[[2,2',2''-[29H,31H-phthalocyaninetriyltris(methylene)]tris[1H-isoindole-1,3(2H)-dionato]](2-)-N29,N30,N31,N32]copper

Endpoint summary

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

basic toxicokinetics in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: non GLP, but well documented.

Reason / purpose for cross-reference:

reference to same study

Objective of study:

other: distribution in plasma and liver

Principles of method if other than guideline:

Copper-content was determined in plasma and liver samples of rats that had been given gavage doses for 14 days at doses of 300 and 1000 mg/kg bw.

GLP compliance:

no

Specific details on test material used for the study:

Purity >= 97%
Batch no. 120001P040
Date of production: 13 March 2012
stability: unlimited
homogenity: given
appearance: solid, blue
storage conditions: ambient (room temperature); dry storage; protect against humidity

Radiolabelling:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sulzfeld, Germany
- Age at study initiation: 42 +/- 1 days
- Weight at study initiation:
- Fasting period before study: none
- Housing: single
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30 - 70%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2014-06-09 To:2014-06-24

Route of administration:

oral: gavage

Vehicle:

other: dnnkmg water containing 0.5% carboxymethylcellulose with about 5 mg/100 ml Cremophor EL

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Daily
The substance is a homogenous suspension.

Duration and frequency of treatment / exposure:

14 days

Dose / conc.:

300 mg/kg bw (total dose)

Dose / conc.:

1 000 mg/kg bw (total dose)

No. of animals per sex per dose / concentration:

5

Control animals:

yes, concurrent vehicle

Positive control reference chemical:

A 14-day gavage study with an acid-labile copper-pigment (CAS 15680-42-9) was performed in the same year and this study showed a strong increase in liver copper content. (Plasma concentrations were not determined.)

Details on study design:

see endpoint study record for repeated dose toxcity

Details on dosing and sampling:

The determination of copper concentrations in EDTA plasma samples after were performed at the test facility.
The total concentrations of the test substance in plasma were calculated indirectly from the determined copper concentrations (inductively-coupled-plasma mass-spectrometry; experimental error + /- 0,2 mg/kg Cu).
This part of the study was carried out in compliance with the Principles of Good Laboratory Practice.
On study day 15, EDTA blood samples (about 200 μL) were collected from fasted animals by puncturing the retro-bulbar venous plexus under isoflurane anesthesia. After plasma preparation, the samples were frozen at about -80°C prior to analysis.

Liver samples were frozen at about -80°C prior to analysis.

Type:

distribution

Results:

No increase in plasma and liver copper concentration was detected.

Details on distribution in tissues:

No increase in plasma and liver copper concentration was detected (see tables 1 and 2).

Conclusions:

Interpretation of results: no bioaccumulation potential based on study results

Executive summary:

Table 1: Copper content in liver samples

	animal no.	g liver	mg Copper	mg/g liver	mean (n = 5)
	1	2.043	0.011	5.38	5.54
males	2	2.021	0.011	5.44
control	3	2.2772	0.013	5.71
	4	1.9791	0.012	6.06
	5	1.9656	0.01	5.09
	6	2.2573	0.014	6.20	6.01
males	7	1.63	0.01	6.13
300 mg/kg bw	8	1.5929	0.008	5.02
	9	2.0888	0.012	5.74
	10	2.0201	0.014	6.93
	11	2.1328	0.011	5.16	5.31
males	12	1.9677	0.011	5.59
1000 mg/kg bw	13	1.2978	0.007	5.39
	14	1.977	0.011	5.56
	15	2.4663	0.012	4.87
	16	1.3527	0.008	5.91	6.15
females	17	1.2401	0.008	6.45
control	18	1.2873	0.008	6.21
	19	1.3878	0.008	5.76
	20	1.2479	0.008	6.41
	21	1.245	0.008	6.43	6.41
females	22	1.1989	0.008	6.67
300 mg/kg bw	23	1.3348	0.009	6.74
	24	1.5306	0.009	5.88
	25	1.2649	0.008	6.32
females	26	1.2009	0.007	5.83	6.16
1000 mg/kg bw	27	1.2749	0.009	7.06
	28	1.2284	0.007	5.70
	29	1.3773	0.008	5.81
	30	1.2447	0.008	6.43

