Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from - to
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study was performed in compliance with the Good Laboratory Practice (GLP) regulations (revised in 1997, ENV/MC/CHEM(98)17). The method followed that described in the OECD Guidelines for Testing of Chemicals, Updated Guideline No 429 Skin Sensitisation LLNA (adopted 24 April 2002).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Principles of method if other than guideline:
None
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymphnode assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent

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 Netherlands
B.V. Postbus 6174
NL - 5960 AD Horst / The Netherlands

- Age at study initiation:
8 - 12 weeks (beginning of treatment)

- Weight at study initiation:
22 +/- 1.5

- Housing:
single

- Diet (e.g. ad libitum):
pelleted standard diet, ad libitum (Harlan Winkelmann GmbH, D-33178 Borchen)

- Water (e.g. ad libitum):
ad libitum

- Acclimation period:
At least 5 days prior to the start of dosing under test conditions after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ( 22 +/- 3) °C
- Humidity (%): 30-65 %
- Air changes (per hr): --
- Photoperiod (hrs dark / hrs light): 6 a.m. - 6 p.m.

Study design: in vivo (LLNA)

Vehicle:
other: water containing 1 % (w/v) Lutrol® F 68
Concentration:
2.5%
5%
10%
No. of animals per dose:
Total 20 animals
3 test groups with 5 animals
1 control group with 5 animals
Details on study design:
according to guideline
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
The mean values and standard deviations were calculated in the body weight tables.
The ANOVA (Dunnett-test) was conducted to assess whether the difference is statistically significant between test item groups and negative control (vehicle) group. Statistical significance was at the five per cent level (p < 0.05). However, both biological and statistical significance were considered together.

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: 2.5%: 0.61 5%: 0.66 10%: 2.24
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: 2.5%: 359.8 5%: 390.4 10%: 1328.8

Any other information on results incl. tables

 Test item concentration % (w/v)  Mean DPM per animal (2 lymph nodes)  S.I.
 0  592.2  1.0
 2.5 359.8 0.61
 5 390.4 0.66
 10  1328.8  2.24

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The test item was found to be NOT a skin sensitiser under the described conditions.
Executive summary:

Purpose

The purpose of this Local Lymph Node assay was to identify the contact allergenic potential of the test item when administered to the dorsum of both ear lobes of mice. This study should provide a rational basis for risk assessment to the sensitising potential of the test item in man.

Study Design

In order to study a possible allergenic potential of the test item, three groups each of four female mice were treated with different concentrations of the test item dissolved in water (1% Lutrol F68) by topical application at the dorsum of each ear lobe (left and right) on three consecutive days. A control group of four mice was treated with the vehicle only. Five days after the first topical application, the mice were intravenously injected into a tail vein with radio-labelled thymidine (3H-methyl thymidine). Approximately five hours after intravenous injection, the mice were sacrificed and the draining auricular lymph nodes excised and pooled per group. Single cell suspensions of lymph node cells were prepared from pooled lymph nodes which were subsequently washed and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was determined by the incorporation of 3H-methyl thymidine measured in a beta-scintillation counter.

Results

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 (SI) of 0.61, 0.66, and 2.24 were determined with the test item at concentrations of 2.5, 5 and 10% (w/v) in water (1% Lutrol F68), respectively.

Conclusion

Based on the results obtained, the test item was not a skin sensitiser in this assay.