2,2'-ethylenedioxydiethyl bis(2-ethylhexanoate)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-compliant OECD guideline study, available as unpublished report, no restrictions, adequate for assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.42 (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:

2,2'-ethylenedioxydiethyl bis(2-ethylhexanoate)

EC Number:

202-319-2

EC Name:

2,2'-ethylenedioxydiethyl bis(2-ethylhexanoate)

Cas Number:

94-28-0

Molecular formula:

C22H42O6

IUPAC Name:

2-(2-{2-[(2-ethylhexanoyl)oxy]ethoxy}ethoxy)ethyl 2-ethylhexanoate

Details on test material:

- Name of test material (as cited in study report): Eastman TEG-EH
- Description: Eastman(TM) TEG-EH (Triethylene Glycol Bis (2-EthylHexanoate)) Plasticizer
- Physical state: clear colourless liquid
- Specific gravity: 0,968 (20/20C)
- Analytical purity: 99,1% (ester by GC)
- Impurities (identity and concentrations): see confidential details on test material
- Composition of test material, percentage of components: see confidential details on test material
- Purity test date: 2007-07-11
- Expiration date of the lot/batch : not provided (production date = 2007-07-11)
- Lot/batch No.: TD-7022702
- Storage condition of test material: room temperature in the dark

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

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 23 g
- Housing: one per cage
- Diet (e.g. ad libitum): ad libitum (Certified Rat and Mouse Diet)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least five days

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

IN-LIFE DATES: From: 2007-08-10 To 2007-08-23

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

25% and 50% v/v in acetone/olive oil 4:1 and 100%.

No. of animals per dose:

5 animals per dose

Details on study design:

RANGE FINDING TESTS:
The preliminary screening test suggested that the test material would not produce systemic toxicity or excessive local irritation at the highest suitable concentration.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: The test material 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.

TREATMENT PREPARATION AND ADMINISTRATION:
Groups of five mice were treated with the undiluted test material or the test material at concentrations of 25% or 50% v/v in acetone/olive oil 4:1. 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 1, 2, 3). The test material formulation was administered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette. A further group of five mice received the vehicle alone in the same manner.

Positive control substance(s):

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

Statistics:

Data was processed to give group mean values for disintegrations per minute and standard deviations where appropriate. Individual and group mean disintegrations per minute values were assessed for dose response relationships by analysis of homogeneity of variance followed by one way analysis of variance (ANOVA). In the event of a significant result from the ANOVA, pairwise comparisons were performed between control and treated groups. For homogenous datasets Dunnett's Multiple Comparison test was used and for non-homogenous datasets Dunnett's T3 Multiple Comparison Method was used.

Results and discussion

Positive control results:

The stimulation indices of a-Hexylcinnamaldehyde, Tech, 85% as a solution in acetone/olive oil 4:1 at concentrations of 5%, 10% and 25% v/v were 2.50, 4.03 and 9.13 respectively.
a-Hexylcinnamaldehyde was considered to be a sensitiser under the conditions of the test.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: A stimulation index of less than 3 was recorded for the three concentrations of the test material (25% and 50% v/v in acetone/olive oil 4:1 and 100%).

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: See Table 1 below

Results 3

Parameter:

SI

Value:

2.33

Test group / Remarks:

5 animals, 25% v/v

Results 4

Parameter:

SI

Value:

1.94

Test group / Remarks:

5 animals, 50% v/v

Results 5

Parameter:

SI

Value:

2.97

Test group / Remarks:

5 animals, 100% v/v

Any other information on results incl. tables

Table 1: Individual Disintegrations per Minute and Stimulation Indices

Concentration (% v/v) in acetone/olive oil 4:1	Animal Number	dpm/ Animal^a	Mean dpm/Animal (Standard Deviation)	Stimulation Index^b	Result
Vehicle	1-1	1384.20	909.29 (± 360.61)	N/A	N/A
	1-2	1104.56
	1 -3	704.98
	1-4	906.81
	1-5	445.90
25	2-1	1486.57	2123.00 (± 1531.08)	2.33	Negative
	2-2	1569.86
	2-3	1541.47
	2-4	1170.16
	2-5	4846.92
50	3-1	2490.75	1767.19 (± 617.87)	1.94	Negative
	3-2	2309.19
	3-3	1008.41
	3-4	1460.92
	3-5	1566.69
100	4-1	2666.68	2703.65* (± 745.62)	2.97	Negative
	4-2	1668.84
	4-3	2447.86
	4-4	3687.54
	4-5	3047.35

dpm = Disintegrations per minute

a = Total number of lymph nodes per animal is 2

b = Stimulation Index of 3.0 or greater indicates a positive result

N/A = Not applicable

* = Significantly different from control group p<0.05

Additional remarks:

There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test.

Bodyweight changes of the test animals between Day 1 and Day 6 were comparable to those observed in the corresponding control group animals over the same period.

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

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

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

Executive summary:

Following a preliminary screening test, three groups, each of five animals, were treated with 50 µl (25 µl per ear) of the undiluted test material or the test material as solution in acetone/olive oil 4:1 at concentrations of 25% or 50% v/v. A further group of 5 animals was treated with acetone/olive oil 4:1 alone. The stimulation index expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group was 2.33, 1.94 and 2.97 for the 25, 50 and 100% solutions. All test groups were therefore negative for sensitization.