Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Copper phthalocyanine pigments are inert high molecular weight blue to green colorants that did not cause treatment-related effects in GLP-compliant subacute gavage OECD 407 (Pigment Blue 16, Pigment Green 7, CAS 14832 -14 -5, CAS 68987-63-3) or subchronic feed application studies (Pigment Green 7 and Pigment Blue 16). The latter two studies were performed by the US National Toxicology programm to serve as a dose-range finder for a chronic study. However, since the subchronic feed application did not result in an increase in liver and kidney copper concentrations, the substances were concluded to be not taken up after ingestion and chronic testing was not performed.

The substance is assessed to be non hazardous from experimental data on related copper phthalocyanine pigments. Details can be found in the read-across justification.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
; no clinical chemistry, hematology, or urinalysis were conducted and no organ weights were taken (as recommended in OECD Guideline 408).
GLP compliance:
no
Limit test:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): Phthalocyanine Green 7
- Analytical purity: 97.8 %
Green 7 (Phthalocyanine Green) manufactured by Dry Color Manufacturers' Association, was obtained from Midwest Research institute on March 28, 1978. The elemental analysis pcrfortied at Midwest Research Institute was low for copper and nitrogen, slightly low for carbon, and slightly high for hydrogen and chlorine; values for all determined elements totaled 97.8 percent.
Species:
mouse
Strain:
B6C3F1
Sex:
male/female
Details on test animals or test system and environmental conditions:
- Source: Harlan Industries
- Age at study initiation: 8.5 weeks
- Weight at study initiation: males: 20 - 24 g; females: 15 - 18 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:
unchanged (no vehicle)
Details on oral 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): 225 g test material and water + 4275 g meal
- Dose level 2.5 % (w/w): 112.5 g test material and water + 4387.5 g meal
- Dose level 1.25 % (w/w): 56.25 g test material and water + 4443.75 g meal
- Dose level 0.6 % (w/w): 27 g test material and water + 44735 g meal
- Dose level 0.3 % (w/w): 13.5 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.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
One analysis was perfomed to determine the accuracy of the mixture concentration. Results were within +- 10 % of the desired dose concentration.
Duration of treatment / exposure:
91 days
Frequency of treatment:
daily
Dose / conc.:
0.3 other: % in the diet
Dose / conc.:
0.6 other: % in the diet
Dose / conc.:
1.25 other: % in the diet
Dose / conc.:
2.5 other: % in the diet
Dose / conc.:
5 other: % in the diet
No. of animals per sex per dose:
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, 2.5 and 5.0 % in feed were used in this study (approx. 0, 1000, 2000, 4000, 8000 or 16000 mg/kg bw/day for males, [based on 8.2 g/d average food consumption, 0.026 kg average bw] and approx. 0, 1200, 2500, 5000, 10000 and 20000 mg/kg bw/day for females [based on 7.7 g/d average food consumption, 0.019 kg average bw]).
- The selected doses were prepared by mixing together weighed portions of Purina Lab Chow in meal form with weighed portions of the chemical.
- Each dosed group received dosed feed mixture on 91 consecutive days.
Observations and examinations performed and frequency:
Animals were observed twice each day for clinical signs, with at least 6 hours between observations. All clinical signs were recorded daily. Additional studies included blood sampling for the animal disease screening program from 10 control mice, 5 males and 5 females.
Sacrifice and pathology:
Mice were necropsied on day 92 and 93.
- Gross examination were performed on all animals from all dosage groups.
- Microscopic examinations were performed on following organs from all animals in the control group and the highest dose treatment group: Kidney, liver, lung, heart, epididymis, stomach, thyroid, skin (only in control group: bone marrow, urinary bladder, testis).
Other examinations:
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. See details and results in endpoint "7.1.1. Basic toxicokinetics"
Clinical signs:
no effects observed
Description (incidence and severity):
No treatment related abnormal clinical signs were observed.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
4 deaths occurred during the study; 3 males at 1.25 % and 1 control male. These deaths were not considered to be test substance related. There were no early dead females during the course of the study.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
A -9 % to +8 % differential weight gain was seen in the dosed females. Animals in the highest dose levels had less weight gain than controls. However, given the normal variability in mouse body weight measurements, this differences did not necessarily reflect a manifestation of toxicity. Dosed male mouse groups showed a positive weight differential in all groups except the 0.6 % dosage group. In this group during week 12 with a corresponding weight loss was observed, which was not fully recovered during the final week of the study.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There were no trends in diet consumption among dosed animals compared with controls in either male or female mice. Average daily consumption
among male mice dosed groups and controls ranged from 7.6-9.5 g; female mice dosed groups and controls ranged from 7.5 to 7.8 g. While male ranges were somewhat erratic, there was no indication of dose-related effects.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No substance related changes were reported on macroscopic and microscopic examination of the animals. Gross examination were performed on all animals from all dosage groups.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No substance related changes were reported on macroscopic and microscopic examination of the animals. The microscopical lesions found were encountered in both the control and experimental high dose group with similar frequency and severity.
Details on results:
A dosage level of 5 % and 2.5 % was recommended to be used test substance in the chronic study, due to lack of compound-related lesions.
Dose descriptor:
NOAEL
Effect level:
ca. 16 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: absence of effects
Remarks on result:
other: calculated from dietary concentrations
Dose descriptor:
NOAEL
Effect level:
ca. 20 000 mg/kg bw/day (nominal)
Sex:
female
Basis for effect level:
other: absence of effects
Remarks on result:
other: calculated from dietary concentrations
Critical effects observed:
no
Conclusions:
Although this study does not determine all parameters required by OECD guideline 408, it convincingly shows absence of systemic via the lack of accumulation of copper in liver and kidney after 90 day feed application. It therefore serves as key study for the endpoint of subchronic oral toxicity.
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
; no clinical chemistry, hematology, or urinalysis were conducted and no organ weights were taken (as recommended in OECD Guideline 408). In accition, copper levels were determined in liver and kidneys
GLP compliance:
no
Limit test:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): Green 7 (C54637)
- Analytical purity: 97.8 %

