Reaction mass of 2-dodecylhexadecyl methacrylate and 2-Tetradecyloctadecyl methacrylate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 February 2019 - 06 February 2019

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

impurity 1

Reference substance name:

Unknown Impurities

IUPAC Name:

Unknown Impurities

impurity 2

Reference substance name:

13-Methylheptacosane

Molecular formula:

C28H58

IUPAC Name:

13-Methylheptacosane

impurity 3

Reference substance name:

Hexadecyl methacrylate

EC Number:

219-672-3

EC Name:

Hexadecyl methacrylate

Cas Number:

2495-27-4

Molecular formula:

C20H38O2

IUPAC Name:

hexadecyl methacrylate

impurity 4

Reference substance name:

Tridecyl methacrylate

EC Number:

219-671-8

EC Name:

Tridecyl methacrylate

Cas Number:

2495-25-2

Molecular formula:

C17H32O2

IUPAC Name:

tridecyl methacrylate

impurity 5

Reference substance name:

Pentadecyl methacrylate

EC Number:

228-126-3

EC Name:

Pentadecyl methacrylate

Cas Number:

6140-74-5

Molecular formula:

C19H36O2

IUPAC Name:

pentadecyl methacrylate

impurity 6

Reference substance name:

Tetradecyl methacrylate

EC Number:

219-835-9

EC Name:

Tetradecyl methacrylate

Cas Number:

2549-53-3

Molecular formula:

C18H34O2

IUPAC Name:

tetradecyl methacrylate

impurity 7

Reference substance name:

Dodecyl methacrylate

EC Number:

205-570-6

EC Name:

Dodecyl methacrylate

Cas Number:

142-90-5

Molecular formula:

C16H30O2

IUPAC Name:

dodecyl methacrylate

impurity 8

Reference substance name:

2-Decyltetradecyl methacrylate

Molecular formula:

C28H54O2

IUPAC Name:

2-Decyltetradecyl methacrylate

Constituent 1

Reference substance name:

2-Tetradecyloctadecyl methacrylate

Cas Number:

1046154-63-5

Molecular formula:

C36H70O2

IUPAC Name:

2-Tetradecyloctadecyl methacrylate

Constituent 2

Reference substance name:

2-Dodecylhexadecyl methacrylate

Cas Number:

536972-42-6

Molecular formula:

C32H62O2

IUPAC Name:

2-Dodecylhexadecyl methacrylate

Test material form:

liquid

In vitro test system

Test system:

human skin model

Remarks:

EpiDerm™ reconstructed human epidermis

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: MatTek Corporation EpidermTM reconstructed tissue model: EPI-200
- Tissue batch number(s): Lot 28682
- Delivery date: 05 February 2019
- Date of initiation of testing: 05 February 2019

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C 5% CO2 - Relative Humidity (%): ≥95%

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- Spectrophotometer: BMG LabTech FluoStar Optima
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: PASS
- Barrier function: PASS
- Morphology: PASS
Tissue thickness: PASS
- Contamination: No contamination reported = PASS

NUMBER OF REPLICATE TISSUES : 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE :
Prior to the assay, the test item was checked for interference (water colouration or MTT interference) and found not to interfere.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3 tissues per condition

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be corrosive to skin if the relative tissue viability after 3 minutes treatment 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 relative tissue viability after 3 minutes treatment exposure is more and after 1 hour is less than or equal to 15%.
- Justification for the selection of the cut-off point(s) : Justification in accordance with Table 5 of testing guideline OECD 431

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Single topical administration of 50ul of neat test item

NEGATIVE CONTROL : Sterile water (tissue grade)
Lot number: RNBG8380
- Amount(s) applied : 50µl
- Concentration (if solution): Neat

POSITIVE CONTROL : Potassium Hydroxide
Lot number: SLBD3295V
- Amount(s) applied : 50µl
- Concentration (if solution): 8N

Duration of treatment / exposure:

3 minutes and 60 minutes

Duration of post-treatment incubation (if applicable):

N/A

Number of replicates:

three tissues per condition (n=3).

