Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with Good laboratory Practice and internationally accepted guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
An LLNA study was not conducted because this method is prone to give false positive results for copper compounds and strong dermal irritants.

Test material

Constituent 1
Reference substance name:
Copper(II) nitrate hydrate
IUPAC Name:
Copper(II) nitrate hydrate
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): Copper(II) nitrate hydrate
- Analytical purity: >= 99% w/w
- Lot/batch No.: 093750
- Storage condition of test material: room temperature

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 (F-69592)
- Age at study initiation: 4 weeks
- Weight at study initiation: 209 - 282 g
- Housing: Either in groups of 2 or individually in polycarbonate containers, the flooring of which was covered in dust-free cuttings and the top fitted with a stainless steel lid.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Minimum 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25°C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 - 15 changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

IN-LIFE DATES: From: To:

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Refer to deatils on study design.
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Refer to deatils on study design.
No. of animals per dose:
Refer to details on study design.
Details on study design:
RANGE FINDING TESTS:

Determination of the Maximal Non-Necrotising Concentration (MNNC):
This test was conducted for the purpose of defining a MNNC of the test item which, on intradermal injection during the induction phase, does not risk causing too great a lesion (non-necrotising concentration), should be well-tolerated systemically and should be the highest to cause mild-to-moderate skin irritation. Following a series of tests carried out at higher concentrations resulting in necrosis, two animals received on both sides of the spine a volume of 0.1 mL of the tet item at 3 concentrations: diluted at 0.02%, 0.01% and 0.005% in isotonic sodium chloride. A macroscopic evaluation of the cutaneous reactions was conducted 24 or 48 hours after injection.

Determination by topical application of the Pre-Maximal Non-Irritant Concentration (Pre-MNIC):
This test, which allowed evaluation of the irritant potential of the test item, defined whether an application of sodium lauryl sulphate would be needed during the topicll induction phase. The test item was applied on the dorso-lumbar zone of two guinea pigs shorn beforehand, with an occlusive dressing for 24 hours, at 4 different concentration: diluted at 90%, 50%, 20% and 10% in distilled water. A macroscopic evaluation of the cutaneous reactions ws conducted 24 hours after removl of the dressing.

Determination by topical application of the Maximal Non-Irritant Concentration (MNIC):
Thi test was carried out in order to determine the MNIC of the test item without rik of an irritant effect during the challenge phase. Three guinea pigs were treated according to te same treatment as animals from GROUP 1 (negative control) for the induction phase (i.e. isotonic sodium chloride and distilled water). During the challenge phase, the animals were treated with the test item placed onto the selected treatment sites and covered with an occlusive dressing for a period of 24 hours at 4 different concentrations: diluted at 10%, 5%, ,2.5% and 1.25% in distilled water. A macroscopic evaluation of the cutaneous reactions was conducted 24 and 48 hours after removal of the occlusive dressing and rinse with distilled water.

MAIN STUDY

GROUP 1 (negative control): 5 female guinea pigs;
GROUP 2 (treated): 10 female guinea pigs.

A. INDUCTION EXPOSURE

1st Intradermal Induction:
Day 0: After shearing the scapular zone, 3 pairs of intradermal injection (ID) of 0.1 mL were performed on the scapular zone on either side as follows:
GROUP 1 (negative control):
2 ID Freund's Complete Adjuvant diluted at 50% in isotonic sodium chloride.
2 ID Isotonic sodium chloride.
2 ID A mixture with equal volumes v/v: Freunds Complete Adjuvant at 50% and isotonic sodium chloride.
GROUP 2 (treated):
2 ID Freund's Complete Adjuvant diluted at 50% in isotonic sodium chloride.
2 ID Test item at 0.005% in Isotonic sodium chloride.
2 ID A mixture with equal volumes v/v: Freunds Complete Adjuvant at 50% and the test item at 0.01% in isotonic sodium chloride.

