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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:
key study
Study period:
Experimental start date for non-GLP Pre-Experiment: 30th March 2007 Experimental start date (GLP main experiment): 25th April 2007 Experimental completion date: 1st May 2007 Study completion date: 11th June 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: - Study generated according to generally valid and/or internationally accepted testing guidelines - Performed according to GLP - Test parameters based on specific testing guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
yes
Remarks:
The relative humidity in the animal room was between approximately 30-82%. This deviation to the study plan, however, did not affect the validity of the study.
GLP compliance:
yes (incl. QA statement)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study report): FP-100
- Physical state: Solid, pale yellow
- Analytical purity: >99%
- Purity test date: No details provided in report
- Lot/batch No.: 060511
- Expiration date of the lot/batch: September 2007
- Storage condition of test material: At room temperature at about 20°C. Protection from light is desirable.
- Other: All test item handling was performed in the dark.

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Netherlands, By. Postbus 6174, NL - 5960 AD Horst/The Netherlands
- Age at study initiation: 7 - 8 weeks (beginning of acclimatisation)
- Weight at study initiation: No details provided
- Housing:
Housing: single
Cage Type: Makrolon Type I, with wire mesh top (EHRET GmbH, D-79302 Emmendingen)
Bedding: granulated soft wood bedding (Harlan Winkelmann GmbH, 0-33178 Borchen)
- Diet (e.g. ad libitum): pelleted standard diet, ad libitum (Harlan Winkelmann GmbH, D-33178 Borchen)
- Water (e.g. ad libitum): tap water, ad libitum, (Gemeindewerke, 0-64380 Rossdorf)
- Acclimation period: 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 (%): relative humidity 30-82%
- Air changes (per hr): no details provided in report
- Photoperiod (hrs dark / hrs light): artificial light 6.00 a.m. - 6.00 p.m

Study design: in vivo (LLNA)

Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
concentration of 6.25, 12.5, 25% (W/W) in acetone/olive oil 4+1 (V/V). the application volume of 25 microlitre was spread over the entire dorsal surface of each ear lobe once daily for three consecutive days.
No. of animals per dose:
Number of animals for the pre-test (non-GLP): 2 females
Number of animals for the main study: 16 females
Number of animals per group: 4 females (nulliparous and non-pregnant)
Number of test groups: 3
Number of control (vehicle) groups: 1
Details on study design:
RANGE FINDING TESTS:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Once daily for 3 consecutive days
- Exposure period:
- Test groups: 3
- Control group: 1
- Site: Dorsal surface of each ear lobe (left and right)
- Frequency of applications: Once daily for 3 consecutive days
- Duration:
- Concentrations:

B. CHALLENGE EXPOSURE
- No. of exposures:
- Day(s) of challenge:
- Exposure period:
- Test groups:
- Control group:
- Site:
- Concentrations:
- Evaluation (hr after challenge):

OTHER:
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
See report

Results and discussion

Positive control results:
Positive controls responded as expected

In vivo (LLNA)

Resultsopen allclose all
Key result
Parameter:
SI
Value:
ca. 0.83
Remarks on result:
other: less than 1
Key result
Parameter:
SI
Value:
ca. 0.62
Remarks on result:
other: less than 1
Key result
Parameter:
SI
Value:
ca. 0.54
Remarks on result:
other: less than 1

Any other information on results incl. tables

See report

Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Conclusions:
DISCUSSION: In order to study a possible contact allergenic potential of FP-100, three groups each of four female mice were treated daily with the test item at concentrations of 6.25, 12.5, 25% (w/v) in acetone:olive oil 4+1 (v:v) by topical application to the dorsum of each ear lobe (left and right) for three consecutive days. A control group of four mice was treated with the vehicle (acetone:olive oil 4+1 (v:v)) only. Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine (3H-methyl thymidine). Approximately five hours after intravenous injection, the mice were sacrificed, 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 p-scintillation counter.

All treated animals survived the scheduled study period and no signs of toxicity were observed.

A test item is regarded as a sensitiser in the LLNA if the exposure to one or more test concentration resulted in 3-fold or greater increase in incorporation of 3HTdR compared with concurrent controls, as indicated by the Stimulation Index (S.I.). The estimated concentration of test item required to produce a S.I. of 3 is referred to as the EC3 value.

In this study Stimulation Indices of 0.83, 0.62, and 0.54 were determined with the test item at concentrations of 6.25, 12.5, 25% in acetone:olive oil 4+1 (v:v). The fact that all S.I. values are below 1 is due to a relatively high DPM value of the vehicle control. Nevertheless, all DPM values are within a normal range.
The EC3 value could not be derived, since none of the tested concentrations induced an S.l. greater than 3.

CONCLUSION: The test item FP-100 was found to be a not a skin sensitiser under the described conditions.
Executive summary:

In the study the test item FP-100 dissolved in acetone:olive oil 4+1 (v:v) was assessed for its possible contact allergenic potential.

 

For this purpose a local lymph node assay was performed using test item concentrations of 6.25, 12.5, 25%.

 

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 0.83, 0.62, and 0.54 were determined with the test item at concentrations of 6.25, 12.5, 25% in acetone:olive oil 4+1 (v:v), respectively. The EC3 value could not be calculated, since all SI’s are below 3.

 

The test item FP-100 was not a skin sensitiser in this assay.