1,1'-(1,3-phenylene)bis-1H-pyrrole-2,5-dione

Endpoint summary

Description of key information

Acute Oral Toxicity: LD50 (male/female) >300-2000 mg/kg bw (OECD 423/GLP)

Acute Inhalational Toxicity (OECD 403):

LC50 Males =    0.055 mg/l (0.015 mg/l - 0.332 mg/l)

LC50 Females =    0.136 mg/l

LC50 Combined =    0.089 mg/l (0.044 mg/l - 0.256 mg/l)

Acute Dermal Toxicity: Waived

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:

From 03-08-2010 to 31-03-2011

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

Purity: 99.3%
Lot No.: 006001

Species:

rat

Strain:

other: Crl:CD(SD)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Co., Ltd
- Age at study initiation: 8-10 weeks
- Weight at study initiation: 184-202 g
- Fasting period before study: 16 hours before administration
- Housing: Aluminum front and floor stainless steel mesh breeding cage (W 19.7 × D 26.3 × H 18.0 cm) containing the animals individually.
- Diet: Radiation sterilized solid feed (CRF-1, Lot No. 100203, 100302) ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.8 to 23.2 °C
- Humidity (%): 53.8 to 64.8%
- Air changes (per hr): 12 times
- Photoperiod (hrs dark / hrs light): 12 hrs

Route of administration:

oral: gavage

Vehicle:

other: 0.5% (w/v) aqueous solution of methylcellulose

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 and 20 mg/mL
- Amount of vehicle (if gavage): 1 ml/100g bw
- Lot no.: 7085417

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Mortality:

At 2000 mg/kg bw, all animals were dead by Day 4. At 300 mg/kg bw, there were no mortalities.

Clinical signs:

other: At 2000 mg/kg bw, mydriasis, mucous stool, soiled fur of anogenital region, ptosis and lacrimation were observed. At 300 mg/g bw, mucous stool and loose stool as the treatment-related slight effects on the digestive system were observed until 4 hours afte

Gross pathology:

At necropsy of the 2000 mg/kg bw group, dilated lumen and liquid contents of stomach were observed in all animals, and red patch of glandular stomach and reddish contents of small intestine were observed. Some findings suggesting digestive injury were considered to be treatment-related effects.

There were no changes noted at necropsy in the 300 mg/kg bw group.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

In an acute oral study in the rat, the LD50 (female) for N,N′-m-phenylenedimaleimide was >300-2000 mg/kgbw.

Executive summary:

In an acute oral toxicity study (C541 (314 ‐022)), groups of Crl:CD(SD) female rats (3/sex) were given single oral doses of N,N′-m-phenylenedimaleimide  (99.3%) in 0.5 w/v% methylcellulose solution at doses of 300 and 2000 mg/kg bw and observed for 14 days.

LD50 (male/female): was >300-2000 mg/kg bw.

At 2000 mg/kg bw, all animals were dead by Day 4. At 300 mg/kg bw, there were no mortalities. At 2000 mg/kg bw, mydriasis, mucous stool, soiled fur of anogenital region, ptosis and lacrimation were observed. At 300 mg/g bw, mucous stool and loose stool as the treatment-related slight effects on the digestive system were observed until 4 hours after administration in one or two animals. At 2000 mg/kg bw, all animals were dead by Day 4 so no final body weights were recorded. At 300 mg/kg bw, there were no effects on body weights noted on Day 14. At necropsy of the 2000 mg/kg bw group, dilated lumen and liquid contents of stomach were observed in all animals, and red patches of glandular stomach and reddish contents of small intestine were observed. Some findings suggesting digestive injury were considered to be treatment-related effects. There were no changes noted at necropsy in the 300  mg/kg bw group.

This acute oral study is classified as acceptable. It does satisfy the guideline requirement for an acute oral study (OECD 401) in the rat.  

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

The key study was the only study available and was assigned a Klimisch score of 2.

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

Remarks:

Non-GLP study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

GLP compliance:

no

Test type:

traditional method

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BASF/8801
- Purity test date:28th calendar week 1988

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: Stability was ensured for at least the study period.

OTHER SPECIFICS: Purity >95%

Species:

rat

Strain:

other: SPF Wistar/Chbb: THOM

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr. K. Thomae GmbH, 0-7950 Biberach,FRG
- Age at study initiation: 8 - 9 weeks
- Weight at study initiation: Males - 267 ± 10.9 g; Females - 185 ± 7.4g;
- Housing: groups of five in cages type DK III of Becker
- Diet: KLIBA rat/mouse laboratory diet 24-343-4 10 mm pellets, Klingentalmuhle AG, CH-4303 Kaiseraugst, Switzerland
- Water: ad libitum

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

Route of administration:

other: dust aerosol

Type of inhalation exposure:

nose/head only

Vehicle:

other: 1 wt% of Aerosil (Groups 1-4, 6). Group 5 - unchanged

Mass median aerodynamic diameter (MMAD):

> 1.3 - < 7.4 µm

Geometric standard deviation (GSD):

>= 2.2 - <= 4.3

Remark on MMAD/GSD:

Group 1: 1.6μm, GSD:3.5;
Group 2: 1.4μm, GSD:2.6;
Group 3: 1.5μm, GSD:3.6;
Group 4: 1.3μm, GSD:4.3;
Group 5: 7.4μm, GSD:2.2;
Group 6: 3.3μm, GSD:2.8;

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: 55L

- Method of holding animals in test chamber: The animais were restrained in tubes and their snouts projected into the inhalation chamber.

