[No public or meaningful name is available]

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-10-14 to 2014-11-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

The department of health of the government of the United Kingdom

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Amides, C18, branched and linear

Molecular formula:

Not applicable as UVCB

IUPAC Name:

Amides, C18, branched and linear

Test material form:

solid: pellets

Details on test material:

- Name of test material (as cited in study report): Amides, C18, branched and linear
- Physical state: extremely pale yellow granular solid
- Lot/batch No.: 202637
- Expiration date of the lot/batch: 20 April 2016
- Storage condition of test material: at RT, protect from light
- Analytical purity: 99.3%

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/CaOlaHsd

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: 2 – 3 months
- Weight at study initiation: 15 – 23 g
- Housing: the animals were caged individually caged in suspended solid floor polypropylene cages furnished with softwood woodflakes.
- Diet: 2014C Teklad Global Rodent diet, Harlan Laboratories UK Ltd., Oxon, UK, ad libitum
- Water: (tap/filtered) water, ad libitum
- Acclimation period: at least five days

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

Study design: in vivo (LLNA)

Vehicle:

other: ethanol/distilled water 7:3

Concentration:

2.5, 5 and 10% (w/w)

No. of animals per dose:

4

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: highest concentration that was suitable for dosing: 10%
- Irritation: No signs of visual local skin irritation or irritation indicated by an equal to or greater than 25% increase in mean ear thickness were noted.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: 3H-methyl thymidine incorporation
- Criteria used to consider a positive response: The test item will be regarded as a sensitizer if at least one concentration of the test item results in a threefold or greater increase in 3HTdR incorporation compared to control values. Any test item failing to produce a threefold or greater increase in 3HTdR incorporation will be classified as a “non-sensitizer”.

TREATMENT PREPARATION AND ADMINISTRATION:
25 µL of the test item was applied to the dorsal surface of each ear of each mouse for three consecutive days; A further group of four mice received the vehicle alone in the same manner. local irritation reactions were assessed. On day 6 an injection of 250 µL phosphate buffered saline (PBS) containing 20 µCi of 3H-methyl thymidine (3H-TdR) was made into the tail vein of each experimental mouse. Five hours later, the draining auricular lymph node of each ear was excised and pooled for each experimental group. For each group 1 mL PBS was added to the pooled lymph nodes. A single cell suspension of pooled lymph node cells was prepared from by mechanical disaggregation through a stainless steel gauze. The pooled lymph node cells were pelleted, the pellet was re-suspended in PBS. Radioactive material was precipitated with 5% trichloroacetic acid at 4 °C for 18 hours. 3HTdR incorporation was measured by β-scintillation counting.

Positive control substance(s):

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

Statistics:

Not applicable

Results and discussion

Positive control results:

A group of five animals was treated with 50 µL of the positive control substance (15% α-hexyl cinnamaldehyde in ethanol/distilled water 7:3). A stimulation index of 8.21 was noted in order that 15% α-hexyl cinnamaldehyde was considered to be a sensitizer under the conditions of the test.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Value:

ca. 0.63

Test group / Remarks:

Test material concentration is 2.5%w/w. Four animals per dose used.

Remarks on result:

other: Based on these results, the test substance can be considered as a non-sensitizer. See more details on the table 1 of "Any other information on results incl. tables"

Results 2

Parameter:

SI

Value:

ca. 0.78

Test group / Remarks:

Test material concentration is 5% w/w. Four animals per dose used.

Remarks on result:

other: Based on these results, the test substance can be considered as a non-sensitizer. See more details on the table 1 of "Any other information on results incl. tables"

Results 3

Parameter:

SI

Value:

ca. 1.68

Test group / Remarks:

Test material concentration is 10% w/w. Four animals per dose used.

Remarks on result:

other: Based on these results, the test substance can be considered as a non-sensitizer. See more details on the table 1 of "Any other information on results incl. tables"

Results 4

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: see Remark

Remarks:

The number of disintegrations per minute (dpm) for the vehicle was 5.699.10, resulting in a dpm/node of 712.39. A concentration of 2.5% of the test substance shows a dpm of 3590.91, and a dpm/node of 448.86. 5% of the test substance shows a dpm of 4423.56, and a dpm/node of 552.95. The highest concentration of 10% test substance shows a dpm of 9585.23 and a dpm/node of 1198.15.

Any other information on results incl. tables

Table 1: Disintegrations per minute, disintegrations per minute/node and stimulation index

Concentration (% w/w) in ethanol/distilled water 7:3	dpm	dpm/node	Stimulation index	Result
Vehicle	5699.10	712.39	NA	NA
2.5	3590.91	448.86	0.63	Negative
5	4423.56	552.95	0.78	Negative
10	9585.23	1198.15	1.68	Negative
Positive control	NA	NA	8.21	Positive

dpm: Disintegrations per minute

NA: Not applicable

CLINICAL OBSERVATIONS AND MORTALITY DATA:

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

BODY WEIGHT:

Body weight change of the test animals between day 1 and day 6 was comparable to that observed in the corresponding control group animals over the same period.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: not classified
DSD: not classified