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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
2010-05-18 to 2010-09-04
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
, 1992-07-17
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
signed 2009-11-12
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): Vanadium oxide sulphate pentahydrate
- Trivial name: Vanadyl sulphate
- Producer: GfE Metalle and Materialien GmbH, Höfener Str. 45, 90431 Nürnberg, Germany
- Molecular formula: VOSO4 * 5H2O
- Molecular weight: 253.0 g/mol
- Physical state: Solid; blue crystals
- Storage condition of test material: Kept dry in tightly sealed containers at room temperature
- Water solubility: Freely soluble [MSDS provided by the producer]
- Relative density: 1.09 [GESTIS database]
No further information on the test material was stated.

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services Germany GmbH, Stolzenseeweg 32 – 36, 88353 Kißlegg, Germany
- Age at study initiation: 32 days
- Weight at study initiation: 323 - 360 g (excluding positive control group); Positive control group: 312 - 381 g
- Housing: The animals were kept in pairs in MAKROLON cages (MZK 80/25). Granulated textured wood (Granulat A2, J. BRANDENBURG, 49424 Goldenstedt, Germany) was used as bedding material in the cages.
- Diet (ad libitum): Commercial diet, ssniff® Ms-H V2233 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany)
- Water (ad libitum): Tap water
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20°C ± 3°C (maximum range)
- Relative humidity: 55% ± 15% (maximum range)
- Photoperiod (hrs dark / hrs light): 12/12
No further information on the test animals was stated.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Remarks:
Aqua ad iniectabilia (Batch nos. 911728 and 002152; Delta Select GmbH, 63303 Dreieich, Germany)
Concentration / amount:
Intracutaneous (Induction): 0.01% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
Topical (induction): 10% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
Topical (challenge): 0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Remarks:
Aqua ad iniectabilia (Batch nos. 911728 and 002152; Delta Select GmbH, 63303 Dreieich, Germany)
Concentration / amount:
Intracutaneous (Induction): 0.01% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
Topical (induction): 10% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
Topical (challenge): 0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
No. of animals per dose:
Preliminary test: 10 animals
Main study: 10 animals (treatment group) plus 5 animals (vehicle control group)
Positive control group: 20
Details on study design:
RANGE FINDING TESTS:
The aim of the preliminary study was to determine the appropriate dose level of the test item following intracutaneous and topical administration. The concentration used in the main study following the induction procedure should produce mild to moderate (grade 1 or grade 2) skin reactions following topical application and be adjusted to the highest level that can be well tolerated locally and generally following intracutaneous injection. For the challenge exposure a subirritating concentration was used which produced no evidence of skin irritation in non-sensitised animals.
The shoulder and the flank region of the animals were shaved or shaved and depilated (approx. 5 cm x 5 cm).
(a) intracutaneous: 0.1 mL of the prepared test item was administered intracutaneously (shoulder region).
Three concentrations of the test item were injected intradermally into four animals, respectively. In all, twelve concentrations were tested intracutaneously.
(b) topical: The test area of 6 animals was shaved (3 animals) or shaved and depilated (3 animals). 2 mL of the test preparation was spread over a filter paper (2 cm x 4 cm) and applied to the test area and held in contact by an occlusive dressing.
Two concentrations each were applied to the shaved or shaved and depilated flanks of 6 animals each.
The occlusive dressing and the filter paper containing the test item were removed after 24 or 48 hours and the application sites were assessed immediately, 24 and 48 (depilated) or immediately and 24 hours (non-depilated) after removal of the filter paper for erythema and oedema using the grading scale of MAGNUSSON/KLIGMAN..
Results:
Twelve concentrations of vanadium oxide sulphate pentahydrate were tested by intracutaneous injection: 0.00001, 0.00005, 0.0001, 0.0005, 0.001, 0.005, 0.01, 0.1, 0.5, 1, 5 or 10% solutions in aqua ad iniectabilia.
Concentrations up to 0.005% did not reveal any skin reaction. A concentration of 0.01% revealed a moderate and confluent erythema 24 hours and an intense erythema and swelling 48 and 72 hours after administration. Concentrations of 0.1% and above revealed an intense erythema and swelling 24 to 72 hours after administration.
Six concentrations of vanadium oxide sulphate pentahydrate were tested by topical application: 0.5, 1, 5, 10, 25 and 50% concentrations in aqua ad iniectabilia.
- Non-depilated skin: A concentration of 0.5% did not reveal any skin reactions. Concentrations of 1 or 5% revealed a discrete or patchy erythema, concentrations of 10% and above a moderate and confluent erythema 48 and 72 hours after start of administration.
- Depilated skin: A concentration of 0.5% did not reveal any skin reactions. A concentration of 1% revealed a discrete or patchy erythema 24 hours after administration. A concentration of 5% revealed an intense erythema and swelling 24 hours and a moderate and confluent erythema 48 and 72 hours after start of administration. Concentrations of 10% and above revealed an intense erythema and swelling 24, 48 and 72 hours after start of administration.
It was decided to use a 0.01% solution for the 1st (intracutaneous) induction stage, a concentration of 10% for the 2nd (topical) induction stage and a 0.5% concentration for the challenge.

