Disodium piperazine-1,4-diethanesulphonate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 Aug 2017 to the 01 Sep 2017

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Disodium piperazine-1,4-diethanesulphonate

EC Number:

278-562-3

EC Name:

Disodium piperazine-1,4-diethanesulphonate

Cas Number:

76836-02-7

Molecular formula:

C8H18N2O6S2.2Na

IUPAC Name:

disodium 2-[4-(2-sulfonatoethyl)piperazin-1-yl]ethane-1-sulfonate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Identification: 1,4 piperazinediethanesulfonic acid disodium
salt
Appearance: White powder
Purity/Composition: No correction factor required
Test item storage: At room temperature
Stable under storage conditions until: 30 November 2018 (retest date)
CAS number: 76836-02-7
EC number: 278-562-3

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis.

Details on animal used as source of test system:

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

Justification for test system used:

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

Vehicle:

unchanged (no vehicle)

Details on test system:

Tissues
On the day of receipt the tissues were kept on agarose and stored in the refrigerator. On the next day, at least one hour before starting the assay the tissues were transferred to 6-well plates with 0.9 ml DMEM.

DMEM (Dulbecco’s Modified Eagle’s Medium)
Supplemented DMEM, serum-free supplied by MatTek Corporation.

MTT medium
MTT concentrate (5 mg/ml) diluted (1:5) with MTT diluent (supplemented DMEM). Both supplied by MatTek Corporation.

Environmental conditions
All incubations, with the exception of the test item incubation of 3 minutes at room temperature, were carried out in a controlled environment, in which optimal conditions were a humid atmosphere of 80 - 100% (actual range 56 - 84%), containing 5.0 ± 0.5% CO2 in air in the dark at 37.0 ± 1.0°C (actual range 36.3 - 37.1°C). Temperature and humidity were continuously monitored throughout the experiment. The CO2 percentage was monitored once on each working day. Temporary deviations from the humidity and CO2 percentage may occur due to opening and closing of the incubator door. Based on laboratory historical data
these deviations are considered not to affect the study integrity.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Test item was applied directly on top of the skin tissue and was spread to match the size of the tissue

Duration of treatment / exposure:

Two tissues were used for a 3-minute exposure to the test item and two for a 1-hour exposure.

Number of replicates:

The test was performed on a total of 4 tissues per test item together with a negative control and positive control.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minutes

Value:

96

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: no indication of corrosion

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 hour

Value:

94

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

12% after the 1-hour exposure

Any other information on results incl. tables

A test item is considered corrosive in the in vitro skin corrosion test if: a) The relative mean tissue viability obtained after 3-minute treatment compared to the negative control tissues is decreased below 50%. b) In addition, a test item considered non-corrosive (viability > 50%) after the 3-minute treatment is considered corrosive if the relative tissue viability after 1-hour treatment with the test item is decreased below 15%. A test item is considered non corrosive in the in vitro skin corrosion test if: a) The relative mean tissue viability obtained after the 3-minute treatment compared to the negative control tissues is not decreased below 50%. b) In addition, the relative tissue viability after the 1-hour treatment is not decreased below 15%.

The test item 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.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, 1,4 piperazinediethanesulfonic acid disodium salt is not corrosive in the in vitro skin corrosion test under the experimental conditions described.

Executive summary:

The objective of this study was to evaluate 1,4 piperazinediethanesulfonic acid disodium salt for its ability to induce skin corrosion on a human three dimensional epidermal model (EpiDerm (EPI-200)). The possible corrosive potential of the test item 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 29040-44484 of the test item was a white powder. Skin tissue was moistened with 25 µl of Milli-Q water and at least 25 mg of the test item was applied directly on top of the skin tissue. The positive control had a mean relative tissue viability of 12% 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 < 17%, 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 the test item compared to the negative control tissues was 96% and 94%, respectively. Because the mean relative tissue viability for the test item was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment the test item is considered to be not corrosive.