2nd Topical induction:
Day 6: The scapular zone of all the animals in each group was shorn.
Day 7: A topical application under occlusive dressing for 48 hours was performed on the injection sites of each animal.
GROUP 1 (negative control): 0.5 mL of distilled water.
GROUP 2 (treated) 0.5 mL of the test item diluted at 50% in distilled water.
Day 9: Occlusive dressing removal and rinse with distilled water.

B. CHALLENGE EXPOSURE

Day 20: The experimental procedure of this phase was idential for both GROUP 1 and GROUP 2. On the previously shorn dorso-lumbar zone, the following applications on either side of the spine, under an occlusive dresing, were performed for a period of 24 hours:
- 1 sample cup containing the test item diluted at 2.5% (MNIC) and 1 sample cup containing the test item diluted at 1.25% (0.5 MNIC).
Day 21: Occlusive dressing removal and rinse with distilled water.
Positive control substance(s):
yes
Remarks:
alpha-Hexylcinnamaldehyde

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
2.5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Spot of scab (in edge of patch), 4 animals. Spot of scab with slight oedema (in edge of patch), 4 animals.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 2.5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Spot of scab (in edge of patch), 4 animals. Spot of scab with slight oedema (in edge of patch), 4 animals..
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
2.5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Spot of scab (in edge of patch), 7 animals. Spot of scab with slight oedema (in edge of patch), 1 animal.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 2.5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Spot of scab (in edge of patch), 7 animals. Spot of scab with slight oedema (in edge of patch), 1 animal..
Reading:
1st reading
Hours after challenge:
72
Group:
test chemical
Dose level:
2.5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Spot of scab (in edge of patch), 6 animals. Spot of scab with slight oedema (in edge of patch), 1 animal.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 72.0. Group: test group. Dose level: 2.5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Spot of scab (in edge of patch), 6 animals. Spot of scab with slight oedema (in edge of patch), 1 animal..
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
2.5%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
Spot of scab (in edge of patch), 2 animals. Spot of scab with slight oedema (in edge of patch), 2 animals.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 2.5%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: Spot of scab (in edge of patch), 2 animals. Spot of scab with slight oedema (in edge of patch), 2 animals..
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
2.5%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
Spot of scab (in edge of patch), 4 animals.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 2.5%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: Spot of scab (in edge of patch), 4 animals..
Reading:
1st reading
Hours after challenge:
72
Group:
negative control
Dose level:
2.5%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
Spot of scab (in edge of patch), 4 animals.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 2.5%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: Spot of scab (in edge of patch), 4 animals..
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
1.25%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Spot of scab (in edge of patch), 3 animals. Spot of scab with slight oedema (in edge of patch), 1 animal.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 1.25%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Spot of scab (in edge of patch), 3 animals. Spot of scab with slight oedema (in edge of patch), 1 animal..
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
1.25%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Spot of scab (in edge of patch), 4 animals.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1.25%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Spot of scab (in edge of patch), 4 animals..
Reading:
1st reading
Hours after challenge:
72
Group:
test chemical
Dose level:
1.25%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Spot of scab (in edge of patch), 4 animals.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 72.0. Group: test group. Dose level: 1.25%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Spot of scab (in edge of patch), 4 animals..
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
1.25%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
Spot of scab (in edge of patch), 2 animals.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 1.25%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: Spot of scab (in edge of patch), 2 animals..
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
1.25%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
Spot of scab (in edge of patch), 2 animals.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 1.25%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: Spot of scab (in edge of patch), 2 animals..
Reading:
1st reading
Hours after challenge:
72
Group:
negative control
Dose level:
1.25%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
Spot of scab (in edge of patch), 2 animals.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 1.25%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: Spot of scab (in edge of patch), 2 animals..

Any other information on results incl. tables

Preliminary Studies:

- MNNC determination: No necrosis was observed with the concentration of 0.005%. The first induction of the Group 2 was carried out by intradermal injection at the maximal non necrosing concentration of 0.005%.

