2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)ethane-1-sulfonic acid-potassium (1/1)

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-06-14 to 2018-09-06

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:

29 July 2016

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EU Method B.40bis (In Vitro Skin Corrosion: Human Skin Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

potassium 2-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)ethane-1-sulfonate

EC Number:

618-797-4

Cas Number:

91893-72-0

Molecular formula:

C10H9NO5S.K

IUPAC Name:

potassium 2-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)ethane-1-sulfonate

Test material form:

solid: particulate/powder

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: no data

Justification for test system used:

As recommended by OECD guideline

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™
- Tissue batch number(s): 28629
- Date of initiation of testing: 19 June 2018

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: not specified
- Temperature of post-treatment incubation (if applicable): 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Observable damage in the tissue due to washing: No


MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3h
- Spectrophotometer: Tecan Sunrise Magellan Version 6.4
- Wavelength: 540 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: OD (540-570 nm) [1.0 -3.0]: Result: 1.884 ± 0.1525
- Barrier function: ET-50 assay, 100 µL 1% Triton X-100, 4 time-points, n = 3, MTT assay, ET-50 [4.77-8.72 h]: Result: 4.79 h
- Contamination: Longe term antibiotic and antimycotic free culture, No contamination: Result: Sterile

NUMBER OF REPLICATE TISSUES: 2

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- N. of replicates : 2

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(s) applied (volume or weight with unit): 25 mg moistened with 25 µL deionized water

VEHICLE
- Amount(s) applied (volume or weight with unit): Deionized water 25 µL
- Lot/batch no. (if required): 180958001

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

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

Duration of treatment / exposure:

1st period: 3 min
2 nd period: 60 min

Duration of post-treatment incubation (if applicable):

3 h incubation with MTT

Number of replicates:

2

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Remarks:

3 minutes exposure

Run / experiment:

tissue 1

Value:

97.8

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Remarks:

3 minutes exposure

Run / experiment:

tissue 2

Value:

109.8

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 3

Irritation / corrosion parameter:

% tissue viability

Remarks:

60 minutes exposure

Run / experiment:

tissue 1

Value:

93.4

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 4

Irritation / corrosion parameter:

% tissue viability

Remarks:

60 minutes exposure

Run / experiment:

tissue 2

Value:

94.8

Vehicle controls validity:

not examined

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: 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
- Range of historical values if different from the ones specified in the test guideline:

Short time incubation – 3 min
Negative control: Mean OD 1.624 (2.5±2.4) Average viability [%] 100 (2.5 ± 2.4), Viability range [%]: 93.7–106.3; % difference 0.14–8.6, unqualified experiments: 0#1
8 N KOH: Mean OD0.126 (8.8±7.4) Average viability [%] 7.6 (8.8 ± 7.4), Viability range [%]: 2.0–15.6; % difference <0.01–23.7, unqualified experiments: 0
Long time incubation – 60 min
Negative control
Mean OD 1.650 (4.0±5.0), Average viability [%] 100 (4.0±5.0), Viability range [%]: 85.6–114.4; % difference 0.13–18.3, unqualified experiments: 0#1
8 N KOH
Mean OD 0.090 (5.7±9.8), Average viability [%] 5.9, Viability range [%]: (5.7 ± 9.8); % difference 2.0–12.6 0.30–38.4, unqualified experiments: 0#2
#1 Unqualified results = if the mean OD of the NC tissues is < 0.8 or > 2.8 if difference in viability for duplicate tissues > 30%
#2 Unqualified results = 8 N KOH: viability > 15% (1-h exposure)

Any other information on results incl. tables

Table 1: Viability after both exposure times

	Viability 3-minutes exposure		Viability 60-minutes exposure
	Optical density [OD540] mean tissue 1 and 2 (percent differences)	Mean viability (Difference of Viability) [%]	Optical density [OD540] mean tissue 1 and 2 (percent differences)	Mean viability (Difference of Viability) [%]
Negative control (deionized water)	1.564	100.0	1.606	100.0
	(2.8)	(2.7)	(0.6)	(0.5)
TA-1	1.62	103.8	1.512	94.1
	(11.6)	(12.0)	(1.5)	(1.4)
Positive control (8 N KOH)	0.068	4.4	0.070	4.4
	(11.8)	(0.5)	(0.0)	(0.0)

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions of the study, the test item tested at two exposure periods of 3 minutes or 1 hour was non-cytotoxic and, hence, predicted to be non-corrosive to skin in an experiment employing an artificial three-dimensional model of human skin. Therefore, the test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS), and is thus considered to be not corrosive to the skin.