Reaction mass of dodecyl methacrylate and tridecyl methacrylate

Endpoint summary

Description of key information

Skin irritation:

- not irritating (OECD TG 431 / OECD TG 439; RL1, GLP)

- not irritating in rabbits (Draize test, RL2, non-GLP); undiluted test material applied to the intact skin of 6 animals for 4 hours; very slight or moderate to severe erythema in five rabbits 30-60 minutes after end of exposure; very slight erythema in one rabbit after one day; very slight or slight oedema in four rabbits 30-60 minutes after end of exposure; fully reversible within 2 days.

 

Eye irritation:

- not irritating (OECD TG 437; RL1, GLP)

- not irritating in rabbits (Draize eye test; non-GLP; RL2); undiluted test material applied to left eye of 3 animals, eyes not rinsed; conjunctival effects in 3/3 rabbits (slight redness, slight chemosis and moderate or severe discharge), no corneal or iridial effects; fully reversible after 2 days

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Principles of method if other than guideline:

According to Draize J H (1959). Appraisal of the safety of chemicals in foods, drugs and cosmetics. Association of Food and Drug Officials of the US, 49.

A single four-hour application was made to the left flank and a total of three four-hour applications, on consecutive days, was made to the right flank.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2404-2879 g

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

Duration of treatment / exposure:

4 h

Observation period:

3 d

Number of animals:

6

Details on study design:

TEST SITE
- Area of exposure: shorn dorso-lumbar region, 2.5 cm²
single applications (left flank)
repeated applications (right flank): The repeated-application sites were assessed approximately 30-60 minutes after the removal of the dressings following each application and 1, 2, 3, 7, 9 and 14 days after removal of the dressings following the final application.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): On removal of the dressings, the skins were cleansed with clean warm water.

OBSERVATION TIME POINTS
30-60 min, 1, 2, 3 days

SCORING SYSTEM:
- Method of calculation: Draize scale (Draize 1959),

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 2 days

Irritation parameter:

erythema score

Basis:

animal: #2-6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal: #1-6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

Single Application: Very slight or moderate to severe erythema was observed in five rabbits 30-60 minutes after decontamination. Very slight erythema was observed in one rabbit one day after decontamination. All
signs of erythema had disappeared two days after decontamination. Very slight or slight oedema was observed in four rabbits 30-60 minutes after decontamination. All signs of oedema had disappeared one day after
decontamination.

Repeated Applications: Repeated (x3) four-hour applications of the undiluted test sample to rabbit skin caused moderate to severe erythema in all six rabbits during the application phase and one, two and three days after decontamination of the final application. All signs of erythema had disappeared seven days after decontamination of the final application. Slight to severe oedema was observed in all six rabbits during the application phase and one day after decontamination of the final application. Slight to severe oedema was observed in four rabbits two days after decontamination of the final application; one rabbit had thickening obscuring any oedema. Moderate or severe oedema was still observed in three rabbits three days after decontamination of the final application.
All signs of oedema had disappeared seven days after decontamination of the final application.
Additional signs of irritation were desquamation, thickening and hardening.

Other effects:

One animal was killed during the study but its illness was not thought to be compound-related.

Single exposure

Score at time point / Reversibility	Erythema	Edema
	Max. score: 4	Max. score: 4
60 min	3/1/0/1/1/1	1/0/0/2/1/2
24 h	1//00/0/0/0	0/0/0/0/0/0
48 h	0/0/0/0/0/0	0/0/0/0/0/0
72 h	0/0/0/0/0/0	0/0/0/0/0/0
Average 24h, 48h, 72h	0.3/0/0/0/0/0	0/0/0/0/0/0
Reversibility	Completely reversible within 48 h	Not applicable

 

Repeated exposures (3)