Table 2: Copper content in plasma samples

	animal no.	mg/kg	mean
	1	1	1.28
males	2	1.3
control	3	1.3
	4	1.1
	5	1.7
	6	2.1	1.48
males	7	1.1
300 mg/kg bw	8	1.5
	9	1.2
	10	1.5
	11	1.5	1.4
males	12	1.1
1000 mg/kg bw	13	1.5
	14	1.4
	15	1.5
	16	1	1.18
females	17	1
control	18	1.3
	19	1.1
	20	1.5
	21	1.5	1.4
females	22	1.1
300 mg/kg bw	23	1
	24	1.5
	25	1.9
females	26	1.3	1.52
1000 mg/kg bw	27	2.3
	28	1
	29	1.1
	30	1.5

Reference 2

Endpoint:

basic toxicokinetics in vivo

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Well documented and acceptable study with the following restrictions: A 91 day subchronic dosed feed toxicity study was conducted in mice, similar to OECD Guideline 408 (Subchronic Oral Toxicity - Rodent: 90-day Study), but not a toxicokinetics study (similar to OECD Guideline 417). However, the study additionally included the examination of systemic copper absorption after exposure to the test material, analyzed in livers and kidneys of the animals.

Reason / purpose for cross-reference:

reference to same study

Objective of study:

distribution

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD Guideline 408 (Subchronic Oral Toxicity - Rodent: 90-day Study)

Principles of method if other than guideline:

This subchronic toxicity study, similar to OECD Guideline 408, was conducted to assist in selecting Maximum Tolerated Dosages (MTD) for a 104 week chronic study in a dosed feed subchronic study. Additionally the study examined, whether or not systemic absorption of copper occured after exposure to the test material. Copper analyses were conducted in livers and kidneys of the animals.

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): Pigment Blue 15B
- Analytical purity: The chemical analysis, performed at Midwest Research Institute indicated that the purity was 104.7 % +- 1.1 % (elemental analysis)
- Elemental analysis indicated that the compound contained less than 0.01 % chlorine.

- Structural formula attached as image file (if other than submission substance): see Fig.

Radiolabelling:

no

Species:

mouse

Strain:

B6C3F1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Industries
- Age at study initiation: 8.5 weeks
- Weight at study initiation: males: 16 - 23 g; females: 16 - 19 g
- Housing: polycarbonate cages: groups of 5 mice per cage
- Diet: weighed portions of Purina Lab Chow in meal form, mixed together with weighed portions of the test material (see details at "doses/concentrations")
- Water: ad libitum
- Acclimation period: 15 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21 - 23 °C
- Humidity: 40 - 60 %
- Air changes: at least 15 per hour
- Photoperiod: 12 hrs dark / 12 hrs light

Route of administration:

oral: feed

Vehicle:

other: 12 % water was added to the test material as a dust control agent

Details on exposure:

Dose levels of 5.0, 2.5, 1.25, 0.6 and 0.3 % (w/w) were selected for both males and females. The selected doses were prepared by mixing weighed portions of purina Lab Chow in meal form with weighed portions of the test material. 12 % water was added to the test material as a dust control agent prior to mixing with the meal. For each dose level, one weekly lot of 4500 g (+ 12 % water compensation) was prepared.

The actual mixtures were composed of the following ingredients:
- Dose level 5.0 % (w/w): 252 g test material and water + 4275 g meal
- Dose level 2.5 % (w/w): 126 g test material and water + 4387.5 g meal
- Dose level 1.25 % (w/w): 63 g test material and water + 4443.75 g meal
- Dose level 0.6 % (w/w): 30.24 g test material and water + 44735 g meal
- Dose level 0.3 % (w/w): 15.12 g test material and water + 4486.5 g meal

Each diet was mixed in a Patterson-Kelly twin shelled V blender for 15 min.
The doses were mixed one or two days prior to the week of their use in the study, and stored at 23 °C.
One analysis was perfomed to determine the accuracy of the mixture concentration.

Duration and frequency of treatment / exposure:

90 days

Dose / conc.:

0.3 other: %

Remarks:

in diet, approx. 1000 mg/kg bw for males [based on 7.3 g/d average food consumption, 0.023 kg average bw] and approx.1100 mg/kg bw for females [based on 7.1 g/d average food consumption, 0.019 kg average bw], respectively).