- Source and rate of air: Test groups 1 - 4 and 6: 2,000 L/hr; Test group 5: 1,500 L/hr

- System of generating particulates/aerosols: A dust aerosol was generated by means of a (i) solid particle generator (brush-generator. Technical University of Karlsruhe/BASF) (test group 5) (b) dosing-wheel dust generator (test groups 1-4 and 6)

- Method of particle size determination:
Equipment
- Stack Sampler Mark III (Andersen)
- Vacuum Compressed Air Pump - Sampling probe (internal diameter 6.9 mm)
- Balance: Mettler AE 240 and Sartorius M3P
- Evaluation unit (IBM-PC with software PGA)

Procedure
Test group 1:
The sampling was started on the same time as the exposure. The dust aerosol generation was stopped after the end of the particle size sampling (after 8 hours).

Test group 2:
The sampling was started on the same time as the exposure and was stopped after exposure.

Test groups 3 - 6:
30 minutes after the beginning of the test at the earliest, one sample was taken per test group for the particle size analysis.

Test groups 1 - 6:
Before the sampling. the impactor was equipped with glass-fiber collecting discs and a backup particle filter. The impactor was connected to the pump and the test apparatus, and one sample (180 - 1,440 L) was taken. The impactor was taken apart, and the collecting discs and the backup particle filter were weighed. The contents of the pre-impactor as well as the amounts of the material adsorbed on the walls of the impactor and in the sampling probe (wall losses) were also determined quantitatively.

- Treatment of exhaust air: By means of an exhaust air system the pressure ratios in the inhalation system were adjusted in such a way that the amount of exhaust air was abaut 5 - 10% lower (excess pressure). This ensured that the mixture of test substance and air was not diluted with laboratory air in the breathing zones of the animais.

- Temperature, humidity, pressure in air chamber: The exposure System was placed in an air-conditioned laboratory. Temperatures in the exposure system were 19- 25°C.

TEST ATMOSPHERE
- Brief description of analytical method used:
Apparatus:

- Vacuum compressed air pump (Millipore)
- Filtration equipment with probe, internal diameter: 7 mm, (Millipore)
- Filter: MN 85/90 Bf (d =4.7 cm)
- Sampling velocity: 1.25 m/s
- Sampling amount: 18 L - 150 L
- Sampling position: immediately adjacent to the animals' noses
- Sampling frequency: 1 sample per concentration group about hourly

Gravimetric determination of the inhalation atmosphere concentration:

Equipment: balance: Mettler AE 240

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
concentrations were corrected for the amount of the added
excipient (test groups 1 - 4 and 6).

- Samples taken from breathing zone: Yes

VEHICLE
- Composition of vehicle (if applicable): Aerosil (Synthetic Amorphous Silica; CAS # 7631-86-9, 112945-52-5, 112926-00-8). The data owner of this study (BASF) provided a document that summarises the vehicle properties (see GPS Safety Summary Silicon Dioxide-EVONIK)
- Concentration of test material in vehicle (if applicable): 1%
- Justification of choice of vehicle: Used to achieve a more uniform dust concentration in air

TEST ATMOSPHERE (if not tabulated)
The test substance used in test groups 1 - 4 and 6 was mixed with 1 wt% of Aerosil in order to achieve a more uniform dust concentration in air and a cyclonic separator was connected down-stream with the generator.
The test substance used in test group 5 was unchanged. The concentration was adjusted by varying the piston feed of the dust reservoir and by varying the brush rotation (test group 5) or the rotation of the metering disc (test groups 1 - 4 and 6).

- Particle size distribution:
Group 1: A respirable dust aerosol fraction that might reach the alveolar region of 95% was obtained.
Group 2: A respirable dust aerosol fraction that might reach the alveolar region of 99% was obtained.
Group 3: A respirable dust aerosol fraction that might reach the alveolar region of 94% was obtained.
Group 4: A respirable dust aerosol fraction that might reach the alveolar region of 93% was obtained.
Group 5: A failure in generation technique produced an insufficient particle size. Therefore, the results of this test group are not used for evaluating the toxicity of the substance.
Group 6: A respirable dust aerosol fraction that might reach the alveolar region of 88% was obtained.
Refer to the test report for detailed information on particle size distribution in Section 3.3.3.

- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.):
Group 1: 1.6μm, GSD:3.5;
Group 2: 1.4μm, GSD:2.6;
Group 3: 1.5μm, GSD:3.6;
Group 4: 1.3μm, GSD:4.3;
Group 5: 7.4μm, GSD:2.2;
Group 6: 3.3μm, GSD:2.8;

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

ca. 4 h

Concentrations:

The nominal concentration (mg/l) was calculated from the amount of substance consumed and the air flow.
Group 1:0.30
Group 2: 0.36
Group 3: 0.89
Group 4:3.02
Group 5: 1.64 (disregarded due to a failure in the generation technique that produced an insufficient particle size)
Group 6: 3.35

Analytical concentrations:
Group 1: 0.006 ± 0.0003
Group 2: 0.012 ± 0.018
Group 3: 0.043 ± 0.003
Group 4: 0.11 ± 0.008
Group 5:0.26 ± 0.013 (disregarded due to a failure in the generation technique that produced an insufficient particle size)
Group 6: 0.29 ± 0.007

No. of animals per sex per dose:

5 male and 5 female per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (all groups except 5)
- Frequency of observations and weighing: The body weight of the animais was checked before the beginning of the test, after 7 days and at the end of the observation period. Clinical findings were recodled several times during exposure and at least once an each workday in the observation period. A check for dead animais was made daily.
- Necropsy of survivors performed: Yes

Sex:

male/female

Dose descriptor:

LC50

Effect level:

0.089 mg/L air

95% CL:

> 0.044 - < 0.256

Exp. duration:

4 h

Sex:

male

Dose descriptor:

LC50

Effect level:

0.055 mg/L air

95% CL:

> 0.015 - < 0.332

Exp. duration:

4 h

Sex:

female

Dose descriptor:

LC50

Effect level:

0.136 mg/L air

Exp. duration:

4 h

Mortality:

Group 1: 0/10 deaths
Group 2: 2/10 deaths
Group 3: 2/10 deaths
Group 4: 7/10 deaths
Group 5: 0/10 deaths (disregarded)
Group 6: 7/10 deaths

Clinical signs:

other: Group 1: After 1 hr, all animals had accelerated respiration. From Day 3 -14, no signs were evident in any animals. Group 2: After 15 mins, all animals had restlessness which ceased after 30 mins; after 30 mins, all animals had accelerated respiration whi

Body weight:

The body weight gain of male rats in test groups 1 – 3 and 5 and female rats in the test groups 1, 5, 6, compared with a historical control collective, was not affected by the substance over the total observation period.

The body weight gain of female rats in the test group 4, compared with a historical control collective, was retarded in the first week of the observation period and adjusted to normal in the second week of the observation period.

The body weight gain of female rats in the test groups 2 and 3, compared with a historical control collective, was retarded in the second week of the observation period.

The body weight gain of male animals in the test groups 4 and 6 could not be evaluated because only one animal survived.

Gross pathology:

Animais that died spontaneously/sacrificed in a moribund state:
General congestion.
Lungs: focal hyperemia, some animals with emphysema.
Adrenais: dark in same animals.

Sacrificed animals: No pathalogic findings noted.

Interpretation of results:

Category 2 based on GHS criteria

Conclusions:

In an acute inhalation study in rats, the LC50 (male/female) of N,N′-m-phenylenedimaleimide was 0.089 mg/l (0.044 mg/l - 0.256 mg/l).

Executive summary:

In an acute inhalation toxicity study (1744), groups of young adult Wistar rats (5/sex) were exposed by inhalation route to N,N′-m-phenylenedimaleimide  (>95%) in 1% Aerosil for 4 hours (head/nose only) at concentrations of  0.006, 0.012, 0.043, 0.11, 0.26 and 0.29 mg/l.  Animals were then were observed for 14 days.

LC50 Males = 0.055 mg/l (0.015 mg/l - 0.332 mg/l)

LC50 Females =  0.136 mg/l

LC50 Combined = 0.089 mg/l (0.044 mg/l - 0.256 mg/l)

The results from the 0.26 mg/l dose were disregarded due to a failure in the generation technique that produced an insufficient particle size. The MMAD for the other tests groups was 1.3 μm – 3.3 μm and GSD was 2.6 – 4.3. A respirable dust aerosol fraction that might reach 88 -99% of the alveolar region was obtained. As the dosing increased, the number of mortalities increased (0.012, 0.043 mg/l, 2/10 deaths by Day 1; 0.11, 0.29 mg/l, 7/10 deaths by Day 2). There was also a dose-dependent increase in clinical signs including effects on respiration, squatting position, ruffled fur, high stepping gait, and deteriorated general state. The body weight gain of male rats in test groups 1 – 3 and 5 and female rats in the test groups 1, 5, 6, compared with a historical control collective, was not affected by the substance over the total observation period. The body weight gain of female rats in the test group 4, compared with a historical control collective, was retarded in the first week of the observation period and adjusted to normal in the second week of the observation period. The body weight gain of female rats in the test groups 2 and 3, compared with a historical control collective, was retarded in the second week of the observation period. The body weight gain of male animals in the test groups 4 and 6 could not be evaluated because only one animal survived. In animals that died spontaneously or were sacrificed in a moribund state, pathological findings were general congestion, focal hyperemia in lungs and some animals with emphysema and dark adrenals. In sacrificed animals there were no pathological findings noted.

