Amides, tall-oil fatty, N,N'-(iminodi-2,1-ethanediyl)bis-, maleated

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23-08-2012 to 04-09-2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study according to the guidelines under GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

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:

Amides, from branched and linear C16-18 and C18-unsatd. fatty acids, diethylenetriamine and tall-oil fatty acids, citrates

Cas Number:

1393571-43-1

IUPAC Name:

Amides, from branched and linear C16-18 and C18-unsatd. fatty acids, diethylenetriamine and tall-oil fatty acids, citrates

Test material form:

other: extremely viscous liquid

Details on test material:

- Name of test material (as cited in study report):Fatty acids, tall-oil, reaction products with fatty acids C16-18 and C18-unsatd, branched and linear, diethylenetriamine and citric acid
- Description : dark amber coloured extremely viscous liquid

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Strain: CBA/CaOlaHsd
- Source: Harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 15 - 23 g,
- Housing: individually in solid-floor polypropylene cages
- Diet: 2014C Teklad Global Rodent diet ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25°C
- Humidity (%): 30-70
- Air changes (per hr): ca 15
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

0, 10, 25 and 50%

No. of animals per dose:

4/dose

Details on study design:

RANGE FINDING TESTS:
- dose: 50%
- Irritation: very slight erythema on day 4 and 5
- Lymph node proliferation response: no increase of ear thickness > 25%
- Toxicity: none

MAIN STUDY
TREATMENT PREPARATION AND ADMINISTRATION:
Exposure during 3 consecutive days at the dorsal site of the ears (50 μl (25 μl per ear)) with test substance or vehicle. On day 6 3H-methyl thymidine(20 μCi/mouse) was injected in the tail vein. Five hours thereafter mice were killed and draining auricular lymph nodes were pooled per dose group and assessed for 3HTdR after 18 hours incubation at 4°C using a Beckman LS6500 scintillation system. The EC3 value is calculated based on the results at the different tested concentrations.

EXAMINATIONS
Clinical signs: twice daily on day 1-3, daily on day 3-6
Body weights: propor to administration on day 1, and prior to termination at day 6

ANIMAL ASSIGNMENT AND TREATMENT
- Criteria used to consider a positive response: a threefold or greater increase in 3HTdR incorporation compared to control values (expressed as DPM/min).

Statistics:

NA

Results and discussion

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

1.02

Test group / Remarks:

10 % w/w

Results 2

Key result

Parameter:

SI

Value:

2.63

Test group / Remarks:

25 % w/w

Results 3

Key result

Parameter:

SI

Value:

4.75

Test group / Remarks:

50 % w/w

Results 4

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: at 50 % > three fold increase compared to vehicle controls

Any other information on results incl. tables

Concentration (% w/w) in acetone/ olive oil 4:1	dpm (pooled)	dpm/Node	Stimulation Index	Result
Vehicle	14866.96	1858.37	na	na
10	15180.10	1897.51	1.02	Negative
25	39147.58	4893.45	2.63	Negative
50	70616.64	8827.08	4.75	Positive

No effects on bodyweight or clinical signs, except for slight redness of the ears at the highest concentration.

Applicant's summary and conclusion

Interpretation of results:

Category 1B (indication of skin sensitising potential) based on GHS criteria

Conclusions:

The test substance is considered sensitizing to the skin

Executive summary:

Exposure of 4 mice/ treatment during 3 consecutive days at the dorsal site of the ears with test substance (10, 25 or 50%) or vehicle. On day 6, 3H-methyl thymidine(20 μCi/mouse) was injected in the tail vein. Five hours thereafter mice were killed and draining auricular lymph nodes were pooled per dose group and assessed for 3HTdR after 18 hours incubation at 4°C using a Beckman LS6500 scintillation system.

At the highest dose tested the stimulation index was 4.75. The calculated EC3 is 27%. The test substance is considered sensitizing to the skin.