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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 442B (Skin Sensitization: Local Lymph Node Assay: BrdU-ELISA)
Version / remarks:
adopted July 2010
Qualifier:
according to guideline
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
adopted May 2008
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
adopted March 2003
GLP compliance:
yes (incl. QA statement)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction products of fatty acids, tall oil and fatty acids, C18 unsaturated, trimers and fatty acids, C18 unsaturated, dimers with (9Z)-octadec-9-en-1-amine
EC Number:
942-330-6
Molecular formula:
Not applicable (UVCB substance)
IUPAC Name:
Reaction products of fatty acids, tall oil and fatty acids, C18 unsaturated, trimers and fatty acids, C18 unsaturated, dimers with (9Z)-octadec-9-en-1-amine
Details on test material:
- Name of test material (as cited in study report): Reaction mass of Fatty acids, tall-oil, compds. with oleylamine and Fatty acids, C18-unsatd., trimers, compds. with oleylamine
- Physical state: viscous light brown to clear liquid
- Analytical purity: The test item is a complex mixture of isomers and homologues components, so no purity can be stated. (For details see analytical report BASF study-No.: 13L00017)
- Purity test date: Feb.-April 2013
- Lot/batch No.: D987-AM-202011
- Expiration date of the lot/batch: 2013-04-18
- Storage condition of test material: 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 Winkelmann GmbH, Borchen, Germany
- Age at study initiation: main test: 8 weeks; pretest: 9-12 weeks
- Weight at study initiation: 19.5-21.6g
- Housing: single in Makrolon type II cages
- Diet (e.g. ad libitum): Stanrab (P) SQC, SDS Special Diets Services, ad lib.
- Water (e.g. ad libitum): tab water ad lib.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 30-70%
- Air changes (per hr): app. 10
- Photoperiod (hrs dark / hrs light): 12h/12h

Study design: in vivo (LLNA)

Vehicle:
propylene glycol
Concentration:
0.5%, 1%, 2%
No. of animals per dose:
2 (pre study)
5 (main study)
Details on study design:
RANGE FINDING TESTS: 3 pre-tests were performed in 6 animals per test (2/concentration) to determine the highest non irritant concentration. Mice were treated with 100%, 50%, 25%, 10%, 5%, and 2.5% (w/w) on three consecutive days.
- Irritation: at 10% and above, mice showed local irritation confirmed by increased ear weights >25%. In addition, concentrations above 25% also led to ear swelling > 25%. Body weight loss >5% was observed at 100%, 50%, and in one animal treated with 25%. Groups 5% also showed increased ear weights >25%, while only one animal treated with 2.5% showed slightly increased ear weight. Thus concentrations of 0.5, 1, and 2% were selected for the main study.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- for better handling, the test substance was heated for app. 10min at 60°C and emulsified in propylene glycol
- 25µL of hand warm preparations were spread over the entire dorsal surface of each ear for three consecutive days
- 4 days after the first administration, 5mg BrdU were i.p. injected per mouse.
- mice were sacrificed after 24h and BrdU incorporation measured using a commercial cell proliferation assay kit (Roche, Mannheim)

- Criteria used to consider a positive response:
At least one concentration of the test item resulted in an incorporation of BrdU at least 1.6-fold or greater than that recorded in control mice, as indicated by the Stimulation Index.
Data are compatible with a conventional dose response, although allowance must be made (especially at high topical concentrations) for either local
toxicity or immunological suppression.
An increase in ear weight exceeding the threshold value of 25% was considered to be indicative for excessive local skin irritation.
For BALB/c mice, a cutoff-value for the lymph node cell count index of 1.55 was reported for a positive response.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:
SI = 5.6

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks:
0.5 %
Value:
1.6
Parameter:
SI
Remarks:
1 %
Value:
1.9
Parameter:
SI
Remarks:
2 %
Value:
4
Cellular proliferation data / Observations:
The animals did not show any relevant signs of systemic toxicity or excessive signs of local irritation. Stimulation Indices (S.I.) of 1.6, 1.9 and 4.0 were determined with the test item at concentrations of 0.5, 1 and 2% (w/w) in propylene glycol, respectively. Thus the EC1.6 value was equal to the lowest concentration of 0.5% (w/w). A clear dose response was observed.

Any other information on results incl. tables

A statistically significant increase in ear weights was observed in the 1% and 2% group, but remained below the threshold of excessive irritation (increase > 25%). Furthermore, the cut-off-value for a positive response regarding the lymph node cell count index of 1.55 reported for BALB/c mice was exceeded in the mid (1%) and high (2%) dose group and reached statistical significance as well.

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information