Fatty acids, C16-18 and C18-unsatd., branched and linear, reaction products with diethylenetriamine and tall-oil fatty acids, Me maleates

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23-08-2012 to 25-09-2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study according to the guidelines under GLP

Qualifier:

according to guideline

Guideline:

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

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)

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

Vehicle:

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

Concentration:

0, 1, 2.5 and 5%

No. of animals per dose:

4/dose

Details on study design:

RANGE FINDING TESTS:
- dose: 1, 5, 25 and 50%
- Irritation: very slight erythema at 5, 10 and 50%
- Lymph node proliferation response: increase of ear thickness > 25% at 10, 25 and 50%
- Toxicity: none

MAIN STUDY
TREATMENT PREPARATION AND ADMINISTRATION:
Exposure during 3 consecutive days at the dorsal site of the ears 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

Parameter:

SI

Remarks on result:

other: at 5% 5.15

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: at 5 % > three fold increase compared to vehicle controls

Concentration (% w/w) in acetone/ olive oil 4:1	dpm (pooled)	dpm/Node	Stimulation Index	Result
Vehicle	23838.77	2979.85	na	na
1	49802.74	6225.34	2.09	Negative
2.5	68597.50	8574.69	2.88	Negative
5	122867.00	15358.38	5.15	Positive

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

Interpretation of results:

sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

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 (1, 2.5 or 5%) 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 5.15. The calculated EC3 is 3%. The test substance is considered sensitizing to the skin.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

In an LLNA test 4 mice/treatment were exposed to the test substance (1, 2.5 or 5%) or vehicle at the dorsal site of the ears during 3 consecutive days. On day 6, 3H-methyl thymidine(20 μCi/mouse) was injected in the tail vein. At the highest dose tested the stimulation index was 5.15. The calculated EC3 is 3%. The test substance is considered sensitizing to the skin (Harlan 2012j).

Migrated from Short description of key information:
The test substance was sensitizing in the local lymphnode assay at a concentration of 5%. The EC3 was calculated to be 3%.

Justification for selection of skin sensitisation endpoint:
Study according to the guideline under GLP

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

In view of the low vapour pressure (2.3E-03 Pa at 25 °C) and the extreme viscosity of the test substance, exposure via the inhalation route is expected to negligible.

Justification for classification or non-classification

The test substance needs to be classified as a skin sensitizer: category 1B H317