Animal number	Erythema			Edema
	Number of applications			Number of applications
	1	1	2	1	2	3
1	2	2	4	0	1	2
2	1	1	2	0	0	2
3	0	1	3	0	1	3
4	2	2	4	2	4	4
5	1	2	4	1	1	4
6	1	3	4	2	2	4

 

Animal number	Erythema						Edema
	Days after final application						Days after final application
	1	2	3	7	9	14	1	2	3	7	9	14
1	4	4	2	-	-	-	2	2	0	-	-	-
2	3	2	2	0	0	0	2	2	0	0	0	0
3	2	2	3	0	0	0	3	3	3	0	0	0
4	4	4	4	0	0	0	4	4	4	0	0	0
5	4	4	2	0	0	0	2	0	0	0	0	0
6	4	4	3	0	0	0	4	?	3	0	0	0

- animal killed, therefore, no observation made

? thickening obscuring possible oedema

Interpretation of results:

GHS criteria not met

Conclusions:

Reaction mass of dodecyl methacrylate and tridecyl methacrylate was not irritating to skin in this study.

Executive summary:

In a primary dermal irritation study 6 female young adult New Zealand White rabbits were dermally exposed to 0.5 mL of undiluted Reaction mass of dodecyl methacrylate and tridecyl methacrylate for 4 hours. Animals then were observed for 3 days. Irritation was scored by the method of Draize.

Very slight or moderate to severe erythema was observed in five rabbits 30-60 minutes after decontamination. Very slight erythema was observed in one rabbit one day after decontamination. All signs of erythema had disappeared two days after decontamination. Very slight or slight oedema was observed in four rabbits 30-60 minutes after decontamination. All signs of oedema had disappeared one day after decontamination.

Additionally, the substance was applied in three repeated 4 hours exposures. Repeated (x3) four-hour applications of the undiluted test sample to rabbit skin caused moderate to severe erythema in all six rabbits during the application phase and one, two and three days after decontamination of the final application. All signs of erythema had disappeared seven days after decontamination of the final application. Slight to severe oedema was observed in all six rabbits during the application phase and one day after decontamination of the final application. Slight to severe oedema was observed in four rabbits two days after decontamination of the final application; one rabbit had thickening obscuring any oedema. Moderate or severe oedema was still observed in three rabbits three days after decontamination of the final application. All signs of oedema had disappeared seven days after decontamination of the final application. Additional signs of irritation were desquamation, thickening and hardening.

Reaction mass of dodecyl methacrylate and tridecyl methacrylate is not classified as skin irritant based on the results of this study according to the GHS criteria.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Reference 2

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

7 March 2018 to 23 March 2018 (Study initiation to Experimental completion)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source of test material: Kyoeisha Chemical Co., Ltd
- Lot No.of test material: 7061301
- Expiration date of the lot/batch: 13 June 2019
- Purity test date: 3 October 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (Ambient) in original container as supplied by the Sponsor. Container was kept tightly closed and away from heat or sunlight
- Stability under test conditions: Assumed stable for the duration of the study
- Solubility and stability of the test substance in the solvent/vehicle: Test item administered as suplied, no solvent used.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: None

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None, used as suplied

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other:

Justification for test system used:

This study addresses the human health endpoint skin corrosion. It uses reconstructed human epidermis (RHE) (human derived non-transformed epidermal keratinocytes) which closely mimics the histological, morphological, biochemical and physiological properties of the upper parts of the human skin. Use of reconstructed human epidermis (RHE) is recommended by the OECD and other regulatory authorities. The SkinEthicTM RHE model has been validated and is part of OECD validated reference methods (VRMs), it is a recommended model for conducting in vitro skin corrosion studies. The results of the study are believed to be of value in predicting the potential of inducing skin corrosivity by the test item in humans.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Reconstructed human epidermis tissues, SkinEthicTM RHE model
- Tissue batch number: Lot N° 18-RHE-029
- Production date:
- Shipping date:
- Delivery date:
- Date of initiation of testing: 10 March 2018

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 3 minutes at room temperature and 60 minutes at 37±1 °C in 5±1% CO2 in a humidified incubator.
- Temperature of post-treatment incubation (if applicable): After washing, inserts were transferred to holding plates containing 300 µL maintenance medium.

