Oxybis(methyl-2,1-ethanediyl) diacrylate

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:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Principles of method if other than guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay) is now describing the LLNA in detail

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Oxybis(methyl-2,1-ethanediyl) diacrylate

EC Number:

260-754-3

EC Name:

Oxybis(methyl-2,1-ethanediyl) diacrylate

Cas Number:

57472-68-1

Molecular formula:

C12H18O5

IUPAC Name:

oxydipropane-1,2-diyl bisacrylate

Details on test material:

- Name of test material (as cited in study report): Dipropylene glycol diacrylate
- Physical state: liquid
- Analytical purity: 87.0 % by GLC
- Lot/batch No.: CEFIC 17012000
- Storage condition of test material: refigerator in the dark

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan UK
- Age at study initiation: young adults
- Housing: 4mice/cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): a minimum of 15 changes/hour
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (LLNA)

Vehicle:

other: acetone

Concentration:

3%, 10% or 30% (25 µl w/v preparation of the test substance in acetone)

No. of animals per dose:

4

Details on study design:

RANGE FINDING TESTS:
no data


MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: The criterion for a positive response is that one or more concentrations of the test substance should
elicit a 3-fold or greater increase in isotope incorporation relative to the vehicle control group. The assay is able to identify those materials that elicit
moderate or greater responses in standard guinea pig tests for skin sensitisation. Consequently, a test substance which does not fulfil the above
criterion is designated as unlikely to be a moderate or strong sensitiser.


TREATMENT PREPARATION AND ADMINISTRATION:
Groups of four male mice were used for this study. Approximately 25µl of a 3%, 10% or 30% w/v preparation of the test substance in acetone was
applied, using a variable volume micro-pipette, to the dorsal surface of each ear. A vehicle control group was similarly treated using acetone alone. The procedure was repeated daily for 3 consecutive days. Three days after the third application, all the animals were injected, via the tail vein, with
approximately 250 µl of phosphate buffered saline (PBS) containing approximately 20µCi of a 2.0 Ci/mmol specific activity 3H-methyl thymidine.
Approximately 5 hours later, the animals were humanely killed by inhalation of halothane vapour followed by cervical dislocation. The draining
auricular lymph nodes were removed from each animal and, together with the nodes from the other animals in the group, were placed in a container of PBS.
A single cell suspension was prepared by mechanical disaggregation of lymph nodes through a 200-mesh stainless steel gauze. The cell suspensions were then washed three times by centrifugation with approximately 10 ml of PBS. Approximately 3ml of 5% w/v trichloroacetic acid (TCA) was added
and after overnight precipitation at 4°C, the samples were pelleted by centrifugation and the supernatant was discarded. The cells were then
resuspended in approximately 1 ml of TCA.
The lymph node suspensions were transferred to scintillation vials and 10ml of scintillant (Optiphase) was added prior to -scintillation counting using a Packard Tri-Carb 2500TR Liquid Scintillation Counter.

Positive control substance(s):

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

Statistics:

no data

Results and discussion

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Value:

19.55

Test group / Remarks:

3%

Results 2

Parameter:

SI

Value:

18.16

Test group / Remarks:

10%

Results 3

Parameter:

SI

Value:

17.58

Test group / Remarks:

30%

Results 4

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: 0 % (vehicle only): 3431 dpm; 3 %: 67078 dpm; 10 %: 62324 dpm; 30 % 60335 dpm

Any other information on results incl. tables

The application of the test substance at concentrations of 3%, 10% and 30% w/v in acetone resulted in an increase in isotope incorporation which was greater than 3-fold at all concentrations. Consequently, the test substance was shown to be a potential skin sensitiser. However, taking into account the observed irritating effects on the ears, and the missing dose-dependency of the lymph node proliferation, the sensitising effect can not be clearly discriminated from at least partly possible lymph node proliferation based

on skin irritancy.

Following the third application, the ears of animals dosed with 10% and 30% w/v preparations were red and, for the following 3 days, the ears were also swollen and sensitive to touch. The ears of animals dosed with the 3% w/v preparation were red on the day following the third application.

Applicant's summary and conclusion

Interpretation of results:

sensitising

Remarks:

Migrated information

Conclusions:

In conclusion, dipropylene glycol diacrylate is considered likely to be a skin sensitiser under the conditions of the test. However, taking into account the observed irritating effects on the ears, and the missing dose-dependency of the lymph node proliferation, the sensitising effect can not be clearly discriminated from at least partly possible lymph node proliferation based on skin irritancy.