[1-[[(2-hydroxyphenyl)imino]methyl]-2-naphtholato(2-)-N,O,O']copper

Endpoint summary

Description of key information

No mortality upon single gavage doses of 5000 and 10000 mg/kg bw was observed in rats. Both studies were performed prior to the introduction of GLP and OECD guidelines, but were performed in a very similar design.  

Dust inhalation caused mortality in the LC₅₀range of 1- 4 mg/L. Two studies are available. A 4h-study performed around 1980 performed with a poorly characterized test material an LC₅₀of 1825 mg/m³. A GLP and OECD 403 guideline conform study with the nanomaterial performed in 2014 showed that rats survive a 4h exposure to 0.97 mg/L, but not to 4.1 mg/L. These findings are consistent with the LC50 calculated from the older study.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1978

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

no

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

Batch: EN 75024
physical state: solid

Species:

rat

Strain:

other: Tif:RAIf

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: raised on the premises of the testing facility
- Age at study initiation: 7 to 8 weeks
- Weight at study initiation: ca 180 g for males and ca 170g for females
- Fasting period before study: overnight
- Housing: groups of five
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 + 1°C
- Humidity (%): 55 + 5 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 14/10

IN-LIFE DATES: From: 1978-10-17 To: 1978-11-8

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10 or 20 ml/kg

DOSAGE PREPARATION (if unusual): Before treatment the suspension was homogeneously dispersed with an Ultra-Turrax and during treatment it was kept stable with a magnetic stirrer.

Doses:

3000, 4000 and 5000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Weighing daily, observations on hour 1,2,4, 6 and 24, then once daily until day 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs (Sedation, Dyspnoea, Dacryorrhoea, Chromodacryorrhoea, Rinorrhoea, Epistaxis, Exophthalmos,Salivation, Ruffled fur, Pallor, Cyanosis, Diarrhoea, Body position (ventral/lateral/curved), Ataxia, Trismus, Tremor, Tonic clonic muscle spasms Convulsion), body weight

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality occurred.

Mortality:

No mortality occurred.

Clinical signs:

other: Effects occurred at all dose levels in a dose dependent manner. The animals recovered within 8 days. Sedation: up to 24h Dyspnoea: up to up to 8 days Exophthalmos: up to 6 days Ruffled fur: up to 8 days Curved body position: up to 8 days

Gross pathology:

No substance related gross organ changes were seen.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

5 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

yes

Remarks:

Two extra animals were exposed under comparable exposure conditions with a concentration of 4.129 mg/L and sacrificed on the day of exposure. Histopathology of the respiratory tract was performed.

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories B.V., 5961 NM Horst, NIEDERLANDE
- Age at study initiation: male animals approx. 8 weeks, female animals approx. 10 weeks
- Weight at study initiation:
- Fasting period before study: no
- Housing: 5 per group, single cages for satellite animals
- 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 (%): 70%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12/+2

IN-LIFE DATES: From: 2013-10-23 To: 2013-12-03

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

other: air, with flowability-increasing agent AEROSIL® R 972 (1% based on test material)

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: For each test group the dusts were produced inside the inhalation system with the above mentioned dust generator and compressed air and passed into the inhalation system. The concentrations were adjusted by varying the apertural width rotation of the dosing wheel of the dust generator For each test group the dusts were produced inside the inhalation system with the above mentioned dust generator and compressed air and passed into the inhalation system. The concentrations were adjusted by varying the apertural width rotation of the dosing wheel of the dust generator
- Exposure chamber volume: 34L
- Method of holding animals in test chamber: restraining tubes
- Source and rate of air: central air conditioning system, 0.8 m³/h
- Method of conditioning air: Central air conditioning system provides cold air of about 15°C. This cold air passes through an activated charcoal filter, be adjusted to room temperature of 20 to 24°C and pass through a second particle filter (H13 (HEPA) Camfil Farr, Germany). The so generated conditioned air is used to generate inhalation atmospheres.
- System of generating particulates/aerosols: The test substance was stirred in its container before a sample for dust generation was taken.
The test substance was desagglomerated in a mixer (mixing for 10 seconds) under addition 1% AEROSIL® R 972 (flowability) before introduction into the dust generator via a dosing wheel dust generator (Gericke/BASF)
- Method of particle size determination: Stack Sampler Mark III (Andersen) . Before sampling, the impactor stages were assembled with preweighed glass fiber collecting discs, and equipped with a backup particle filter. The impactor was connected to the vacuum pump, and for each test group samples were taken from the breathing zone of the animals according to the following tables. Sampling occurred 30 minutes (or later) after the beginning of the exposure.
- Treatment of exhaust air: The exhaust air is filtered and conducted into the exhaust air of the building.