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: After exposure, tissues were rinsed then dried with cotton buds. Treated tissues were rinsed 20 times in a constant stream of 1 mL DPBS at a 5-8 cm distance from the insert to remove all residual test item from the epidermal surface. Mesh (applied on test item treated, negative and positive control tissues) was removed by washing. The bottom of tissue inserts were dried on sterile absorbent paper (Kim wipes) for 1-2 seconds. The surface of the stratum corneum was gently swept using both ends of a cotton tip (5-6 turns per end). After washing, inserts were transferred to holding plates (at room temperature) containing 300 µL maintenance medium.
- Observable damage in the tissue due to washing: None
- Modifications to validated SOP: None

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 180 minutes
- Spectrophotometer: SynergyHT Microplate Reader: BioTek
- Wavelength: 570 nm.
- Filter: not specified
- Filter bandwidth: not specified
- Linear OD range of spectrophotometer: not specified

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: All Optical Density (OD) values (corrected OD) for negative control replicates were between 0.896 and 1.076, against the guideline requirement of ≥ 0.8 and ≤ 3.0 (the acceptance criteria for the SkinEthicTM RHE model).
- Barrier function: Not specidfied
- Morphology: It makes use of reconstructed human epidermis (RHE) (human derived non-transformed epidermal keratinocytes) which closely mimics the histological, morphological, biochemical and physiological properties of the upper parts of the human skin.
- Contamination: None
- Reproducibility: Within acceptable historical parameters

NUMBER OF REPLICATE TISSUES: Triplicate per tissue

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
The test item did not produce direct MTT reduction when compared to the concurrent negative control (maintenance medium).

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: One main test

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 or equal to 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%.
- Justification for the selection of the cut-off point(s) if different than recommended in TG 431 and 439: not applicable

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

40 µL/0.5 cm2 of sterile distilled water (negative control)
40 µL/0.5 cm2 of C12-13 alkyl methacrylate (test item)
40 µL/0.5 cm2 of 8N KOH (positive control)

Nylon mesh was applied to assist the uniform spreading of liquid materials.

Duration of treatment / exposure:

3 minutes at room temperature and 60 minutes at 37±1 °C in 5±1% CO2 in a humidified incubator.

Duration of post-treatment incubation (if applicable):

After washing, inserts were transferred to holding plates containing 300 µL maintenance medium.

Number of replicates:

Triplicate per tissue

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

one / 3 minute exposure

Value:

72.1

Negative controls validity:

valid

Remarks:

Sterile distilled water

Positive controls validity:

valid

Remarks:

8N KOH

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

one / 60 minute exposure

Value:

101.1

Negative controls validity:

valid

Remarks:

Sterile distilled water

Positive controls validity:

valid

Remarks:

8N KOH

Treatment	Viability
	3 Minutes Exposure	60 Minutes Exposure
Negative control (Sterile distilled water)	100%	100%
C12-13 alkyl methacrylate	72.1%	101.1%
Positive control (8N KOH)	-	0.10%

Interpretation of results:

GHS criteria not met

Conclusions:

It is concluded that C12-13 alkyl methacrylate is non-corrosive in accordance with the United Nations Globally Harmonized System of Classification and Labelling of Chemicals as indicated in OECD Test Guideline 431 under the specified conditions of this study.

Reference 3

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

7 March 2018 to 23 March 2018 (Sudy initiation to experimental completion)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source of test material: Kyoeisha Chemical Co., Ltd
- Lot/batch No.of test material: 7061301
- Expiration date of the lot/batch: 13 June 2019
- Purity test date: 3 October 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (Ambient) in original container as supplied by the Sponsor. Container was kept tightly closed and away from heat or sunlight
- Stability under test conditions: Assumed stable for the duration of the study
- Solubility and stability of the test substance in the solvent/vehicle: Test substance is a liquid and was administered undiluted, as supplied, to the test system. Assumed stable for the duration of the study
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: None

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: obtained from human derived non-transformed epidermal keratinocytes

Justification for test system used:

the study addresses the human health endpoint skin irritation in accordence with REACH (Annex VII) requirements. It utilises reconstructed human epidermis (RhE) (from human derived non-transformed epidermal keratinocytes) which closely mimics the histological, morphological, biochemical and physiological properties of the upper parts of the human skin. The use of RhE is recommended by the OECD and other regulatory authorities. The SkinEthicTM RHE model has been validated and is part of the OECD validated reference methods and is a recommended model for conducting in vitro skin irritation studies. The results of the study are believed to be of value in predicting the potential of inducing skin irritation by the test item in humans.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: In Vitro Skin Irritation: Reconstructed Human Epidermis (RHE) Test (SkinEthicTM RHE model)
- Tissue batch number: 18-RHE-029
- Production date:
- Shipping date:
- Delivery date:
- Date of initiation of testing: 10 March 2018 (Experimental start)

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: Room temperature for 42 minutes
- Temperature of post-treatment incubation: Post treatment, after washing and drying, tissues were incubated in a 6-well plate containing 2 mL growth medium at 37± 1°C in 5± 1% CO2 in a humidified incubator for 42 hours.

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: After exposure, tissues were rinsed and dried. The test item was removed from the epidermal surface by rinsing 25 times in a constant stream of 1 mL DPBS at a distance of 5-8 cm. Mesh (applied on test item treated tissues, negative and positive control tissues) was removed by washing. The bottom of tissue inserts were dried on a sterile absorbent paper (Kim wipes) for 1-2 seconds. The surface of the stratum corneum was gently swept using both ends of a cotton tip. After washing, inserts were transferred to holding plates containing 300 µL maintenance medium.
- Observable damage in the tissue due to washing: None
- Modifications to validated SOP: None

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL solution
- Incubation time: Incubated at 37 ± 1 °C in 5± 1% CO2 in a 95% humidified atmosphere for 180 minutes (3-hours), in the dark.
- Spectrophotometer: SynergyHT Microplate Reader : BioTek Instruments
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: yes, cell viability in tissues of the negative control, test item and positive control was compared against laboratory historical data.
- Barrier function: Not specified
- Morphology: The cells form a multi-layered, highly differentiated and stratified epidermis model of the human epidermis that consists of main basal, supra basal, spinous and granular layers and a functional stratum corneum.
- Contamination: None
- Reproducibility: yes, standard deviation of % viability for the negative control (0.01), test item (0.06) and positive control (0.00).

NUMBER OF REPLICATE TISSUES: 3 (triplicate per tissue)

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
The test item did not produce direct MTT reduction when compared to the concurrent negative control (Maintenance medium).

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: One main study

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- Justification for the selection of the cut-off point(s) if different than recommended in TG 431 and 439: Criteria in accordence ith OECD 439.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume with unit): 16 µL of test item per 0.5 cm2 (insert of 0.5 cm2 polycarbonate filter at the air-liquid interface in a chemically defined growth medium).
- Concentration (if solution):Test item is a liquid and was administered undiluted, as suplied by the Sponsor.

NEGATIVE CONTROL
- Amount(s) applied (volume): 16 µL/0.5 cm2 of sterile dulbecco’s phosphate buffered saline (DPBS)

POSITIVE CONTROL
- Amount(s) applied (volume): 16 µL/0.5 cm2 of sodium dodecyl sulphate solution
- Concentration (if solution): 5% aqueous solution

Duration of treatment / exposure:

42-minute exposure time

Duration of post-treatment incubation (if applicable):

After washing and drying, tissues were incubated in a 6-well plate containing 2 mL growth medium at 37± 1°C in 5± 1% CO2 in a humidified incubator for 42 hours.