Dose / conc.:

0.6 other: %

Remarks:

in diet, approx. 2000 mg/kg bw for males [based on 7.3 g/d average food consumption, 0.023 kg average bw] and approx. 2200 mg/kg bw for females [based on 7.1 g/d average food consumption, 0.019 kg average bw], respectively).

Dose / conc.:

1.25 other: %

Remarks:

in diet, approx. 4000 mg/kg bw for males [based on 7.3 g/d average food consumption, 0.023 kg average bw] and approx. 4700 mg/kg bw for females [based on 7.1 g/d average food consumption, 0.019 kg average bw], respectively).

Dose / conc.:

2.5 other: %

Remarks:

in diet, approx. 8000 mg/kg bw for males [based on 7.3 g/d average food consumption, 0.023 kg average bw] and approx. 9400 mg/kg bw for females [based on 7.1 g/d average food consumption, 0.019 kg average bw], respectively).

Dose / conc.:

5 other: %

Remarks:

in diet, approx. 16000 mg/kg bw for males [based on 7.3 g/d average food consumption, 0.023 kg average bw] and approx. 18700 mg/kg bw for females [based on 7.1 g/d average food consumption, 0.019 kg average bw], respectively).

No. of animals per sex per dose / concentration:

10 males and 10 females per dose

Control animals:

yes, plain diet

Details on study design:

- 10 animals were used per sex and dose group.
- Five dose levels of 0.0, 0.3, 0.6, 1.25 and 5.0 % in feed were used in this study (approx. 1000, 2000, 4000, 8000 and 16000 mg/kg bw for males [based on 7.3 g/d average food consumption, 0.023 kg average bw] and approx. 1100, 2200, 4700, 9300 and 18700 mg/kg bw for females [based on 7.1 g/d average food consumption, 0.019 kg average bw], respectively).
- The selected doses were prepared by mixing together weighed portions of Purina Lab Chow in meal form with weighed portions of the chemical. 12% water was added to the chemical as a dust control agent prior to mixing with the meal.
- Each dosed group received on 91 consecutive days of dosed feed mixture.
- Mice were necropsied on day 92 and 93.

Copper analyses were completed in the liver and kidney tissues and the formalin preserving those tissues from male mice in the highest dose group (5 % w/w) and control groups:
- Tissue samples were prepared for analysis by digesting in 10 ml of concentrated nitric acid until most of the organic material was destroyed. Perchloric acid was then added and the solutions were evaporated to strong fumes, additional nitric acid being added as required. The solutions were then fumed to dryness, the residues were dissolved in 5 % nitric acid and the solutions were diluted to 10 ml.
- Formalin samples were filtered through a Millex-GS 0.22 µm filter unit and 5 ml portions of each sample were prepared for analysis by the procedure used to prepare the tissue samples.
- The samples were then subjected to atomic absorption spectrophotometry to determine copper content:
A Perkin-Elmer Model 5000 atomic absorption spectrophotometer was utilized for the work. A series of 10 ml standard solutions, ranging from 0.05 to 2.0 ppm were prepared in 5 % nitric acid by dilution of a certified standard copper stock solution. These solutions were used to calibrate the instrument, which was programmed to print out data as total microgramms of copper per sample. The prepared sample solutions were used in the same manner as the standards. Concentrations of copper in the tissue samples were calculated by dividing the total microgramms found by the weight of the sample. Concentrations of copper in the formalin samples were calculated on a volume basis.

Statistics:

Student´s T-test (alpha = 0.05) was used to compare the highest dose group results with control results.

Preliminary studies:

No data given.

Details on absorption:

No data given.

Details on distribution in tissues:

Slight, but statistically significant increases of copper incorporation were reported in the liver tissues (3.98 ppm +- 1.16 ppm; p < 0.05) and kidney (7.47 ppm +- 2.86 ppm; p < 0.05) tissues of treated male animals of the highest dose group, compared to controls (liver: 3.0 ppm +- 0.34 ppm; kidney: 4.66 ppm +- 0.6 ppm). From all the formalin analyses performed, the authors drawed the conclusion that no detectable levels of copper were leached from the preserved tissue into the formalin bath.
However, an evaluation of the results was conducted by expert judgement and the author draw the following conclusions:
The results of the study do not provide unequivocal evidence for a lack of absorption of the test material. However, the total lack of findings during the 13 week study, coupled with the insolubility of the test material and the minimal changes in tissue copper residues strongly suggests that the test material was not appreciably absorbed.

