Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

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)
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:
- Batch No.: Tk253_20090901
- BASF Test Item No.: 08/0016-3
- Purity: 99.7 area-%; dose calculation not adjusted to purity
- Stability in solvent: The stability of the test substance in the vehicle was verified indirectly by a concentration control.
- Storage: At room temperature

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 Laboratories B.V., AD Horst / The Netherlands
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: 17.7 - 21.8 g
- Housing: Single caging
- Diet (e.g. ad libitum): Pelleted standard diet, ad libitum
- Water (e.g. ad libitum):Tap water, ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): 21-65
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours

Study design: in vivo (LLNA)

Vehicle:
methyl ethyl ketone
Remarks:
for the test item; vehicle for the positive control: Acetone:olive oil (4+1)
Concentration:
25, 50 or 100% test
item concentration
No. of animals per dose:
5 animals
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: A solubility experiment was performed according to the recommendations given by OECD 429. The highest test item concentration, which can be technically used was 100 % of the undiluted test item.
- Irritation: To determine the highest test item concentration that does not induce local signs of skin irritation or systemic toxicity, a pre-test was performed in two animals. Two mice were treated with test item concentrations of 50 and 100% each on three consecutive days. In the pre-test clinical signs were recorded within 1 hour and 24 ± 4 hours after each application as well as on day 6. At the tested concentrations the animals did not show signs of excessive local irritation as confirmed by the ear thickness measurements. An increase in ear weight of about 13.4 % was observed in the animal treated with the undiluted test item (100 %) in comparison to the animal treated with 50% test item concentration. However, this was not considered as a severe sign of local irritation. Also, the animals did not show any signs of systemic toxicity.

TREATMENT PREPARATION: The test item was placed into a volumetric flask on a tared balance and methyl ethyl ketone was quantitatively added. Test item solution at different concentrations was prepared by serial dilution. The preparations were made freshly before each dosing occasion.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
For all statistical calculations SigmaStat for Windows (Version 2.0) was used. A One- Way-Analysis-of-Variance was used as statistical method. In case of significant results of the One-Way-ANOVA, multiple comparisons were performed with the Dunnett test and the Student Newman Keuls test.

Results and discussion

Positive control results:
Mean DPM per group:
Vehicle: 988.7
25 % Positive Control: 7155.5

SI:
25 % Positive Control alpha-Hexylcinnamaldehyde (mean): 8.24 (Statistically significant increase in comparison to the control (vehicle) group (p=0.008))

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: see Remark
Remarks:
25 % Oxooel 13 (mean): 2.98 (Statistically significant increase in comparison to the control (vehicle) group (p<0.001)) 50 % Oxooel 13 (mean): 4.86 (Statistically significant increase in comparison to the control (vehicle) group (p<0.001)) 100 % Oxooel 13 (mean): 9.92 (Statistically significant increase in comparison to the control (vehicle) group (p<0.001))
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: 0 (mean): 1124.3 25 % (mean): 3348.1 50 % (mean): 5468.1 100 % (mean): 11155.5

Any other information on results incl. tables

Calculation of the EC 3 value:

  Test item concentration % S.I.
Test Group 2 25 (a) 2.98 (b)
Test Group 3 50 (c) 4.86 (d)
EC3 = (a-c)[(3-d)/(b-d)] + c = 25.3 % (w/v)

EC3 = Estimated concentration for a S.I. of 3.

a,b,c,d = Co-ordinates of the two pairs of data lying immediately above and below the S.I. value of 3 on the LLNA dose response plot

Applicant's summary and conclusion