Soybean oil, epoxidized, acrylate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June-August 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been conducted according to OECD Guideline No. 429 and under GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

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:

Soybean oil, epoxidized, acrylate

EC Number:

294-415-6

EC Name:

Soybean oil, epoxidized, acrylate

Cas Number:

91722-14-4

Molecular formula:

C63H108O15

IUPAC Name:

Soybean oil, epoxidized, acrylate

Details on test material:

- Name of test material (as cited in study report): Epoxidized Soybean Oil Acrylate
- Substance type: UVCB
- Physical state: yellow viscous liquid
- Analytical purity: 100 %
- Lot/batch No.: S702300037
- Expiration date of the lot/batch: 31 December 2012
- Storage condition of test material: Room temperature (15-25 oC, below 70 RH%)

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Species and strain: CBA/J Rj mice
- Source: ELEVAGE JANVIER
Route des Chènes Secs B.P. 4105
53940 LE GENEST-ST-ISLE, France
- Age at study initiation: 11 weeks old
- Weight at study initiation: 20.3 – 22.5 grams
- Housing: Group caging / mice were provided with glass tunnel-tubes. Cage type: Type II. polypropylene/ polycarbonate
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 27 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 15-20 air exchange/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From: 20 June 2012 To: 26 June 2012

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

The Preliminary Irritation/Toxicity Test was performed in CBA/J Rj mice using two doses (test item concentrations of 50 and 25% (w/v)) in the selected vehicle. The observations recorded in the preliminary test suggest that the 50% (w/v) formulation is a suitable maximum dose level for a valid LLNA.

In the main test, groups of four female CBA/J Rj mice were treated with: 50, 25 and 10% (w/v) of the test substance in AOO.

No. of animals per dose:

4 females per dose

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: acetone/olive oil (4:1 v/v) used as vehicle
- Irritation: The Preliminary Irritation/Toxicity Test was performed in CBA/J Rj mice using two doses (test item concentrations of 50 and 25% (w/v)) in the selected vehicle. The observations recorded in the preliminary test suggest that the 50% (w/v) formulation is a suitable maximum dose level for a valid LLNA.

MAIN TEST
TREATMENT PREPARATION AND ADMINISTRATION:
In the main test, groups of four female CBA/J Rj mice were treated with: 50, 25 and 10% (w/v) of the test substance in AOO. The solutions of the test substance were applied on the dorsal surface of ears of experimental animals (25 µl/ear) for three consecutive days (Days 1, 2 and 3). There was no treatment on Days 4, 5 and 6. On Day 6, the cell proliferation in the local lymph nodes was measured by incorporation of tritiated methyl thymidine (3HTdR) and the values obtained were used to calculate stimulation indices (SI).

Positive control substance(s):

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

Results and discussion

Positive control results:

α-Hexylcinnamaldehyde (25% (w/v) dissolved in AOO) was used as a positive control. A significant lymphoproliferative response (stimulation index value of 6.8) was noted for the positive control chemical and this result confirmed the validity of the assay.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: Stimulation index values of the test substance were 2.6, 1.0 and 0.4 at treatment concentrations of 50, 25 and 10% (w/v), respectively.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: see Remark

Remarks:

Test Group Measured Group No. of DPN Stimulation Name DPM/group DPM Nodes Index Values Background (5 (w/v) % TCA ) 35.5 - Negative control AOO 1891 1855.5 8 231.9 1.0 Epoxidized Soybean Oil Acrylate 50% (w/v) in AOO 4825 4789.5 8 598.7 2.6 Epoxidized Soybean Oil Acrylate 25% (w/v) in AOO 1882 1846.5 8 230.8 1.0 Epoxidized Soybean Oil Acrylate 10% (w/v) in AOO 727 691.5 8 86.4 0.4 Positive control 25 % HCA in AOO 12703 12667.5 8 1583.4 6.8

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test substance did not have a sensitisation potential (non-sensitizer) in the Local Lymph Node Assay.

Executive summary:

The aim of this study was to determine the skin sensitisation potential of test substance following dermal exposure.

The test subtance was soluble in acetone/olive oil (4:1 v/v) (abbreviation: AOO).The test item formed an applicable solution in AOO at 50% (w/v),therefore it was chosen as vehicle for the test. The test item is very viscousliquid at room temperature, which is not applicable undiluted. The Preliminary Irritation/Toxicity Test was performed in CBA/J Rj mice using two doses (test item concentrations of 50 and 25% (w/v)) in the selected vehicle. The observations recorded in the preliminary test suggest that the 50% (w/v) formulation is a suitable maximum dose level for a valid LLNA.

In the main test, groups of four female CBA/J Rj mice were treated with: 50, 25 and 10% (w/v) of the test substance in AOO. The solutions of the test substance were applied on the dorsal surface of ears of experimental animals (25 µl/ear) for three consecutive days (Days 1, 2 and 3). There was no treatment on Days 4, 5 and 6. On Day 6, the cell proliferation in the local lymph nodes was measured by incorporation of tritiated methyl thymidine (³HTdR) and the values obtained were used to calculate stimulation indices (SI).

No mortality or sign of systemic toxicity or local irritation were observed during the study. No treatment related effects were observed on animal body weights in any treated groups. The observed clinical signs are summarized in Appendix 3. Stimulation index values of the test substance were 2.6, 1.0 and 0.4 at treatment concentrations of 50, 25 and 10% (w/v), respectively.

α-Hexylcinnamaldehyde (25% (w/v) dissolved in AOO) was used as a positive control to demonstrate the appropriate performance of the assay [1]. A significant lymphoproliferative response (stimulation index value of 6.8) was noted for the positive control chemical and this result confirmed the validity of the assay.

In conclusion, under the conditions of the present assay, the test substance did not have a sensitisation potential (non-sensitizer) in the Local Lymph Node Assay.