29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32 copper

Endpoint summary

Description of key information

Oral: LD50 (rat) > 6400 mg/kg bw (pre-GLP study similar to OECD TG 401)

Dermal: LD50 (rat) > 5000 mg/kg bw (non-GLP study accroding to OECD TG 402)

Inhalation: LC50 (rat) > 2 mg/L bw (read-across, GLP study accroding to OECD TG 403)

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Meets generally accepted scientific standards, well documented study with the following restriction: In some cases in the highest dose group the application volume per animal slightly exceeded the volume of 2 ml/100 g bw, which is recommended in OECD guideline 401 as maximum application volume for aqueous solutions.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Principles of method if other than guideline:

BASF-Test:
In principle, the methods described in the OECD Guideline 401 were used. Young adult laboratory rats were purchased from breeder. Several groups of 5 rats per sex and dose were treated simultaneously by gavage with preparations of the test substance in suitable vehicle. The concentrations of these preparations were used to achieve comparable volumes per kg body weight. Group-wise documentation of clinical signs was performed over the 14 day study period. The LD50 value was estimated on the basis of the observed mortalities.

GLP compliance:

no

Remarks:

GLP was not compulsory at the time the study was conducted

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

- Analytical purity: 100 %

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Firma Gassner, Ottobrunn; SPF breeding
- Average weight at study initiation: males: ca. 170 g; females: ca. 145 g
- Diet: Altromin-R (Altrogge, Lage/L., Germany), ad libitum
- Water: ad libitum

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

Concentration of the test material in vehicle:
- 30 % (6400 and 3200 mg/kg bw)
- 16 % (1600 mg/kg bw),
- 2 % (200 mg/kg bw)

Amount of test material applied per gavage:
- 21.3 ml/kg bw for 6400 mg/kg bw;
- 10.6 ml/kg bw for 3200 mg/kg bw;
- 10.0 ml/kg bw for 1600 mg/kg bw;
- 10.0 ml/kg bw for 200 mg/kg bw

Doses:

200, 1600, 3200, 6400 mg/kg bw

No. of animals per sex per dose:

5 animals per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: Animals were observed and examined for clinical signs of toxicity during the first hour following application, after 4 hours and further on day 1, 4, 5, 6, 7, 8, 11, 12, and 13 after dosing.
- The body weights of the individual animals were gathered prior to application of the test material.
- Necropsy of survivors performed: yes. Deceased animals and those sacrificed at the end of the observation period (on day 14 after dosing) were necropsied.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 6 400 mg/kg bw

Mortality:

No mortality was observed at any dose group.

Clinical signs:

other: - 6400 - 3200 mg/kg bw: Ca. 4 hours after application the animals were found in a crouched down position with intermittent breath. On the following morning slight apathy,a crouched down position and intermittent breath were observed. The stool of the anim

Gross pathology:

Autopsy revealed no relevant findings.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the BASF test (similar to OECD guideline 401) for acute toxicity after oral application, the LD50 for the test material is > 6400 mg/kg body weight for male and female rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

6 400 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

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

Remarks:

non-GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

- Analytical purity: 100 %

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding Farm VELAZ
- Age at study initiation: 7-9 weeks
- Weight at study initiation: 236-278 g
- Housing: individually in plastic breeding cage Velaz T4
- Diet: DOS 2b - standard pelleted diet, ad libitum
- Water: drinking tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature: 22 +/- 3 °C
- Humidity: 30-70 %
- Photoperiod: 12 hrs dark / 12 hrs light

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

The amount of the test substance for each animal was weighed (acc. to its body weight) and moistened with water for injection immediately before application.
Approx. 24 h before application of the test substance, the fur on the back of the animals was shaved at area about 6 cm x 6 cm.
Aliquot part of the test material was applied on the depilated area of skin. The application site was covered by mull, plastic foil and held in contact by plaster (strapping). After 24 h the occlusive dressing and the remains of the test substance were removed.

Duration of exposure:

24 h

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

5 males

Control animals:

no

Details on study design:

Body weight:
Animals were weighed before application, at 8th day and before euthanasia of animals. Average body weight in a group was calculated from individual body weights.

Clinical examination:
After application of the test material the animals were observed individually. Observations included changes of health conditions. Observations were performed daily during the 14 days observation period.

