Copper, diammine[5,5'-bi-1H-tetrazolato(2-)-kN1, kN1']

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June-July 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

dated 28 july 2015
method B40bis of the Council regulation No.440/2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Copper, diammine[5,5'-bi-1H-tetrazolato(2-)-kN1, kN1']

EC Number:

611-056-6

Cas Number:

538313-26-7

Molecular formula:

C2H6CuN10

IUPAC Name:

Copper, diammine[5,5'-bi-1H-tetrazolato(2-)-kN1, kN1']

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Provider : AUTOLIV ASP, 16700 W. Hwy 83, Promontory, UTAH 84307, USA
- batch No. P3656461
- Expiration date of the lot/batch: august 25th 2020
- Purity: 97.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in a restricted area dedicated to explosive products, at outside temperature and humidity
- Solubility and stability of the test substance in the solvent/vehicle: not soluble in water

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: no treatment

FORM AS APPLIED IN THE TEST (if different from that of starting material)
not different

In vitro test system

Test system:

artificial membrane barrier model

Source species:

human

Cell type:

other: 0.6 cm2 reconstituted epidermis

Cell source:

other: epiCS, Cell Systems

Justification for test system used:

corrosion assay on animals is forbiden

Vehicle:

unchanged (no vehicle)

Details on test system:

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature
- Temperature of post-treatment incubation (if applicable): no
REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: 20 times

DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer: Elx80 absorbance microplate reader Biotek
- Wavelength:570 nm

NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION:
PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if viability measured is <50% after 3 min of exposure or if viability measured is >=50% after 3 min and <15% after 60 minutes of exposure
- The test substance is considered to be non-corrosive to skin if viability measured is >= 50% after 3 min and >=15% after 60 minutes of exposure

Control samples:

yes, concurrent negative control

yes, concurrent no treatment

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied 25 mg

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): PBS PAN BIOTECH GmbH, batch No. 23802016


POSITIVE CONTROL
- Amount(s) applied (volume or weight): 8N KOH, Sigma, batch No. SLBD3295V

Duration of treatment / exposure:

3 minutes
30 minutes

Duration of post-treatment incubation (if applicable):

no applicable

Number of replicates:

2

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

test item: after 3 minutes: 95.11% of viability after 60 minutes: 94.25% of viability

Value:

> <

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system:
- Direct-MTT reduction: the intrinsic colour of the test item is blue, wich did not allow to conclude on its direct >MTT reduction potential. 2 killed skin models were added.
- Colour interference with MTT: not interfere with MTT assay

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control:
- Acceptance criteria met for positive control:
- Acceptance criteria met for variability between replicate measurements:
- Range of historical values if different from the ones specified in the test guideline:

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In accordance with the regulation (EC) No. 1272/2008, the results obtained under these experimental conditions enable to conclude taht the test item Copper Dianmmine Bitetrazole (CuDABT) does not have to be classified in category 1 "corrosive". The hasard statement "H314: causes severe skin burns and eye damage" with the signal word "Danger" are not required.

Executive summary:

The aim of the study was to evaluate the possible corrosive effects of the test item after topical administration on in vitro human reconstituted epidermis (epiCS(R), CellSystems(R)).

The test item Copper Diammine Bitetrazole (CuDABT) was applied as supplied, at the dose of 25 mg, to 2 living and 2 killed Human skin model surfaces (0.6 cm2, epiCS(R), CellSystems(R)) during 3 minutes and 1 hour, followed by a rince with 20 mL of PBS. Cell viability was then measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that was quantitatively measured after extraction from tissues. The experimental protocol was established in accordance with O.E.C.D. test Guideline No. 431 dated 28 July 2015 and the method B.40bis of the Council regulation No.440/2008.

3 minutes and 1 hour after the test item application, the mean corrected percents viability of epidermis skins treated with the test item were 95.11% and 94.25%, versus 5.13% and 0.39%, respectively, with the positive control item (potassium hydroxide 8N).

In accordance with the Regulation (EC) No. 1272/2008, the results obtained under these experimental conditions enable to conclude that the test item Copper Diammine Bitetrazole (CuDABT) does not have to be classified in Category 1 'Corrosive". The hazard statement "H314: Causes severe skin burns and eye damage" with signal word "Danger" are not required.