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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:
information on start of stdy not yet available
Reliability:
1 (reliable without restriction)

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
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:
Balb/c
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeding farm VELAZ S.LO., Kolec u Kladna, Czech Republic, RCH CZ 21760118
- Age at study initiation: 8-10 weeks
- Weight at study initiation: 16.6 - 19.22 g
- Housing: 6/cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5d

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

Study design: in vivo (LLNA)

Vehicle:
other: DAB 433 - mixture of40% dimethylacetamide, 30% acetone and 30% ethanol
Concentration:
0.2, 2, 20 % w/v
No. of animals per dose:
5/dose
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: 20% technically highest concentration
- Irritation: no effects or clinical symptoms
- Lymph node proliferation response:

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: SI >=3

TREATMENT PREPARATION AND ADMINISTRATION: 25 ul of the appropriate dilution to the dorsum of each ear once a day morning for 3 consecutive days. The application was performed very slowly by micropipette. Losses caused by draining from the ear must be minimized.

Day 1:
Open application of 25f.lL (in the morning, by pipette) of appropriate suspensions of the test substance, the vehicle alone or the positive control to the dorsum of each ear.

Days 2 and 3:
The application procedure repeated as carried out on day 1.

Days 4 and 5:
No treatment.

Day 6:
Injection 250 ul of phosphate-buffered saline (PBS) containing 7.425 x10^5 Bq of 3H-methyl thymidine into all test and control mice via the tail vein. Five hours later, the animals were killed.
Positive control substance(s):
other: Dinitrochlorobenzene (DNCB)
Statistics:
For statistical calculations the software Statgraphic ® Centurion (version XV, USA) was used. Statistical evaluation of measured parameters was performed at first by applying the non-parametric Kruskal-Wallis test for the comparison of the measured effect in all treatment groups with the vehicle control group, as global test, and then the non-parametric two-group Mann-Whitney rank test (probability level 0.05) for all two-group comparisons.

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
Key result
Parameter:
SI
Value:
1.58
Variability:
concentration 20 % w/w
Test group / Remarks:
6 animals
Remarks on result:
other:
Remarks:
negative control: 1, positive control: 9.56
Key result
Parameter:
SI
Value:
1.05
Variability:
concentration 2 % w/w
Test group / Remarks:
6 animals
Remarks on result:
other:
Remarks:
negative control: 1, positive control: 9.56
Key result
Parameter:
SI
Value:
1.18
Variability:
concentration 0.2 % w/w
Test group / Remarks:
6 animals
Remarks on result:
other:
Remarks:
negative control: 1, positive control: 9.56

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the given test conditions, the test substance does not elicit sensitising rl)~ponse in LLNA assay.