Pathological examination:
All test animal survivors to the end of the study were sacrificed on the 15th day and gross necropsy was carried out. Normally the nutritious state, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated.

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Mortality:

All animals survived the 14 day observation period and the day of sacrifice.

Clinical signs:

other: No clinical signs of toxicity were observed during the study period.

Gross pathology:

No individual records of macroscopic changes were found.

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

Additional information

Oral:

There are valid data available for the assessment of the acute oral toxicity of test substance. Five male and five female Sprague-Dawley rats were treated with 200, 1600, 3200, 6400 mg/kg bw under standardized conditions; the test method is comparable to OECD guideline 401. The animals were observed for 14 d, necropsy was performed even with the survivors. The LD50 was > 6400 mg/kg bw for male and female rats. No mortality was observed at any dose group and autopsy revealed no relevant findings. Clinical signs were reported for all groups. In the 6400 - 3200 mg/kg bw groups, ca. 4 hours after application the animals were found in a crouched down position with intermittent breath. On the following morning slight apathy, a crouched down position and intermittent breath were observed. The stool of the animals was bluish discoloured until day 4. On the following days the animals were without any findings. In the 1600 - 200 mg/kg bw groups, immediately after application, irregular breath and masticatory movement were observed, after 4 hours the animals were found in a crouched down position. On the following days until day 3, the stool of the animals was bluish discoloured but later on the animals were without any findings (BASF AG, 1971).

In another supporting study, conducted with five male and five female mice per group, the animals were treated with 0 or 16000 mg/kg bw under standardized conditions and were observed for 14 d, necropsy was performed with all animals. The LD50 was > 16000 mg/kg bw for male and female mice. No mortality was observed and autopsy revealed no relevant findings. The following clinical signs were reported shortly after dosing in all treated mice: pilo-erection and abnormal body carriage (hunched posture) and abnormal gait (waddling). These findings were accompanied by lethargy (4 animals), ptosis (1 animal), diarrhoea, coloured blue (1 animal). Recovery of all treated mice as judged by external appearance and behaviour, was apparently complete within 4 days of dosing (Huntingdon 1981, Val. 2).

Other studies with partially limited reliability provided a LD50 range of > 10000 to > 15000 mg/kg bw in rats (Kurlandsky 1984, Val. 4; Webb 1984 Val. 4), the LD 50 value for another common test species (mouse) was > 10000 mg/kg bw (Gosselin 1976, Val. 4).

Dermal

There are valid data available for the assessment of the acute dermal toxicity of Copper phthalocyanine. Five male Wistar rats were treated with 5000 mg/kg bw under standardized conditions; the test method was acc. to OECD guideline 401 (limit test). The animals were observed for 14 d, necropsy was performed even with the survivors. The LD50 was > 5000 mg/kg bw for male rats. No mortality was observed, autopsy revealed no relevant findings and no clinical signs of toxicity were reported. Body weight gain was within the normal range (Synthesia 2009, Val. 2).

