Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2009-07-27 to 2009-08-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (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 of test material (as cited in study report): Fluidized Bed Combustion (FBC) Fly Ash
- Physical state: Light grey
- Impurities (identity and concentrations): Metals: As, Be, Cd, Co, Cr, Cu, Hg, Mo, Ni, Pb, Sb, Se, Tl, V, Zn, sum < 0.1%
- Lot/batch No.: FBC/230309/T1
- Expiration date of the lot/batch: 15 years/ 03/2024
- Storage condition of test material: The substance will be stored in PE container at room temperature.

In vivo test system

Test animals

Species:
mouse
Strain:
Balb/c
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeding farm VELAZ s.r.o., Kolec u Kladna, Czech Republik
- Age at study initiation: 8-10 weeks
- Weight at study initiation: 19.6-22.6 g
- Housing: in groups of max. 6 animals
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (LLNA)

Vehicle:
other: DAE 433 (mixture of 40% Diethylacetamid, 30% Acetone and 30% Ethanol)
Concentration:
0.3, 3 and 30% (w/v)
No. of animals per dose:
Pilot experiment: 3 female
Exposure experiment: 15 females (5 in 3 groups)
Positive control: 5 females
Negative control: 5 females
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: 30% (w/v) was the maximum technically practicable concentration derived from the pilot experiment
- Irritation: When the ear weights are statistically significant increased the subsatnce may cause irritation to the skin. Positive control inducd ear swelling, negative control and test material showed no irritating effects.
- Lymph node proliferation response:

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response:
Incorporation of ³H-methyl-thymidine: When SI ≥ 3 and dose response is recorded, the subsatnce is regared as skin sensitiser

TREATMENT PREPARATION AND ADMINISTRATION:
Day 1: open application of 25 µL of the three test concentrations, the positive and the negative control to the dorsum of the ear
Day2 and 3: repeated application as on day 1
Day 6: Injection of 250 µL of PBS containing 7.8x10^5 Bq (c.21 µCi) of ³H-methyl-thymidine to all animals via the tail vein. Five hours later all animals were sacrificed.
Positive control substance(s):
other: DNBC (Dinitrochlorobenzene, CAS: 97-00-7)
Statistics:
Software Statgraphic centurion (Version XV, USA) was used:
- Non parametric Kruskal-Wallis test
- Non parametric two group Mann-Whitmey rank test

Results and discussion

Positive control results:
8756 DPM and SI of 12.1

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: 0.3%-group: 2.65, 3%-group: 1.23, 30%-group: 1.61
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: negative control: 711.51, 0.3%-group: 1886.5, 3%-group: 872.74, 30%-group: 1144.06

Any other information on results incl. tables

No animals died during the study. No symptomes of toxicity were observed in the treatment and negative control groups. Positive control group animals showed hyperaemia of skin and clonospasm.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU