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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:
11 August 2010 - 18 October 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted under GLP conditions and OECD Guideline 429

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): MTDID 21721, TFEE-5
- Substance type: clear colorless liquid
- Physical state: liquid
- Lot/batch no.: TFEE5-276/12/09-1
- Expiration date of the lot/batch: 12 Dec 2012
- Stability under test conditions: stable
- Storage condition of test material: room temperature in the dark under nitrogen

In vivo test system

Test animals

Species:
mouse
Strain:
other: CBA/J
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River France, L'Arbresle Cedex, France
- Age at study initiation: approximate 12 weeks
- Weight at study initiation: 24 - 27 g
- Housing: individually in Macrolon cages containing sterilized sawdust bedding material and paper as cage enrichment
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.9 - 23.2
- Humidity (%): 46 - 84
- Air changes (per hr): approximately 15
- Photoperiod (hrs dark / hrs light): 12 / 12
IN-LIFE DATES: From: 11 August 2010 To: 30 August 2010

Study design: in vivo (LLNA)

Vehicle:
unchanged (no vehicle)
Concentration:
0% (naive sham control) and 100% (undiluted)
No. of animals per dose:
5
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: Results of trial formulations at the test facility showed that no suitable formulation could be prepared using the guideline vehicles (acetone/olive oil, N,N-dimethylformamide, MEK, propylene glycol, or DMSO). In addition, toluene/olive oil, 1% watery pluronic L92, and 2-propanol did not prove to be suitable vehicles.
- Irritation: No irritation and no signs of systemic toxicity were observed in one animal treated with the undiluted test article
- Lymph node proliferation response: not evaluated
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT: 5 animals each were assigned to TFEE-5 (100%) and naive (sham) control groups
- Name of test method: reduced mouse local lymph node assay (LLNA)
- Criteria used to consider a positive response: a stimulation index greater than or equal to 3 relative to the naive control group
TREATMENT PREPARATION AND ADMINISTRATION: TFEE-5 was administered undiluted by pipette to the dorsal surface of both ears (25 microliters/ear) of animals in the 100% treatment group at approximately the same time each day for 3 consecutive days. Naive control animals were treated in the same manner as the animals in the 100% treatment group except that no test article or vehicle was administered.
Positive control substance(s):
other: A concurrent positive control was not evaluated in this study. See section "Any other info on materials and methods"
Statistics:
none

Results and discussion

Positive control results:
A concurrent positive control was not evaluated in this study. A reliability check on the test system was last performed in April 2010 using the positive control substance alpha-hexyl cinnamaldehyde (HCA). In this test, stimulation indices for the positive control substance at 5%, 10% and 25% concentrations were 1.3, 1.7, and 4.1, respectively, relative to the vehicle control (acetone/olive oil 4:1 v/v) and the EC3 was 18.1%. The EC3 was in the acceptable range of 2% to 20%. EC3 values for HCA in seven previous 6-month reliability checks ranged from 11.9 to 16.0.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: 100% Test Substance: 0.8 +/- 0.2 (mean +/- SEM) Naive (sham) Control: 1.0 +/- 0.2 (mean +/- SEM)
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: 100% Test Substance: 269 +/- 57 (mean +/- SEM) Naive (sham) Control: 328 +/- 50 (mean +/- SEM))

Any other information on results incl. tables

No mortality occurred and no signs of systemic toxicity were observed among the study animals. No irritation of the ears was observed in any of the test or control animals. All auricular lymph nodes examined from the test and control animals were considered to be normal in size except for one node of one control animal which was considered to be small. Body weights and body weight gains of the test animals remained in the same range as the controls over the study period.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
TFEE-5 is not a skin sensitizer in this test system.
Executive summary:

OBJECTIVE: The dermal sensitizing potential of TFEE-5 (MTDID 21721, batch TFEE5-276/12/09-1) was evaluated in a local lymph node assay (LLNA) in female CBA/J mice.

METHODS: This study was performed according to OECD GLP guidelines (1997). The homogeneity, stability and concentration of the test substance were not analyzed. The study design was based on OECD Guideline 429 (2002), EC No. 440/2008 B42 L142 (2008), and USEPA OPPTS 870.2600 (2003). TFEE-5 was tested as received. Mice (5/treatment) received sham treatment or 100% (undiluted) TFEE-5 on three consecutive days by open application on the ears. Three days after the last exposure all animals were subjected to a 3H-methyl thymidine treatment and the auricular lymph nodes were removed to measure the amount of proliferative DNA by disintegrations per minute (DPM). The stimulation index (SI) was calculated for each group and, if possible, the concentration at which TFEE-5 could elicit an SI greater than or equal to 3 was determined (EC3). Hexylcinnamic aldehyde (HCA) is tested in this study design every 6 months at the test facility as a positive control and the most recent results indicated that model was valid.

RESULTS: Mean DPM/animal value was 269 for the 100% TFEE-5 group. The mean DPM/animal value for the vehicle control group was 328. The SI value was 0.8 for 100% TFEE-5. The EC3 could not be calculated because the 100% concentration produced less than a 3.0 SI.

CONCLUSION: Based on the results of this study, TFEE-5 showed no evidence of a dermal sensitization potential.