TEST ATMOSPHERE
- Brief description of analytical method used: gravimetric,filtration equipment with probe, internal diameter: 7 mm
- Samples taken from breathing zone: yes

VEHICLE
In technical trial runs, various modifications of the dust generation procedure were tested to produce the limit concentration of 5 mg/L. To improve the flowability, 1% AEROSIL® R 972 was added to the test substance. AEROSIL® is a brand name of various amorphous silicas produced by Evonik Industries, Frankfurt am Main, Germany. AEROSIL® R 972 is a hydrophobic fused silica, which is used to improve free flow and anti-caking characteristics of powders.


TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: see attached figure
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): see table M3

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: limit dose for GHS

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4

Concentrations:

0.971 and 4.710 mg/L (main groups)
0.971 and 4.13 mg/L (satellite groups)

No. of animals per sex per dose:

5 (main group)
1 (satellite group)

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days (main group), 1 day (satellite group)
- Frequency of observations and weighing:
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Details on sacrifice:
Main group: On the last day of the observation period, the surviving animals were sacrificed by CO2-inhalation in a chamber with increasing concentration over time, followed by necropsy and gross-pathological examination. Animals which die prematurely were necropsied and assessed by gross pathology.

Satellite group: The low dose animals assigned for light microscopic examination (Satellite groups SG) were sacrificed under pentobarbital anesthesia by exsanguination from the abdominal aorta and vena cava. The exsanguinated animals were necropsied and assessed by gross pathology. Animals of the high-dose satellite died and were necropsied and assessed by histopathology and gross pathology immediately after death.

Sex:

male/female

Dose descriptor:

LC100

Effect level:

4.13 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: All animals died on the day of exposure.

Sex:

male/female

Dose descriptor:

LC0

Effect level:

0.97 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

Main group (4.71 mg/L): All of the five male animals and all of the five females died during the study day 0.
Satellite group (4.13 mg/L): Both animals died during the study day 0.

Clinical signs:

other: At the lethal dose group, animals showed labored respiration from the first hour until death (hour 3 or 4). At the non-lethal dose, clinical symptoms were obsered as listed in table 1. Adverse symptoms resolved within 12 days in males and 7 days in fema

Body weight:

Test group 1 (0.971 mg/L)
The mean body weights of the animals decreased during the first post exposure observation day and increased from study day 3 onward.

Test group 2 (4.710 mg/L)
No body weight data present because all animals died.

Gross pathology:

Low dose group:
No gross pathology findings were observed at the low dose animals at scheduled sacrifice on day 14. The 2 satellite animals that were sacrificed on the day of exposure showed few foci in the lung, dark-red discolored in all lobes, partly sunken with a diameter of 5 mm.

High dose group:
See table R2

Other findings:

Histopathology of satellite animals, sacrificed on the day of exposure

Animal 1, high dose satellite group
Nasal cavity: Pigment deposition (yellow, clumped material), diffuse, severe
Larynx/Pharynx: Pigment deposition (yellow, clumped material), diffuse, moderate
Trachea: pigment deposition (yellow, clumped material), moderate
Lung: Pigment deposition (yellow, clumped material), bronchial, bronchiolar and alveolar, severe, with obstruction; acute diffuse emphysema