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 Minutes

Value:

104.23

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 minutes

Value:

101.94

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: None reported
- Direct-MTT reduction: No direct reduction
- Colour interference with MTT: No colour interference

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD570 of the negative control tissues must be ≥0.8. results for 3 minute = 1.94 and for 1 hour = 2.07 therefore passed.
- Acceptance criteria met for positive control: The mean of the positive control relative percentage viability, after 1hour exposure must be < 15% of the mean of the negative control. The result was a pass at 5.51
- Acceptance criteria met for variability between replicate measurements:should not exceed 0.3 (30%) results= Pass.

Any other information on results incl. tables

3 Minute Endpoint		NC	PC	TA1
Tissue 1	Aliquot 1 OD	1.74	0.29	2.10
	Aliquot 2 OD	1.79	0.28	2.07
Tissue 2	Aliquot 1 OD	1.90	0.34	1.85
	Aliquot 2 OD	1.88	0.33	1.85
Tissue 3	Aliquot 1 OD	2.15	0.32	2.12
	Aliquot 2 OD	2.17	0.31	2.12
All tissues	Average OD	1.94	0.31	2.02
	Average % Viability	100.00	15.96	104.23
	Average % SD	8.37	1.09	6.25
	Average % CV	8.37	6.83	5.99

1 Hour Endpoint		NC	PC	TA1
Tissue 1	Aliquot 1 OD	2.08	0.11	2.23
	Aliquot 2 OD	2.09	0.09	2.25
Tissue 2	Aliquot 1 OD	2.12	0.12	2.05
	Aliquot 2 OD	2.08	0.12	2.09
Tissue 3	Aliquot 1 OD	2.01	0.12	2.01
	Aliquot 2 OD	2.03	0.13	2.01
All tissues	Average OD	2.07	0.11	2.11
	Average % Viability	100.00	5.51	101.94
	Average % SD	1.69	0.54	4.74
	Average % CV	1.69	9.90	4.65

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The skin corrosion potential of Reaction mass of 2-Propenoic acid, 2-methyl-, 2-dodecylhexadecyl ester and 2-Propenoic acid, 2-methyl-, 2-tetradecyloctadecyl ester was evaluated. The tissue viability of the test item treated tissues were assessed and compared to a negative control. The percentage viability obtained after 3 minutes was 104.23% and after 60 minutes was 101.94%.The substance did not meet the criteria for classification as a corrosive in accordance with Regulation (EC) No. 1272/2008.

Executive summary:

The skin corrosion potential of Reaction mass of 2-Propenoic acid, 2-methyl-, 2-dodecylhexadecyl ester and 2-Propenoic acid, 2-methyl-, 2-tetradecyloctadecyl ester was evaluated using OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method). The test was GLP compliant.

Triplicate EpiDerm skin model tissues were treated with a single topical application of 50µl of Reaction mass of 2-Propenoic acid, 2-methyl-, 2-dodecylhexadecyl ester and 2-Propenoic acid, 2-methyl-, 2-tetradecyloctadecyl ester. Additional triplicate tissues were treated with 50µl of Sterile Water (Negative control) and 50µl of Potassium Hydroxide (Positive control). All tissues were exposed for 30 minutes and 60 minutes at 37°C, 5% CO2, ≥95% Relative Humidity.

All controls were valid and demonstrated the reliability of the test system. The test substance did not interfere with MTT.

Mean tissue viability (as a percentage of the negative control), was 104.23% and 101.94 % after 3 and 60 minutes of exposure, respectively. Therefore, 2-Propenoic acid, 2-methyl-, 2-dodecylhexadecyl ester and 2-Propenoic acid, 2-methyl-, 2-tetradecyloctadecyl ester does not meet the criteria for classification according to CLP Regulation (EC) No. 1272/2008.