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Description of key information

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
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
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)
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

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
Positive control results:
8756 DPM and SI of 12.1
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

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.

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

A local lymph node assay (LLNA) according to OECD guideline 429 was performed with the read across substance Fluidized Bed Combustion (FBC) Fly Ash in mice (Brabnikova, 2009). 5 animals per dose group were exposed to 0.3, 3 and 30% FBC in DAE 433 (mixture of 40% Diethylacetamid, 30% Acetone and 30% Ethanol) at 3 consecutive days. On day six radiolabled ³H-Methyl-Thymidine was injected and after 5 hours the measurements were performed. The stimulation index (SI) was 2.65 for the 0.3% group, 1.23 for the 3% group, 1.61 for the 30% group and 12.1 for the high control DNBC (Dinitrochlorobenzene). FBC has no sensitising potential because the SI of the treatment groups were < 3 and the positive control was valid.


Migrated from Short description of key information:
LLNA with read across substance Fluidized Bed Combustion (FBC) Fly Ash: not sensitising

Justification for selection of skin sensitisation endpoint:
Hazard assessment is conducted by means of read-across from a structural analogue. The selected study is the most adequate and reliable study based on the identified similarities in structure and intrinsic properties between source and target substance and overall quality assessment (refer to the endpoint discussion for further details).

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:
Justification for selection of respiratory sensitisation endpoint:
Study not required according to Annex VII-X of Regulation (EC) No 1907/2006.

Justification for classification or non-classification

Testing with the read across substance Fluidized Bed Combustion (FBC) Fly Ash revealed no sensitising potential in a LLNA. Therefore the test substance has not to be classified.