Reaction mass of 2-hydroxy-1,3-propanediyl bismethacrylate and 101525-90-0

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10.08.2016 - 21.12.2016

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 429 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

adopted 22. July 2010

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 2-hydroxy-1,3-propanediyl bismethacrylate and 101525-90-0

EC Number:

931-227-1

Cas Number:

28497-59-8

Molecular formula:

C11H16O5

IUPAC Name:

Reaction mass of 2-hydroxy-1,3-propanediyl bismethacrylate and 101525-90-0

impurity 1

Reference substance name:

2,3-dihydroxypropyl methacrylate

EC Number:

227-642-6

EC Name:

2,3-dihydroxypropyl methacrylate

Cas Number:

5919-74-4

Molecular formula:

C7H12O4

IUPAC Name:

2,3-dihydroxypropyl methacrylate

impurity 2

Reference substance name:

Glycerol trimethacrylate

EC Number:

231-004-2

EC Name:

Glycerol trimethacrylate

Cas Number:

7401-88-9

Molecular formula:

C15H20O6

IUPAC Name:

propane-1,2,3-triyl tris(2-methylacrylate)

impurity 3

Reference substance name:

oxybis(propane-2,1,3-triyl) tetrakis(2-methylacrylate)

Molecular formula:

C22H30O9

IUPAC Name:

oxybis(propane-2,1,3-triyl) tetrakis(2-methylacrylate)

impurity 4

Reference substance name:

Benzyl methacrylate

EC Number:

219-674-4

EC Name:

Benzyl methacrylate

Cas Number:

2495-37-6

Molecular formula:

C11H12O2

IUPAC Name:

benzyl methacrylate

impurity 5

Reference substance name:

Methacrylic acid

EC Number:

201-204-4

EC Name:

Methacrylic acid

Cas Number:

79-41-4

Molecular formula:

C4H6O2

IUPAC Name:

2-methylprop-2-enoic acid

additive 1

Reference substance name:

Mequinol

EC Number:

205-769-8

EC Name:

Mequinol

Cas Number:

150-76-5

Molecular formula:

C7H8O2

IUPAC Name:

4-methoxyphenol

Test material form:

liquid

Specific details on test material used for the study:

Batch No. : 20104-24D
Reactiveester content: 96.07 %
Diester content (isomer mix): 86.4%
Triester content. 1.9 %
Methacrylic acid: 0.86%
Expiry Date: 15 January 2017
Storage Conditions: At room temperature

In vivo test system

Test animals

Species:

mouse

Strain:

other: Mice, CBA/CaOlaHsd

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- CBA/CaOlaHsd mice, nulliparous, non-pregnant
- Age at study initiation (pre-test and main study): 9 - 10 weeks (beginning of treatment)
- Weight at study initiation (main test): 17.8 - 21.1 g
- Housing: single; Makrolon Type II (pre-test)/ III (main study, with wire mesh top
- Bedding: granulated soft wood bedding
- Diet (e.g. ad libitum):2018C Teklad Global 18% protein rodent diet (certified), ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: at least 5 days prior to start of dosing
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 2°C
- Humidity (%): 45-65% (except for deviations)
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12

Study design: in vivo (LLNA)

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

1 (Control ) 0
2 (Low Dose) 0.5 % (w/v)
3 (Mid Dose) 2.5 % (w/v)
4 (High Dose) 5 % (w/v)

No. of animals per dose:

Main study: 5 females (nulliparous and non-pregnant)
3 Pre-tests: 2 females (each)

Details on study design:

