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

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

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

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)

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

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)

Positive control results:

SI = 5.6

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.

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.

Interpretation of results:

sensitising

Remarks:

Migrated information

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

Three groups of five female mice each were treated once daily with the test item at concentrations of 0.5, 1, and 2% (w/w) in propylene glycol by topical application to the dorsum of each ear for three consecutive days. Concentrations at and above 5% led to excessive irritation as determined by ear weight increase > 25% in the pretests performed. Four days after the first topical application the mice were intraperitoneally injected with BrdU. Approximately 24 hours after intraperitoneally injection, the mice were sacrificed and the draining auricular lymph nodes excised, pooled per animal and immediately weighed. Furthermore, after excision of the lymph nodes, both ears of the mice were punched at the apical area using a biopsy punch and were immediately weighed pooled per animal using an analytical balance. Afterwards, single cell suspensions of lymph node cells were prepared from lymph nodes pooled per animal. An aliquot of each cell suspension was used for determination of lymph node cell count. The proliferative capacity of pooled lymph node cells was determined by the incorporation of BrdU. 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.

Migrated from Short description of key information:
LLNA: EC 1.6 = 0.5% (BASF 2013, OECD442B, GLP)

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

In the LLNA theReaction products of Fatty acids, tall-oil, compds. with oleylamine and Fatty acids, C18-unsatd., trimers, compds. with oleylaminewas demonstrated to be a skin sensitizer due to a dose dependent increase in lymphocyte proliferation and an S. I. value greater or equal to 1.6 at all concentration tested. The EC1.6 value was 0.5%. Based on these resultsReaction products of Fatty acids, tall-oil, compds. with oleylamine and Fatty acids, C18-unsatd., trimers, compds. with oleylamineis required to be classified with R43 according to 67/548/EEC and as a skin sensitizer cat.1A according to CLP/EU-GHS requirements.

 

There are no data available for classification as a sensitizer of the respiratory tract.