Details on excretion:

No data given.

Metabolites identified:

not measured

Details on metabolites:

No data given.

Table 1: Copper determinations in tissues and formalin of male mice from the subchronic study, treated with the test material for 91 days

male animal #	ppm copper
	liver	kidney	formalin	remark
highest dose group (5 % w/w)
1	7.0	14.5	--
2	3.2-3.6 *	6.8	--	* results of 2 analyses
3	3.4	7.7	--
4	3.4	4.8	--
5	3.5	6.3	--
6	3.7	7.3	--
7	3.9	6.4	--
8	4.1	8.2	--
9	3.3-3.6 *	5.2	--	* results of 2 analyses
control
1	3.5	4.2	0.1
2	3.1	3.8	0.1
3	3.1	4.6	0.1
4	2.8	4.6	0.1
5	2.9	4.2	0.1
6	3.2	4.7	0.1
7	3.2	5.8	0.1
8	2.9	5.3	0.1
9	2.3	4.7	0.1

Conclusions:

Interpretation of results: no bioaccumulation potential based on study results

Reference 3

Endpoint:

basic toxicokinetics in vivo

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Well documented and acceptable study with the following restrictions: A 91 day subchronic dosed feed toxicity study was conducted in rats, similar to OECD Guideline 408 (Subchronic Oral Toxicity - Rodent: 90-day Study), but not a toxicokinetics study (similar to OECD Guideline 417). However, the study additionally included the examination of systemic copper absorption after exposure to the test material, analyzed in livers and kidneys of the animals.

Reason / purpose for cross-reference:

reference to same study

Objective of study:

distribution

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD Guideline 408 (Subchronic Oral Toxicity - Rodent: 90-day Study)

Deviations:

yes

Principles of method if other than guideline:

This subchronic toxicity study, similar to OECD Guideline 408, was conducted to assist in selecting Maximum Tolerated Dosages (MTD) for a 104 week chronic study in a dosed feed subchronic study. Additionally the study examined, whether or not systemic absorption of copper occured after exposure to the test material. Copper analyses were conducted in livers and kidneys of the animals.

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): Pigment Blue 15B
- Analytical purity: The chemical analysis, performed at Midwest Research Institute indicated that the purity was 104.7 % +- 1.1 % (elemental analysis)
- Elemental analysis indicated that the compound contained less than 0.01 % chlorine.
- Structural formula attached as image file (if other than submission substance): see Fig.

Radiolabelling:

no

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Industries
- Weight at study initiation: males: 70 - 105 g; females: 70 - 100 g
- Housing: polycarbonate cages: groups of 5 rats per cage
- Diet: weighed portions of Purina Lab Chow in meal form, mixed together with weighed portions of the test material (see details at "doses/concentrations")
- Water: ad libitum
- Acclimation period: 15 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21 - 23 °C
- Humidity: 40 - 60 %
- Air changes: at least 15 per hour
- Photoperiod: 12 hrs dark / 12 hrs light

Route of administration:

oral: feed

Vehicle:

other: 12 % water was added to the test material as a dust control agent

Details on exposure:

Dose levels of 5.0, 2.5, 1.25, 0.6 and 0.3 % (w/w) were selected for both males and females. The selected doses were prepared by mixing weighed portions of purina Lab Chow in meal form with weighed portions of the test material. 12 % water was added to the test material as a dust control agent prior to mixing with the meal. For each dose level, one weekly lot of 4500 g (+ 12 % water compensation) was prepared.

