Copper, diamminebis(nitrato-kO)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vitro

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

For the skin sensitisation in vitro, only one study has been conducted on the substance: the Keratinosens test (OECD 442D).
The two other tests which are part of the Adverse Outcome Pathway for skin sensitisation, i.e. DPRA and h-CLAT tests, have not been performed. Indeed, the DPRA test is not applicable to metal elements such as the test item.
Regarding the h-CLAT test, it is needed to know the log Kow of the substance before performing the test to conclude correctly. Indeed, substances having a log Kow > 3.5 tend to produce false negative results. Here, the log Kow is not provided for the substance because it is not stable in the water. Therefore, the h-CLAT test would be only interpretable in case of a positive result. It was decided not to conduct this test due to this uncertainty.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)

GLP compliance:

yes

Type of study:

activation of keratinocytes

Test material

Test material information

Constituent 1

Reference substance name:

Copper, diamminebis(nitrato-kO)

EC Number:

604-942-9

Cas Number:

15414-04-7

Molecular formula:

CuH6N4O6

IUPAC Name:

Copper, diamminebis(nitrato-kO)

In vitro test system

Details on the study design:

Test item
The test item was tested at 12 concentrations according to a geometric progression of ratio 2 from
0.2 µg/ml to 400 µg/ml.

Reference item
Negative control: 6 wells of solvent control (1% DMSO in treatment medium) with cells and 1 well of solvent control without cell by culture plate.
Positive control: 5 concentrations of cinnamaldehyde on each culture plate. The concentration varies from 4 to 64 µM according to a geometric progression of ratio 2.

The study was composed of three independent repetitions. For each repetition the test item and the reference items were replicated on three independent plates for the measurement of induction and two plates for the measurement of cytotoxicity. Each repetition were performed with fresh stock solution.

Cells: KeratinoSens™, cultured in maintenance medium at 37°C, 5% CO2.
Cells are exempt of mycoplasma. Cells were used at passage 16 in repetition 1 and passage 18 in repetition 2 and 3.

Results and discussion

Positive control results:

Cinnamaldehyde:
Repetition 1, EC1.5 is higher than the historical data value and OECD validation interval. A third repetition was therefore performed.
Repetition 2 and 3, EC1.5 are within historical data or within the OECD validation interval.
All other validation criteria are met which validates the test.

In vitro / in chemico

Results

Results 1

Key result

Run / experiment:

mean

Parameter:

other: Imax

Value:

101.05

Positive controls validity:

valid

Results 2

Key result

Run / experiment:

mean

Parameter:

other: EC1.5 (mg/mL)

Value:

200

Positive controls validity:

valid

Other effects / acceptance of results:

For the test item, Imax are higher than 1.5, the EC1.5 are lower than 200 mg/ml and, at the EC1.5 concentration, viability is higher than 70% (see below).

Any other information on results incl. tables

Test item

	VIABILITY	INDUCTION
	IC70  µg/ml	Imax	Linear EC1.5  µg/ml	EC1.5 Lin/Log  µg/ml
Rep 1	124.31	92.63	<0.20	0.17
Rep 2	113.95	131.77	0.66	0.64
Rep 3	162.86	78.76	1.18	1.12
Mean	-	101.05	-	-
Geometric mean	132.13	-	-	-

Applicant's summary and conclusion

Interpretation of results:

Category 1B (indication of skin sensitising potential) based on GHS criteria

Conclusions:

Under the retained experimental conditions MNP-363H code ID-17/10295 may be classified as sensitizer.
The test method KeratinoSensTM is considered scientifically valid to be used as part of an integrated approaches to testing and assessment, to support the identification of the sensitization potential of test item for hazard classification and labeling purposes.