Dutscher, Batch No. 5691217).
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 h (at 37 ± 1 °C)
- Spectrophotometer: plate spectrophotometer
- Wavelength: 570 nm
- Filter: not specified
- Filter bandwidth: max. ± 30 nm

PREDICTION MODEL / DECISION CRITERIA:
See Table 1 in box ‘Any other information on materials and methods incl. tables’

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL KOH
- Concentration (if solution): 8N

Negative CONTROL
- Amount(s) applied (volume or weight): 50 µL distilled water
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The test item was applied as supplied at the dose of 25 mg to the epidermal surface of the 2 living human skin models previously moistened with 25 μL of distilled water, during 3 minutes at room temperature and during 1 hour at 37°C ± 1°C, 5% ± 1% CO2.

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL
- Concentration (if solution): 8N

Duration of treatment / exposure:

3 minutes and 1 hour

Duration of post-treatment incubation (if applicable):

3 h

Number of replicates:

2

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Remarks:

3 min exposure

Run / experiment:

Mean of two replicates

Value:

100.51

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Remarks:

60 min exposure

Run / experiment:

Mean of two replicates

Value:

99.87

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: not specified
- 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

For individual results see Table 2 in box "Any other information on results incl. tables".

Any other information on results incl. tables

Table 2: Results of main experiment

	Mean viability (%)
	3 min exposure	1 hour exposure	Conclusion
Positive control	6.08	0.39	Category 1A
Test item	100.51	99.87	Non Corrosive

Applicant's summary and conclusion

Interpretation of results:

other: non-corrosive

Conclusions:

In this study under the given conditions the test item showed no corrosive effects. The test item is therefore not classified as “non-corrosive“ in accordance with UN GHS Criteria "Category 1".

Executive summary:

In a primary skin corrosion study conducted according to guideline OECD 431, two EpiDerm tissues per dose group were exposed to 25 mg of Phenyl 4-hydroxybenzoate, chloride (99.1 % purity) for 60 min and 3 min each and cytotoxicity was measured in comparison to the concurrent negative controls. Irritation was scored by the method of mean relative tissue viability. No corrosive effects were observed after treatment with the test item. The mean relative tissue viability (% negative control) of the epidermis skins treated with the test item were 100.51% (considered as 100%) and 99.87%. In accordance with the Regulation (EC) No. 1272/2008, the results obtained under these experimental conditions enable to conclude that the test item does not have to be classified for Category 1 “Corrosive”. The hazard statement “H314: Causes severe skin burns and eye damage” with the signal word “Danger” are not required.