Animal 2, high dose satellite group
Nasal cavity: Pigment deposition (yellow, clumped material), diffuse, severe
Larynx/Pharynx: Pigment deposition (yellow, clumped material), diffuse, moderate
Trachea: Pigment deposition (yellow, clumped material), moderate
Lung: Pigment deposition (yellow, clumped material), bronchial, bronchiolar and alveolar, severe, with obstruction; acute diffuse emphysema

Animal 1: low dose satellite group
Nasal cavity: only level IV: Pigment deposition (yellow, clumped material), focal, slight
Larynx: only level II: Pigment deposition (yellow, clumped material), focal, slight
Pharynx/Trachea: without findings
Lung: Pigment deposition (yellow, clumped material), bronchial, bronchiolar and alveolar, multifocal, moderate; pigment-associated mild to moderate purulent inflammation with necrosis, multifocal

Animal 2, low dose satellite group
Nasal cavity: Pigment deposition (yellow, clumped material), multifocal, slight, with epithelial necrosis
Larynx: only level I + II: Pigment deposition (yellow, clumped material), multifocal, slight, with epithelial necrosis
Pharynx /Trachea: without findings
Lung: Pigment deposition (yellow, clumped material), bronchial, bronchiolar and alveolar, multifocal, moderate to severe; pigment-associated moderate to severe purulent inflammation with necrosis, multifocal

Table R1: Clinical symptoms at the non-lethal dose group (0.97 mg/L) (d=day, h = hour)

Test group 1 (0.971 mg/L)	Male animals	Female animals
Fur, substance contaminated	d0 - d5	d0 - d3
Fur, yellow discolored	d6 - d13	d3 - d12
Piloerection	d0 - d2	d0 - d2
Respiration, labored	h1 - h4	h1 - h4
Respiration, abdominal	d0 - d12	d0 - d7
Respiration, sounds	d4	-

Table R2: Necropsy findings of animals that died during the study

Findings	4.7 mg/L	4.13 mg/L (high dose satellite group)
Number of animals	5 males + 5 females	1 male + 1 female
Organs without particular findings	1 male + 2 females	-
Lung: few dark-red foci in all lobes, diameter 3 mm	3 males + 2 females	-
Lung: few red foci in all lobes, diameter 2 mm	1 male	-
Lung: few brown foci in lobus sinister diameter 1 mm	1 female	-
Lung: multifocal foci, partly sunken	-	1 female
Pharynx and trachea: greenish discoloration	-	1 male

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Exposure to a dust atmosphere of 4.71 mg/L is lethal to rats. No mortality was observed at the measured concentration of 0.97 mg/L.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

1 825 mg/m³ air

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The acute oral toxicity was investigated performed with rats prior to the introduction of OECD guidelines and GLP (Ciba 1978). Since the study design included the basic elements of the current OECD testing guidelines, the study is considered adequate for hazard assessment. The study was performed with dose levels of up to 5000 mg/kg bw.

The purity of the test item is not described, but the test item is identified via the trade name indicating that the pigment is contained at a concentration of >80%. Considering the high doses, purity of the test item is not crucial. The test material was not characterized on its content of particles of the nano-size-dange.

A single treatment with 5000 mg/kg bw did not cause mortality or gross findings at necropsy at the end of the 14 -day observation period.

To determine the acute inhalation toxicity (single 4-hour exposure, head-nose only) as a dust, a study was performed in male and female Wistar rats according to OECD-Guideline method 403, as well as EC and EPA guidelines (BASF 2014).

The following measured concentrations were tested: 0.971 and 4.710 mg/L. To examine the cause of death at the high concentration, a satellite group (one male and one female animal) were exposed under comparable exposure conditions with a concentration of 4.129 mg/L. Cascade impactor measurements resulted in particle size distributions with mass median aerodynamic diameters (MMADs) between 1.8 and 2.6 μm, which are well within the respirable range.