Green 7 (Phthalocyanine Green) manufactured by Dry Color Manufacturers' Association, was obtained from Midwest Research institute on March 28, 1978. The elemental analysis pcrfortied at Midwest Research Institute was low for copper and nitrogen, slightly low for carbon, and slightly high for hydrogen and chlorine; values for all determined elements totaled 97.8 percent.
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:
unchanged (no vehicle)
Details on oral 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. For each dose level, one weekly lot of 4500 g was prepared.

The actual mixtures were composed of the following ingredients:
- Dose level 5.0 % (w/w): 225 g test material and water + 4275 g meal
- Dose level 2.5 % (w/w): 112.5 g test material and water + 4387.5 g meal
- Dose level 1.25 % (w/w): 56.25 g test material and water + 4443.75 g meal
- Dose level 0.6 % (w/w): 27 g test material and water + 44735 g meal
- Dose level 0.3 % (w/w): 13.5 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.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
One analysis was performed to determine the accuracy of mixture concentrations; results were within +- 10 % of the desired dose concentration.
Duration of treatment / exposure:
91 days
Frequency of treatment:
daily
Dose / conc.:
0.3 other: % in the diet
Dose / conc.:
0.6 other: % in the idiet
Dose / conc.:
1.25 other: % in the diet
Dose / conc.:
2.5 other: % in the diet
Dose / conc.:
5 other: % in the diet
No. of animals per sex per dose:
10 males and 10 females per dose
Control animals:
yes, plain diet
Details on study design:
- Five dose levels of 0.3, 0.6, 1.25, 2.5 and 5.0 % in feed were used in this study (approx. 300, 600, 1200, 2300 and 4600 mg/kg bw for males [based on 15.4 g/d average food consumption, 0.167 kg average bw] and approx. 300, 600, 1250, 2500 and 5000 mg/kg bw for females [based on 12.3 g/d average food consumption, 0.123 kg average bw], respectively).
- All dose levels were prepared on a weight per weight basis. There were five dose level groups and one group of controls per sex, with ten individuals of each sex in each dosage and control group. Each dosed group received dosed feed mixture on 91 consecutive days.
Positive control:
not required
Observations and examinations performed and frequency:
Animals were observed twice each day for clinical signs, with at least six hours between observations. All clinical signs were recorded daily. Additional studies included blood sampling for the animal disease screening program from ten control rats, five males and five females.
Sacrifice and pathology:
- Rats were necropsied on day 92 and 93.
- Gross examination were performed on all animals from all dosage groups.
- Microscopic examinations were performed on all tissues from all animals in the control group and the highest dose treatment group: Kidney, liver, lung, heart, testis, epididymis, prostate, stomach, thyroid, skin, cecum, pancreas and spleen.
Other examinations:
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. See details and results in endpoint "7.1.1. Basic toxicokinetics" in endpoint study record "Batelle 76-34-106002, mouse".
Clinical signs:
no effects observed
Description (incidence and severity):
No treatment related abnormal clinical signs were observed.
Mortality:
no mortality observed
Description (incidence):
No substance related mortality was reported.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
There were dose-related trends in body weights among both male and female rats, when body weights are expressed in percent differentials of weight gains compared with untreated controls. Male rats at the highest doses of 5.0 % and 2.5 % had differential weight gains of -11 % and -5 % respectively. Female rats at the top doses of 5.0 and 2.5 % had differential weight gains of -9.0 %. These changes in dosed group body weights are not severe and may not be a meaningful sign of toxicity.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There were slight trends in patterns of diet consumption between dosed and control rats of both sexes, with dosage groups eating about 1 g more of the higher concentrations of the dosed feed. This may be an adjustment in diet intake to offset the relatively high portion of non-caloric dyestuff present in the diet. Daily average diet consumption for female rats ranged from 11.5 g (controls) to as much as 12.7 g (1.25 %); for male rats, the range was from 14.6 g (controls) to 16.6 g (2.5 %).
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
No substance related changes were reported on macroscopic and microscopic examination of the animals. Gross examination were performed on all animals from all dosage groups.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No substance related changes were reported on macroscopic and microscopic examination of the animals. The microscopical lesions found were encountered in both the control and experimental high dose group with similar frequency and severity.
Details on results:
The authors recommended an dosage level of 5 % and 2.5 % test substance to be used in the chronic study, due to lack of compound-related lesions.