MAIN STUDY
A. INDUCTION EXPOSURE

- No. of exposures: 2 (intradermal injection and topical administration)

- Test groups:
Stage 1:
Day 0
Three pairs of intradermal injections of 0.1 mL were given in the shoulder region which was cleared of hair so that one of each pair lay on each side of midline.
(1) Freund's complete adjuvant (Batch no. 049K8700; SIGMA-ALDRICH Chemie GmbH, 82024 Taufkirchen, Germany)(diluted 1 : 1 with 0.9% NaCl (Batch no. 005632; Delta Select GmbH, 63303 Dreieich, Germany))
(2) the test item (vanadium oxide sulphate pentahydrate) (0.01 % in aqua ad injectabilia)
(3) the test item in a 1:1 mixture (v/v) FCA/physiological saline
In injection 3, the final concentration of the test item was equal to that in injection 2.
Stage 2:
Day 7
7 days after the intracutaneous injection, the shoulder region of the same animals was shaved again but not depilated and treated topically using the patch-test technique (as described above under "Range finding tests") (exposure time: 48 hours). The patch was removed after 48 hours. No cleaning of the skin was necessary.

- Control group:
The vehicle control animals were treated in the same way as the animals of the test group, but received aqua ad iniectabilia instead of the test item.

- Skin observations and scoring: The skin reaction results of the first induction exposure (intradermal injection) were evaluated at 24 and 48 hours, of the second induction (topical administration) at 48 and 72 hours after start of exposure.

B. CHALLENGE EXPOSURE

- No. of exposures: 1 (topical administration)

- Test groups:
Stage 3:
Two weeks (Day 21) after the topical application (corresponds to a monitoring period of 21 days) the flanks of the same animals were shaved and depilated for a further topical application using the patch-test technique (as described above under "Range finding tests"). The filter paper containing the test item was applied to the left flank, the filter paper with the vehicle to the right flank of the animal (exposure time: 24 hours). 21 hours after the filter paper had been removed the treated skin was cleaned and fur removed, if necessary.

- Control group:
The left flank was treated with the test item, the right flank with the vehicle i.e. in the same way as the test group.

- Evaluation (hr after challenge):
21 hours after removing the filter paper the challenge area was cleaned and cleared of hair if necessary. Three hours later (at 48 hours from the start of challenge application) the skin reaction was observed and recorded. 24 hours after this observation a second observation (72 hours) was made and recorded. The skin reactions were graded following the grading scale of MAGNUSSON/KLIGMAN.

OTHER OBSERVATIONS:
Mortality: daily during the observation period
Clinical signs: daily during the observation period
Body weight: at start of study and at study termination
Pathology: No necropsy was performed. No animal was found dead in the main study or was sacrificed in extremis.