No lethality was observed in male and female animals at 0.971 mg/L. At 4.710 mg/L all of the male and female animals died during or after the exposure on study day 0. Animals of the satellite group died during exposure (hour 2 and 3) at a measured concentration of

4.129 mg/L. Clinical signs of toxicity in animals exposed to 0.971 mg/L comprised labored respiration, abdominal respiration, respiration sounds, piloerection, substance contaminated and yellow discolored fur. These findings were observed from hour 1 of exposure until study day 13. No clinical signs and findings were observed on study day 14. The mean body weights of the

animals decreased during the first post exposure observation day and increased from study day 3 onward. No gross pathological abnormalities were noted during the necropsy at the termination of the post exposure observation period of the main group animals.

To evaluate morphological changes, a satellite group (one male animal and one female animal) was included in main group 1. The satellite animals were sacrificed on the day after exposure (study day 1). The animals of the satellite group showed few dark-red discolored foci, partly sunken in all lung lobes (sacrificed scheduled one day after exposure). Histopathological examination of satellite animals of this group revealed the following microscopic changes:

Animal No. 821 (scheduled sacrifice on study 1):

Nasal cavity: only level IV: Pigment deposition (yellow, clumped material), focal, slight

Larynx: only level II: Pigment deposition (yellow, clumped material), focal, slight

Pharynx/Trachea: without findings

Lung: Pigment deposition (yellow, clumped material), bronchial, bronchiolar and alveolar, multifocal, moderate; pigment-associated mild to moderate purulent inflammation with necrosis, multifocal

Animal No. 822 (scheduled sacrifice on study 1):

Nasal cavity: Pigment deposition (yellow, clumped material), multifocal, slight, with epithelial necrosis

Larynx: only level I + II: Pigment deposition (yellow, clumped material), multifocal, slight, with epithelial necrosis

Pharynx /Trachea: without findings

Lung: Pigment deposition (yellow, clumped material), bronchial, bronchiolar and alveolar, multifocal, moderate to severe; pigment-associated moderate to severe purulent inflammation with necrosis, multifocal

Clinical signs of toxicity in animals exposed to 4.710 mg/L comprised labored respiration. Findings were observed from hour 1 of exposure until death of the animals. Lethality occurred during exposure from hour 2 onward. Only one animal was found dead after

exposure. Therefore, no body weight data could be collected. During gross necropsy of the dead animals, few foci (red, dark red or brown) were observed in the lung of 3 females and 4 males. In 1 male and 2 female dead animals no gross pathological abnormalities were noted during the necropsy.

To examine the cause of death, a second satellite group (one male and one female animal) were exposed under comparable exposure conditions to dust aerosol of the test substance at a measured concentration of 4.129 mg/L. A concurrent control group (1 male and 1 female) was exposed to conditioned air and was sacrificed after the death of the exposed animals. Histological examination of the respiratory tract of satellite animals reveals several, partly severe findings in exposed animals (as indicated below), while no abnormalities were observed in the concurrent controls.

Animals 781 and 782 (died during or shortly after exposure on study 0): Nasal cavity: only level IV: Pigment deposition (yellow, clumped material), diffuse, severe

Larynx: Pigment deposition (yellow, clumped material), diffuse, moderate

Pharynx/Trachea: Pigment deposition, moderate

Lung: Severe pigment deposition (yellow, clumped material), bronchial, bronchiolar and alveolar, with obstruction, acute diffuse emphysema

The pathological findings in the lung are indicative for a total obstruction of airways with subsequent death by asphyxiation.

The test material was characterized on its content of particles of the nano-size-dange.

Experimental data on acute dermal toxicity is not available.

Justification for classification or non-classification

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. No mortality was observed at doses exceeding 2000 mg/kg bw. As a result the substance is not considered to be classified for acute oral toxicity under Regulation (EC) No. 1272/2008, as amended for the thirteenth time in Regulation (EC) No. 2018/1480.

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.

The LC50 for dust was between 1 and 5 mg/L.

As a result the substance is considered to be classified for acute inhalation toxicity (Category 4) under Regulation (EC) No. 1272/2008, as amended for the thirteenth time in Regulation (EC) No. 2018/1480.

No experimental data is available for acute dermal toxicity. However, the substance is of poor solubility in water, not irritating and unlikely to penetrate the skin. Considering the absence of acute oral toxicity, no hazard label is assigned for acute dermal toxicity.