Dose descriptor:
NOAEL
Effect level:
4 600 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: no effects observed
Remarks on result:
other: calculated from dietary concentrations
Dose descriptor:
NOAEL
Effect level:
5 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: absence of effects
Remarks on result:
other: calculated from dietary concentrations
Critical effects observed:
no
Conclusions:
Due to lack of compound-related lesions, a NOAEL of ca. 4600 mg/kg bw per day was determined for male rats and a NOAEL of ca. 5000 mg/kg bw was determined for female rats under the test conditions chosen.
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
; no clinical chemistry, hematology, or urinalysis were conducted and no organ weights were taken (as recommended in OECD Guideline 408). Extra investigations on copper contents in liver and kidney were included.
GLP compliance:
no
Limit test:
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.
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 oral 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.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
One analysis was perfomed to determine the accuracy of the mixture concentration. Results were within +- 10 % of the desired dose concentration.
Duration of treatment / exposure:
91 days
Frequency of treatment:
daily
Dose / conc.:
0.3 other: % in the diet
Dose / conc.:
0.6 other: % in the diet
Dose / conc.:
1.25 other: % in the diet
Dose / conc.:
2.5 other: % in the diet
Dose / conc.:
5 other: % in the diet
No. of animals per sex per dose:
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, 2.5 and 5.0 % in feed were used in this study (approx. 0, 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. 0, 1100, 2200, 4700, 9400 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.
Observations and examinations performed and frequency:
Animals were observed twice each day for clinical signs, with at least 6 hours between observations. All clinical signs (or negative observations) were recorded daily. Additionally, blood sampling was conducted from 10 control mice, 6 treated males and 4 treated females.
Sacrifice and pathology:
- Mice were necropsied on day 92 and 93.
- Gross examination were performed on all animals from all dosage groups.
- Microscopic examinations were performed on following organs from all animals in the control group and the highest dose treatment group: Kidney, liver, lung, (only in control group: heart).
Other examinations:
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. See details and results in endpoint "7.1.1. Basic toxicokinetics" in endpoint study record "Batelle 76-34-106002, mouse".
Clinical signs:
no effects observed
Description (incidence and severity):
During the course of the 91-day study, neither treatment related signs of toxicity, nor abnormal clinical signs were observed. Cannibalism was observed within one cage group of control females and among four different dose levels in male mice, but was not considered to be drug-related or significant.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
There were seven early deaths during this study. Four male mice from four different dose or control groups died during weeks 3, 7, and 10, while 3 control female mice died during the week 3.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The body weight data ranged from -7 to +11 % differential weight gain in the dosed females, without any dose-relation. Dosed male mouse groups showed a positive weight differential in all groups except the lowest dosage group of 0.3 %. No trends coinciding with dosage levels or food consumption are evident in these data.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There were no trends in diet consumption among dosed animals compared with controls in either male or female mice. Female control mice consumed 2 to 3 grams more feed than dosed animals. However, diet consumption data in this group were based on seven survivors during the last ten weeks of the study and have a higher value than expected. The dosed female mouse groups had average and expected levels of diet consumption.
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
No substance related changes were reported on macroscopic and microscopic examination of the animals. At necropsy there were no consistent lesions noted.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No substance related changes were reported on macroscopic and microscopic examination of the animals. Microscopically there were no compound related lesions noted.
Details on results:
The authors recommended a dosage level of 5 % and 2.5 % test substance to be used in the chronic study, due to lack of compound-related lesions.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 16 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: no adverse findings
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 18 700 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: no adverse findings
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Critical effects observed:
no
Conclusions:
