Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

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: Guideline study with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report date:
2009

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
BASF AG, Experimental Toxicology and Ecology
Type of study:
mouse local lymph node assay (LLNA)

Test material

Constituent 1
Chemical structure
Reference substance name:
9-Octadecenoic acid (Z)-, reaction products with 2-[(2-aminoethyl)amino]ethanol
EC Number:
272-379-2
EC Name:
9-Octadecenoic acid (Z)-, reaction products with 2-[(2-aminoethyl)amino]ethanol
Cas Number:
68815-51-0
Molecular formula:
C18 H34 O2 . C4 H12 N2 O
IUPAC Name:
2-(2-aminoethylamino)ethanol; (Z)-octadec-9-enoic acid
Details on test material:
- Name of test material (as cited in study report): Kerocom F 100
- Analytical purity: approx. 100 %
- Lot/batch No.: 09001529U0
- Storage condition of test material: The stability under storage conditions over the study period was guaranteed by the manufacturer, and the manufacturer holds this responsibility.

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, 97633 Sulzfeld
- Age at study initiation: 6-12 weeks
- Weight at study initiation: 18.0 g – 21.6 g
- Diet (e.g. ad libitum): Kliba-Labordiet
- Water (e.g. ad libitum): Tap water
- Acclimation period: 14 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (LLNA)

Vehicle:
methyl ethyl ketone
Concentration:
3%, 10% and 30%
No. of animals per dose:
5
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: MEK (methyl ethyl ketone) was used as the vehicle because good solubility of the preparation was achieved


MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
Groups of 5 females were treated epicutaneous with 25 µl of 3, 10 & 30 % of the test substance preparation at the dorsal part of both ears for 3 consecutive applications (day 0 – day 2) to the same application site. Individual body weights on day 0 prior to the first application and on day 5 prior to the sacrifice of the animals. All animals were observed for signs of systemic toxicity and local changes. Mortality was observed twice on each working day.
³H-thymidine injection: On study day five (about 66 to 72 hours after the last application of test substance to the ears) the mice were injected intravenously with 20 μCi of 3H-thymidine in 250 μl of sterile saline into a tail vein.

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: ³H-thymidine incorporation lead to SI values in the vehicle, 3%, 10% and 30% test substance of 1.00, 5.85, 7.30 and 10.76, respectively (Table 1).
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: ³H-thymidine incorporation lead to values (DPM /Lymph Node Pair) in the vehicle, 3%, 10% and 30% test substance of 434.3, 2539.5, 3171.7 and 4672.7, respectively (Table 1).

Any other information on results incl. tables

No signs of systemic toxicity were noticed. When applied as 30%, 10% and 3% preparations in methyl ethyl ketone (MEK), the test substance induced a biologically relevant response (increase to 1.5 fold or above of control value = stimulation index (SI) ≥ 1.5) in the auricular lymph node cell counts (Table 1). There was an increase in lymph node weights, as well. Concomitantly, the increase of 3H-thymidine incorporation into the cells was biologically relevant (increase above the cut off stimulation index of 3) at these concentrations. Some or severe increases in ear weights were observed in mice treated with the 10% and 30% test-substance preparation as indication of ear skin irritation. On the day of lymph node removal scaling was observed on the ears of the 30% test-substance group. As increases of lymph node parameters occurred at all concentrations, the lymph node effects cannot be attributed to ear skin irritation.

Table 1. Cell count, 3H-thymidine incorporation and lymph node weight: test group mean values and stimulation indices

Treatment

Cell counts

Counts/Lymph Node Pair

Stimulation Index

Vehicle

6,694,667

1.00

3%

14,210,667

2.12

10%

18,744,000

2.80

30%

23,901,333

3.57

 

Treatment

³H-thymidine incorporation

DPM/Lymph Node Pair

Stimulation Index

Vehicle

434.3

1.00

3%

2,539.5

5.85

10%

3,171.7

7.30

30%

4,672.7

10.76

 

Treatment

Lymph Node Weight

mg/Lymph Node Pair

Stimulation Index

Vehicle

4.7

1.00

3%

8.2

1.77

10%

10.0

2.15

30%

13.3

2.85

 

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information