Hydrogen [tris[[[3-[(2-ethylhexyl)oxy]propyl]amino]sulphonyl]-29H,31H-phthalocyaninesulphonato(3-)-N29,N30,N31,N32]cuprate(1-), compound with 3-[(2-ethylhexyl)oxy]propylamine (1:1)

Endpoint summary

Description of key information

The substance is not irritating to rabbit eye and skin. It did not significantly reduce the viability of culured human corneocytes (EPIOCULAR).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1972

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: scientifically acceptable study report

Qualifier:

according to guideline

Guideline:

other: Company-internal protocol

Principles of method if other than guideline:

20h occlusive dermal exposure of 2 rabbits to 1g of moistened test substance on a cotton patch of 2.5 x 2.5 cm

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: purchased externally
- Weight at study initiation:2,87 kg and 3.17 kg

IN-LIFE DATES: From: 1972-03-27 To: 1972-04-04

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

other: untreated skin of the same animal

Amount / concentration applied:

ca. 1 g of a 50% aqueous test substance preparation

Duration of treatment / exposure:

20 hours

Observation period:

8 days

Number of animals:

2

Details on study design:

Application area: 2.5 x 2.5 cm
Application site: back
The test substance was not washed off.

Scoring for redness, oedema, scaring, scabbing, necrosis. (0 = no finding; then four categories "questionable", "mild", "strong", "severe")

Time points scored: After 20h, day 2, day 3, day 4 and day 8

Irritation parameter:

erythema score

Basis:

other: both animals

Time point:

other: 20 h, days 2,3,4 and 8

Score:

0

Max. score:

4

Remarks on result:

other: blue test substance residues; inflammation may not have been visible.

Irritation parameter:

edema score

Basis:

other: both animals

Time point:

other: 20 h, days 2,3,4 and 8

Score:

0

Max. score:

4

Irritant / corrosive response data:

The animals did not show signs of inflammation.

Other effects:

Animal one had one lost 90g of weight from the beginning of treatment to the end of the observation period.
Animal two had gained 50g of weight from the beginning of treatment to the end of the observation period.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Executive summary:

20h occlusive exposure of shaved rabbit skin did not cause local effects other than bluish coloration by the blue substance.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1972

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: scientifically acceptable study report, performed prior to the introduction of GLP and OECD guidelines. Minor deviations to current guideline (eg 2 instead of 3 rabbits, 8 day observation period).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

8 day observation period, 2 animals, 50 mg applied

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: purchased externally
- Weight at study initiation: 2.96 kg and 3.17 kg


IN-LIFE DATES: From: 1972-04-24 To: 1972-05-02

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated eye of the same animal

Amount / concentration applied:

ca. 50 mg (one spoon full)

Duration of treatment / exposure:

single treatment

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

2

Details on study design:

The test substance was not washed out.
Eyes were examined using fluorescein on the last observation day.
Scoring was performed after 1 and 3h, then after 1, 2, 3, 4 and 8 days.
Scoring for redness, oedema, scaring, cloudiness, necrosis. (0 = no finding; then four categories 1 = "questionable", 2 = "mild", 3 = "strong", 4 = "severe")

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72h

Remarks on result:

other: With talcum powder (used as comparison), the same grade of redness was obtained.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 48h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: 24h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 48h

Irritant / corrosive response data:

No clouding was noted and no effects on iris are reported. Specific scores for iris and cornea were not part of the reporting procedure.

Other effects:

Blue substance residues were observed until the 48h oberservation time point. They were no longer present at the 72h observation.

Eyes were investigated using fluorescein after 5 days. No adverse findings were noted.

Both rabbits gained weight during the observation period.

(2.9gk g t o 3.2 kg for animal 1 and 3.17kg to 3.51 kg for animal 2)

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The substance was tested for skin and eye irritation in rabbits prior to the existence of OECD testing guidelines. The standard company-specific protocols were applied. In case of skin irritation, the procedure differs in various parameters. Instead of 4h with semiocclusive exposure, the animal is treated for 20 hours with occlusive exposure. Only two animals are used and the observation period ended after 8 instead of 14 days. The scoring system consists of the same number of categories as the Draize scores and redness and chemosis are described. Also the scoring time points match the OECD guideline.