This acute inhalation toxicity test in rats is acceptable and satisfies the guideline requirement for an OECD 403 study.

 

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

0.089 mg/m³ air

Quality of whole database:

The key study was the only study available and was assigned a Klimisch score of 2.

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute oral toxicity

The acute oral toxicity of N,N′-m-phenylenedimaleimide has been evaluated in one study in rats.

In an acute oral toxicity study (OECD 423/GLP), groups of Crl:CD(SD) female rats (3/sex) were given single oral doses of N,N′-m-phenylenedimaleimide  (99.3%) in 0.5 w/v% methylcellulose solution at doses of 300 and 2000 mg/kg bw and observed for 14 days. At 2000 mg/kg bw, all animals were dead by Day 4. At 300 mg/kg bw, there were no mortalities. At 2000 mg/kg bw, mydriasis, mucous stool, soiled fur of anogenital region, ptosis and lacrimation were observed. At 300 mg/g bw, mucous stool and loose stool as the treatment-related slight effects on the digestive system were observed until 4 hours after administration in one or two animals. At 2000 mg/kg bw, all animals were dead by Day 4 so no final body weights were recorded. At 300 mg/kg bw, there were no effects on body weights noted on Day 14. At necropsy of the 2000 mg/kg bw group, dilated lumen and liquid contents of stomach were observed in all animals, and red patches of glandular stomach and reddish contents of small intestine were observed. Some findings suggesting digestive injury were considered to be treatment-related effects. There were no changes noted at necropsy in the 300  mg/kg bw group. The LD50 (male/female) was >300-2000 mg/kg bw.

Acute inhalational toxicity

The acute inhalational toxicity of N,N′-m-phenylenedimaleimide has been evaluated in one study in rats.

In an acute inhalation toxicity study (OECD 403), groups of young adult Wistar rats (5/sex) were exposed by inhalation route to N,N′-m-phenylenedimaleimide  (>95%) in 1% Aerosil for 4 hours (head/nose only) at concentrations of  0.006, 0.012, 0.043, 0.11, 0.26 and 0.29 mg/l.  Animals were then were observed for 14 days. The results from the 0.26 mg/l dose were disregarded due to a failure in the generation technique that produced an insufficient particle size. The MMAD for the other tests groups was 1.3 μm – 3.3 μm and GSD was 2.6 – 4.3.A respirable dust aerosol fraction that might reach 88 - 99% of the alveolar region was obtained.

As the dosing increased, the number of mortalities increased (0.012, 0.043 mg/l, 2/10 deaths by Day 1; 0.11, 0.29 mg/l, 7/10 deaths by Day 2). There was also a dose-dependent increase in clinical signs including effects on respiration, squatting position, ruffled fur, high stepping gait, and deteriorated general state. The body weight gain of male rats in test groups 1 – 3 and 5 and female rats in the test groups 1, 5, 6, compared with a historical control collective, was not affected by the substance over the total observation period. The body weight gain of female rats in the test group 4, compared with a historical control collective, was retarded in the first week of the observation period and adjusted to normal in the second week of the observation period. The body weight gain of female rats in the test groups 2 and 3, compared with a historical control collective, was retarded in the second week of the observation period. The body weight gain of male animals in the test groups 4 and 6 could not be evaluated because only one animal survived. In animals that died spontaneously or were sacrificed in a moribund state, pathological findings were general congestion, focal hyperemia in lungs and some animals with emphysema and dark adrenals. In sacrificed animals there were no pathological findings noted. The LC50 values were as follows:

LC50 Males = 0.055 mg/l (0.015 mg/l - 0.332 mg/l)

LC50 Females = 0.136 mg/l

LC50 Combined = 0.089 mg/l (0.044 mg/l - 0.256 mg/l)

The results from both of these studies are acceptable to use in the human health risk assessment.

Justification for classification or non-classification

Based on the available information in the dossier, the substance N,N′-m phenylenedimaleimide (CAS No. 3006-93-7) should be classified for Acute Oral Toxicity Category 4 and Acute Inhalation Toxicity Category 2 when the criteria outlined in Annex I of 1272/2008/EC are applied.