2,9-bis(p-methoxybenzyl)anthra[2,1,9-def:6,5,10-d'e'f']diisoquinoline-1,3,8,10(2H,9H)-tetrone

Endpoint summary

Description of key information

Oral: LD50 (m/f) > 5000 mg/kg bw, rat, similar to OECD TG 401, no GLP

Inhalation: LC50 (m/f) > 5.4 mg/L air, rat, similar to OECD TG 403, no GLP

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:

1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Principles of method if other than guideline:

A test group consisting of 5 animals/sex was treated by single gavage application with an aqueous solution of the test substance. The animals were observed for mortality and for clinical symptoms of toxicity. At the end of the observation period of 14 days, the surviving animals were sacrificed for the purpose of necropsy; animals that died during the observations period also were subjected to necropsy.

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr. K. Thomae GmbH, Biberach, FRG
- Age at study initiation: About 12 weeks
- Weight at study initiation: 209 g (males) and 187 g (females)
- Fasting period before study: 16 hours before administration
- Housing: Type DK-111 stainless steel wire mesh cages (supplied by Becker L Co., Castrop-Rauxel, FRG), 5 animals/cage
- Diet (e.g. ad libitum): Ssniff R; supplied by Ssniff, Versuchstierdiäten, D-4470 Soest, FRG
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Acclimatisation for at least one week in the animal care unit

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Air changes (per hr): Fully airconditioned rooms
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.5%

Details on oral exposure:

- Concentration in vehicle: 25 % (w/v)
- Administration volume: 20 ml/kg

Doses:

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
- Cage side observations: recording of signs several times on the day of administration, at least once each workday. Check for moribund and dead animals twice each workday and once daily on weekends and public holidays.
- Body weights: beginning of the test, day 3, 7 and 13
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

None of the animals died during the observation period.

Clinical signs:

other: Blue faeces at day 1 and 2.

Gross pathology:

No abnormal findings.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for the tested substance is above the tested dose of 5000 mg/kg bw.

Executive summary:

In an oral toxicity study comparable to OECD guideline 401, five Wistar rats per sex were treated with the test article at a single dose of 5000 mg/kg bw by gavage followed by a 14-day observation period. None of the animals died during the exposure period. At day 1 and 2 of the observation period the feces were blue. The animals showed a normal body weight gain. No abnormal findings were observed at necropsy. Therefore, based on the results of this study, the LD50 was determined at greater than 5000 mg/kg body weight.

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:

13 Oct 1982 - 27 Oct 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: THOM; Zuchtbetrieb: Dr. K. Thomae GmbH, 7950 Biberach
- Age at study initiation: 8 weeks
- Weight at study initiation: 245 g (males) and 179 g (females)
- Housing: type DIII (Fa. Becker)
- Diet: SSNIFF R 10 mm Pellet, Ssniff-Versuchstierdiäten GmbH, 4770 Soest, ad libitum
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20- 24
- Humidity (%): 30 - 70
- Air changes (per hr): Fully airconditioned room
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose/head only

Vehicle:

clean air

Mass median aerodynamic diameter (MMAD):

1.9 µm

Geometric standard deviation (GSD):

3.9

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Head-nose inhalation system INA 20 (glass-steel construction, BASF Aktiengesellschaft)
- Exposure chamber volume: ca. 55 L
- Method of holding animals in test chamber: the animals were restrained in tubes and their snouts projected into the inhalation chamber
- Source and rate of air: 1500 L/h, supply air
- Method of conditioning air: The exposure system was placed in an air-conditioned laboratory. Temperatures in the exposure system were 19-25 °C
- System of generating particulates/aerosols: A dust aerosol air mixture was generated by means of a dust generator.
- Method of particle size determination: Stack Sampler Mark III (Andersen)

TEST ATMOSPHERE
- Brief description of analytical method used: The preweighed filter was placed into the filtration equipment. By means of a vacuum compressed air pump a volume of the dust aerosol was drawn through the filter. The dust concentration in mg/l was calculated from the difference between the preweight of the filter and the weight of the filter after sampling, with reference to the sample volume of the inhalation atmosphere. The concentration was corrected for the amount of the added excipient.
- Samples taken from breathing zone: yes

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gravimetric determination

Duration of exposure:

4 h

Concentrations:

5.4 mg/L

No. of animals per sex per dose:

10

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: daily
- Body weights: at the beginning of the study, after 7 days and at the end of the study
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.4 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

None of the animals died during the observation period

Clinical signs:

other: Erratic breathing, slight nasal excretion, ruffled fur during the first 4 days of the observation period, no abnormal findings were observed from day 5 onward.

