Diallyl phthalate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 17 December 2002 and 06 January 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline Study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

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:

Diallyl phthalate

EC Number:

205-016-3

EC Name:

Diallyl phthalate

Cas Number:

131-17-9

Molecular formula:

C14H14O4

IUPAC Name:

1,2-bis(prop-2-en-1-yl) benzene-1,2-dicarboxylate

Test material form:

other: liquid

Details on test material:

- Name of test material (as cited in study report): Diallyl Phthalate
- Physical state: clear colorless liquid
- Lot/batch No.: 12092
- Storage conditions: approximately 4°C in the dark

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/Ca

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: B & K Universal Ltd, Hull, UK.
- Age at study initiation: eight to twelve weeks old
- Weight at study initiation: 15 to 23g
- Housing: solid-floor polypropylene cages furnished with softwood woodflakes.
- Free access to mains tap water and food (Certified Rat and Mouse Diet (Code 5LF2) supplied by International Product Supplies Limited, Wellingborough, Northants, UK) was allowed throughout the study.
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25
- Humidity (%): 30 to 70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (06.00 ta 18.00) and twelve hours darkness.

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

0.5, 5, or 50 % w/v

No. of animals per dose:

2 mice for preleminary test
4 mice per dose in the main test

Details on study design:

RANGE FINDING TESTS:
In order to establish that dose levels chosen for use in the main study were non-toxic a range-finder test was performed using two mice. Each mouse was treated by daily application of 25µl of the undiluted test material or a concentration of 50% w/v in acetone/olive oil 4:1, to the dorsal surface of each ear for up to three consecutive days (Days 0, 1, 2). The animals were observed daily for up to 5 days. Any signs of toxicity or signs of ill health during this period were recorded. The bodyweight of each mouse was recorded on Day 0 (prior to dosing) and the surviving mouse on Day 5. The highest concentration producing no signs of systemic toxicity or skin irritation plus two lower concentrations were chosen for the main study.

- Compound solubility: acetone/olive oil 4:1 vehicle was chosen as it produced the most suitable formulation at the required concentration.
- Irritation / Lymph node proliferation response: The highest concentration producing no signs of systemic toxicity or skin irritation plus two lower concentrations were chosen for the main study.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Criteria used to consider a positive response: The test substance will be regarded as a sensitiser if at least one concentration of the test material results in a threefold or greater increase in 3HTdR incorporation compared to control values. Any test material failing to produce a threefold or greater increase in 3HTdR incorporation, will be classified as a "non-sensitiser".

TREATMENT PREPARATION AND ADMINISTRATION:
Groups of four mice were treated with the test material at concentrations of 0.5%, 5% or 50% w/v in acetone/olive ail 4:1. Results from the range-finder test indicated that the test material would not produce systemic toxicity or excessive local irritation at the maximum concentration of 50% w/v.
The mice were treated by daily application of 25 µl of the appropriate concentration of the test material to the dorsal surface of each ear for three consecutive days (Days 0, 1, 2). The test material formulation was adrninistered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette.

Positive control substance(s):

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

Results and discussion

Positive control results:

The Stimulation Index (SI) expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group are as folIows: 5.7, 5.5, and 33.5, respctively for 5, 10, and 50% w/v in Acetone/Olive Oil 4:1.

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

0.98

Test group / Remarks:

0.5%

Results 2

Key result

Parameter:

SI

Value:

3.23

Test group / Remarks:

5%

Results 3

Key result

Parameter:

SI

Value:

10.74

Test group / Remarks:

50%

Any other information on results incl. tables

Dpm, Dpm/Node and Stimulation Index (SI)

Concentration (% w/v) in acetone/olive oil 4:1	Dpm	Dpm/Node	Stimulation Index (SI)	Result
Vehicle	9005.91	1125.74	N/A	N/A
0.5	8838.93	1104.87	0.98	Negative
5	29087.39	3635.92	3.23	Positive
50	96725.08	12090.64	10.74	Positive

Dpm/Node obtained by dividing the Dpm value by 8 (total number of lymph nodes)

Stimulation Index of 3.0 or greater indicates a positive result.

N/A = Not Applicable

The latest positive control was α-Hexylcinnamaldehyde. Five positive substances have previously been investigated.

Applicant's summary and conclusion

Interpretation of results:

sensitising

Remarks:

Migrated information Criteria used for interpretation of results: other: The test material was classified as a sensitiser according to EU labelling regulations Commission Directive 2001/59/EC.

Conclusions:

The test material was considered to be a sensitiser under the conditions of the test.