The actual mixtures were composed of the following ingredients:
- Dose level 5.0 % (w/w): 252 g test material and water + 4275 g meal
- Dose level 2.5 % (w/w): 126 g test material and water + 4387.5 g meal
- Dose level 1.25 % (w/w): 63 g test material and water + 4443.75 g meal
- Dose level 0.6 % (w/w): 30.24 g test material and water + 44735 g meal
- Dose level 0.3 % (w/w): 15.12 g test material and water + 4486.5 g meal

Each diet was mixed in a Patterson-Kelly twin shelled V blender for 15 min.
The doses were mixed one or two days prior to the week of their use in the study, and stored at 23 °C.
One analysis was perfomed to determine the accuracy of the mixture concentration.

Duration and frequency of treatment / exposure:

90 days

Dose / conc.:

0.3 other: %

Remarks:

in diet; approx. 250 mg/kg bw for both sexes [based on 16.4 g/d average food consumption, 0.182 kg average bw for males and on 11.55 g/d average food consumption, 0.130 kg average bw] for females)

Dose / conc.:

0.6 other: %

Remarks:

in diet; approx. 500 mg/kg bw for both sexes [based on 16.4 g/d average food consumption, 0.182 kg average bw for males and on 11.55 g/d average food consumption, 0.130 kg average bw] for females)

Dose / conc.:

1.25 other: %

Remarks:

in diet; approx. 1100 mg/kg bw for both sexes [based on 16.4 g/d average food consumption, 0.182 kg average bw for males and on 11.55 g/d average food consumption, 0.130 kg average bw] for females)

Dose / conc.:

2.5 other: %

Remarks:

in diet; approx. 2200 mg/kg bw for both sexes [based on 16.4 g/d average food consumption, 0.182 kg average bw for males and on 11.55 g/d average food consumption, 0.130 kg average bw] for females)

Dose / conc.:

5 other: %

Remarks:

in diet; approx. 4500 mg/kg bw for both sexes [based on 16.4 g/d average food consumption, 0.182 kg average bw for males and on 11.55 g/d average food consumption, 0.130 kg average bw] for females)

No. of animals per sex per dose / concentration:

10 males per dose and 10 females per dose

Control animals:

yes, plain diet

Details on study design:

The concentrations of the chemical mixture were the same for male and female rats. All dose levels were prepared on a weight per weight basis. There were 5 dose level groups with 10 individuals of each sex in each dosage and control group. Each dosed group received 90 consecutive days of dosed feed mixture. After one day of observation, the animals were necropsied. Animals were observed twice each day for clinical signs, with at least 6 hours between observations. All observations were recorded daily. Additionally, blood sampling was conducted from 10 control rats, 6 males and 4 females.

Copper analyses were completed in the liver and kidney tissues and the formalin preserving those tissues from male rats in the highest dose group (5 % w/w) and control groups:
- Tissue samples were prepared for analysis by digesting in 10 ml of concentrated nitric acid until most of the organic material was destroyed. Perchloric acid was then added and the solutions were evaporated to strong fumes, additional nitric acid being added as required. The solutions were then fumed to dryness, the residues were dissolved in 5 % nitric acid and the solutions were diluted to 10 ml.
- Formalin samples were filtered through a Millex-GS 0.22 µm filter unit and 5 ml portions of each sample were prepared for analysis by the procedure used to prepare the tissue samples.
- The samples were then subjected to atomic absorption spectrophotometry to determine copper content:
A Perkin-Elmer Model 5000 atomic absorption spectrophotometer was utilized for the work. A series of 10 ml standard solutions, ranging from 0.05 to 2.0 ppm were prepared in 5 % nitric acid by dilution of a certified standard copper stock solution. These solutions were used to calibrate the instrument, which was programmed to print out data as total microgramms of copper per sample. The prepared sample solutions were used in the same manner as the standards. Concentrations of copper in the tissue samples were calculated by dividing the total microgramms found by the weight of the sample. Concentrations of copper in the formalin samples were calculated on a volume basis.

Details on dosing and sampling:

PHARMACOKINETIC STUDY (distribution)
- Tissues and body fluids sampled: liver and kidneys

Statistics:

Student´s T-test (alpha = 0.05) was used to compare the highest dose group results with control results.

Details on absorption:

No data given.

