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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Current Guideline study according to GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Netherlands; B.V. Postbus 6174; NL - 5960 AD Horst / The Netherlands
- Age at study initiation: 6-8 weeks
- Weight at study initiation: 20,2 +/- 1,2 g
- Housing: individual
- Diet (e.g. ad libitum): ad lib.
- Water (e.g. ad libitum): ad lib.
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-3
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (LLNA)

Vehicle:
acetone/olive oil (4:1 v/v)
Remarks:
in a repeat experiment: propylene glycol
Concentration:
0-5-10-25 %
No. of animals per dose:
4
Details on study design:
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: Si >/= 3 and positie dose/response relation

TREATMENT PREPARATION AND ADMINISTRATION:
Each test group of mice was treated by topical (epidermal) application to the dorsal surface of each ear lobe (left and right) with different test item concentrations of 5, 10 and 25% (w/v) in acetone:olive oil, 4:1 (v/v) in the main experiment, and 25% (w/v) in propylene glycol in the repeat experiment. The application volume, 25 μl, was spread over the entire dorsal surface (∅ ∼ 8 mm) of each ear lobe once daily for three consecutive days. A further group of mice was treated with an equivalent volume of the relevant vehicle alone (control animals). A hair dryer was used to dry the ear's surface as quickly as possible to avoid loss of test item applied.

Results and discussion

In vivo (LLNA)

Results
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Tables below

Any other information on results incl. tables

Table 1 Result from Experiment 1

 

Group/Test item conc. (%)

DPM

DPM - Background

Number of LN

DPM per LN

Stimulation Index (SI)

Back ground 1

0

 

 

 

 

Back ground 2

2,4

 

 

 

 

Aceton/Olive oil

3059,2

3058

8

382,3

1

5%

14134

14130

8

1766,6

4,6

10

13366

13365

8

1670,6

4,4

25

11291

11289

8

1411,2

3,7

 

 

Table 1 Result from Experiment 2

 

Group/Test item conc. (%)

DPM

DPM - Background

Number of LN

DPM per LN

Stimulation Index (SI)

Back ground 1

34,5

 

 

 

 

Back ground 2

34,1

 

 

 

 

Propylene glycol

2360,5

2326,2

8

290,8

1

25

9412,3

9378

8

1172,3

4,0

 

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
As the stimulation index was above 3 in all tested concentrations the test material is regarded as a sensitiser under the conditions of the LLNA.
Executive summary:

In the study the test item dissolved in acetone:olive oil, 4:1 (v/v) in the main experiment and in propylene glycol in the repeat experiment was assessed for its possible contact allergenic potential. For this purpose a local lymph node assay was performed using test item concentrations of 5, 10 and 25% in the main experiment and 25% in the repeat experiment.

The animals did not show any clinical signs during the course of the study and no cases of mortality were observed.

In this study Stimulation Indices (S.I.) of 4.6, 4.4 and 3.7 were determined with the test item at concentrations of 5, 10 and 25% (w/v) in acetone:olive oil, 4:1 (v/v), respectively.

In the repeat experiment a Stimulation Index of 4.0 was determined with the test item at a concentration of 25% (w/v) in propylene glycol.

An EC3 value could not be calculated since the S.I. was above 3 even at the lowest tested concentration of 5 % (w/v).

The test item was found to be a skin sensitiser under the described conditions.