Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study in compliance with guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name: "Glucose Syrups Wheat hydrolized".
Trade name: Dried wheat glucose syrup.
Batch No.: 5095187.
Appearance: White, slightly hygroscopic powder.
Purity : 97.8 % dry solids
Solubility: Good soluble in water.
pH: Neutral.
Conditions of storage: Room temperature, no protection from light.
Stability at conditions of storage: Stable.
Expiry date: June 2011.

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
Animals: Mice, CBA/Ca, females, young adult nulliparous.

Justification for the selection of the species: Mice are recommended by the guidelines for LLNAs.

Age at the first administration: About 8 – 12 weeks.

Body mass range: At the initiation of the study a maximum deviation of 20 % of any individual body weight from the mean is accepted.

Hygiene: Optimal hygienic conditions.

Room No.: EI 1-13.

Room temperature: About 22 °C (continuous monitoring and recording).

Relative humidity: Between 30% and 70% (continuous monitoring and recording).

Lighting: Only artificial light from 6.00 a.m. to 6.00 p.m.

Cages: Single caging. Makrolon cages type II, (22 cm x 16,5 cm ground area, 15 cm high).

Feed: Maintenance diet for rats and mice R/M-H (item V1534-300), autoclavable, ad libitum.

Water: Tap water from Makrolon-bottles with stainless steel canules or from an automatic watering system, ad libitum.

Bedding material: Aspen wood chips, ABEDD®, LAB & VET Service GmbH, Hasnerstraße 84/6, 1160 Wien (item LTE E-001). Germ reduction by autoclaving; Changed 1/week.

Environmental Enrichment: Nibbling wood bricks and/or nesting material, same material and source as the bedding material, are offered once a week. A "mouse house" (red polycarbonate shelter, 9.8 x 8.8 cm ground area, 5.5 cm high) is offered per cage. Germ reduction by autoclaving.

Acclimatisation: At least 5 days.

Identification of the animals: By shearing a defined fur region on the back of the animal:
Group A: top left
Group B: top right
Group C: bottom left
Group P: top left and top right
Group K: unshorn
and by cage tag.

Study design: in vivo (LLNA)

Vehicle:
dimethylformamide
Concentration:
Solubility testing of the test substance in the guideline-recommended vehicles showed that the highest concentrations suitable for application of the test substance can be achieved with DMF (14.9%, w/w).
No. of animals per dose:
Test substance concentrations:
Group A (low dose), 5 animals, 5% test substance in DMF (w/w)
Group B (mid dose), 5 animals, 10% test substance in DMF (w/w)
Group C (high dose), 5 animals, 14.9% test substance in DMF (w/w)

Control substance concentrations:
Group K (negative control), 5 animals, DMF
Group P (positive control), 5 animals, 25% HCA in AOO (v/v)
Details on study design:
The individual animals are allocated to their groups by random numbers (5 mice/group). The test substance is administered in 3 concentrations to the dorsal surfaces of the ears of the animals of the test substance groups. In a manner identical to that of animals in the treatment groups animals of one negative control group and one positive control group are treated with DMF and HCA respectively. The positive and negative control groups might be simultaneously used for other, concurrently performed studies. Each animal is treated for 3 consecutive days. 3 days after the last administration the proliferation of the lymphocytes of the draining lymph nodes is measured by the determination of the amounts of incorporated radioactive ³H-methyl thymidine (3HTdR). If the dpm data of the negative or positive control differ very strong from the historical data the main study will be repeated.

Time schedule
Day 1,2,3: Epicutaneous administration of the appropriate dilutions of the test or control substances to the dorsum of both ears.
Day 6: Administration of 20 μCi 3HTdR to all mice via a tail vein.
5 hours later, the draining auricular lymph nodes are excised and pooled for each experimental group.
Day 7: Measurement of 3HTdR incorporation by ß-scintillation counting.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
The allocation of the animals to the groups is performed by randomisation.

Results and discussion

Positive control results:
Positive control is valid. SI was higher than 3.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: The substance is regarded as non sensitizer; since the SIs of all tested concentrations were clearly lower than 3.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see below

Any other information on results incl. tables

 

dpm

SI

group K(negative control)

494

1

group A(low dose)

360

0.7

group B(mid dose)

320

0.6

group C(high dose)

327

0.7

group P(positive control)

2315

4.7

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information