Body weight:

Normal weight gain.
Day 7, mean weight (g): males: 267; females 201
Day 14, mean weight (g): males: 316; females 211.

Gross pathology:

No abnormal findings.

Results of analytical measurements:

Sample No	Analyt. Conc. (mg/l)
1	5.31
2	5.07
3	5.40
4	5.65
Mean	5.4
standard deviation of the mean	0.33
Nominal concentration	39

Particle size analysis:

Stage	EACD 50% [µm]	[mg]	percentage distribution [%]	cummulative distribution [%]
pre-impactor	26.6	9.35	2.502	97.498
0	29.5	0.618	2.032	95.466
1	18.2	0.502	6.133	89.333
3	8.5	1.515	13.209	76.124
4	5.5	3.263	18.22	57.904
5	2.8	4.501	18.755	39
7	1.2	4.633	39.149	149
backup filter	< 1.2	9.671
Sum		34.053

The MMAD 50% = 1.9 µm (geometrical standard deviation =3.9) was calculated from the results of the particle size analysis.

A respirable dust aerosol fraction that might reach the alveolar region of 89% was obtained from the results of the particle size analysis.

Interpretation of results:

GHS criteria not met

Conclusions:

According to the EU and GHS classification criteria, no classification is warranted as to acute inhalation toxicity for the test substance.

Executive summary:

In an acute inhalation toxicity study, 10 male and 10 female Wistar rats were exposed to the test substance as a dust aerosol at a concentration of 5.4 mg/L for 4 hours. The animals were observed for 14 days after exposure.

No mortality occurred at the tested concentration. Clinical signs observed after treatment were erratic breathing, slight nasal excretion, ruffled fur during the first 4 days of the observation period, no abnormal findings were observed from day 5 onward. The body weight gain of the test group was normal and compareable with the control group. No gross pathological abnormalities were noted during the necropsy at termination of the post exposure observation period.

Cascade impactor measurements resulted in particle size distributions with mass median aerodynamic diameters (MMADs) of 1.9 µm.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

5 400 mg/m³ air

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Oral toxicity

In an oral toxicity study comparable to OECD guideline 401, five Wistar rats per sex were treated with the test article at a single dose of 5000 mg/kg bw by gavage followed by a 14-day observation period (BASF AG, 1982). None of the animals died during the exposure period. At day 1 and 2 of the observation period the feces were blue. The animals showed a normal body weight gain. No abnormal findings were observed at necropsy. Therefore, based on the results of this study, the LD50 was determined at greater than 5000 mg/kg body weight.

 

Inhalation toxicity

In an inhalation toxicity study comparable to OECD guideline 403, Wistar rats (10/sex) were exposed to the test article as dust for 4 hours at a measured concentration of 5.4 mg/L followed by a 14-day observation period (BASF, 1983). The mass median aerodynamic diameter (MMAD) was 1.9 µm (GSD: 3.9). None of the animals died during the study period. Clinical signs included erratic breathing, slight nasal excretion and ragged fur during the first 4 days of the observation period, no abnormal findings were observed from day 5 onward. No abnormal findings were noted at necropsy. Therefore, based on the results of this stud, the LC50 was determined to be greater than 5.4 mg/L.

 

Further toxicological data of category members:

The test article belongs to the "perylene based organic pigments" category (see attached document for details on category members and for read across justification). According to the category approach, missing toxicity endpoints can be addressed with data available for other category members. Regarding acute toxicity, reliable data are available for the test article and for other members of the "Perylene based pigments category". All of these data are taken into account for the evaluation and assessment of the acute toxicity of the test article.

In several studies performed with other substances from the Perylene category the potential for oral toxicity was found to be very low. None of these studies raised any concerns regarding acute toxicity after oral application and therefore none of the substances requires classification. The LD50 values observed for these compounds were greater than 5000 mg/kg bw in alle studies performed.

To assess the acute inhalation toxicity, other category members were tested in studies according or similar to OECD 403 guideline. In all studies, rats were exposed to analytically verified dust aerosols for a duration of 4 hours. Except for one study with a single case of mortality all animals survived the procedure. The observed clinical signs included accelerated respiration, pulmonary respiration sounds, squatting posture, piloerection, flight behavior and smeared fur. No pathological abnormalities of the organs were observed at termination in all animals in any of the studies. The LC50 was always greater than tested limit dose respectively.

There are also studies concerning the acute dermal toxicity. In the studies similar to OECD testing guideline 402 no signs for local or systemic toxicity could be found.

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 occurred at the limit dose of 2000 mg/kg bw. As a result, the substance is not considered to be classified for acute oral or inhalation toxicity under Regulation (EC) No. 1272/2008, as amended for the fourteenth time in Regulation (EC) No. 2020/217.