To determine the highest non-irritant test concentration that at the same time did not induce signs of systemic toxicity, a pre-test was performed in two animals and stated in raw data and report. Two mice were treated by (epidermal) topical application to the dorsal surface of each ear with test item concentrations of 50 and 100% once daily each on three consecutive days. Prior to the first application of the test item and before sacrifice the body weight was determined. Clinical signs were recorded at least once daily. Eventual signs of local irritation were documented and a score was used to grade a possible erythema of the ear skin. Furthermore, prior to the first application of the test item (day 1), on day 3 and before sacrifice (day 6) the ear thickness was determined using a micrometer. Additionally, for both animals, the ears were punched after sacrifice (day 6) at the apical area using a biopsy punch (Ø 8 mm corresponding to 0.5 cm2) and were immediately pooled per animal and weighed using an analytical balance. Eventual ear irritation was considered to be excessive if an erythema of the ear skin of a score value ≥3 was observed at any observation time and/or if an increase in ear thickness of ≥25% was recorded on day 3 or day 6.
At the tested concentrations the animals showed an erythema of the ear skin (Score 3) and a decrase of body weight >5% from day 1 to day 3. Furthermore, the animal treated with 50% of the test item showed beginning eschar formation.
Therefore, the animals were sacrificed and a second pre-test performed with concentration of 10 and 25% was performed. At these concenrations, the animals showed an erythema of the ear skin (Score 1 to 2), slight erythema of scalp and slight eschar formation.The animal treated with 25% of the test item showed visible swelling of the ears and a transient decrease in body weight. Furthermore, for the animal treated with 25%, an increase of ear thickness and ear weight above the recommended threshold was also observed.
Therefore, a third pre-test was performed using concentrations of 2.5 and 5%. Both animals showed an erythema of the ear skin (Score 1 to 2) as well as scaly ears and slight eschar formation on day 6. Additionally, the animal treated with 5% test item concentration showed slight erythema of the scalp and increased spontaneous activity.
Despite the signs of irritation observed in the pre-test, the main study was performed at test item concentrations of 0.5, 1, 2.5, and 5 to cover as far as possible the concentration range in which the substance is used in practice.


MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method:
- Criteria used to consider a positive response:

TREATMENT PREPARATION AND ADMINISTRATION:

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

Within the program the Dean-Dixon-Test and Grubb’s Test were used for identification of possible outliers. An outlier (DPM value determined for animal 8) was detected in the Dean Dixon Test, but not in the Grubb’s Test, and was therefore not excluded from calculations.

Results and discussion

Positive control results:

A positive control performed with alpha-Hexylcinnamaldehyd in October 2013 resulted in an S.I. of 1.8 at 5% (w/v)
alpha-Hexylcinnamaldehyd in acetone/olive oil (4+1, v/v), 3.2 at 10 % and 5.8 at 25 %. An EC3 of 9.3 % (w/v) was calculated.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Value:

1

Test group / Remarks:

Vehicle control

Results 2

Parameter:

SI

Value:

1.1

Test group / Remarks:

0.5 % Glycerol dimethacrylate

Results 3

Parameter:

SI

Value:

1.1

Test group / Remarks:

1.0 % Glycerol dimethacrylate

Results 4

Parameter:

SI

Value:

1

Test group / Remarks:

2.5 % Glycerol dimethacrylate

Results 5

Parameter:

SI

Value:

0.7

Test group / Remarks:

5 % Glycerol dimethacrylate

Any other information on results incl. tables

Calculation of Stimulation Indices per Dose Group

Test item concentration	Group calculation
	Mean DPM per animal (2 lymph nodes)a)	SD	S.I.
Vehicle Control Group (acetone/olive oil (4+1, v/v))	2554.1	1263.1	1.0
0.5% Glycerol dimethacrylate	2808.5	1558.9	1.1
1% Glycerol dimethacrylate	2705.7	964.5	1.1
2.5% Glycerol dimethacrylate	2618.7	1672.6	1.0
5% Glycerol dimethacrylate	1913.1	284.6	0.7

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In this vaild Local Lymph Node Assay according to OECD 429 Glycerol dimethacrylate was not a skin sensitizer.
CLP EU GHS (Regulation (EC) No 1272/2008) classification: not sensitizing

Executive summary:

In this vaild dermal sensitisation study according to OECD Guideline 429 (adopted July 2010) Glycerol dimethacrylate in acetone:olive oil (4+1, v/v), groups of 5 female CBA/CaOlaHsd mice were tested using the LLNA method with the individual approach.

In this Local Lymph Node Assay Glycerol dimethcrylate was not a sensitizer.

 

The animals did not show any signs of systemic toxicity during the course of the study and no cases of mortality were observed. From day 2 to 6 the animals treated with test item concentrations of 2.5 and 5% showed an erythema of the ear skin (Score 1), as well as scaly ears on test days 5 and 6. Animals treated with 0.5 and 1% test item concentrations did not show any signs of local skin irritation. In this study Stimulation Indices (S.I.) of 1.1, 1.1, 1.0, and 0.7 were determined with the test item at concentrations of 0.5, 1, 2.5, and 5% in acetone/olive oil (4+1, v/v), respectively.

CLP EU GHS (Regulation (EC) No 1272/2008) classification: not sensitizing