Inhalation

In a procedure following OECD guideline 403 and GLP, five male and five female albino rats per group were exposed to the test substance by nose-only inhalation for four hours at gravimetrically determined mean concentrations of 1.1 and 5.2 mg/L air. All animals were observed for clinical signs and mortality during the inhalation exposure and the subsequent 14-day observation period or until they were found dead. Body weights were recorded prior to exposure on test day 0 (acclimatization period), and during the observation period on test days 1, 3 and 7 before necropsy and weekly thereafter, or before the sacrifice of the animals at the end of the observation period. At the end of the observation period, all animals were sacrificed and necropsied. Animals which died were necropsied as soon as they were found dead. All animals exposed to 5.2 mg/l air died within two days after exposure start. All animals exposed to 1.1 mg/L air survived the scheduled observation period. Clinical signs of toxicity in animals exposed to 1.1 mg/L comprised accelerated respiration, intermittent respiration, abdominal respiration, respiration sounds, feces substance like discolored, piloerection, substance-discolored fur and substance-contaminated fur. Findings were observed in the males from hour 1 of exposure through study day 11. No findings were detected in the male animals during the postexposure observation period from study day 12 onwards. Findings were observed in the females from hour 1 of exposure until the end of the post-exposure observation period. No gross pathological findings were noted during the necropsy of the animals at the termination of the post-exposure observation period. Clinical signs of toxicity in animals exposed to 5.2 mg/L comprised accelerated respiration, depressed respiration, abdominal respiration, no feces, activity: attention reduced, piloerection, substancediscolored fur and substance-contaminated fur. Findings were observed from hour 1 of exposure until the death of the animals. The mean body weights of the surviving animals decreased during the first post-exposure observation days. During necropsy of the dead five male and five female animals, many black foci were seen in the lung lobes with sunken surface. Blue discoloration of content of the stomach was seen in four males and three females and blue depositions in the trachea were present in four males and two females. For further evaluation, histopathological examinations of the respiratory tract (nasal cavity, larynx, pharynx, trachea, and lung) from three animals were performed. The lung showed large amount of blue pigment within the bronchi, bronchioles and terminal bronchioles, leading to obstruction of the airways in two animals with one of them also showing emphysema. The trachea of all examined animals was dilated and contained blue pigment. The larynx at level I - II showed obstruction by blue pigment and large amounts of blue pigment at level III in 2 animals. The third animal presented with large amounts of blue pigment at larynx level I-II and small amounts at the third level. The nasal cavity at level I - IV contained small to moderate amounts of blue pigment. The histopathological findings in the lung, the trachea, and the larynx of animal Nos. 793, 798 and 800 indicate an airway obstruction caused by the inhaled blue pigment as cause of death.

MPPD modulation (rat vs. human) The Multiple Path Particle Dosimetry (MPPD) model was used to predict aerosol deposition patterns representative of a guideline limit test acute inhalation toxicity study in the Sprague Dawley rat. Specifically, a 4-hr nose-only inhalation exposure to generic aerosol particles having a mass median aerosol diameter (MMAD) of 2.75 µm and a geometric standard deviation (GSD) of 1.0 (i.e., monodisperse distribution), a unit density of 1 g/cm3, and an airborne concentration of 5000 mg/m3 (5 mg/L) was simulated. Human simulations for a variety of hypothetical activity and breathing conditions for the same exposure were also conducted to highlight cross-species differences, although it is unlikely that such high aerosol exposures would be considered tolerable, even for non-toxic materials. Rats are obligate nose breathers and with over 30% of inhalable aerosol depositing in the head region, anterior nasal tissues and laryngeal regions are likely to receive a significant amount of deposition based upon a computational fluid-particle dynamics (CFPD)-informed analysis of MPPD simulations. In general, greater deposition of aerosols occurs in the head region of humans following nasal than with oral breathing. However, oral breathing and increased activity (increased bpm and minute volume) can lead to deeper penetration of aerosols within the pulmonary region. Given the smaller airways and higher breathing frequencies and minute volumes per unit body weight in rats vs. humans, greater amounts of aerosol mass are deposited in the tracheobronchial region in rats, while in humans, a greater mass is deposited in the pulmonary region. However, when normalized to airway surface areas, both species receive a relatively low overall pulmonary regional dose. Mucociliary clearance rates are also slower in humans than in the rat and the time to clear tracheobronchial airways varies between ~48 and 72 hr depending upon breathing mode and activity level vs. ~12 hr in the rat. Both species have slow macrophage clearance rates in the pulmonary region with neither species clearing much material (<87%-97% of deposited mass) by 7 days.

However, given the smaller airways in the rat and the potential for significant amounts of deposited and retained mass of aerosols in individual airways, the potential for airway obstruction and resulting decreases in airflows and pulmonary function was assessed in individual airways using a geometric analysis of MPPD simulations. This analysis showed that a significant percentage of pulmonary airways and to a lesser extent, bronchiolar airways, may be at risk of occlusion and impairment of lung function and gas exchange in the rat. Such a potential for physical impact on lung function for certain materials that are presumed to be of low toxicity raises questions for assessing acute toxicity risks for humans based upon guideline acute toxicity studies in the rat at limit test concentrations.