Due to lack of compound-related lesions, a NOAEL of ca.16000 mg/kg bw per day was determined for male mice and a NOAEL of ca.18700 mg/kg bw per day was determined for female mice under the test conditions chosen.
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
; no clinical chemistry, hematology, or urinalysis were conducted and no organ weights were taken. Extra investigation on copper content in liver and kidney included.
GLP compliance:
no
Limit test:
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.
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: 72 - 94 g; females: 70 - 90 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 oral 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.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
One analysis was performed to determine the accuracy of mixture concentrations; results were within +- 10 % of the desired dose concentration.
Duration of treatment / exposure:
91 days
Frequency of treatment:
daily
Dose / conc.:
0.3 other: % in the diet
Dose / conc.:
0.6 other: % in the diet
Dose / conc.:
1.25 other: % in the diet
Dose / conc.:
2.5 other: % in the diet
Dose / conc.:
5 other: % in the diet
No. of animals per sex per dose:
10 males and 10 females per dose
Control animals:
yes, plain diet
Details on study design:
- Five dose levels of 0.0, 0.3, 0.6, 1.25, 2.5 and 5.0 % in feed were used in this study (approx. 0, 250, 500, 1100, 2200 and 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).
- 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 91 consecutive days of dosed feed mixture.
Observations and examinations performed and frequency:
Animals were observed twice each day for clinical signs, with at least six hours between observations. All clinical signs were recorded daily. Additional studies included blood sampling for the animal disease screening program from 10 control rats, 5 males and 5 females.
Sacrifice and pathology:
- Rats were necropsied on day 92 and 93.
- Gross examination were performed on all animals from all dosage groups.
- Microscopic examinations were performed on all tissues from all animals in the control group and the highest dose treatment group: Kidney, liver, lung, heart, pancreas and pituitary.
Other examinations:
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. See details and results in endpoint "7.1.1. Basic toxicokinetics" in endpoint study record "Batelle 76-34-106002, rat".
Clinical signs:
no effects observed
Description (incidence and severity):
During the course of the 91-day study, neither treatment related signs of toxicity, nor abnormal clinical signs were observed.
Mortality:
no mortality observed
Description (incidence):
No substance related mortality was reported.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The differential weight gains in dosed rats were no greater than +/- 10 % except in the highest dose level females which had a +/- 12 % differential gain. However, no trend was established through the groups in either males or females. The greatest range between any two female groups was 12 % to -5 % between the two highest doses (5% and 2.5%). The greatest range between any two male groups was 9 % to -9 % between the highest dose and the third highest dose (5 and 1.25 %).
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Diet consumption among both males and females did not vary widely, nor showed any dose related trends. The average daily consumption of male rats showed a difference not greater than 2 g between any 2 groups. In female rats the difference was not greater than 1.6 g.
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
No substance related changes were reported on macroscopic and microscopic examination of the animals.
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
No substance related changes were reported on macroscopic and microscopic examination of the animals. The microscopical lesions found were encountered in both the control and experimental high dose group with similar frequency and severity. The high incidence of pulmonary lesions in both groups (control and experimental high group) was, according to the authors, thought to be associated with Sendai virus infection. No histopathologic lesions considered to be compound related were encountered in the tissues examined.
Details on results:
The authors recommended an dosage level of 5 % and 2.5 % test substance to be used in the chronic study, due to lack of compound-related lesions.
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 4 500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse findings
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
No accumulation of copper in liver or kidney
Critical effects observed:
no
Conclusions:
Due to lack of compound-related lesions, a NOAEL of ca. 4500 mg/kg bw per day was determined for male and female rats under the test conditions chosen.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated dose oral toxicity has been studied in two subchronic feeding studies each in mice and rats with Pigment Blue 15 and Pigment Green 7. Subacute GLP-compliant gavage studies were performed with Pigment Blue 15, Pigment Green 7, CAS 14832-14-5 and CAS 68987-63-3. All studies consistently show absence of adverse effects at the limit dose.

