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Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010-05-18 to 2010-09-04
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
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
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
, 1992-07-17
Deviations:
yes
Remarks:
, guinea pigs were housed at a temperature of 22°C +/- 3°C instead at a temperature of 20 +/- 3°C
GLP compliance:
yes (incl. QA statement)
Remarks:
signed 2009-11-12
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
see attached justification document in the endpoint summary for skin sensitisation

Test material

Constituent 1
Reference substance name:
Sodium metavanadate
IUPAC Name:
Sodium metavanadate
Constituent 2
Chemical structure
Reference substance name:
Sodium metavanadate
EC Number:
237-272-7
EC Name:
Sodium metavanadate
Cas Number:
13718-26-8
Molecular formula:
NaVO3
IUPAC Name:
237-272-7
Constituent 3
Reference substance name:
13718-26-8
Cas Number:
13718-26-8
IUPAC Name:
13718-26-8
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Sodium metavanadate
- Trivial name: Sodium metavanadate
- Molecular formula: NaVO3
- Molecular weight: 121.93 g/mol
- Physical state: Solid; white powder
- Storage condition of test material: Kept cool and dry in tightly sealed containers
- pH: Approx. 9 (2.5% solution)
-Melting point: 630°C
- Water solubility: 211 g/L
- Relative density: 2.9 [MSDS provided by the producer]
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: 325 - 361 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: 22°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 sodium metavanadate in aqua ad iniectabilia
Topical (induction): 1% solution of sodium metavanadate in aqua ad iniectabilia
Topical (challenge): 0.05% solution of sodium metavanadate 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 sodium metavanadate in aqua ad iniectabilia
Topical (induction): 1% solution of sodium metavanadate in aqua ad iniectabilia
Topical (challenge): 0.05% solution of sodium metavanadate in aqua ad iniectabilia
No. of animals per dose:
Preliminary test: 16 animals
Main test: 10 animals (treatment group) plus 5 animals (vehicle control group)
Positive control group: 20 animals
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 12 animals was shaved (6 animals) or shaved and depilated (6 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.
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 of MAGNUSSON/KLIGMAN.
Results:
Twelve concentrations of sodium metavanadate 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, 48 and 72 hours after administration. A concentration of 0.1% revealed a moderate and confluent erythema 24 hours and an intense erythema and swelling 48 and 72 hours after administration. A concentration of 0.5% revealed an intense erythema and swelling 24, 48 and 72 hours after administration. The animal employed with the concentrations of 1, 5 and 10% died prematurely within 24 hours after administration.
Twelve concentrations of sodium metavanadate were tested by topical application: 0.0005, 0.001, 0.005, 0.01, 0.05, 0.1 0.5, 1, 5, 10, 25 and 50% concentrations in aqua ad iniectabilia.
Non-depilated skin: Concentrations up to 0.05% did not reveal any skin reactions. Concentrations of 0.1 or 0.5% revealed a discrete or patchy erythema, a concentrations of or 5% revealed a moderate and confluent erythema, concentrations of 10% and above revealed an intense erythema and swelling 48 and 72 hours after start of administration, respectively.
Depilated skin: Concentrations up to of 0.05% did not reveal any skin reactions. A concentration of 0.1% revealed a discrete or patchy erythema 24, 48 and 72 hours after start of administration. A concentration of 0.5% revealed a moderate and confluent erythema 24 hours and an intense erythema 48 and 72 hours after start of administration. Concentrations of 1% 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 1% for the 2nd (topical) induction stage and a 0.05% 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 (sodium metavanadate) (0.01 % in aqua ad iniectabilia)
(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 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 were evaluated at 24 and 48 hours, of the second induction at 48 and 72 hours after start of exposure. The skin reactions were graded following the grading scale of MAGNUSSON/KLIGMAN.

B. CHALLENGE EXPOSURE
- No. of exposures:

- Test groups:
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 study. The study was performed during March 2010.
No further information on the study design was stated.

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

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 chemical
Dose level:
0.05% solution of sodium metavanadate in aqua ad iniectabilia
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No skin irritation was observed.
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
0.05% solution of sodium metavanadate in aqua ad iniectabilia
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No skin irritation was observed.
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0.05% solution of sodium metavanadate 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:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0.05% solution of sodium metavanadate 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:
no indication of skin sensitisation
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:
positive indication of skin sensitisation
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:
positive indication of skin sensitisation

Any other information on results incl. tables

A 0.01% solution of sodium metavanadate 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 1% concentration of sodium metavanadate in aqua ad iniectabilia per 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 0.05% solution of sodium metavanadate in aqua ad iniectabilia per 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 sodium metavanadate 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, sodium metavanadate revealed no sensitising properties in guinea pigs in a test model according to MAGNUSSON and KLIGMAN.
Therefore, the test item must not be classified and labeled according to regulation (EC) No.: 1272/2008 and subsequent regulations.
Also, the test item must not be classified and labeled according to 67/548/EC and subsequent regulations.

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