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 Replicates/Tissue and 3 Tissue/Run.

Value:

97.4

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: None
- Direct-MTT reduction: None
- Colour interference with MTT: None

DEMONSTRATION OF TECHNICAL PROFICIENCY:

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: Historical data values detailed below;
Optical Density at 570±30 nm
Negative Control Positive Control
(Dulbecco's phosphate-buffered saline) (Sodium dodecyl sulfate, 5% aqueous)

Exposure time 42-minute exposure time
Mean 2.081 0.028
Standard deviation 0.208 0.007
Minimum 1.964 0.021
Maximum 2.776 0.053

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of this study, the classification for C12-13 alkyl methacrylate is as follows:

Globally Harmonized System of Classification and : No Category (Non Skin Irritant)
Labelling of Chemicals

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Principles of method if other than guideline:

Draize eye test

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2311-2720 g

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

eyes were not rinsed

Observation period (in vivo):

4 d

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

SCORING SYSTEM: Draize scale; a modified form of the system described by Kay and Calandra (1962) was used to interpret and classify the numerical scores

Irritation parameter:

cornea opacity score

Basis:

animal: #1-3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: #1-3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: #1-3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal: #2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4
24 h	0/0/0	0/0/0	1/1/1	1/0/0
48 h	0/0/0	0/0/0	0/0/0	0/0/0
72 h	0/0/0	0/0/0	0/0/0	0/0/0
Average 24h, 48h, 72h	0/0/0	0/0/0	0.33/0.33/0.33	0.33/0/0

Interpretation of results:

GHS criteria not met

Conclusions:

Reaction mass of dodecyl methacrylate and tridecyl methacrylate is not irritating to the eyes in this study.

Executive summary:

In a primary eye irritation study 0.1 mL of Reaction mass of dodecyl methacrylate and tridecyl methacrylate was instilled into the conjunctival sac of the left eye of 3 female young adult New Zealand White rabbits. The eyes were not rinsed afterwards. The animals then were observed for 3 days.  Irritation was scored by the method of Draize.

Instillation into the rabbit eye caused no initial pain (class 0 on a 0-5 scale). Conjunctival effects were observed in all three rabbits and consisted of slight redness, slight chemosis and moderate or severe discharge. There were no corneal or iridial effects. All signs of irritation had disappeared two days after instillation.

As a result of these effects, Reaction mass of dodecyl methacrylate and tridecyl methacrylate is not classified as eye irritant.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 January 2018 to 24 February 2018 ( Study Initiation to Experimental Completion)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and of test material: Kyoeisha Chemical Co., Ltd
- Lot/batch No.of test material: 7061301
- Expiration date of the lot/batch: 13 June 2019
- Purity test date: 3 October 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (Ambient). Containers were kept tightly closed and away from heat or sunlight
- Stability under test conditions: Assumed stable for the duration of the study
- Solubility and stability of the test substance in the solvent/vehicle: Test item is a liquid and was administered as supplied
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: None

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Deonar Abattoir slaughter house, Mumbai, Maharashtra
- Number of eyes: Minimum of nine. Three sets, each consisting of three corneas were tested.
- Characteristics of donor animals (e.g. age, sex, weight): Between 1 to 5 years (the age of the animals was determined based on the teeth count and horn ring count in addition to the Horizontal Diameter of corneas and central corneal thickness)
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Transported (in a sealed plastic container) under cold condition in Hanks’ Balanced Salt Solution containing antibiotics [e.g., penicillin at 100 IU/mL and streptomycin at 100 μg/mL]
- Time interval prior to initiating testing: Eyes were used within 24 hours of slaughter
- indication of any existing defects or lesions in ocular tissue samples: None
- Indication of any antibiotics used: Penicillin at 100 IU/mL and Streptomycin at 100 μg/mL

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent vehicle

yes, concurrent positive control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume with unit): 750 μL of C12-13 alkyl methacrylate, as supplied

Duration of treatment / exposure:

Corneas were exposed for approximately 10 minutes ± 30 seconds at 32 ± 1 °C.