POSITIVE CONTROL GROUP:
The animals of the positive control group (same origin (strain) as those used in the study) were treated with a 2% (w/v) benzocaine solution intracutaneously in stage 1, a 5% (w/v) benzocaine solution topically in stage 2 and in stage 3.
The positive control data are obtained from the historical background of the laboratory that conducted this test. The study was performed during March 2010.
No further information on the study design was stated.
Challenge controls:
Vehicle control group: Aqua ad iniectabilia
5 males were used for the vehicle control group.
Positive control substance(s):
yes
Remarks:
Benzocaine (dissolved in 40% ethanolic 0.9% NaCl solution)

Study design: in vivo (LLNA)

Statistics:
The body weight was analysed statistically using STUDENT's t-test (p ≤ 0.01).

Results and discussion

Positive control results:
Animals of this strain treated with benzocaine in 40% ethanolic 0.9% NaCl solution exhibited a sensitising reaction in all animals in form of a moderate and confluent erythema or an intense erythema and swelling.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No skin irritation was observed.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No skin irritation was observed..
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No skin reaction was observed.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No skin reaction was observed..
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
The vehicle control revealed no skin reactions.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: The vehicle control revealed no skin reactions..
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
The vehicle control revealed no skin reactions.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0.5% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: The vehicle control revealed no skin reactions..
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
5% (w/v) benzocaine solution (Benzocaine was dissolved in 40% ethanolic 0.9% NaCl solution)
No. with + reactions:
20
Total no. in group:
20
Clinical observations:
Animals exhibited a sensitising reaction in form of a moderate and confluent erythema or an intense erythema and swelling.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 5% (w/v) benzocaine solution (Benzocaine was dissolved in 40% ethanolic 0.9% NaCl solution). No with. + reactions: 20.0. Total no. in groups: 20.0. Clinical observations: Animals exhibited a sensitising reaction in form of a moderate and confluent erythema or an intense erythema and swelling. .
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
5% (w/v) benzocaine solution (Benzocaine was dissolved in 40% ethanolic 0.9% NaCl solution)
No. with + reactions:
20
Total no. in group:
20
Clinical observations:
Animals exhibited a sensitising reaction in form of a moderate and confluent erythema or an intense erythema and swelling.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 5% (w/v) benzocaine solution (Benzocaine was dissolved in 40% ethanolic 0.9% NaCl solution). No with. + reactions: 20.0. Total no. in groups: 20.0. Clinical observations: Animals exhibited a sensitising reaction in form of a moderate and confluent erythema or an intense erythema and swelling. .

Any other information on results incl. tables

A 0.01% solution of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia chosen for the 1st (intracutaneous) induction stage revealed in all 10 animals a moderate and confluent erythema 24 hours and a discrete or patchy erythema 48 hours after administration.

2 mL of a 10% concentration of vanadium oxide sulphate pentahydrate in aqua ad iniectabilia/animal chosen for the 2nd (topical) induction stage revealed in all 10 animals a discrete or patchy erythema 48 and 72 hours after start of exposure.

The challenge with 2 mL of a 0.5% solution of vanadium oxide sulphate pentahydrate inaqua ad iniectabilia/animal revealed no skin irritation in any animal and, thus, the test item had no sensitising properties.

The body weight gain of the animals treated with vanadium oxide sulphate pentahydrate was within the range of the vehicle control during the experiment.

Behaviour remained unchanged.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the present test conditions, vanadium oxide sulphate pentahydrate revealed no sensitising properties in guinea pigs in a test model according to MAGNUSSON and KLIGMAN.
Therefore, vanadium oxide sulphate pentahydrate must not be classified and labeled according to regulation (EC) No.: 1272/2008 and subsequent regulations.
Also, vanadium oxide sulphate pentahydrate must not be classified and labeled according to 67/548/EC and subsequent regulations.