Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
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 methylhydrogen maleate
- Description : 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, 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

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
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

Any other information on results incl. tables


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.

Applicant's summary and conclusion

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.