Further information on read-across is given in the attachment of the endpoint study record.

CAS No. 147-14-8 (Pigment Blue 15):

A GLP-compliant 28 day subacute study was conducted, according to the Japanese guideline for 28 Day Repeated Dose Toxicity Test of Chemicals with Wistar rats, which received 0, 40, 200 and 1000 mg/kg/day copper phthalocyanine by gavage (JETOC 1995). The test material is described to be of technical grade. Food consumption and body weight were determined. The animal’s state of health was checked by cage side observations as well as by detailed clinical observations. Clinicochemical and hematological examinations were carried out at the end of the administration period and after a recovery period of 14 days. All animals were subjected to gross-pathological assessment, followed by histopathological examination. No changes in general condition, body weight gain or food consumption were detected in any of the groups. After the 28 days of administration, a slight, but significant decrease in red blood cell count (RBC) and decrease of hemoglobin (Hb) and packed cell volume (PCV) were detected in the 200 and 1000 mg/kg male groups. These slight changes were dose dependent, but within the normal variation range. After the recovery period, a significant increase of erythroblasts was detected in the 1000 mg/kg female group. Also after the recovery period, slight increases of absolute organ weights of lung, spleen, adrenal and salivary gland and a tendency for increased relative organ weights of the spleen were evident in the 1000 mg/kg male group in comparison to the control group. These differences were of statistical, but not of biological relevance. No histopathological changes due to administration of phthalocyanine blue were detected. Therefore, a NOAEL of 1000 mg/kg bw per day was determined for male and female rats under the test conditions chosen.

