Dodecanedioic acid, 1,12-bis[2-[4-(4,6-diphenyl-1,3,5-triazin-2-yl)-3-hydroxyphenoxy]ethyl] ester

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

5th February 2019 - 12th March 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2010

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EC No 640/2012, Part B: "Skin Sensitization: Local Lymph Node Assay".

Version / remarks:

2012

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Version / remarks:

2003

Deviations:

no

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

1,12-bis({2-[4-(4,6-diphenyl-1,3,5-triazin-2-yl)-3-hydroxyphenoxy]ethyl}) dodecanedioate

EC Number:

939-042-8

Cas Number:

1482217-03-7

Molecular formula:

C58H56N6O8

IUPAC Name:

1,12-bis({2-[4-(4,6-diphenyl-1,3,5-triazin-2-yl)-3-hydroxyphenoxy]ethyl}) dodecanedioate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): T-1620L
- Analytical purity: 99.9%
- Lot/batch No.:OF1211
- Batch expiry date: 30 April 2014
- Storage condition of test material: RT in the dark
- Other:

Specific details on test material used for the study:

- Physical Description: Pale yellow powder
- Batch (Lot) Number: 10145
- Expiry date: 29 November 2019
- Purity/Composition: 97.3%
- Storage Conditions: At room temperature

In vivo test system

Test animals

Species:

mouse

Strain:

CBA:J

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Females nulliparous and non-pregnant: yes
- Age at study initiation: approximately 11 weeks old
- Weight at study initiation: 20.2 to 24.3 g.
- Housing: in polycarbonate cages containing sterilized sawdust as bedding material equipped with water bottles.
- Diet: ad libitum, pelleted rodent diet
- Water: ad libitum, municipal tap-water
- Acclimation period: at least 5 days
- Number of Animals: 20 Females. Five females per group.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24
- Humidity (%): 40 to 70
- Air changes (per hr): >=10, with 100% fresh air (no air recirculation)
- Photoperiod : A 12-hour light/12-hour

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

10, 25 or 50% w/w

No. of animals per dose:

5

Details on study design:

PRE-SCREEN TESTS:
- Compound solubility: 50% is the highest concentration that could be prepared homogeneously
- Irritation: slight
- Systemic toxicity: none
- Ear thickness measurements: slight increase (7-9%, Day 6)
- Erythema scores: 25%: 1 (Day 2-3)
0 (Day 6);
50%: 1 (Day 2-5)
0 (Day 6);

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: SI ≥ 3,



Irritation
Erythema and eschar formation observations were performed once daily on Days 1-6 (on
Days 1-3 within 1 hour after dosing), according to the following numerical scoring system.
Furthermore, a description of all other (local) effects was recorded.
Erythema and eschar formation:
No erythema ......................................................................................................................................................... 0
Very slight erythema (barely perceptible) ............................................................................................................ 1
Well-defined erythema ......................................................................................................................................... 2
Moderate to severe erythema (beet redness) to slight eschar formation (injuries in depth) .............................. 3
Severe erythema (beet redness) to eschar formation preventing grading of erythema ..................................... 4

Positive control substance(s):

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

Results and discussion

Positive control results:

Performed not more than 6 months previously. For both scientific and animal welfare reasons, no concurrent positive control group was included in the study.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Value:

ca. 1

Variability:

±0.2

Test group / Remarks:

Group 1 (control)

Results 2

Parameter:

SI

Value:

ca. 0.8

Variability:

±0.1

Test group / Remarks:

Group 2 (10% w/w)

Results 3

Parameter:

SI

Value:

ca. 0.6

Variability:

±0.1

Test group / Remarks:

Group 3 (25% w/w)

Results 4

Parameter:

SI

Value:

ca. 1

Variability:

±0.2

Test group / Remarks:

Group 4 (50 %w/w)

Any other information on results incl. tables

Skin Reactions / Irritation

The very slight erythema of the ears as shown by all test item treated animals between Days 1 and 4 was considered not to have a toxicologically significant effect on the activity of the nodes.

