Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 Sept 2013 to 30 Jan 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report date:
2014

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
In accordance with Annex VII, Section 8.3, Column 2 the use of the Guinea-pig maximisation test (GMPT) of Magnusson and Kligmman (OECD 406) as opposed to the Local Lymph Node Assay (LLNA) is justified on the basis that LLNA studies have been shown to give false positive results for copper and copper compounds and as such the GMPT study is considered to be the most reliable for determining the true sensitisation potential of a copper-based material.

Test material

Constituent 1
Chemical structure
Reference substance name:
Dicopper hydroxide phosphate
EC Number:
235-285-2
EC Name:
Dicopper hydroxide phosphate
Cas Number:
12158-74-6
Molecular formula:
Cu2HO5P
IUPAC Name:
copper(2+) hydroxide phosphate (2:1:1)

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Germany
- Age at study initiation: young adult
- Weight at study initiation: 304-327 g
- Housing: in groups up to 10 animals per cage
- Diet: commercial feeding mixture (Muhle Knull, Rostock, Germany) ad libitum
- Water: tap water (drinking quality, supplemented with 1 g/l vitamin C) ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20±3 °C
- Humidity: 30-70 %
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
The test material was not soluble in distilled water and was injected as suspension. For topical application the test material was moistened with distilled water (100%) or applied as a suspension (50%, 25%).
Challenge
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
The test material was not soluble in distilled water and was injected as suspension. For topical application the test material was moistened with distilled water (100%) or applied as a suspension (50%, 25%).
No. of animals per dose:
Test group: 10
Control group: 5
Preliminary test: 6
Details on study design:
RANGE FINDING TESTS:
The appropriate concentrations of the test material and the appropriate vehicle for the intradermal induction phase, topical induction phase and challenge phase were determined on additional 6 Freund complete adjuvant (FCA) treated animals.
Intradermal injection:
The irritation response to intradermal injection of various concentrations of the test substance was examined in three guinea pigs. An area of the flanks was clipped free from hair with electric clippers. Amounts of 0.1 ml of the selected test concentrations (5%; 2.5%; 1%. and 0.5% suspensions of the test material in distilled water) were applied by intradermal injection. 24 and 48 hours after injection the animals were examined for signs of irritation according to the MAGNUSSON AND KLIGMAN GRADING SCALE.
Topical treatment:
The irritation response to topical treatment of various concentrations of the test substance was examined in three further guinea pigs. The flanks of the animals were clipped. Filter paper fully-loaded with the test substance (100%; 50% 25% in distilled water) was attached to the skin of the guinea-pigs and held in contact by an occlusive dressing for 24 hours.
The animals were observed and examined for signs of irritation according to the MAGNUSSON AND KLIGMAN GRADING SCALE approximately 24 hours and 48 hours after removing the test material.

MAIN STUDY
A1. INDUCTION EXPOSURE: intradermal injection (day 0)
- Test groups:
A) 1:1 mixture (v/v) FCA/water
B) Test substance suspension of 0.5% in distilled water
C) Test substance at the selected concentration formulated in a 1:1 mixture (v/v) FCA/water
- Control group: vehicle
- Site: dorsal skin from the scapular region (approximately 4 cm x 6 cm)
- Frequency of applications: once

A2. INDUCTION EXPOSURE: topical application (day 7)
- Exposure period: 48 hours
- Test groups: a patch loaded with 100% finely pulverised test substance moistened with distilled water
- Control group: vehicle
- Site: same as at intradermal injection
- Frequency of applications: once
Because the agent was non-irritating at this concentration, the region was pretreated with 10% sodium lauryl sulphate in vaseline for 24h.

B. CHALLENGE EXPOSURE (day 21)
- Exposure period: 24 hours
- Test groups: a patch loaded with 100% finely pulverised test substance moistened with distilled water
- Control group: vehicle
- Site: one flank of the animals
- Evaluation (hr after challenge): 24 and 48
Positive control substance(s):
no
Remarks:
tested on regularly in the perfomring laboratory

Results and discussion

Positive control results:
The reliability checks are performed regularly with hexyl cinnamic aldehyde (HCA -CAS No. 101-86-0) in the test laboratory (every six months).
The latest reliability check was performed from 04/09/2013 - 11/10/2013.
90% of animals treated with the positive control (HCA - 55% in vaseline) showed a skin reaction with numeral grading from 0-1 up to 2 according to "MAGNUSSON AND KLiGMAN GRADING SCALE FOR THE EVALUATION OF CHALLENGE PATCH TEST REACTIONS". The positive control showed a sensitization effect and the validity of the test system.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
100% of the test material moistened with distilled water
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
100% of the test material moistened with distilled water
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No
Remarks on result:
no indication of skin sensitisation
Group:
positive control
Remarks on result:
positive indication of skin sensitisation
Remarks:
tested on regularly in the perfomring laboratory

Applicant's summary and conclusion

Interpretation of results:
other: CLP/EU GHS criteria are not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
The test material Copper(ll)-hydroxide Phosphate is considered to be not skin sensitizing. The test material Copper(ll)-hydroxide Phosphate did not induce any skin reactions in intradermally and topically induced guinea pigs after challenge treatment. Therefore, the test material does not fulfil the criteria for classification according to Regulation (EC) No. 1272/2008 (CLP).