 

In two 90 day subchronic feeding studies with F344 rats and B6C3F1 mice, male and female animals received dose levels of 5.0, 2.5, 1.25, 0.6 and 0.3 % (w/w) copper phthalocyanine, administered in the food (approx. 0, 250, 500, 1100, 2200 and 4500 mg/kg bw for rats of both sexes, approx. 0, 1000, 2000, 4000, 8000 and 16000 mg/kg bw for male mice, and approx. 0, 1100, 2200, 4700, 9400 and 18700 mg/kg bw for female mice, calculated on average food consumption for 91 consecutive days (Batelle 1979). Animals were observed twice each day for clinical signs, with at least 6 hours between observations. All clinical signs were recorded daily. Gross examinations were performed on all animals from all dosage groups. Microscopic examinations were performed from kidney, liver, lung, heart, pancreas and pituitary from all animals in the control group and the highest dose treatment group. No clinical chemistry, hematology, or urinalysis were conducted and no organ weights were taken. No substance related mortality was reported for rats. There were seven early deaths for mice. Four male mice from four different dose or control groups and 3 control female mice died during the study. During the course of the 91-day study, neither treatment related signs of toxicity, nor abnormal clinical signs were observed in both rats and mice. Diet consumption among both male and female rats did not vary widely, nor showed any dose related trends. In mice, there were also no trends in diet consumption among dosed animals compared with controls in either male or female mice. No substance related changes were reported on macroscopic and histopathological examination of both rats and mice.

This study was originally performed to serve as a range-finder for a chronic feeding study. It included determination of copper levels in kidneys and livers to find out if there was indication of uptake after ingestions. Since there was no significant increase in copper concentration, Pigment Blue 16 was considered to be inert and not absorbed, and the chronic toxicity study was not performed.

 

CAS No. 1328-53-6 (Pigment Green 7):

Valid experimental data were available to assess the toxicity of the tested material after repeated administration.

oral

A GLP-compliant 28 day subacute study was conducted, according to the Japanese guideline for 28 Day Repeated Dose Toxicity Test of Chemicals with Wistar rats, which received 0, 100, 300 and 1000 mg/kg/day polychloro copper phthalocyanine by gavage using corn oil as vehicle (MHLW 2000). Food consumption and body weight were determined. The animal’s state of health was checked by cage side observations as well as by detailed clinical observations. Clinicochemical and hematological examinations as well as urinalysis were carried out at the end of the administration period and after a recovery period of 14 days. No deaths occurred in either sex. No effects of polychloro copper phthalocyanine were detected in terms of general condition, body weights, food consumption, hematological or blood chemistry parameters, urinalysis, organ weights, or pathological examination in males or females.

 

In two 90 day subchronic studies with F344 rats and B6C3F1 mice, male and female animals received dose levels of 5.0, 2.5, 1.25, 0.6 and 0.3 % (w/w) copper phthalocyanine, administered in the food (approx. 300, 600, 1200, 2300 and 4600 mg/kg bw for male rats, approx. 300, 600, 1250, 2500 and 5000 mg/kg bw for female rats as well as approx. 0, 1000, 2000, 4000, 8000 or 16000 mg/kg bw/day for male mice and approx. 0, 1200, 2500, 5000, 10000 and 20000 mg/kg bw/day for female mice, calculated on average food consumption, for 91 consecutive days (Batelle 1979).Animals were observed twice each day for clinical signs, with at least 6 hours between observations. All clinical signs were recorded daily. Gross examinations were performed on all animals from all dosage groups. Microscopic examinations were performed from kidney, liver, lung, heart, pancreas and pituitary from all animals in the control group and the highest dose treatment group. No clinical chemistry, hematology, or urinalysis were conducted and no organ weights were taken.

