Propan-2-yl-oxocyclobutane carboxylate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

03-12-2018 to 07-12-2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

propan-2-yl 3-oxocyclobutane-1-carboxylate

EC Number:

834-970-9

Cas Number:

130111-95-4

Molecular formula:

C8H12O3

IUPAC Name:

propan-2-yl 3-oxocyclobutane-1-carboxylate

Test material form:

liquid

Details on test material:

Batch (Lot) Number: GR13224
Expiry date: 31 January 2020 (expiry date)
Physical Description: Colourless to pale yellow liquid
Purity/Composition: 99.6%
Storage Conditions: At room temperature

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

skin obtained from plastic surgery from multiple donors

Source strain:

not specified

Details on animal used as source of test system:

All cells used to produce EpiDerm are purchased or derived from tissue obtained by Matiek Corporation from accredited institutions. In all cases, consent was obtained by these institutions from the donor or the donor's legal next of kin, for use of the tissues or derivatives of the tissue for research purposes.

Justification for test system used:

Recommended test system in international guidelines (OECD and EC).

Vehicle:

unchanged (no vehicle)

Details on test system:

EpiDerm Skin Model (EPI-200, Lot no.: 29642, kit J and K, Appendix 4).
The model consists of normal, human-derived epidermal keratinocytes which have been
cultured to form a multilayered, highly differentiated model of the human epidermis.
It consists of organized basal, spinous and granular layers, and a multi-layered stratum
corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those
found in vivo. The EpiDerm tissues (surface 0.6 cm2) were cultured on polycarbonate
membranes of 10 mm cell culture inserts.

Source
MatTek Corporation, Ashland MA, U.S.A.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

yes, concurrent MTT non-specific colour control

Amount/concentration applied:

The liquid test item was applied undiluted (50 µL) directly on top of the tissue.

Duration of treatment / exposure:

3 minutes and 1 hour

Number of replicates:

duplicate (two)

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

experiment 1 3minute exposure

Value:

84

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

experiment 1: 1 hour exposure

Value:

75

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The in vitro skin corrosion test is considered acceptable if it meets the following criteria:
a) The absolute mean OD570
of the two tissues of the negative control should reasonably be
within the laboratory historical control data range.
b) The mean relative tissue viability following 1-hour exposure to the positive control
should be <15 %.
c) In the range 20 - 100% viability, the Coefficient of Variation (CV) between tissue
replicates should be <= 30%.
The absolute mean OD570 (optical density at 570  nm)  of the negative control tissues was
within the acceptance limits of OECD 431 (lower acceptance limit ≥0.8 and upper acceptance
limit <_ 2.8) and the laboratory historical control data range. The mean
relative tissue viability following the 1-hour exposure to the positive control was 4.2%.
In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was
<=11%, indicating that the test system functioned properly

Any other information on results incl. tables

PF-06328566 was checked for color interference in aqueous conditions and possible direct

MTT reduction by adding the test item to MTT medium. Because the solutions did not turn

blue / purple nor a blue / purple precipitate was observed it was concluded that the test item

did not interfere with the MTT endpoint.

Skin corrosion is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability

obtained after the 3-minute and 1-hour treatments with  PF-06328566 compared to the

negative control tissues was 84% and 75% respectively. Because the mean relative tissue

viability for  PF-06328566 was not below 50% after 3 minutes treatment and not below 15%

after 1 hour treatment  PF-06328566 is considered to be not corrosive.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, PF-06328566 is not corrosive in the in vitro skin corrosion test under the
experimental conditions described in this report.

Executive summary:

The objective of this study was to evaluate  PF-06328566 for its ability to induce skin

corrosion on a human three dimensional epidermal model (EpiDerm (EPI-200)). The

possible corrosive potential of  PF-06328566 was tested through topical application for

3 minutes and 1 hour.

The study procedures described in this report were based on the most recent OECD and EC

guidelines.

Batch GR13224 of  PF-06328566 was a colourless to pale yellow liquid.  PF-06328566 was

applied undiluted (50 µL) directly on top of the skin tissue.

The positive control had a mean relative tissue viability of 4.2% after the 1-hour exposure.

The absolute mean OD570

(optical density at 570  nm)  of the negative control tissues was

within the acceptance limits of OECD 431 (lower acceptance limit ≥0.8 and upper acceptance

limit <_ 2.8) and the laboratory historical control data range. In the range of 20 - 100%

viability the Coefficient of Variation between tissue replicates was <_ 11%, indicating that the

test system functioned properly.

Skin corrosion is expressed as the remaining cell viability after exposure to the test item. The

relative mean tissue viability obtained after 3-minute and 1-hour treatments with

PF-06328566 compared to the negative control tissues was 84% and 75%, respectively.

Because the mean relative tissue viability for  PF-06328566 was not below 50% after the

3-minute treatment and not below 15% after the 1-hour treatment  PF-06328566 is considered

to be not corrosive.

In conclusion,  PF-06328566 is not corrosive in the in vitro skin corrosion test under the

experimental conditions described in this report.