Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Feb 13th to Mar 7th 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study, performed in conformity with GLP-principles
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

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
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymphnode assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): LZ1780 (identical with L-Carnitine-L-Tartrate)
- Physical state: white, crystalline powder
- Lot/batch No.: 21205
- Stability under test conditions: stable
- Storage condition of test material: ambient temperature
- Expiry date: 09 December 2007

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River France, L'Arbresle Cedex, France
- Number of animals: 25 females (nulliparous and non-pregnant), five females per group
- Age and bodyweight: Young adult animals (approx. 10 weeks old) were selected. Body weight variation was within +I- 20% of the sex mean
- Housing: Individual housing

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +/- 3°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day.

Study design: in vivo (LLNA)

Vehicle:
other: 1% watery pluronic L92 (BASF, New Jersey, USA)
Concentration:
TEST GROUP:
- 10%
- 25%
- 50%

POSITIVE CONTROL:
- 25%
No. of animals per dose:
Three groups of five animals were treated with one test substance concentration per group. One group of five animals was treated with vehicle and one group of five animals was with a positive control substance.
Details on study design:
Test substance concentrations selected for the main study were based on the results of a preliminary study. In the main study, three groups of five experimental animals were treated with test substance concentrations of 10%, 25% or 50% on three consecutive days, by open application on the ears.
Five vehicle control animals were similarly treated, but with vehicle alone (1% watery pluronic L92) and five positive control animals were treated with 25% hexylcinnamic aldehyde.
Three days after the last exposure, all animals were injected with 3H-methyl thymidine and after five hours the draining (auricular) lymph nodes were excised. After precipitating the DNA of the lymph node cells, radioactivity measurements were performed. The activity was expressed as the number of Disintegrations Per Minute (DPM) and a stimulation index was subsequently calculated for each group.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:
A reliability check is carried out at regular intervals to check the sensitivity of the test system and the reliability of the experimental techniques as used by the test lab. In this study, performed in August 2005, females of the CBA mouse strain (from Charles River France, L'Arbresle Cedex, France) were checked for the sensitivity to alpha-hexylcinnamicaldehyde, tech. 85%. The females were approx. 11 weeks old at commencement of the study. Concentrations used for this study were 5, 10 and 25% in Acetone:Olive oil (4:l).

The SI values calculated for the substance concentrations 5, 10 and 25% were 2.1, 3.6 and 7.5 respectively. An EC3 value of 8.0% was calculated using linear interpolation. The calculated EC3 value was found to be in the acceptable range of 2 and 20%. The results of the 6 monthly HCA reliability checks of the recent years were 8.8, 5.5, 7.3, 10.3, 9.5 and 10.5%.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: see Remark
Remarks:
The SI values calculated for the substance concentrations 10, 25 and 50% were 2.0, 1.3 and 1.9 respectively. There was no indication that the test substance could elicit a SI >= 3. An SI value of 3.0 was calculated for the positive control substance, indicating that it is possible to elicit a positive response with hexylcinnamic aldehyde using 1% watery pluronic L92 as vehicle in the Local Lymph Node Assay as performed at NOTOX.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
Mean DPM/animal values for the experimental groups treated with test substance concentrations 10, 25 and 50% were 234, 154 and 21 7 respectively.The mean DPM/animal value for the vehicle control group was 11 6 and a mean DPM/animal value of 352 was obtained from the positive control group.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The test item was not a skin sensitiser under the described test conditions. Based on these results, the test item does not have to be classified and hasno obligatory labelling requirement for skin irritation according to the European CLP-Regulations (GHS).
Executive summary:

The test item was tested for skin sensitization in a Local Lymph Node Assay (LLNA) according to OECD-Guideline No. 429, EU-Testing Method B.42 and in conformity with GLP-principles. The study was performed from 13 February - 07 March 2006. The SI values calculated for the substance concentrations 10, 25 and 50% were 2.0, 1.3 and 1.9 respectively. There was no indication that the test substance could elicit a SI >= 3.

In conclusion, the test item was not a skin sensitiser under the described test conditions. Based on these results, the test item does not have to be classified and has no obligatory labelling requirement for skin sensitization according to the European CLP-Regulations (GHS).