Fats and Glyceridic oils, vegetable, winterized, reaction products with ammonia-ethanolamine reaction by-products

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30-07-2014 to 17-09-2014

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:

2014

Report date:

2014

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:

2-(2-hexadecyl-4,5-dihydro-1H-imidazol-1-yl)ethyl heptadecanoate; 2-(4-heptadecanoylpiperazin-1-yl)ethyl heptadecanoate; 2-heptadecyl-1-[2-(2-hexadecyl-4,5-dihydro-1H-imidazol-1-yl)ethyl]-4,5-dihydro-1H-imidazole

EC Number:

800-253-4

Cas Number:

1419212-73-9

IUPAC Name:

2-(2-hexadecyl-4,5-dihydro-1H-imidazol-1-yl)ethyl heptadecanoate; 2-(4-heptadecanoylpiperazin-1-yl)ethyl heptadecanoate; 2-heptadecyl-1-[2-(2-hexadecyl-4,5-dihydro-1H-imidazol-1-yl)ethyl]-4,5-dihydro-1H-imidazole

Constituent 2

Reference substance name:

Fats and Glyceridic oils, vegetable, winterized, reaction products with ammonia-ethanolamine reaction by-products

IUPAC Name:

Fats and Glyceridic oils, vegetable, winterized, reaction products with ammonia-ethanolamine reaction by-products

Constituent 3

Reference substance name:

Reaction products of C16 saturated and C18 unsaturated fatty acids and fatty acid glycerides with by-products from 2 aminoethanol and ammonia reaction

IUPAC Name:

Reaction products of C16 saturated and C18 unsaturated fatty acids and fatty acid glycerides with by-products from 2 aminoethanol and ammonia reaction

Test material form:

liquid: viscous

Details on test material:

- Name of test material (as cited in study report):Fats and Glyceridic oils, vegetable, winterized, reaction products with ammonia-ethanolamine reaction products
- Substance type: UVCB
- Physical state: extremely viscous liquid
- Storage condition of test material: room temperature in dark

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, 2.5, 5.0 and 10%

No. of animals per dose:

4/dose

Details on study design:

RANGE FINDING TESTS:
- dose: 10, 25, 50 and 100%
- Irritation: very slight erythema at 25 and 50%
- Lymph node proliferation response: increase of ear thickness > 25% at 25, 50 and 100%
- 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: prior 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:

2.52

Test group / Remarks:

2.5 % w/w

Results 2

Key result

Parameter:

SI

Value:

2.89

Test group / Remarks:

5 % w/w

Results 3

Key result

Parameter:

SI

Value:

4.56

Test group / Remarks:

10% w/w

Remarks on result:

other: at 10% 4.56

Results 4

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: at 10 % > 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	9409.87	1176.23	NA	NA
2.5	23681.98	2960.25	2.52	Negative
5	27150.29	3393.79	2.89	Negative
10	42901.62	5326.70	4.56	Positive

No effects on bodyweight or clinical signs

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

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 (2.5, 5 or 10%) 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 4.56. The calculated EC3 is 5.3%. The test substance is considered sensitizing to the skin.