 

Systemic Toxicity

No mortality occurred and no clinical signs of systemic toxicity were observed in the animals. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

 

Macroscopic Examination of the Lymph Nodes and Surrounding Area

The majority of auricular lymph nodes were considered normal in size, except for one node in one animal treated at 50% which was considered to be enlarged. No macroscopic abnormalities of the surrounding area were noted for any of the animals.

 

Radioactivity Measurements and SI Values

Mean DPM/animal values for the experimental groups treated with test item concentrations 10, 25 and 50% were 992, 835 and 1294 DPM, respectively. The mean DPM/animal value for the vehicle control group was 1297 DPM. The SI values calculated for the test item concentrations 10, 25 and 50% were 0.8, 0.6 and 1.0, respectively.

Table 2 Main Study: Skin Reactions

Group	test item (% w/w)	animal	Day 1		Day 2		Day 3		Day 4		Day 5		Day 6
			Erythema^1		Erythema		Erythema		Erythema		Erythema		Erythema
			left	right	left	right	left	right	left	right	left	right	left	right
1	0	1	0	0	0	0	0	0	0	0	0	0	0	0
		2	0	0	0	0	0	0	0	0	0	0	0	0
		3	0	0	0	0	0	0	0	0	0	0	0	0
		4	0	0	0	0	0	0	0	0	0	0	0	0
		5	0	0	0	0	0	0	0	0	0	0	0	0
2	10	6	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f	0	0
		7	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f	0	0
		8	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f	0	0
		9	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f	0	0
		10	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f	0	0
3	25	11	1f	1f	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f
		12	1f	1f	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f
		13	1f	1f	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f
		14	1f	1f	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f
		15	1f	1f	1f	1f	1f	1f	1f	1f	0f	0f	0f	0f
4	50	16	1f	1f	1f	1f	1f	1f	0f	0f	0	0	0	0
		17	1f	1f	1f	1f	1f	1f	0f	0f	0	0	0	0
		18	1f	1f	1f	1f	1f	1f	0f	0f	0	0	0	0
		19	1f	1f	1f	1f	1f	1f	0f	0f	0	0	0	0
		20	1f	1f	1f	1f	1f	1f	0f	0f	0	0	0	0

f. White staining of test item remnants on the dorsal surface of the ears which did not hamper scoring for erythema.

1 Grading erythema and eschar formation (Left = dorsal surface of left ear; right = dorsal surface of right ear):

0 = No erythema

1 = Very slight erythema (barely perceptible)

Table 3

Main Study: Relative Size Lymph Nodes, Radioactivity Counts (DPM) and Stimulation Index (SI)

Group	Test item (% w/w)	Animal	Size nodes^1		DPM^2/ animal	mean DPM ± SEM3	mean SI ± SEM
			left	right
1	0	1	n	n	843	1297 ± 299	1.0 ± 0.2
		2	n	n	575
		3	n	n	2006
		4	n	n	2001
		5	n	n	1059
2	10	6	n	n	591	992 ± 116	0.8 ± 0.1
		7	n	n	966
		8	n	n	1303
		9	n	n	996
		10	n	n	1103
3	25	11	n	n	630	835 ± 75	0.6 ± 0.1
		12	n	n	952
		13	n	n	681
		14	n	n	918
		15	n	n	995
4	50	16	n	n	1148	1294 ± 312	1.0 ± 0.2
		17	n	n	637
		18	n	n	1143
		19	n	+	2483
		20	n	n	1060

1 Relative size auricular lymph nodes (-, -- or ---: degree of reduction, +, ++ or +++: degree of enlargement, n: considered to be normal).

2 DPM = Disintegrations per minute.

3 SEM = Standard Error of the Mean.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

There was no indication that the test item elicits a SI ≥ 3 when tested up to 50%. Based on these results, T-1620L would not be regarded as a skin sensitizer and has no obligatory labelling requirement for sensitization by skin contact according to the GHS and the CLP.

Executive summary:

Although the results of this study indicate that T-1620L is regarded as a potential skin sensitizer, the response was borderline and there was no evidence of a dose relationship.  For these reasons in a later registration to US TSCA  a repeat LLNA was requested  by the US EPA. The repeat study gave an unequivocal result and is therefore assigned as a key study and used for classification and labelling. Consequently, this study is assigned as supporting information only.