Copper dinitrate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

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 guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

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: 104.1%
- Lot/batch No.: 093750
- Storage condition of test material: room temperature in the dark.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: Relative mean viability

Remarks on result:

other: Reconstructed human epidermis model. Time point: 240 minute exposure period.

Remarks:

The relative mean viability of the test item treated tissues was 18.2% after a 240 minute exposure period.

Results 2

Irritation / corrosion parameter:

other: Relative mean viability

Remarks on result:

other: Reconstructed human epidermis model. Time point: 60 minute exposure period.

Remarks:

Time point: 60 minute exposure period. Remarks: The relative mean viability of the test item treated tissues was 19.9% after a 60 minute exposure period.

Results 3

Irritation / corrosion parameter:

other: Relative mean viability

Remarks on result:

other: Reconstructed human epidermis model. Time point: 3 minute exposure period.

Remarks:

The relative mean viability of the test item treated tissues was 53.4% after a 3 minute exposure period.

Any other information on results incl. tables

RESULTS

Direct MTT Reduction

The MTT solution containing the test item did not turn blue which indicated that the test item did not directly reduce MTT. Quantitative correction of the results was therefore unnecessary.

Test Item, Positive Control Item and Negative Control Item

Mean OD540 values and viabilities for the negative control, positive control and test item are given in Table 1 (attached).

The relative mean viability of the test item treated tissues was as follows:

240 minutes exposure: 18.2%

60 minutes exposure: 19.9%

3 minutes exposure: 53.4%

The 240 and 60 minute exposures induced tissue viabilities less than 35%. Therefore according to the prediction model a corrosive classification was triggered.

Quality Criteria

The relative mean tissue viability for the positive control treated tissues was 17.0% relative to the negative control treated tissues following the 240-Minute exposure period. The positive control acceptance criterion was therefore satisfied.

The mean OD540 for the negative control treated tissues was 0.176. The negative control acceptance criterion was therefore satisfied.

The test item was classified as corrosive to the skin.

Applicant's summary and conclusion

Interpretation of results:

Category 1B (corrosive)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Classification according to Directive 67/548/EEC: Corrosive (C). R34, Causes burns.
Classification according to CLP/GHS: Category 1B Corrosive, H314, Causes severe skin burns and eye damage.
UN Packing Group II "Substance Presenting Medium Danger"

Executive summary:

A GLP-compliant study was conducted in accordance with OECD Guideline 431 and EU Method B.40 BIS. Duplicate tissues were treated with the test item for exposure periods of 3, 60 and 240 minutes. At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTT-Ioading. After MTT loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT -loaded tissues. At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200 µl samples were transferred to the appropriate wells of a pre-Iabelled 96-well plate. The optical density (OD) was measured at 540 nm (OD540). Data were presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues). The relative mean viability of the test item treated tissues was as follows:

240 minutes exposure: 18.2%
60 minutes exposure: 19.9%
3 minutes exposure: 53.4%

The quality criteria required for acceptance of results in the test were satisfied. The test item was classified as corrosive to the skin. The following classification criteria apply:

EU DSD (67/548/EEC) Corrosive requires symbol "C" risk phrase R34 "CAUSES BURNS".
EU CLP (1272/2008/EC) and UN GHS Hazard statement H314 "Causes severe skin burns and eye damage" Category 1 B.

UN Packing Group II "Substance Presenting Medium Danger"