-Pre MNIC determination: 24 hours after removal of the occlusive dressings, severe erythem was recorded on the treated area at 90%, moderate to severe erythema was recorded on the treated area at 50% and moderate erythema wa noted on the treated area at 20%. 24 hour after removl of the occlusive dressings, no macroscopic cutaneous reaction was recorded on the treated area at 10%. In view of these results, the concentration selected was 50% for the 2nd induction of the Group 2 and the MNIC determination began at the concentration of 10%.

- MNIC determination: 24 hours after the removal of the occlusive dressings, slight to moderate erythema wa recorded on the treated areas at 10% and 5%. 24 and 48 hours after the removal of the occlusive dressings, no macroscopic cutaneous reaction was recorded on the treated areas at 2.5% and 1.25%. In view of this result, the concentrations selected were 2.5% (MNIC) and 1.25% (0.5 MNIC) for the challenge phase.

Main study:

- Induction phase Group 1: The induction phase was performed by intradermal injection on D0 with the test item at 0.005% in isotonic sodium chloride and by topical application on D7 with distilled water. No cutaneous reaction was recorded after the induction phase. See Table 1 (attached).

- Induction phase Group 2: The induction phase was performed by intradermal injection on D0 with the test item at 0.005% in isotonoc sodium chloride and by topical application on D7 with the test item diluted at 50% in distilled water. No cutaneous reaction was recorded after the first induction. During the second induction, scab was noted in eight animals (8/10), 24 hours after the removal of the occlusive dressing. See Table 1 (attached).

- Challenge phase Groups 1 and 2: The test item was used diluted at 2.5% and 1.25% in distilled water.

Sensitising potential assesment:

See Tables 2 - 4 (attached).

No cutaneous reaction attributable to allergy (erythema or oedema) was recorded in animals from the treated group after the challenge phase, on the treated area with the test item at 2.5% and 1.25%.

No cutaneous intolerance reaction (erythema or oedema) ws recorded in animals from the negative control group after the chalenge phase, on the treated area with the test item at 2.5% and 1.25%.

A spot of scab in the edge of the patch, associated or not with a very slight oedema, was noted in four animals (4/5) and two animals (2/5) from the negatve control group on the treated areas at 2.5% and 1.25% respectively.

Similar reactions were noted in eight animls 8/10 and four animals 4/10 from the treated group on the treated areas at 2.5% and 1.25% respectively.

Weight evolution:

The weight gain of negative control animals (Group 1) and treated animals (Group 2) shows no abnormalities. See Tables 5 and 6 and Figure 1 (attached).

Mortality:

No mortality was seen during the main test.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test item is not classified as a skin sensitiser.
Executive summary:

A GLP compliant guinea pig maximisation test was carried out in comliance with OECD Guideline 406 (EU mentod B6) without significant deviation. After induction (intradermal injection at 0.005% and topical application at 50%) of 10 guinea pigs and a 10 -day rest phase, the challenge phase under occlusive dressing for 24 hours consisted of a single topical application of the test item diluted at 2.5% and 1.25% in distilled water.

No cutaneous reaction attributable to allergy (erythema or oedema) was recorded in animals from the treated group after the challenge phase, on the treated area with the test item at 2.5% and 1.25%. No cutaneous intolerance reaction (erythema or oedema) ws recorded in animals from the negative control group after the challenge phase, on the treated area with the test item at 2.5% and 1.25%. A spot of scab in the edge of the patch, associated or not with a very slight oedema, was noted in four animals (4/5) and two animals (2/5) from the negatve control group on the treated areas at 2.5% and 1.25% respectively. Similar reactions were noted in eight animls 8/10 and four animals 4/10 from the treated group on the treated areas at 2.5% and 1.25% respectively.

On the basis of these results,it is concluded that the test item should not be classified as a sensitiser.