2,9-bis(3,5-dimethylphenyl)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

LD50 oral in rats > 5000 mg/kg bw
LC50 inhalation in rats > 5200 mg/m³
LD50 dermal in rats > 2500 mg/kg

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:

06.04.1982 - 20.04.1982

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)

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Stamm:Hoe WISKf(SPF71); Eigenzucht
- Weight at study initiation: 201.2 g
- Fasting period before study: 16 h
- Housing: in groups in plastic cages with soft wooden granules
- Diet (e.g. ad libitum): ALTROMIN 1324 (Altromin GmbH, Lage/Lippe), ad libitum
- Water (e.g. ad libitum): tap water, ad libitum

Route of administration:

oral: gavage

Vehicle:

other: sesame oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 25% (25 g/100 mL)

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

10 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality observed

Mortality:

None of the animals died during the observation period.

Clinical signs:

other: No abnormal observations. The pigment was excreted via the faeces.

Gross pathology:

No abnormal findings.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according 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: strain: SPF Wistar/Chbb: THOM; breeding facility: Dr. K. Thomae GmbH, D-7950 Biberach, FRG)
- Age at study initiation: 8 - 9 weeks
- Weight at study initiation: 251 g (males) and 172 g (females)
- Housing: cages type D III of Becker, without bedding, 5 animals per cage
- Diet (e.g. ad libitum): KLIBA rat/mouse laboratory diet 24-343-4 10 mm pellets, Klingentalmühle AG, CH-4303 Kaiseraugst, Switzerland, ad libitum
- Water (e.g. ad libitum): ad libitum

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

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose/head only

Vehicle:

other: Aerosil

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 dosing-wheel dust generator (Gericke/BASF).
- 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

VEHICLE
- Composition of vehicle (if applicable): 1 wt% of Aerosil

TEST ATMOSPHERE (if not tabulated)
- MMAD (Mass median aerodynamic diameter): 3.0 µm (GSD: 3.6)

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gravimetric determination

Duration of exposure:

4 h

Concentrations:

5.2 mg/L

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The body weight of the animals was checked before the beginning of the test, after 7 days and at the end of the observation period. Clinical findings were recorded several times during exposure and at least once an each workday in the observation period. A check for dead animals was made daily.
- Necropsy of survivors performed: yes

Statistics:

The statistical evaluation of the concentration/effect relationship was carried out on the basis of the binomial test (Wittig, H.: Mathematische Statistik 1974, pp. 32 - 35) in accordance with tables of the BASF Computer Center.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.2 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: no mortality observed

Mortality:

None of the animals died during the observation period

Clinical signs:

other: Irregular, accelerated and/or intermittent respiration, flight behaviour and discoloured fur. From day 7 of the observation period onward, no abnormalities, except discoloured fur, were detected in the animals.

Body weight:

The body weight gain of male and female rats in the test group, compared with a historical control collective, was not affected by the substance over the total observation period.

Gross pathology:

No pathologic findings were noted.

Results of analytical measurements:

Sample No	Analyt. Conc. (mg/l)
1	5.01
2	5.5
3	5
4	5.32
Mean	5.26
mean corrected for 1 % additive	5.22
mean rounded	5.2
standard deviation of the mean	0.2
Nominal concentration	16.8

Particle size analysis:

Cascade Impactor
Stage	EACD 50% [µm]	[mg]	percentage distribution [%]	cummulative distribution [%]
pre-impactor	26.6	0.97	4	96
0	29.5	0.79	3.3	92.7
1	18.2	2.67	11.1	81.6
3	8.5	2.69	11.2	70.5
4	5.5	6.58	27.2	43.3
5	2.8	4.45	18.4	24.9
7	1.2	6.01	24.9	-
backup filter	< 1.2
Sum		24.15

The MMAD 50% = 3.0 µm (geometrical standard deviation =3.6) was calculated from the results of the particle size analysis.

A respirable dust aerosol fraction that might reach the alveolar region of 82% was obtained from the results of the particle size analysis.

Interpretation of results:

GHS criteria not met

Conclusions:

In the study presented, acute toxicity of the test substance has been investigated using rats exposed via inhalation. No animal died during the study period. The LC50 was concluded to be greater than 5.2 mg/l.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 200 mg/m³ air

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

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 402 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

other: D.N.NOAKES und D.M.SANDERSON (A Method for Determining the Dermal Toxicity of Pesticides; Brit.Journ.Ind.Med. 26, 59, 1969

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Wiga, SPF-bred
- Weight at study initiation: males: 141 g; females: 132 g

Type of coverage:

not specified

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: back, area of 50 cm^2

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5 ml/kg
- Concentration (if solution): 50% solution

Duration of exposure:

Not specified

Doses:

2500 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: daily
- Necropsy of survivors performed: yes

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 2 500 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality was observed

Mortality:

None of the animals died during the observation period.