Duration of post- treatment incubation (in vitro):

After rinsing, the corneas were incubated for an additional period of approximately 2 hours ± 10 minutes at 32 ± 1ºC.

Number of animals or in vitro replicates:

Three sets, each consisting of three corneas (negative control, test item and positive control) were tested.

Details on study design:

SELECTION AND PREPARATION OF CORNEAS: The eyes were used within 24 hours of slaughter. For the pre-test procedure a baseline reading for each cornea holder was taken with medium prior to loading of the cornea on to the cornea holder. Corneas, free from defects, were dissected so they had a 2-3 mm rim of sclera and were transferred to a container containing Hanks’ Balanced Salt Solution containing antibiotics.

Selected corneas were mounted on the corneal holders with the endothelial side against the O-ring of the posterior half of the holder. The anterior half of the holder was then placed on the top of the cornea and fixed in place. Both chambers were then filled to excess with pre-warmed phenol red free Eagle's Minimum Essential Medium (EMEM) (the posterior chamber filled first to allow the cornea to return to its natural concave position), ensuring no bubbles were present within the holders. The device was then equilibrated at 32 ± 1°C for at least one hour to allow the corneas to equilibrate with the medium and achieve normal metabolic activity, to the extent possible. Following equilibration, the medium was removed from both chambers and fresh pre-warmed phenol red free EMEM added to both chambers. A baseline opacity reading was then taken for each cornea.

Corneas having an opacity value <7 opacity units for the opacitometer were used in the study. Three corneas were tested in each group.

QUALITY CHECK OF THE ISOLATED CORNEAS : Eyes were examined prior to use and corneas free from any visible defects were used. Corneas possessing an opacity less than seven opacity units or equivalent for the opacitometer were used in the study.

NUMBER OF REPLICATES : Three sets, each consisting of three corneas were tested

NEGATIVE CONTROL USED : Saline

POSITIVE CONTROL USED : Dimethylformamide

APPLICATION DOSE AND EXPOSURE TIME : 750 μL, corneas were treated and exposed for approximately 10 minutes ± 30 seconds at 32 ± 1 °C.

TREATMENT METHOD: closed chamber.

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: At the end of exposure period, the test item, positive control and negative control were removed from their respective anterior chamber and the corneal epithelium washed until no visual evidence of test item, positive control or negative control was observed using EMEM (containing phenol red).
- POST-EXPOSURE INCUBATION: After rinsing, the corneas were incubated for an additional period of approximately 2 hours ± 10 minutes at 32 ± 1ºC.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: yes, any damage by the test item is assessed by quantitative measurements of changes in corneal opacity
- Corneal permeability: yes, passage of sodium fluorescein dye measured with the aid of an Absorbance Microplate Reader (OD490)
- Others (e.g, pertinent visual observations, histopathology): None

SCORING SYSTEM: In Vitro Irritancy Score (IVIS) , see below table

DECISION CRITERIA: please specify if the decision criteria as indicated in the TG was used. yes

Irritation parameter:

in vitro irritation score

Run / experiment:

one experiment

Value:

0.5

Negative controls validity:

valid

Remarks:

Saline

Remarks on result:

no indication of irritation

Irritation parameter:

in vitro irritation score

Run / experiment:

one experiment

Value:

0.08

Remarks on result:

no indication of irritation

Irritation parameter:

in vitro irritation score

Run / experiment:

one experiment

Value:

129.5

Positive controls validity:

valid

Remarks:

Dimethylformamide

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: None

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes, the mean In-Vitro Irritancy Score (IVIS) of normal saline (control) treated corneas was 0.50 which confirmed the reliability of the test procedure.
- Acceptance criteria met for positive control: yes, the mean In-Vitro Irritancy Score (IVIS) of N,N-dimethylformamide (positive control) treated corneas was 129.50 which confirmed the reliability of the test procedure. The mean In-Vitro Irritancy Score (IVIS) of dimethylformamide (positive control) treated corneas were within the range of two standard deviation of the mean of the historical control data, confirming the reliability of test procedure.
- Range of historical values if different from the ones specified in the test guideline: Not applicable, historical values as per OECD guidence.

