Montan wax

Skin sensitisation

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: Study according to OECD test guideline 429 under 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)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Montan wax

EC Number:

232-313-5

EC Name:

Montan wax

Cas Number:

8002-53-7

Molecular formula:

As montan wax is a UVCB, a molecular formular cannot be specified.

IUPAC Name:

Montan wax

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories B.V., Postbus 6174, 5960 AD Horst / The Netherlands
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 17.5 -22.1 g
- Housing: single caging
- Diet (e.g. ad libitum): pelleted standard diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum,
- Acclimation: At least 5 days prior to the start of dosing under test conditions after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 + 3°C
- Humidity (%): 45-85%
- Photoperiod (hrs dark / hrs light): 12h/12h

Study design: in vivo (LLNA)

Vehicle:

methyl ethyl ketone

Concentration:

2.5, 5.0, 10.0 %

No. of animals per dose:

4

Details on study design:

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: A test item is regarded as a sensitiser in the LLNA if the exposure to one or more test
concentration resulted in 3-fold or greater increase in incorporation of 3HTdR comparedwith concurrent controls, as indicated by the Stimulation Index (S.I.).

TREATMENT PREPARATION AND ADMINISTRATION:
Each test group of mice was treated by topical (epidermal) application to the dorsal surface of each ear lobe (left and right) with different test item concentrations of 2.5, 5, and 10% (w/w) in methyl ethyl ketone

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:

Estimated concentration for a S.I. of 3: 12.9

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: test group 1 (2.5%): 0.99 test group 2 (5%): 0.91 test group 3 (10%): 0.86

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: control group: 2308 test group 1 (2.5%): 2280 test group 2 (5%): 2095 test group 3 (10%): 1995 Background: 16

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

Based on the results of a study according to OECD test guideline 429 (LLNA) under GLP, the test substance is not skin sensitising.

Executive summary:

In order to study a possible contact allergenic potential of the test substance, three groups each of four female mice were treated daily with the test item at concentrations of 2.5, 5, and 10% (w/w) in methyl ethyl ketone by topical application to the dorsum of each ear lobe (left and right) for three consecutive days acc. to OECD test guideline 429 (under GLP). A control group of four mice was treated with the vehicle (methyl ethyl ketone) only. Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine ( 3H-methyl thymidine). Approximately five hours after intravenous injection, the mice were sacrificed, the draining auricular lymph nodes excised and pooled per group. Single cell suspensions of lymph node cells were prepared from pooled lymph nodes, which were subsequently washed and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was determined by the incorporation of 3H-methyl thymidine measured in a G3-scintillation counter. All treated animals survived the scheduled study period and no signs of toxicity were observed. In this study Stimulation Indices of 0.99, 0.91, and 0.86 were determined with the test substance at concentrations of 2.5, 5, and 10% in methyl ethyl ketone.

Therefore, the test substance is not skin sensitising.