Clinical signs:

other: No abnormal findings. Red-brown staining at the application site observed.

Gross pathology:

No abnormal findings.

Interpretation of results:

GHS criteria not met

Conclusions:

In the study presented, acute toxicity of the test substance was investigated using rats exposed via dermal route. No animals died during the study period and thus, the LD50 was concluded to be 2500 mg/kg bw.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Justification for type of information:

see attached justification.

Reason / purpose for cross-reference:

read-across source

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 2 500 mg/kg bw

Based on:

other: Read-across

Remarks on result:

other: no mortality assumed

Interpretation of results:

GHS criteria not met

Conclusions:

Based on an acute dermal toxicity study performed with the read-across substance, the target substance was assumed not to induce mortality. Thus, the LD50 was concluded to be greater than 2500 mg/kg bw/day.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 500 mg/kg bw

Additional information

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.

Oral toxicity

In an oral toxicity study comparable to OECD guideline 401, ten female albino rats were treated with the test article at 15,000 mg/kg bw by single oral dose (gavage) followed by a 7-day observation period (Hoechst AG, 1967). None of the animals died during the exposure period. No abnormal clinical observations were observed; the pigment was excreted via the feces. The LD50 in this study was > 15000 mg/kg bw.

In another study comparable to OECD guideline 401, female Wistar rats (n=10) were treated with the test article at 5000 mg/kg bw by single oral dose (gavage) followed by a 14-day observation period (Hoechst AG, 1982). None of the animals died during the exposure period. No abnormal clinical observations were observed; the pigment was excreted via the feces. A normal weight gain was observed and no abnormal findings were made at necropsy. Based on the results of this study, the LD50 was determined to be greater than 5000 mg/kg bw.

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 ranged from 5,000 to 15,000 mg/kg body weight (maximum doses).

Conclusion: Based on the available data for the test substance and taking the data of category members into account, no classification for acute toxicity is warranted. The LD50 after oral administration in rats was determined to be greater than 10,000 mg/kg.

Inhalation toxicity

In an inhalation toxicity study according to OECD guideline 403, Wistar rats (5/sex) were exposed to the test article as dust for 4 hours at a measured concentration of 5.2 mg/L followed by a 14-day observation period (BASF AG, 1989). Cascade impactor measurements resulted in a particle size distribution with a mass median aerodynamic diameter (MMAD) of 3 µm (GSD: 3.6), which is well within the respirable range. None of the animals died during the study period. Clinical signs included irregular, accelerated and/or intermittent respiration, flight behaviour and discoloured fur. From day 7 of the observation period onward, no abnormalities, except discoloured fur, were detected in the animals. The body weight gain of male and female rats in the test group, compared with a historical control collective, was not affected by the substance over the total observation period. No pathologic findings were noted during gross pathology. The LC50 was >5.2 mg/L.

Two other category members were tested in OECD 403 guideline tests and a third substance was tested in a study similar to guideline 403. In all studies rats were exposed to dust aerosols analytically verified for a duration of 4 hours. Except for one study with a single case of mortality all animals survived the procedures. 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 analytically determined concentration of the test articles was greater than 5.1 mg/l in all of the studies (5.1 - 5.4 mg/l). Therefore, all studies resulted in an LC50 value of greater than 5.1 mg/l.

Conclusion: Based on the data obtained with the test article and taking the results of tests performed with other members of the Perylene category into account, no classification for acute toxicity is required.

Dermal toxicity

No data regarding acute toxicity after dermal exposure is available for the test substance. However, two studies performed with a category member are available. In the key study comparable to OECD guideline 402, 5 Sprague-Dawley rats of each sex were treated with the test substance at 2500 mg/kg bw by single dose followed by a 14-day observation period (BASF AG, 1976). None of the animals died during the exposure period. No abnormal clinical observations were observed and no abnormal findings were reported during necropsy. The LD50 was >2500 mg/kg bw.

In a supporting dermal toxicity study comparable to OECD guideline 402, Sprague-Dawley rats (5/sex) were administered a mixture containing 18.5% of the test substance at 5 ml/kg bw by single dose followed by a 14-day observation period (BASF, 1976). None of the animals died during the exposure period. No abnormal clinical observations were observed and no abnormal findings were reported at necropsy.

Conclusion: Based on the available data no classification for the test substance regarding dermal toxicity is warranted. The result obtained with the category member is used to set the dermal LD50 of the test substance at 2500 mg/kg bw.

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for the purpose of classification under Regulation (EC) No.1272/2008. Based on the data, classification for acute toxicity is not warranted under Regulation (EC) No.1272/2008.