Mixture of reaction products of substituted copper phthalocyanine, hydrogenated resin acids, alkaline earth metal chlorides, and copper phthalocyanine

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 July 2008 - 24 July 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to OECD and EC guidlines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

(1996)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Version / remarks:

(1992)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Version / remarks:

(1998)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

B508

IUPAC Name:

B508

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material: B508
- Physical state: powder

Test animals

Species:

rat

Strain:

other: Crl:WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: 13 weeks old
- Weight at study initiation: 330-380 g for males, 215-238 g for females
- Fasting period before study: no
- Housing: Group housing of 5 animals per sex per cage in labelled Macrolon cages (type IV; height 18 cm) containing sterilised sawdust as bedding material and paper as cage-enrichment. After exposure - group housing, maximally 5 animals per sex per cage in labelled stainless steel wire mesh cages. Nine days after exposure the animals were housed as described.
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF®) except during exposure to the test substance.
- Water (e.g. ad libitum): Free access to tap water except during exposure to the test substance.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.7 – 21.6
- Humidity (%): 45 – 77
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

inhalation

Type of inhalation exposure:

nose only

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION

The test substance was grinded with a pestle and mortar and sieved before use.

Exposure chamber
Animals were exposed to the test material via the inhalatory route. For this purpose the animals were placed in restraining tubes, which were connected to the exposure chamber. The design of the exposure chamber was based on the flow past nose-only inhalation chamber (Am. Ind. Hyg. Assoc. J. 44(12): 923-928, 1983). The chamber consisted of 3 animal sections with 8 animal ports each. The number of open animal ports was adapted to the air flow in such a way that at each animal port the theoretical air flow was approximately 2.0 L/min, which ensures an adequate oxygen supply to the test animals. The inlet of the test atmosphere was located in the top section and the outlet was located in the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal.

Test atmosphere generation
An aerosol was generated by administering the test substance to a stream of pressurized air by means of a spiral feeder and an air mover with a large exhaust opening (to prevent clogging).

Treatment
For exposure the animals were restrained in polycarbonate restraining tubes. After restraining the tubes were connected to the exposure chamber. Fifteen minutes after the last animal was placed the generation of the test atmosphere was started. The exposure lasted 4 hours.


Test atmosphere characterization
Nominal concentration: The nominal concentration was calculated by dividing the amount of test substance used by the volume of pressurized air (average air flow times exposure time) entering the exposure chamber used for exposure of the animals.

Actual concentration: The actual concentration was determined 13 times during the exposure period. Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber. Samples were drawn through a glass fiber filter. The collected amount of test substance in the air sample was measured gravimetrically. Sample volumes were measured by means of a dry gas meter.

Subsequently the mean concentrations and the standard deviations were calculated.


- Particle size distribution: was characterized twice during the exposure period. The samples were drawn (2 L/min) from the test atmosphere through a tube mounted in one of the free animal ports the middle section of the exposure chamber. The samples were collected with an 8 stage Marple personal cascade impactor, fiber glass filters and a fiber glass back-up filter. Amounts of test substance collected were measured gravimetrically. Subsequently the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD) were determined.

- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.):
Group 1st sample 2nd sample
MMAD (µm) GSD MMAD (µm) GSD
1 2.6** ** 2.9 3.4
** particle size distribution not log-normal distributed due to the presence of a large amount (approximately 8%) of particles with an aerodynamic diameter equal to or less than 0.25 µm. The MMAD was estimated by linear interpolation of the data at cut-off points 2.0 and 3.5 µm.

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gravimetrically

Duration of exposure:

4 h

Concentrations:

An average exposure concentration of 4.0 mg/L was attained instead of the target concentration of 5 mg/L.

No. of animals per sex per dose:

5

Control animals:

other: not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
During exposure - Three times during exposure for mortality, behavioural signs of distress and effects on respiration.
Mortality/Viability - Twice daily.
Body weights - Days 1 (pre-administration), 8 and 15.
Clinical signs - Twice (at 1 and at 3 hours after exposure) on the day of dosing (day 1) and once daily thereafter, until day 15.
- Necropsy of survivors performed: yes

Statistics:

Not applicable.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 4 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: The mean actual concentration was 4.0 ± 0.8 mg/L, which was the maximum attainable concentration that could be generated in air for 4 hours.

Mortality:

One male was found dead approximately 2½ hours after initiation of exposure.

Clinical signs:

other: During exposure laboured respiration was noted among all survivors. After exposure hunched posture, lethargy, laboured respiration and irregular breathing were noted among the surviving animals. Rales were noted among the majority of the females. The

Body weight:

Body weight loss was shown by all survivors during the first week after exposure while body weight gain was similar to that expected of normal untreated animals of the same age and strain during the second week after exposure.

Gross pathology:

No abnormality was found at macroscopic post mortem examination of the animal that died during the study. Macroscopic post mortem examination of the surviving animals at termination revealed many bluish foci in the lungs of all animals and bluish discolouration of the skin of the tail.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In an acute inhalation toxicity study with rats, performed according to OECD 403 guideline and GLP Principles, it was considered that the 4-hour LC50 value of B508 exceeds 4 mg/L.

Executive summary:

B508 was administered by inhalation for 4 hours at a limit concentration to rats according to OECD 403 guideline and GLP Principles.

The actual concentration of 4.0 ± 0.8 mg/L was considered the maximum attainable concentration which could be generated in air for 4 hours.

One male was found dead approximately 2½ hours after initiation of exposure. Clinical signs included hunched posture, lethargy, laboured respiration, irregular breathing and rales. Body weight loss was shown by all survivors during the first week after exposure only. No abnormality was found at macroscopic post mortem examination of the animal that died during the study. Macroscopic post mortem examination of the surviving animals at termination revealed many bluish foci in the lungs of all animals and bluish discolouration of the skin of the tail.

The inhalatory 4 -hour LC50 of B508 in rats was found to exceed the maximum attainable concentration of 4.0 mg/L. Although one mortality was observed at this exposure level and clinical signs indicative of respiratory difficulties (laboured respiration and rales) were noted during a significant part of the observation period, it was considered that the 4-hour LC50 of B508 exceeds 4 mg/L.