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

Acute Toxicity: inhalation

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

239-763-1

EC Name:

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

Cas Number:

15680-42-9

Molecular formula:

C17H11CuNO2

IUPAC Name:

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

Test material form:

solid: nanoform

Details on test material:

- Substance type: pigment
- Physical state: Solid / greenish yellow
- Name of test material (as cited in study report): Irgazin Yellow L 0800
- Lot/batch No.: 0005629903
- Analytical purity: 99.0 %
- Purity test date: 2013
- Expiration date of the lot/batch: unlimited stability under storage conditions
- Stability under test conditions: stable
- Storage condition of test material: room temperature
- Date of manufacture: 01 Dec 2011

Test animals

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

Administration / exposure

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.

Results and discussion

Effect levels

Effect levels 1

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.

Effect levels 2

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

Any other information on results incl. tables

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

Applicant's summary and conclusion

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.