Reaction mass of 29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32 copper and Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, (dimethylamino)methyl derivs.

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 July 2016 - 15 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

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

The purity reported is different to the Study Plan due to a new Certificate of Analysis being provided by the sponsor.

Deviations:

yes

Remarks:

this fact has no impact on the results or integrity of the study.

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Commission Regulation (EC) No 440/2008, Annex Part B, B.40.Bis: “In Vitro Skin Corrosion: Human Skin Model Test”, Official Journal of the European Union No. L142 (31 May 2008)

Version / remarks:

The purity reported is different to the Study Plan due to a new Certificate of Analysis being provided by the sponsor.

Deviations:

yes

Remarks:

this fact has no impact on the results or integrity of the study.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2λ⁵,11,20,29,38,39λ⁵,40,41-octaaza-1-cupradodecacyclo[19.17.1.1¹,¹².1³,³⁷.0²,¹⁰.0⁴,⁹.0¹³,¹⁸.0²²,²⁷.0²⁸,³⁹.0³⁰,³⁸.0³¹,³⁶.0¹⁹,⁴¹]hentetraconta-2,4,6,8,10,12,14,16,18,20,22,24,26,28(39),29,31,33,35,37(40)-nonadecaene-2,39-bis(ylium)-1,1-diuide

EC Number:

908-081-2

IUPAC Name:

2λ⁵,11,20,29,38,39λ⁵,40,41-octaaza-1-cupradodecacyclo[19.17.1.1¹,¹².1³,³⁷.0²,¹⁰.0⁴,⁹.0¹³,¹⁸.0²²,²⁷.0²⁸,³⁹.0³⁰,³⁸.0³¹,³⁶.0¹⁹,⁴¹]hentetraconta-2,4,6,8,10,12,14,16,18,20,22,24,26,28(39),29,31,33,35,37(40)-nonadecaene-2,39-bis(ylium)-1,1-diuide

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Batch number: 150205
Description: Blue powder
Manufacture date: 05 February 2015
Expiry date: 05 February 2019
Purity: As a UVCB, purity is considered as 100%
Storage conditions: Controlled room temperature (15-25°C, below 70% RH%).

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: main basal, supra basal, spinous and granular layers and a functional stratum corneum

Source strain:

not specified

Details on animal used as source of test system:

Manufactured tissues, reconstructed epidermis

Justification for test system used:

The model has been validated for corrosivity testing in an international trial (Fentem, 1998) and its use is recommended by the relevant OECD guideline for corrosivity testing (OECD No. 431); therefore, it was considered to be suitable for this study.

Vehicle:

physiological saline

Details on test system:

SKIN DISC PREPARATION
- Procedure used: supplied by manufacturer
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 25.2-26.4°C
REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: 1
- Observable damage in the tissue due to washing: no
DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer: yes
- 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 both disks have mean viability of <35% and the variability is less than 50%
- The test substance is considered to be non-corrosive to skin if both disks have mean viability of ≥35% and the variability is less than 50%

Control samples:

yes, concurrent negative control

yes, concurrent positive control

yes, concurrent MTT non-specific colour control

Amount/concentration applied:

- 20 mg of test item was applied evenly to the epidermal surface of each of two test units and then 100 μL physiological saline was added to the test item to ensure good contact with the epidermis.
- 50 μL of physiological saline was added to each of the two negative control skin units.
- 50 μL of glacial acetic acid was added to each of the two positive control skin units.

Duration of treatment / exposure:

4 hours (±10 min)

Number of replicates:

2

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1

Value:

108.8

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2

Value:

112.5

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

After receipt, the two indicators of the delivered kits were checked. Based on the observed colours, the epidermis units were in proper conditions in each case.
The mean OD value of the two negative control tissues was in the recommended range (0.834).
The positive control treated tissues showed 1.9% viability demonstrating the proper performance of the assay.
The difference of viability between the two test item-treated tissue samples in the MTT assay was 3.3%.
The mean OD value of the blank samples (acidified isopropanol) was 0.046.
All these parameters were within acceptable limits and therefore the study was considered to be valid.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Following exposure with the test item, the mean cell viability was 110.6% compared to the negative control (after adjustment for non-specific MTT reduction). This is above the threshold of 35%, therefore the test item was considered as being non-corrosive. The experiment met the validity criteria, therefore the study was considered to be valid.
In conclusion, in this in vitro model test with the test item, the results indicate that the test item is non-corrosive to the skin.