In this case, the blue test substance resulted in bluish discoloration of the treated site and a potential reddening could not be scored. No erythema was noted at any scoring time point. This clear shows that the substance does not cause local effects so that the evaluation of the skin irritation hazard is possible despite the differences in the study design.

The study for eye irritation in rabbits was performed with only two rabbits, and the applied volume of one spoon of ca 50 mg may have been lower than required by the OECD guideline. The bulk volume is not known, maybe the amount applies to the required 100 mg or 0.1 ml. Therefore, a weight-of-evidence assessment is performed and that includes the eye irritation study of the structural analogue (CAS 81457 -65 -0) and an vitro assay (EPIOCULAR) on the substance itself. In case of the analogue substance, 100 mg were applied and three animals were treated. The analogue substance is of shorter chain length ( 1-methylethoxy versus 2 -ethylhexyl) of the sulfonyl subsitutent which reduces the molecular weight of the large copper phthalocyanine dye only slightly. The side chains affect dispersibility in organic solvents, but not the (in)solubility in water. Therefore, this difference in chemical structure is considered to be of no relevance for eye irritation.

The study with the actual substance was performed with Vienna White rabbits that did not show clinical signs or effects on body weight during the treatment. No rinsing of the treated eyes were performed. Accordingly, residues of the blue test substances were observed during the first days after treatment. Mild redness and chemosis resolved within 72 and 48 hours, respectively. The other eye of the rabbit were treated with talcum poweder as "negative control". Both talcum powder and the test substance caused the same grade of redness.

The analogue substance was tested for irritating properties alo prior to the introduction of OECD testing guidelines and GLP.

Experimental procedures and reporting details are similar to current requirements, so both studies were assigned

a validity score of 2 (valid with restrictions.) The study design for the eye irritation is identical to OECD testing guideline 405, but less information on the animals and housing conditions are given. As no indication of irritation to cornea, iris or conjunctivae was observed, the observation period was terminated after the third day.

In the in-vitro assay (BASF 2015) a single topical application of 50 μL bulk volume (about 23 mg) of the undiluted test substance to a

reconstructed three dimensional human cornea model (EpiOcular™). Two EpiOcular™ tissue samples were incubated with the test substance for 6 hours followed by a 18-hours post-incubation period. Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test. The reduction of mitochondrial dehydrogenase activity, measured by reduced formazan production after incubation with a tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the test substance treated epidermal tissues is compared to that of negative control tissues. The ratio of the values indicates the relative tissue viability. Due to the intense color of the test substance it could not be determined whether the test

substance is able to reduce MTT directly. Therefore an additional MTT reduction control (freeze-killed control tissues (KC)) was introduced. Also, in a pretest for color control it was demonstrated that the blue color of the test substance did not interfere with the colorimetric test. The mean viability of the test-substance treated tissues was 81.7. No compound residues remained on the tissues after the washing procedure. Based on the observed results and applying the evaluation criteria it was concluded, that the test substance does not show an eye irritation potential in the EpiOcular™ eye irritation test.

The procedure followed the OECD (2014a) Draft Proposal for a New Test Guideline: Reconstructed Human Cornealike Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage (available at http://www.oecd.org/chemicalsafety/testing/RhCE-Test-Method-for-Identifying-Chemicals- Not-Requiring%20Classification-Labelling-for-Eye-Irritation-Draft-New-TG-2014-07-25.pdf; accessed 04 Aug 2014).

In conclusion, since both the in vivo eye irritation studies and the in-vitro eye irritation studies show effects below the threshold for classification and labelling as an eye irritant, the substance is considered to be non irritating to eyes in a weight-of-evidence approach.

Justification for selection of eye irritation endpoint:
Arbitrary, weight-of-evidence assessment

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. Mild findings were reversible were no longer observed at the 72h observation time point. As a result the substance is not considered to be classified for skin or eye irritation under Directive 67/548/EEC, as amended for the 31st time in Directive 2009/2/EG.

 

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008.

Mild findings were reversible were no longer observed at the 72h observation time point.

As a result the substance is not considered to be classified for skin or eye irritation under Regulation (EC) No. 1272/2008, as amended for the fifth time in Directive EC 944/2013.