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Toxicological information

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:
April - June 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well-documented GLP-Guideline Study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
other: OECD no406
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The study was conducted in 2007 before the LLNA-method has become prefered method.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: microgranulat
Details on test material:
Name: Fe(III)IDHA
CAS No: 666828-40-6

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hodowla Zwierząt Laboratoryjnych Ilkowice 41, 32-218 Slaboszew, Poland
- Weight at study initiation: averge 376 g
- Housing:
- Diet (e.g. ad libitum): conventional laboratory diet (TOP DOVO Dobra Voda, supplier)
- Water (e.g. ad libitum): water with addition of ascorbic acid in concentration 0.05%
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/- 2
- Humidity (%): 40-70
- Photoperiod (hrs dark / hrs light): artificial, 12h light/ 12h dark

Cages
Housed in groups of 5 in cages in conformity with laboratory welfare legislation.

IN-LIFE DATES:
Deliver: 27.04.2007
Aclimatisation: 4.05.2007
Pilot study: 9-14.05.2007
Division of animals into groups and weighting: 14.05.2007
Induction phase: 1. intradermal injection: 15.05.2007
2. topical appliation: 22.05.2007
Rest period: 23.05.2007 - 4.06.2007
Challenge: 5.06.2007
Observations: 7-8.06.2007
End study: 8.06.2007

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
other: Freund's complete adjuvant (FCA), saline, aqua pro injectione
Concentration / amount:
Induction:
- intradermal injections - 5% Fe(III)IDHA in aqua pro injectione
- topical application - 75% Fe(III)IDHA in aqua pro injectione

Chellenge:
- topical application - 10% Fe(III)IDHA in in aqua pro injectione
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Freund's complete adjuvant (FCA), saline, aqua pro injectione
Concentration / amount:
Induction:
- intradermal injections - 5% Fe(III)IDHA in aqua pro injectione
- topical application - 75% Fe(III)IDHA in aqua pro injectione

Chellenge:
- topical application - 10% Fe(III)IDHA in in aqua pro injectione
No. of animals per dose:
treated group:20
control group: 10
Details on study design:
Pilot study
The ocncentration of test substance used for each induction exposure was selected well-tolerated systemically and the highest to cause mild - to - moderate skin irritation. The concentration for the challenge exposure was selected the highest non-irritant dose. The appropriate concentrations was determined using two animals for intradermal injections and three animals for topical application (occlusive dressing for 24 h).

Induction phase
1. Intradermal injections: three pairs of intradermal injection of 0.1 ml volume were givenin the shoulder region
Day 1 - treated group
injection 1: mixture FCA/saline 1:1
injection 2: the test substance in water at the selected concentration in pilot study
injection 3: mixture injection 2 and injection 1
Day 1 control group
injection 1: mixture FCA/saline 1:1
injection 2: the undiluted vehicle
injection 3: mixture injection 2 and injection 1

Topical application
Day 7 - treated and control groups
Because the substance is a skin irritant, the tet area, is not painted with 0.5 ml of 10% natrii lauryl sulfas in vaseline, in order to cerate a local irritation.
Day 8 - treated group
The test area was cleared of hair. A gauze 2x4 cm was fully loaded with vehicle and was applied to the test area and held in contact by an occlusive dressing for 48 h. During at least 14 days, no treatment was applied.

Challenge
Day 22 - treated and control groups
The flank of treated and control animals was cleared of hair. A patch loaded with the substance was applied to one flank of the animals and patch loaded with the vehicle was applied ti the other flank. The patch was held in contact by an occlusivve dressing for 24 h.

Obsrvations performed
Approximately 21 hours after removing the patch, the challenge area was cleaned and shaved. Approximalety 3 hours lated (48 h from the start of the challenge application) the skin reaction was observed and recorded according to the grades. Approximately 48 h after removing of the patch (72 h from the start of the challenge), the second observation was made and recorded.
Individual starting body weight and ending body weight were statistically evaluated by Statgraphics statistical program. To identify homogenity groups the Bartell test was used. In the case of homogenity One-Way Analysis of Variance was applied.
Positive control substance(s):
no

Results and discussion

Positive control results:
not applicable as no positive controls were used

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
75%
No. with + reactions:
13
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 75%. No with. + reactions: 13.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
75%
No. with + reactions:
10
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 75%. No with. + reactions: 10.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
4
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. No with. + reactions: 4.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0.

Any other information on results incl. tables

Please see attachment.

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Topical application of Fe(III)IDHA incuced the sensitisation rate 50%. According to this value the tet article Fe(III)IDHA belongs to "moderate sensitizer" (III).
Executive summary:

The skin sensitisation potential of Fe(III)IDHA was tested in the Guinea pig maximisation test (Magnusson and Kligman). 20 animals were used in the treated group and 10 animals in the control group. For induction treatment of animals treated group were intradermally injected 5% Fe(III)IDHA in Aqua pro injectione with complete FCA followed one week lated by epidermal application of 75% Fe(III)IDHA in Aqua pro injectione. For induction treatment of animals control group were intradermally injected with vehivle (Aqua pro injectione) with complete FCA followed one week lated by epidermal application of vehicle. Two weeks after epidermal application all animals were challenged with 10% Fe(III)IDHAa in Aqua pro injectione. Fe(III)IDHA induced the sensitisation rate 50%. According to this value the substnce Fe(III)IDHA belongs to "moderate sensitizer" (III) ( Skin Sens 1B, H3317 acc. to Regulation 1272/2008).