Interpretation of results:

GHS criteria not met

Conclusions:

The mean IVIS score for the corneas treated with C12-13 alkyl methacrylate was found to be 0.08.

Based on the results of this study, the classification for C12-13 alkyl methacrylate is as follows:
Classification (OECD 437 UN GHS) : No Category

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

Reaction mass of dodecyl methacrylate and tridecyl methacrylate was not corrosive to skin in an OECD TG 431 study and not irritating to skin in an OECD TG 439 study.

In a primary dermal irritation study 6 female young adult New Zealand White rabbits were dermally exposed to 0.5 mL of undiluted Reaction mass of dodecyl methacrylate and tridecyl methacrylate

for 4 hours. Animals then were observed for 3 days. Irritation was scored by the method of Draize.

Very slight or moderate to severe erythema was observed in five rabbits 30-60 minutes after decontamination. Very slight erythema was observed in one rabbit one day after decontamination. All signs of erythema had disappeared two days after decontamination. Very slight or slight oedema was observed in four rabbits 30-60 minutes after decontamination. All signs of oedema had disappeared one day after decontamination.

Additionally, the substance was applied in three repeated 4 hours exposures. Repeated (x3) four-hour applications of the undiluted test sample to rabbit skin caused moderate to severe erythema in all six rabbits during the application phase and one, two and three days after decontamination of the final application. All signs of erythema had disappeared seven days after decontamination of the final application. Slight to severe oedema was observed in all six rabbits during the application phase and one day after decontamination of the final application. Slight to severe oedema was observed in four rabbits two days after decontamination of the final application; one rabbit had thickening obscuring any oedema. Moderate or severe oedema was still observed in three rabbits three days after decontamination of the final application. All signs of oedema had disappeared seven days after decontamination of the final application. Additional signs of irritation were desquamation, thickening and hardening.

Reaction mass of dodecyl methacrylate and tridecyl methacrylate is not classified as skin irritant based on the results of this study according to the GHS criteria.

 

Eye irritation

Reaction mass of dodecyl methacrylate and tridecyl methacrylate was not irritating to the eyes in an OECD TG 437 study.

In a primary eye irritation study 0.1 mL of Reaction mass of dodecyl methacrylate and tridecyl methacrylate was instilled into the conjunctival sac of the left eye of 3 female young adult New Zealand White rabbits. The eyes were not rinsed afterwards. The animals then were observed for 3 days.  Irritation was scored by the method of Draize.

Instillation into the rabbit eye caused no initial pain (class 0 on a 0-5 scale). Conjunctival effects were observed in all three rabbits and consisted of slight redness, slight chemosis and moderate or severe discharge. There were no corneal or iridial effects. All signs of irritation had disappeared two days after instillation.

As a result of these effects, Reaction mass of dodecyl methacrylate and tridecyl methacrylate is not classified as eye irritant.

 

Respiratory irritation

No data on the respiratory irritation of Reaction mass of dodecyl methacrylate and tridecyl methacrylate are available.

 

There are no data gaps for the endpoint irritation/corrosion. No human information is available for this endpoint. However, there is no reason to believe that these results would not be applicable to humans.

Justification for classification or non-classification

Based on the available data, Reaction mass of dodecyl methacrylate and tridecyl methacrylate does not need to be classified for eye irritation or skin irritation according to regulation (EC) 1272/2008. Thus, no labelling is required.