2-propylheptyl methacrylate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

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:

2-propylheptyl methacrylate

EC Number:

810-760-2

Cas Number:

149855-64-1

Molecular formula:

C14 H26 O2

IUPAC Name:

2-propylheptyl methacrylate

Constituent 2

Reference substance name:

2-Propylheptyl methacrylate

IUPAC Name:

2-Propylheptyl methacrylate

Details on test material:

- Name of test material (as cited in study report): 2-Propylheptyl methacrylate
- Physical state:liquid, colorless, clear
- Analytical purity: 99.4 g/100g
- Lot/batch No.: S731900107

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories B.V., Horst/ The Netherlands
- Age at study initiation: main test: 9 - 10 weeks (pretest: 8 - 9 weeks / 10 - 11 weeks)
- Housing: group (Makrolon Type III, with wire mesh top)
- Diet (e.g. ad libitum): ad libitum (2018C Teklad Global 18% protein rodent diet (certified))
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2 °C
- Humidity (%): 35-65 %
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (LLNA)

Concentration:

5, 10, 25 %

No. of animals per dose:

5

Details on study design:

Each test group of mice was treated by (epidermal) topical application to the dorsal surface of each ear with test item concentrations of 5, 10 and 25% in acetone:olive oil (4+1, v/v). The application volume, 25 μL/ear/day, was spread over the entire dorsal surface (∅ ∼ 8 mm) of each ear once daily for three consecutive days. A further group of mice (control animals) was treated with an equivalent volume of the relevant vehicle alone (control animals).
Five days after the first topical application (day 6) 250 μL of phosphate-buffered saline containing 19.7 μCi of 3H-methyl thymidine (equivalent to 78.9 μCi/mL 3HTdR) were injected into each test and control mouse via the tail vein.
Approximately five hours after treatment with 3HTdR all mice were euthanised by using CO2, which was, after harvesting of the lymph nodes, followed by cervical dislocation to ensure death.
After excision, the lymph nodes were pooled per animal and weighed immediately using an analytical balance.

Positive control substance(s):

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

Statistics:

The mean values and standard deviations were calculated in the body weight tables, for the ear weights, the lymph node weights and lymph node cell count, and for the DPM values (group mean DPM ± standard deviation). The Dean-Dixon-Test and the Grubb’s test were used for detection of possible outliers (performed with Microsoft Excel 2007).

Results and discussion

Positive control results:

The sensitivity and reliability of the experimental technique employed was assessed by use of α-hexyl cinnamaldehyde dissolved in acetone/olive oil (4+1, v/v) (compound listed in OECD 429 Guideline) which is known to have skin sensitisation properties in mice. The periodic positive control experiment was performed using CBA/CaOlaHsd mice in October 2013

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: test substance: 1.17 (5% 2-PHMA); 1.24 (10% 2-PHMA); 1.34 (25% 2-PHMA)

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: test substance: 1485.2 (5% 2-PHMA); 1579.4 (10% 2-PHMA); 1703.8 (25% 2-PHMA); vehicle control: 1271.5

Any other information on results incl. tables

Lymph Node Weights and Cell Counts

The measured lymph node weights and –cell counts of all animals treated were recorded after sacrifice. A statistically significant or biologically relevant increase in lymph node weights or –cell counts was not observed in any of the test item treated groups in comparison to the vehicle control group. For BALB/c mice, a cut-off value for the lymph node cell count index of 1.55 was reported for a positive response (See Ref. 8). The indices determined for the lymph node cell count did not exceed this threshold.

Ear Weights

The measured ear weight of all animals treated was recorded on test day 6 (after necropsy). A statistically significant increase in ear weights was observed in the high dose group in comparison to the vehicle control group (p<0.05). Furthermore, the cut-off value (1.1) of the ear weight index for a positive response regarding ear skin irritation reported for BALB/c mice (see Ref. 9) was slightly exceeded in the high dose group (index of 1.12).

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))	1271.5	518.5	1.00
5% 2-Propylheptyl methacrylate	1485.2	509.2	1.17
10% 2-Propylheptyl methacrylate	1579.4	537.8	1.24
25% 2-Propylheptyl methacrylate	1703.8	692.7	1.34

a) Mean DPM/animal was determined by dividing the sum of the measured values from lymph nodes of all animals within a group by the number of animals in that group (5 animals; except for the vehicle control group with 4 animals, as animal 3 was identified as an outlier and excluded from the calculation)

Applicant's summary and conclusion

Executive summary:

The test item 2-Propylheptyl methacrylate was not a skin sensitiser under the test conditions of this study.