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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2009-08-31 to 2009-10-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: 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
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

Constituent 1
Chemical structure
Reference substance name:
Diniobium pentaoxide
EC Number:
215-213-6
EC Name:
Diniobium pentaoxide
Cas Number:
1313-96-8
Molecular formula:
Nb2O5
IUPAC Name:
diniobium(5+) pentaoxidandiide

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Age at study initiation: 8-9 weeks
- Weight at study initiation: 15-22 g
- Housing: in groups of 5 in IVC cagews, type II L, polysulphone cages on Altromin saw fibre bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+-3
- Humidity (%): 55+-10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light):12/12

IN-LIFE DATES: From: 2009-09-11 To: 2009-10-01

Study design: in vivo (LLNA)

Vehicle:
other: AOO (3+1 (v/v) acetone/olive oil
Concentration:
12.5 %, 25 % and 50 % (w/v) [Based on the results of the preliminary test]
No. of animals per dose:
5
Details on study design:
PRELIMINARY TEST
To determine the highest tolerated and non-irritant test concentration a preliminary test was performed.
For this purpose, two animals were treated by topical application with the test item on three consecutive days at a concentration of 50% (suspended in AOO) to the entire dorsal surface of each ear.
One further animal was treated with 100% AOO and served as negative control.
From day 1 to day 4 the ear thickness of each animal was measured.
During this period also all clinical signs were recorded.
Cageside observations included spontaneous activity, lethargy, recumbent position, convulsions, tremors, apnoea, asphyxia, vocalisation, diarrhoea, changes in the skin and fur, eyes and mucous membranes (salivation, discharge).
Neither signs of systemic toxicity nor signs of irritation at the application site could be detected in the animal.

MAIN TEST
- Topical Application
Each mouse was treated by topical application of 25 μL of the selected solution to the entire dorsal surface of each ear.
Topical applications were performed once daily over three consecutive days.
- Administration of 3H-methyl thymidine
Five days after the first topical application all mice were dosed with 20 μCi 3H-methyl thymidine by intravenous injection (tail vein) of 250μL of 3H-methyl thymidine, diluted to a working concentration of 80μCi/mL.
- Preparation of cell suspension
Approximately 5 hours after 3H-methyl thymidine-injection all mice were sacrificed by cervical dislocation. The draining “auricular lymph nodes” were excised, individually pooled for each animal (2 lymph nodes per animal) and collected in phosphate buffered saline (PBS). A single cell suspension of pooled lymph node cells was prepared by gentle mechanical disaggregation through polyamide gauze (200 mesh size). After washing the gauze with PBS the cell suspension was pelleted in a centrifuge. The supernatant was discarded and the pellets were resuspended with PBS. This washing procedure was repeated.
After the final wash each pellet was resuspended in approx. 1 mL 5% TCA at approx. 4 °C for approximately 18 hours for precipitation of macromolecules. Each precipitate was once washed again, resuspended in 1 mL 5% TCA and 7 mL scintillation fluid was added. Then this solution was transferred into scintillation vials and stored at room temperature overnight.
- Determination of incorporated 3H-methyl thymidine
The 3H-methyl thymidine – incorporation was measured in a ß-counter and expressed as the number of disintegrations per minute (DPM). Similarly, background 3H-methyl thymidine levels were also measured (5% TCA). Determination of radioactivity was performed individually for each animal.

OTHER:
Reliability checks with a positive control substance was performed periodically in the performing laboratory.
Positive control substance(s):
other: P-Phenylenediamine (CAS 106-50-3)

Results and discussion

Positive control results:
Reliability checks with a positive control substance was performed periodically in the performing laboratory.
The mean SI-value of the latest reliability check was 13.8

In vivo (LLNA)

Resultsopen allclose all
Key result
Parameter:
SI
Value:
0.7
Test group / Remarks:
12.5
Key result
Parameter:
SI
Value:
1.3
Test group / Remarks:
25%
Key result
Parameter:
SI
Value:
0.9
Test group / Remarks:
50%
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: DPM (mean values) DPM for negative control: 1251.5 DPM at an extract concentration of 12.5%: 870.3 DPM at an extract concentration of 25%: 1562.2 DPM at an extract concentration of 50%: 1107.5
Cellular proliferation data / Observations:
The stimulation index at an extract concentration of 12.5% was 0.7 The stimulation index at an extract concentration of 25% was 1.3 The stimulation index at an extract concentration of 50% was 0.9.
DPM (mean values) DPM for negative control: 1251.5 DPM at an extract concentration of 12.5%: 870.3 DPM at an extract concentration of 25%: 1562.2 DPM at an extract concentration of 50%: 1107.5.

Any other information on results incl. tables

All animals survived throughout the test period without showing any clinical signs.

All animals gained weight as expected.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
According to OECD guideline 429 and EU criteria the tested substance should not be classified for dermal sensitization.