1,1-Bis(acryloyloxymethyl)ethyl isocyanate

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:

05 Aug - 02 Sep 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

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 Sep 2014

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Behörde für Gesundheit und Verbraucherschutz, Hamburg, Germany

Test material

Test material information

Constituent 1

Reference substance name:

2-isocyanato-2-methyl-3-(prop-2-enoyloxy)propyl prop-2-enoate

Cas Number:

886577-76-0

Molecular formula:

C11H13NO5

IUPAC Name:

2-isocyanato-2-methyl-3-(prop-2-enoyloxy)propyl prop-2-enoate

Test material form:

liquid

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL- Storage condition of test material: Cool and dark place- Stability: 1 year

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: EpiDerm™ (EPI-200)

Source strain:

other: Keratinocyte strain 00267

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE- Model used: EpiDerm™ (EPI-200-SCT) (MatTek Corporation, Bratislava, Slovakia)- Tissue batch number(s): 21691- Shipping date: 2 Sept 2015 - Delivery date: 2 Sept 2015- Date of initiation of testing: 2 Sep 2015TEMPERATURE USED FOR TEST SYSTEM- Temperature used during treatment / exposure: 37 °CREMOVAL OF TEST MATERIAL AND CONTROLS-Volume and number of washing steps: The test item was carefully washed from the skin surface with Dulbecco's phosphate buffered saline (D-PBS)MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE- MTT concentration: 1 mg/mL- Incubation time: 3 h- Spectrophotometer: Tecan Sunrise Magellan Version 6.4 (Tecan Deutschland GmbH, Crailsheim, Germany)- Wavelength: 540 nmFUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA- Viability: The quality of the EpiDerm™ tissue was assessed by an MTT cell viability test.- Barrier function: The barrier function was assessed by determination of the exposure time required to reduce tissue viability by 50% (ET-50) upon application of 100 µL of 1% Triton X-100. The ET-50 value was determined to be 4.91 h.- Contamination: All biological components of the epidermis and the culture medium were tested by the manufacturer for viral, bacterial, fungal and mycoplasma contamination.NUMBER OF REPLICATE TISSUES: Duplicate tissues; from each tissue, 3 absorbance measurements after MTT incubation were performed.CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCEThe test substance did not directly reduce the MTT solution.PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)- 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- Amounts applied: 50 µLNEGATIVE CONTROL- Amounts applied: 50 µLPOSITIVE CONTROL- Amounts applied: 50 µL- Concentration: 8.0 N

Duration of treatment / exposure:

3 and 60 min

Number of replicates:

Duplicate tissues; from each tissue, 3 absorbance measurements after MTT incubation were performed.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Remarks:

mean value of 2 tissues

Run / experiment:

3 min exposure

Value:

79.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Remarks:

mean value of 2 tissues

Run / experiment:

60 min exposure

Value:

73.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:- Direct-MTT reduction: The test substance did not interfere with the MTT assay (no reducing capacity).ACCEPTANCE OF RESULTS: - Acceptance criteria met for negative control: The mean negative control OD, both for the 3 and 60 min exposure period, was in the range of ≥ 0.8 and ≤ 2.8.- Acceptance criteria met for positive control: Exposure to the positive control induced a decrease in the relative cell viability as compared to the negative control, both for the 3 min exposure period (7.1%) and for the 60 min exposure period (5.4%) thus the values were below the cut-off value at 60 min exposure confirming the validity of the test system and the specific batch of tissue models.- Acceptance criteria met for variability between replicate measurements: The SD of all replicates determined was below the limit of acceptance of 30%.- Range of historical values if different from the ones specified in the test guideline: the results of the negative and positive control fell within the historical control range (see Table 1 under "Any other information on results incl. tables").

Any other information on results incl. tables

Table 1: Historical data of negative and positive controls

Material	Criteria	Average viability (%)		OD (mean±SD)		Range of OD		Coefficient of variation (%)
		3 min	60 min	3 min	60 min	3 min	60 min	3 min	60 min
Negative control	Non-corrosive	100	100	1.78± 0.40	1.82± 0.52	1.10 - 2.29	1.02 - 2.42	22.2	28.7
8 N KOH	Corrosive	10.0	5.0	0.18± 0.09	0.09± 0.03	0.03 - 0.33	0.04 - 0.13	50.5	38.6

OD = Optical Density

Table 2: Results after treatment with the test substance and controls

Test group	OD (n = 2 tissues)	SD	% OD540compared to the control % viability	OD (n = 2 tissues)	SD	% OD540compared to the control % viability
	3 min			60 min
Negative control	1.573	0.185	100	1.507	0.035	100
Test substance	1.251	0.061	79.5	1.112	0.252	73.8
Positive control	0.111	0.10	7.1	0.082	0.006	5.4

Applicant's summary and conclusion

Interpretation of results:

other: the results of this study as a stand-alone study are not suitable for classification according to CLP/EU GHS criteria; the results may be used for classification purposes in a weight of evidence approach

Conclusions:

Under the conditions of the RHE test method the test substance showed no corrosive properties.