Details on distribution in tissues:

No statistically significant increase of residual copper incorporation was neither reported for the liver tissues (2.82 ppm +- 0.34 ppm) nor for the kidney tissues (5.62 ppm +- 0.49 ppm) of the treated male rats of the highest dose group, compared to residual copper incorporation found in controls (liver 2.78 ppm +- 0.51 ppm; kidney 5.30 ppm +- 0.83 ppm). From all the formalin analyses performed, the authors drawed the conclusion that no detectable levels of copper were leached from the preserved tissue into the formalin bath.

Details on excretion:

No data given.

Metabolites identified:

not measured

Details on metabolites:

No data given.

Table 1: Copper determinations in tissues and formalin of male rats from the subchronic study, treated with the test material for 90 days

male animal #	ppm copper
	liver	kidney	formalin	remark
highest dose group (5 % w/w)
1	4.6 - 4.3*	8.4	0.1	* results of 2 analyses
2	5.9	12.3	0.1
3	4.1 - 4.4*	8.6 - 10.1*	0.1	* results of 2 analyses
4	6.4	7.7	0.1
5	3.2	6.7	0.1
6	3.5	5.8	0.1
7	3.7	8.1	0.1
8	3.5 - 4.1	8.1	0.1
9	3.1	8.7 - 8.6*	0.1	* results of 2 analyses
10	4.6 - 4.5	7.2	0.1
control
1	3.1	3.7 - 4.0*	0.1	* results of 2 analyses
2	3.3	4.1	0.1
3	3.2	4.9 - 4.6*	0.1	* results of 2 analyses
4	3.7 - 4.0*	5.1	0.1	* results of 2 analyses
5	3.0	4.1	0.1
6	2.8	5.4	0.1
7	2.9	2.9 - 3.4*	0.1	* results of 2 analyses
8	2.6	5.5	0.1
9	2.6	5.4	0.1
10	3.3 - 3.6*	5.4	0.1	* results of 2 analyses

Conclusions:

Interpretation of results: no bioaccumulation potential based on study results

Description of key information

The substance is an inert high molecular weight organic copper complex. It fulfills the criteria of the definition of a nanomaterial in the European Union.

Is not systemically available after ingestion as indicated by lack of increased plasma and liver copper concentrations and absence of toxicity after 14 day gavage dosing of rats with 300 or 1000 mg/kg bw.

Key value for chemical safety assessment

Bioaccumulation potential:

no bioaccumulation potential

Additional information

The substance consists of a mixture of copper phthalocyanine with 0 to 3 1 H-isoindole-dione covalently bound substituents. The average degree of substitution is 2.48 which results in an average mass of 970 g/mol. The substance insoluble in water as examined via total organic carbon and copper content (3 μg/L at loading rate of 0.1 g/L

) and poorly soluble in octanol (1 mg/L at a loading rate of 0.1 g/L). All these properties indicate that transport through biological membranes is hindered. Absence of toxicity upon ingestion in subacute toxicity studies in rats has been observed for related copper phthalocyanine pigments and the copper phthalocyanine core (CAS nos 147-14-8, 75247-18 -6, 81457-65 -0) and inert organic pigments in general.

In support of this, a 14 -day study originally intended as a dose-range-finder study was performed with additional investigations on copper concentrations in liver and plasma at the end of treatment (BASF 2015). This study was performed with each 5 male and female rats and included clincal chemistry and haematology parameters. A similar 14 -day study with an acid-labile and lower molecular weight organic copper pigment (Pigment Yellow 129) had shown that such exposure resulted in a significant increase in liver copper concentrations. In contrast, the target substance did not cause increased copper concentrations in liver and plasma after 14 days of oral dosing at the limit dose. In addition, no blue discoloration of internal tissues were observed and there was no greenish urine discoloration. Clinical chemistry and haematology parameters were not affected and overall no adverse effects were noted. From this it is concluded that the target substance is not taken up by the body after ingestion.

For the weight-of-evidence approach, the same type of information obtained with the copper phthalocyanine core (CAS 147 -14 -8) after subchronic feed application of high doses is also provided. It shows that no copper accumulation in liver results from subchronic high dose exposure.

A read-across justification document with structures and data matrices is attached. Target and structural analogues fulfill the definition of a nanomaterial in the European Union.

Considering the insolubility and the high molecular weight, it is expected that the substance is not able to penetrate the skin.

Inhalation is expected to result in local effects related to inert particles.