Justification for classification or non-classification

WOE justification for classification Current OECD guidelines for acute inhalation toxicity assessments incorporate a limit test aerosol concentration for poorly soluble particles that are either known or expected to be virtually non-toxic (OECD, 2009). Depending upon regulatory requirements, the limit test concentration for aerosols can be as high as 5 mg/L (or the maximum attainable concentration) if the United Nations (UN) Globally Harmonized System of Classification and Labelling of Chemicals (GHS) is used. During the acute dust exposure study (GLP, OECD 403), rats exposed to 5 mg/L of the test substance died of suffocation after agglutinated pigment blocked the airways, whereas all rats survived the 4h dust exposure at the low dose of 1 mg/L. It is widely recognized that external exposure concentrations of airborne particulates are not directly equivalent to the amounts that are inhaled, deposited, and retained in airways of laboratory animals and humans due to major differences in airway anatomy and physiology. In line with this, with regard to dose adjustment, the ECHA R.8 document on “Characterisation of dose[concentration]-response for human health” (Version 2.1, 2012) suggests to account for differences in respiratory rates of the experimental animal (at rest) and the human as well as for the increased respiratory rate of a worker, when inhalation DNELs are derived. Without factoring in these species differences, results from inhalation toxicity studies are not directly translatable to human health risk. Accordingly, the results from the acute inhalation study on rats alone are regarded as insufficient to derive a potential human health hazard posed by the test substance. Therefore, additional computational approaches that incorporate species-specific anatomy, physiology, and the physics of aerosol transport and deposition were utilized to assess whether the results from the acute inhalation study is relevant for humans. Focusing upon acute toxicity testing guidelines, the MPPD model was applied to provide a generic (not chemical-specific) respiratory dosimetry assessment of rats exposed in nose-only inhalation chambers for 4-hr to aerosols with a respirable particle size of 2.75 µm mass median aerosol diameter (MMAD) and a geometric standard deviation (GSD) of 1.0 at a limit test concentration of 5 mg/L – similar to the settings of the in vivo study and analog to the particle properties of the test item used. Particular emphasis was placed upon particle load and retention in individual airways to assess potential vulnerabilities for physical obstruction of bronchial and pulmonary airways in the rat due to high rates of local aerosol deposition. In addition, human simulations for a variety of hypothetical activity and breathing conditions for the same exposure were also conducted to highlight cross-species differences, although it is unlikely that such high aerosol exposures would be considered tolerable, even for nontoxic materials. As a result, the MPPD simulations indicate that the results from the high dose group of the acute dust exposure study (GLP, OECD 403) in rats are not relevant for resting or light exercising humans, since deviating internal exposure patterns of rat and humans were observed. The major difference between both species is that 50% of the material is deposited in the head-region in humans (only 30% in rats) and can be blown or spit out. Moreover, at tracheobronchial and bronchial level, the rat receives ~5-10-fold higher dose/cm2 airway surface (when cumulative deposited mass is normalized to respective airway surface areas) than the human under resting nasal breathing ventilation conditions. The pulmonary region of humans receives a greater fraction of inhaled aerosol than the rat, though when normalized to airway surface areas, both species receive a relatively low overall pulmonary regional dose.

As to the applicability of limit test concentrations for aerosols with presumed low toxicity in guideline acute inhalation toxicity studies with rats, the potential for airway obstruction which could lead to decreases in airflows and pulmonary function was assessed using a geometric analysis of MPPD simulations. Given the smaller airways in the rat and the potential for significant amounts of deposited and retained mass of aerosols during a 4-hr nose-only exposure to aerosols at 5 mg/L atmospheric concentrations, the size (diameter) of retained masses in each airway segment were compared with their corresponding airway diameter. In this analysis, which assumed that the total retained mass in each airway at the end of the 4-hr exposure formed one spherical particle, a significant a percentage of pulmonary airways and to a lesser extent bronchiolar airways may be at risk of occlusion and impairment of gas-exchange. Such a potential for physical impact on pulmonary function under limit test exposure conditions for certain materials that are presumed to be of low toxicity raises questions for assessing acute toxicity risks for humans. Finally, in comparison to the rat, more material is deposited in the head regions in human. Second, the wider airways in human hamper fast agglutination of the test material and therefore obstruction of the airways. At last, a 4h-exposure of worker to such high concentrations is rather unlikely even in case of an accident. The suffocation at 5 mg/L as observed in rats is therefore not regarded as relevant for humans and a classification as acute cat. 4 H332 not justified.

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for acute toxicity (oral, dermal, inhalation) under Regulation (EC) No. 1272/2008.