Copper bis(dimethyldithiocarbamate)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 December 2017 - 13 February 2018

Reliability:

1 (reliable without restriction)

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 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

solid: crystalline

Details on test material:

Batch number 37160906202

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Supplied from romachem batch number 37160906201
- Expiration date of the lot/batch:September 2018




STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None
- Preliminary purification step (if any):none
- Final dilution of a dissolved solid, stock liquid or gel:Neat
- Final preparation of a solid:



OTHER SPECIFICS:

In vitro test system

Test system:

human skin model

Cell type:

other: Noormal human-derived epidermal keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

SKIN DISC PREPARATION
- Procedure used: tested for its potential for MTT interference and water colouration
- Quality control for skin discs: Electrical resistance obtained with two of the isolated skin discs was [complete, e.g. 10 kΩ]
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 10minues +/-1 minute (25mines at RT and 35 mins at 37C, 5% carbon dioxide >= 95% relative humidity)
- Temperature of post-treatment incubation (if applicable): 42 +/- 4hours
REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps:
- Observable damage in the tissue due to washing:
- Modifications to validated SOP:
DYE BINDING METHOD
- Dye used in the dye-binding assay: [none / MTT / Sulforhodamine B / other:] MTT
- Spectrophotometer: BMG Latech Fluostar Optima
- Wavelength: 570nm
- Filter:no
-
NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3
PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be non-corrosive to skin if the skin model viability after exposure and post treatment incubation >50%
- Justification for the selection of the cut-off point(s) if different than recommended in TG 430:

Control samples:

yes, concurrent positive control

other: concurrent negative control 30uL DPBS

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25mg
- Concentration (if solution):

VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

NEGATIVE CONTROL
- Amount(s) applied (volume or weight):30uL
- Concentration (if solution): neat

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30uL
- Concentration (if solution): 5%

Duration of treatment / exposure:

60 +/-1 minutes (25 minutes at room temperature and 35 minutes at 37C, 5% CO2, >=95% Relative humidity

Duration of post-treatment incubation (if applicable):

42 +/- 4 hours post does incubation at 37C, 5% CO2, >= 95% relative humidity.

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

In vitro assessment of skin irritation potential of copper dimethyldithiocarbamate according to OECD Test Guideline 439 (reconstructed Human Epidermis Test Method)

Value:

>= 50 - < 100

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Remarks:

93.91%

Any other information on results incl. tables

Condition	Tissue	Raw data	Blank corrected date	Mean OD	% of Viability
		Aliquot 1/Aliquot 2	Aliquot 1/Aliquot 2
Negative conrol NC	Tissue 1	2.041/2.995	1.889/1.843	1.866	111.226
NC	Tissue 2	1.759/1.79	1.607/1.638	1.623	96.712
NC	Tissue 3	1.701/1.692	1.549/1.540	1.545	92.062
Positive control PC	Tissue 1	0.25/0.235	0.098/0.083	0.091	5.394
PC	Tissue 2	0.217/0.22	0.065/0.068	0.067	3.964
PC	Tissue 3	0.216/0.225	0.064/0.073	0.069	4.083
TA1	Tissue 1	1.772/1.72	1.620/1.568	1.594	95.013
TA1	Tissue 2	1.666/1.662	1.514/1.510	1.512	90.125
TA1	Tissue 3	1.82/1.771	1.658/1.619	1.639	97.665

Name/Code	Mean of OD	SD of OD	Mean of viability(%)	SD of viability (%)	CV (%)	Classification
DPBS/NC	1.678	0.168	100.000	9.996	9.996	Non-irritant
SDS 5%/PC	0.075	0.013	4.480	0.794	I7.716	Irritant
Copper dithiodicarbamate/TA1	1.582	0.064	94.268	3.825	4.058	Non-irritant