No substance related mortality was reported in rats, in mice 4 deaths occurred during the study which were not considered to be test substance related. No treatment related abnormal clinical signs were observed in either rats or mice. In mice, there were no trends in diet consumption among dosed animals compared with controls in either male or female mice. In rats there were slight trends in patterns of diet consumption between dosed and control rats of both sexes, with dosage groups eating about 1 g more of the higher concentrations of the dosed feed. In rats there were dose-related trends in body weights among both male and female rats, when body weights are expressed in percent differentials of weight gains compared with untreated controls. Male rats at the highest doses of 5.0 % and 2.5 % had differential weight gains of -11 % and -5 % respectively. Female rats at the top doses of 5.0 and 2.5 % had differential weight gains of -9.0 %. In mice, a -9 % to +8 % differential weight gain was seen in the dosed females. Animals in the highest dose levels had less weight gain than controls. Dosed male mouse groups showed a positive weight differential in all groups except the 0.6 % dosage group. In this group during week 12 with a corresponding weight loss was observed, which was not fully recovered during the final week of the study. However, given the normal variability in rodent body weight measurements, this differences did not necessarily reflect manifestation of toxicity.

No substance related changes were reported on macroscopic and microscopic examination of the animals.

 

CAS No. 14832 -14 -5

A GLP-compliant 28 day subacute study was conducted, also according to the Japanese guideline for 28 Day Repeated Dose Toxicity Test of Chemicals with Wistar rats, which received 0, 20, 140 and 1000 mg/kg/day copper perchloro phthalocyanine by gavage using olive oil as vehicle (JETOC 1997). The animals were observed for clinical signs, body weights and food consumption; hematological and blood chemical examinations, urinalysis, and padiological examination were conducted. No deaths occurred in either sex. No effects of copper perchloropolychloro phthalocyanine were detected in terms of general condition, body weights, food consumption, hematological or blood chemistry parameters, urinalysis, organ weights, or pathological examination in males or females.

 

CAS No. 68987-63-3

A GLP-compliant 28 day subacute study was conducted following OECD guideline 407 (Huntington Life Sciences 1997). Oral administration at dosages of 40, 200 and 1000 mg/kg/day in corn oil was generally well tolerated. No target organ was identified.

The blue staining seen in the intermediate and high dosage groups, and the abnormal contents of the gastro-intestinal tract in high dosage animals were considered to be due to the test material, since it was a blue colour, and was therefore of no toxicological significance. The NOAEL was determined to be 1000 mg/kg bw.

 

A few old literature publications mentioned in the OECD SIDS document were not included in the REACH registration dossier since they contained little information and also in some cases it was found that the test items were indeed not pigments but soluble dyes.

 

Repeated dose inhalation toxicity

Repeated dose inhalation toxicity was not performed as the substances are inert poorly soluble particles when manufactured as powder. They are non volatile and not inhalable if imbedded in a matrix such as a coating, a plastic article or a suspension.

A 5-day inhalation study with three-week recovery was performed with Pigment Blue 15 (BASF 2016). This study examined both systemic and local effects. The NOAEC was determined to be the highest tested dose of 30 mg/m3.

 

Repeated dose dermal toxicity

No studies on repeated dose dermal toxicity are available. The molecular weights of the copper phthalocyanine pigments is greater than 500 g/mol which in combination with poor solubility is an indication of low skin permeability. Considering that no hazard was identified at the limit dose for the oral route, there is no concern for a hazard via the dermal route. Testing for the dermal route is not needed.

Justification for classification or non-classification

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

The available experimental test data fpr the analogue substances are reliable and suitable for classification purposes under Regulation 1272/2008. There were no significant toxic effects at doses of less than 300 mg/kg bw upon subacute oral exposure in rats. As a result the substance is not classified for repeated dose toxicity under Regulation (EC) No. 1272/2008,as amended for the seventh time in Regulation (EC) No 2015/1221.